{"id":5635,"date":"2024-08-20T18:35:30","date_gmt":"2024-08-20T16:35:30","guid":{"rendered":"https:\/\/www.alveolelab.com\/ressources\/publications\/"},"modified":"2024-08-20T18:35:31","modified_gmt":"2024-08-20T16:35:31","slug":"publications","status":"publish","type":"page","link":"https:\/\/www.alveolelab.com\/fr\/ressources\/publications\/","title":{"rendered":"Publications"},"content":{"rendered":"
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Journal of Fluid Mechanics, 2026<\/div>

Microstreaming induced by a micro-cantilever vibrating elliptically in a viscous fluid<\/h2>\r\n
Jules Ghesquiere, Gustav K. Modler, Saeid Mollaei, David Gu\u00e9rin, Olivier Bou Matar, Henrik Bruus, Michael Baudoin, Sarah Cleve<\/div>
The acoustically excited vibrations of a micrometric object in a viscous liquid induce a net fluid flow known as microstreaming. This phenomenon can be harnessed for a variety of microscale applications, including particle transport, fluid mixing and the propulsion of micro-swimmers. Acoustic propulsion holds significant promise for\u00a0in vivo<\/span> manipulation due to its inherent biocompatibility and remote actuation capability, eliminating the need for an onboard energy source. However, designing steerable swimmers powered by vibrating tails requires a detailed understanding of the relationship between the input acoustic signal and the resulting streaming flow..<\/div> Voir l'article<\/a><\/div>
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Jove, 2026<\/div>

A Novel Platform for In Vitro Cellular Stretching and Imaging<\/h2>\r\n
Benjamin M. Goykadosh, Benjamin M. Goykadosh, Suzanne E Stasiak, Vasuretha Chandar, Samuel R. Polio, Harikrishnan Parameswaran<\/div>
Cells respond to mechanical cues from their environment, such as changes in extracellular matrix (ECM) stiffness and cyclic strain, which regulate cellular processes including cell fate determination, intercellular communication, and development. Alterations in these forces contribute to or drive disease progression in conditions like asthma, hypertension, and cancer. While existing\u00a0in-vitro<\/em> stretching devices can impose uniaxial or isotropic strains, they often use non-physiological stiffnesses, limit live imaging, or cannot achieve high strain amplitudes relevant to physiological and pathological conditions. Here, we present a compact..<\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2026<\/div>

KELPE: knock-in exchangeable dual landing pad embryonic stem cells enable efficient screening of synthetic gene circuits<\/h2>\r\n
Aisling Fairweather, Yana Slavova, Mattias Malaguti<\/div>

The establishment of genetic circuits in pluripotent stem cells (PSCs) allows to model and manipulate developmental events. However, prototyping complex circuitry remains challenging, due to limitations in screening circuit components and transgene silencing. Here, we introduce KELPE: PSCs with two silencing-resistant insulated genomic landing pads targeted to genomic safe harbour sites. KELPE cells enable the stable integration of multiple transgenes into the same genomic region, facilitating fair comparisons of genetic circuit components. We demonstrate this by fine-tuning \u201csynthetic neighbour-labelling\u201d technologies. We first generate optimised PUFFFIN PSCs, which report on cell-cell interactions by fluorescently labelling wild-type neighbours...<\/p><\/div> Voir l'article<\/a><\/div>

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Biorxiv, 2026<\/div>

Tissue-scale mechanics controls differentiation strategy and dynamics of epithelial multilayering<\/h2>\r\n
Cl\u00e9mentine\u00a0Villeneuve, Somiealo Azote Epse Hassikpezi, Marga Albu, Matthias R\u00fcbsam, Leah C. Biggs, Sabrina Vinzens, Kai Kruse, Anubhav Prakash, Peter Zentis, Elizabeth Lawson- Keister, Gautier Follain, Johanna Ivaska, Carien M. Niessen, M. Lisa Manning, Sara A. Wickstr\u00f6m<\/div>
Generating and maintaining multilayered epithelia requires coordinated cell division, differentiation, and tissue architecture, yet the precise mechanisms of multilayering remain unclear. Using the developing mouse epidermis, we show that basal stem cells adopt distinct multilayering strategies depending on tissue mechanics. Combining quantitative morphometry, embryo live imaging and physical modeling, we observe that early in development, the epidermis is fluid-like, allowing undifferentiated cells to move suprabasally through perpendicular divisions or basal detachment before differentiating. As the tissue matures and rigidifies, a mechanical barrier is established that only allows upward movement of basal cells that have committed to differentiation. The final..<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2026<\/div>

StrataChip: a microphysiological system capturing dynamic keratinocyte fate and mechanical transitions during human epidermal morphogenesis<\/h2>\r\n
Justin K. Amakor, Arvind Arul Nambi Rajan, Mageshi Kamaraj, Kyle A. Jacobs, Erica J. Hutchins,Torsten Wittmann, Matthew L. Kutys<\/div>
Epidermal development and homeostasis require precise coordination between keratinocyte differentiation and mechanics. Still, the mechanisms integrating these processes remain poorly understood in part due to limitations of existing experimental systems. Here, we introduce StrataChip, a tractable microphysiological system that enables dynamic, multimodal interrogation of human epidermal morphogenesis. The platform integrates a media perfused dermal tissue with human epidermal keratinocytes within a microfluidic device and supports rapid epidermal stratification following establishment of an air-liquid interface. High-resolution confocal imaging and single-cell RNA-sequencing demonstrate that the StrataChip recapitulates key..<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2026<\/div>

Cryo-ET reveals nanoscale thick filament disorganization in MYH7 P710R hypertrophic cardiomyopathy cardiomyocytes<\/h2>\r\n
Magda Zaoralov\u00e1, Joseph Yoniles, Prerna Giri, Richard G. Held, Alison Vander Roest, Peter D. Dahlberg, Daniel Bernstein, Alexander R. Dunn, Leeya Engel<\/div>
Hypertrophic cardiomyopathy (HCM) is the most common monogenic inherited heart disease and is a major cause of sudden death in individuals under 35 years of age. HCM is associated with progressive tissue-level disarray and subcellular disorganization in individual cardiomyocytes. Mutations in \u03b2-cardiac myosin (MYH7), the second most common genetic cause of HCM, commonly result in changes in sarcomeric force production, but how this leads to altered cell- and tissue-level organization is unclear. Here, we use cryo-electron tomography (cryo-ET) to bridge the molecular and cellular scales by..<\/div> Voir l'article<\/a><\/div>
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Nature protocols, 2026<\/div>

Integrated fluorescence light microscopy-guided cryo-focused ion beam-milling for in situ montage cryo-ET<\/h2>\r\n
Jie E. Yang, Veronika Vrbovsk\u00e1, Joshua M. Mitchell, Tilman Franke, Bryan S. Sibert, Matt R. Larson, Alexander S. Hall, Alex Rigort, Deane F. Mosher, John Mitchels, Elizabeth R. Wright<\/div>
Cryogenic-electron tomography (cryo-ET) permits the in situ visualization of biological macromolecules at the molecular level. Owing to the variable thickness of cells, tissues and organisms, frozen specimens may need to be thinned by cryo-focused ion beam (FIB) milling to produce thin (<500 nm) cryo-lamellae suitable for cryo-ET. Locating regions of interest remains a challenge because untargeted milling can lead to inadvertent ablation and removal of regions of interest. Correlative light and electron microscopy, combined with cryo-FIB milling, can guide the identification of labeled targets in the cellular milieu. Multiple ..<\/div> Voir l'article<\/a><\/div>
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arXiv, 2026<\/div>

Inferring three-body interactions in cell migration dynamics<\/h2>\r\n
Nadav Jacobsen, Noa Ben-Asher, Leeya Engel<\/div>
In active matter and living matter, such as clusters of migrating cells, collective dynamics emerges from the underlying interactions. A common assumption of theoretical descriptions of collective cell migration is that these interactions are pairwise additive. It remains unclear, however, if the dynamics of groups of cells is solely determined by pairwise interactions, or if higher-order interaction terms come into play. To investigate this question, we use time-lapse microscopy to record the dynamics of three cells interacting together in a linear three-site geometry. We collect a large number of cellular trajectories and develop an inference scheme to infer both pairwise and potential three-body cell-cell interactions...<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

In-cell cryo-electron tomography reveals differential effects of type I and type II kinase inhibitors on LRRK2 filament formation and microtubule association<\/h2>\r\n
Tamar\u00a0Basiashvili, Joshua Hutchings, Siyu Chen, Eva P. Karasmanis, W. Alexander Flaherty, Andres E. Leschziner, Elizabeth\u00a0Villa<\/div>
Mutations in leucine-rich repeat kinase 2 (LRRK2) are a leading contributor to developing familial and idiopathic Parkinson\u2019s disease (PD). Most PD-causing LRRK2 mutations increase the kinase activity, leading to abnormal phosphorylation of Rab GTPases, disrupting vesicular trafficking, cytoskeletal dynamics, and autophagy. When expressed in cells, LRRK2 is distributed throughout the cytosol. However, exogenously expressed LRRK2 can form microtubule-associated filaments that have been shown to affect molecular transport along microtubules in vitro. While LRRK2\u2019s association with microtubules has not been detected at endogenous levels, inhibitors being designed and tested as therapeutics have been shown to either promote or prevent filament formation of LRRK2...<\/div> Voir l'article<\/a><\/div>
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Small, 2025<\/div>

Effect of Load on Non-Muscle Myosin 2 Paralog Filaments in a Biomimetic Contractile Actin Array<\/h2>\r\n
Philip Bleicher, David Han,\u00a0\u00a0Neil Billington,\u00a0\u00a0Ryan Hart,\u00a0\u00a0Christian A. Combs,\u00a0\u00a0Shureed Qazi,\u00a0\u00a0Indra Chandrasekar,\u00a0\u00a0Jay R. Knutson, James R. Sellers <\/div>
Cells express three non-muscle myosin 2 (NM2) paralogs that form bipolar filaments of \u224830 motors each. Of these, NM2A and NM2B are best studied and show distinct enzymatic and mechanical properties. Although they can colocalize in cells, they also have unique localization patterns, suggesting functional differences. Most studies have examined these proteins interacting with actin under very low loads, but in cells they likely contribute to generating and maintaining cytoskeletal tension. Inspired by sarcomere-like tension generation in non-muscle cells, a minimal platform is reconstituted to study NM2 activity under load. Using micropatterned formin to align actin filaments in anti-parallel bundles, phosphorylated NM2 filaments are introduced to observe their interactions via TIRF (Total Internal Reflection Fluorescence) microscopy. NM2B contracts and bundles the actin filaments to form a tensed system...<\/div> Voir l'article<\/a><\/div>
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Nature methods, 2025<\/div>

Light-induced extracellular vesicle and particle adsorption<\/h2>\r\n
Colin L. Hisey, Xilal Y. Rima, Jacob Doon-Ralls, Chiranth K. Nagaraj, Sophia Mayone, Kim Truc Nguyen, Sydney Wiggins, Kalpana Deepa Priya Dorayappan, Xin Huang, Mangesh D. Hade, Karuppaiyah Selvendiran, James N. Higginbotham, Oleg Tutanov, Jeffrey L. Franklin, Robert J. Coffey, David Wood, Chunyu Hu, Divya S. Patel, Setty M. Maga\u00f1a, Andre F. Palmer, Derek Hansford, Eduardo Re\u00e1tegui<\/div>
The role of extracellular vesicles (EVs) and particles (EPs\/EVPs) in human health and disease has garnered considerable attention over the past two decades. However, while several types of EVPs are known to interact dynamically with the extracellular matrix and there is great potential value in producing high-fidelity EVP micropatterns, there are currently no label-free, scalable and tunable platform technologies with this capability. We introduce light-induced extracellular vesicle and particle adsorption (LEVA) as a powerful solution to study surface-bound EVPs...<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

A nematic framework for sprouting angiogenesis<\/h2>\r\n
Sara\u00a0Barrasa-Ramos, Carles\u00a0Blanch-Mercader, Abdul I.\u00a0Barakat <\/div>
Sprouting angiogenesis is critical for embryogenesis, wound healing, and tumor growth. Here we show that the liquid crystal framework, which has recently been leveraged to study various morphogenetic events, provides unique insight into angiogenic sprouting. In response to vascular endothelial growth factor, a potent pro-angiogenic factor, endothelial cells cultured on the surfaces of soft collagen hydrogels form elongated cell streams that coexist with more polygonal cell regions. Angiogenic sprouting initiates preferentially at cell stream tips, where nematic order gradients are most pronounced..<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

Cryogenic Electron Tomography Redefines Herpesvirus Capsid Assembly Intermediates Inside the Cell Nucleus<\/h2>\r\n
Stefan L.\u00a0Oliver, Muyuan\u00a0Chen,\u00a0Leeya\u00a0Engel,\u00a0Corey W.\u00a0Hecksel,\u00a0Xueting\u00a0Zhou,\u00a0Michael F.\u00a0Schmid,\u00a0Ann M.\u00a0Arvin, Wah\u00a0Chiu<\/div>
Herpesviruses encapsulate their double-stranded DNA (dsDNA) genomes within an icosahedral nucleocapsid formed in the infected cell nucleus. Four biochemically purified nucleocapsids have been characterized, but their roles in herpesvirus replication remain controversial. The status of the capsid vertex-specific component (CVSC), essential for capsid stability and dsDNA packaging and retention, is also unclear. By integrating cryogenic focused ion beam milling with electron tomography and subtomogram averaging, we derived atomic models for all protein components, including the CVSC, across different herpesvirus capsid types within infected cell nuclei.<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

LigHTS: Massively Parallel Biomimetic Photo-Functionalization for Imaging-Based Ultra-High-Throughput Screening<\/h2>\r\n
Alessandro\u00a0Enrico, Sara\u00a0Rigolli, Julius\u00a0Zimmermann,\u00a0Melissa\u00a0Pezzotti,\u00a0Eloisa\u00a0Torchia,\u00a0Moises\u00a0Di Sante,\u00a0Ferdinando\u00a0Auricchio, Francesco S.\u00a0Pasqualini<\/div>
Imaging-based ultra-high-throughput screening (UHTS) in pharma and biotech still runs on 384\/1536-well plates whose stiff, flat substrates limit biological fidelity and screening efficiency. Highly biomimetic organs-on-chips and organoids improve relevance but lack reproducibility and plate-scale throughput. Biomimetic hydrogel scaffolds can be produced at scale through photopolymerization, which yet uses focused optics to define micrometer-resolved geometries, constraining scalability. To address the technical challenge of truly scalable biomimetic substrates featuring anisotropies, this study presents LigHTS, an all-optical, in-well method that replaces focused with collimated illumination to photofabricate structured hydrogels in standard 384\/1536-well plates.<\/div> Voir l'article<\/a><\/div>
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Analytical chemistry, 2025<\/div>

Multiplex and Spatiotemporal Detection of Single-Cell Antibody Secretions Using Protein-Patterned Microwells<\/h2>\r\n
Julie Van Lent, Karen Ven, Maya Imbrechts, Nick Geukens, Paul Declerck, Karen Vanhoorelbeke, Jeroen Lammertyn<\/div>
Antibodies (Abs) are pivotal components in a myriad of applications including therapeutics and diagnostics. To discover novel target-specific Abs, screening and isolating Ab-expressing cells have proven to be an interesting route. Nevertheless, current single-cell technologies still come with some limitations, as they, for example, do not enable combined temporal and multiplex screening. To overcome these limitations, we present a microwell-based microfluidic device that enables in-depth analysis of Ab-secreting cells..<\/div> Voir l'article<\/a><\/div>
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BMC Biology, 2025<\/div>

An integrated 2D framework for quantifying cellular mechanics reveals the impact of juxtacrine Notch signalling on directed collective migration of endothelial cells<\/h2>\r\n
Janine Grolleman, Carlijn V. C. Bouten, Cecilia M. Sahlgren, Vito Conte<\/div>
Collective migration is the coordinated movement of a group of cells\u2014a fundamental process in health and disease. Many models have been developed to study the molecular and physical mechanisms of collective migration. However, the aim of this study is to engineer a flexible in vitro framework that allows for mechanobiological quantification of the separate and combined contributions of individual cell mechanics to the directed migration of a collective. We utilised this framework to understand the role of juxtacrine Notch signalling during collective endothelial migration\u2014an essential process during the formation of new blood vessels (known as angiogenesis).<\/div> Voir l'article<\/a><\/div>
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ALP Bioengineering, 2026<\/div>

Vertically Integrated System for Tracking and Assessing cell-cycle aware phenotypes under confinement<\/h2>\r\n
Melissa Pezzott, Eloisa Torchia, Julius Zimmermann, Sara Rigolli, Alessandro Enrico, Moises Di Sante, and Francesco S. Pasqualini<\/div>
Quantitative cell biology often examines migration and cell-cycle (CC) progression separately, limiting insights into their interplay under spatial constraints. Here, we present a vertically integrated platform combining multiplexed fluorescent reporters for CC phases, actin, and tubulin with photopatterned extracellular matrix islands of defined sizes, alongside an automated imaging pipeline (Fab2Mic) for high-throughput, live-cell tracking of migration and CC dynamics under planar confinement. Using HT1080 fibrosarcoma cells, we observed that planar confinement progressively reduced cell area and cytoskeletal spread, altered CC phase distributions, and increased abnormal CC events, including..<\/div> Voir l'article<\/a><\/div>
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Nature physics, 2025<\/div>

The actin cortex acts as a mechanical memory of morphology in confined migrating cells<\/h2>\r\n
Yohalie Kalukula, Marine Luciano, Gleb Simanov, Guillaume Charras, David B. Br\u00fcckner and Sylvain Gabriele<\/div>
Cell migration in narrow microenvironments is a hallmark of numerous physiological processes, involving successive cycles of confinement and release that drive significant morphological changes. However, it remains unclear whether migrating cells can retain a memory of their past morphological states, which could potentially enhance their navigation through confined spaces. By combining cell migration assays on standardized microsystems with biophysical modeling and biochemical perturbations, we demonstrate that local geometry governs these morphological switches, thereby facilitating cell passage through long and narrow gaps..<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

In situ Structure of the Human Gap Junction<\/h2>\r\n
Evans Eshriew, Esa-Pekka Kumpula, Shiv K. Sah-Teli, Amiel Abettan, Amina Djurabekova, Vivek Sharma, Juha T. Huiskonen<\/div>
Gap junction plaques (GJPs) enable direct intercellular communication and consist of connexin channels arranged into two-dimensional lattices. While structures of purified connexin channels have informed models of gating, they omit key intracellular regions and lack native context. Here, we use cryogenic electron tomography and focused ion beam milling to determine the in situ structure of human connexin-43 (Cx43) GJPs in HEK293 cells at 14 \u00c5 resolution. We discover a previously unresolved structural role for the large C-terminal domain in mediating lateral channel\u2013channel interactions critical for plaque assembly...<\/div> Voir l'article<\/a><\/div>
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Cell Biomaterials, 2025<\/div>

Mechanosensitive interactions of tumoroids with an engineered environment promote cell proliferation and enhance drug response detection<\/h2>\r\n
Emna OunEmna Ouni, Alexis Peaucelle, Rasta Ghasemi, Francesco Facchinetti, Matthieu Opitz, Ludovic Bigot, Allan Sauvat, Oliver Kepp, Fanny Jaulin, Yohann Loriot, Kristine Schauer<\/div>
Three-dimensional (3D) \u201cmini-tumors\u201d or tumoroids, grown in the lab, are avatars for tumors in patients because they can be directly used to study individual cancer progression and drug responses. Until now, their use was limited through inconsistent animal-derived materials, complex and slow development, limited scalability, and poor compatibility with advanced imaging.<\/div>\r\n
This work turns a biocompatible and synthetic gel (polyethylene glycol [PEG]-acrylate) into an active, tunable habitat for these mini-tumors...<\/div><\/div> Voir l'article<\/a><\/div>
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Frontiers in Cell and Developmental Biology, 2025<\/div>

Fabrication of microcompartments with controlled size and shape for encapsulating active matter<\/h2>\r\n
Benoit Vianay, Christophe Gu\u00e9rin, Laur\u00e8ne Gressin, Magali Orhant-Prioux, Laurent Blanchoin, Manuel Th\u00e9ry, Alexandra Colin<\/div>
In all living systems, the cytoplasm is separated from the external environment by membranes. This confinement imposes spatial constraints on the self-organization of internal components, filaments and organelles. While reconstituted systems are instrumental for understanding fundamental biological principles, traditional experiments often utilize volumes vastly larger than actual cells. In recent studies, water-in-oil droplets or giant unilamellar vesicles have been widely used to impose confinement. However..<\/div> Voir l'article<\/a><\/div>
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Nature Communication, 2025<\/div>

Adherent cells sustain membrane tension gradients independently of migration<\/h2>\r\n
Juan Manuel Garc\u00eda-Arcos, Amine Mehidi, Julissa S\u00e1nchez Vel\u00e1zquez, Pau Guillamat, Caterina Tomba, Laura Houzet, Laura Capolupo, Giovanni D\u2019Angelo, Adai Colom, Elizabeth Hinde, Charlotte Aumeier and Aur\u00e9lien Roux<\/div>
Tension propagates in lipid bilayers over hundreds of microns within milliseconds, seemingly precluding the formation of tension gradients. Nevertheless, plasma membrane tension gradients have been reported in migrating cells and along growing axons. Here, we show that the mechanosensitive, fluorescent membrane probe Flipper-TR visualizes membrane tension gradients in artificial and cellular membranes. Images of tension gradients allow their quantitative characterization, showing that they are long-ranged and linear in all migratory adherent cells. Using this tool, we unexpectedly reveal that tension gradients also exist in non-migrating adherent cells while they are absent in non-adherent migrating cells..<\/div> Voir l'article<\/a><\/div>
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Current Opinion in Structural Biology, 2025<\/div>

Cool and collected: Advances in sample preparation for cryo-electron microscopy<\/h2>\r\n
Shani Tcherner Elad, Leeya Engel, Noa Ben-Asher<\/div>
Cryo-electron microscopy (cryo-EM) has emerged as a transformative tool in structural biology, enabling high-resolution visualization of macromolecules in their native states. Cryo-focused ion beam milling (cryo-FIB) and other advances in sample preparation have expanded the range of biological samples that can be studied with cryo-EM to include cells and tissues. While the dream of high-resolution structural analysis of proteins within their native, cellular context is now being realized, sample..<\/div> Voir l'article<\/a><\/div>
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Nature Communications, 2025<\/div>

Frictiotaxis underlies adhesion-independent durotaxis<\/h2>\r\n
Adam Shellard, Kai Wei\u00dfenbruch, Peter A. E. Hampshire, Namid R. Stillman, Christina L. Dix, Richard Thorogate, Albane Imbert, Guillaume Charras, Ricard Alert, Roberto Mayor<\/div>
Cells move directionally along gradients of substrate stiffness \u2014 a process called durotaxis. In the situations studied so far, durotaxis relies on cell-substrate focal adhesions to sense stiffness and transmit forces that drive directed motion. However, whether and how durotaxis can take place in the absence of focal adhesions remains unclear. Here, we show that confined cells can perform durotaxis despite lacking focal adhesions. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

Fabrication of microcompartments with controlled size and shape for encapsulating active matter<\/h2>\r\n
Benoit Vianay, Christophe Gu\u00e9rin, Laur\u00e8ne Gressin, Magali Orhant-Prioux, Laurent Blanchoin, Manuel Th\u00e9ry, Alexandra Colin<\/div>
In all living systems, the cytoplasm is separated from the external environment by membranes. This confinement imposes spatial constraints on the self-organization of internal components, filaments and organelles. In recent studies, water-in-oil droplets or giant unilamellar vesicles have been widely used to impose confinement. However, these compartments present imaging challenges and make precise protein content control difficult. To address these limitations, we have developed versatile microwells that are straightforward to implement, compatible with different types of imaging, suitable for long-term experiments, and capable of generating large amounts of data. ...<\/div> Voir l'article<\/a><\/div>
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Methods in Molecular Biology, 2025<\/div>

Synaptogenic Assays Using Primary Neurons Cultured on Micropatterned Substrates<\/h2>\r\n
Katalin Cz\u00f6nd\u00f6r, Nathalie Piette, B\u00e9atrice Tessier, Vincent Studer, Olivier Thoumine<\/div>
One of the difficulties for studying the mechanisms of synaptogenesis stems from the spatial unpredictability of contact formation between neurons, and the involvement of many parallel adhesive pathways mediating axon-dendrite recognition. To circumvent these limitations, we describe here a method allowing for the investigation of biomimetic synaptic contacts at controlled locations with high precision and statistics. Specifically, primary neurons are cultured on micropatterned substrates comprising arrays of micron-scale dots coated with purified synaptogenic adhesion molecules. ...<\/div> Voir l'article<\/a><\/div>
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Respiratory Research , 2026<\/div>

A new tunable 3D alveolospheres model from human alveolar epithelial type 2 cells (AEC2) with reduced heterogeneity for studying cigarette smoke extract exposure<\/h2>\r\n
Marina Guecamburu, Arthur Pavot, Caroline Jeanni\u00e8re, Yaniss Belaroussi, Matthieu Thumerel, Emma Sammaniego, Hugues Begueret, Guillaume Maucort, Fanny Decoeur, Jean-William Dupuy, Anne-Aur\u00e9lie Raymond, Pauline Esteves, Leo Grassion, Gael Dournes, Patrick Berger, Elise Maurat, Katharina Raash, Elo\u00efse Latouille, Vincent Studer, Isabelle Dupin, Pauline Henrot, Ma\u00e9va Zysman<\/div>
Rationale<\/strong>: Three-dimensional (3D) organoid models, such as alveolospheres, are unique tools for investigating the mechanisms underlying emphysema. However, high inter-organoid heterogeneity hampers consistent results in emphysema research and drug testing. Objectives<\/strong>: To develop a tunable 3D alveolosphere derived from human primary type II alveolar epithelial cells (AEC2) for modeling alterations linked to cigarette smoke exposure. Methods<\/strong>: AEC2 (HTII-280+) were isolated from 52 lung samples from both COPD and non-COPD patients, then cultured in 3D, comparing Matrigel to preformed photopolymerized hydrogel microwells of adjustable size and stiffness. Topological and phenotypic characterization were performed on days (D)1, 7, and 14. Lamellar bodies (LBs) were quantified using artificial intelligence (AI) analysis of transmission electron microscopy (TEM) serial block-face images. Chronic exposure to 1% or 5% cigarette smoke extract (CSE) was performed for 5 consecutive days...<\/div> Voir l'article<\/a><\/div>
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Lab on a Chip, 2025<\/div>

Stimulus-induced mechanical compaction of biological polymer networks via smart hydrogel microstructures<\/h2>\r\n
Vicente Salas-Quiroz, Katharina Esch and Katja Zieske<\/div>
The remodeling of the extracellular matrix by mechanical forces plays a crucial role in organizing cellular microenvironments. To study these mechanical perturbations, various methods have been developed to modify the cellular microenvironment and to apply controlled forces. However, most existing approaches rely either on instruments that cannot be integrated into lab-on-chip systems or on small probes with limited spatiotemporal precision. In this work, a lab-on-chip system enables spatially and temporally controlled mechanical perturbations of biological polymer networks..<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

Theory of multiscale epithelial mechanics under stretch: from active gels to vertex models<\/h2>\r\n
Adam Ouzeri, Sohan Kale, Nimesh Chahare, Alejandro Torres-Sanchez, Daniel Santos-Olivan, Xavier Trepat, Marino Arroyo<\/div>
Epithelial monolayers perform a variety of mechanical functions, which include maintaining a cohesive barrier or developing 3D shapes, while undergoing stretches over a wide range of magnitudes and loading rates.\u00a0 While the molecular understanding and ability to manipulate cytoskeletal components within cells is rapidly increasing, how these components integrate to control tissue mechanics is far less understood, partly due to the disconnect between theoretical models of sub-cellular dynamics and those at a tissue scale. To fill this gap, here we propose a formalism bridging active-gel models of the actomyosin cortex and 3D vertex-like models at a tissue scale. ...<\/div> Voir l'article<\/a><\/div>
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Science, 2025<\/div>

Interphase cell morphology defines the mode, symmetry, and outcome of mitosis<\/h2>\r\n
Holly E. Lovegrove, Georgia E. Hulmes, Sabrina Ghadaouia, Christopher Revell, Marta Giralt-Pujol, Zain Alhashem, Andreia Pena, Damian D. Nogare, Ellen Appleton, Guilherme Costa, Richard L. Mort, Christoph Ballestrem, Gareth W. Jones, Cerys S. Manning, Ajay B. Chitnis, Claudio A. Franco, Claudia Linker, Katie Bentley, and Shane P. Herbert<\/div>
As tissues assemble, the dynamic shape changes that define their form occur coincident with the asymmetric cell divisions that generate cellular diversity. Lovegrove\u00a0et al<\/i>. used morphometric analyses of tissue formation in multiple contexts, including zebrafish, human, and mouse blood vessel and neural crest development, finding that these morphogenetic events are fundamentally co-dependent. Distinct shifts in shape-switched cells to a so-called \u201cisomorphic\u201d mode of division, which preserves premitotic shape and the unequal distribution of identity determinants throughout division...<\/div> Voir l'article<\/a><\/div>
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Cell Reports, 2025<\/div>

ARHGEF18 is a flow-responsive exchange factor controlling endothelial tight junctions and vascular leakage<\/h2>\r\n
Surya Prakash Rao Batta, Marc Rio, Corentin Lebot, C\u00e9line Baron-Menguy, Maxence Bodet, Reda Moutaoukil, Robin Le Ruz, Ibtissam Babahnini, Gervaise Loirand, Anne-Cl\u00e9mence Vion<\/div>
The shear stress resulting from blood flow is a major regulator of endothelial cell (EC) biology and morphology. Rho protein-mediated cytoskeleton remodeling is an early and essential step of EC responses to flow. However, how Rho protein signaling is controlled by shear stress remains unclear. Here we demonstrate that phosphorylation, activity, and expression of the Rho nucleotide exchange factor (RhoGEF) ARHGEF18 in ECs are modulated by the magnitude of shear stress. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2025<\/div>

Structure of the Thin Filament in Human iPSC-derived Cardiomyocytes and its Response to Heart Disease<\/h2>\r\n
Rahel A. Woldeyes, Masataka Nishiga, Alison S. Vander Roest, Leeya Engel, Prerna Giri, Gabrielle C. Montenegro, Alexander R. Dunn, James A. Spudich, Daniel Bernstein, Michael F. Schmid, Joseph C. Wu and Wah Chiu<\/div>
Cardiovascular diseases are a leading cause of death worldwide, but our understanding of the underlying mechanisms is limited, in part because of the complexity of the cellular machinery that controls the heart muscle contraction cycle. Cryogenic electron tomography (cryo-ET) provides a way to visualize diverse cellular machinery while preserving contextual information like subcellular localization and transient complex formation, but this approach has not been widely applied to the study of heart muscle cells (cardiomyocytes). Here, we deploy an optimized cryo-ET platform that enables cellular-structural biology in human induced pluripotent stem cell\u2013derived cardiomyocytes (hiPSC-CMs). \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Spotiflow: accurate and efficient spot detection for fluorescence microscopy with deep stereographic flow regression<\/h2>\r\n
Albert Dominguez Mantes, Antonio Herrera, Irina Khven, Anjalie Schlaeppi, Eftychia Kyriacou, Georgios Tsissios, Evangelia Skoufa, Luca Santangeli, Elena Buglakova, Emine Berna Durmus, Suliana Manley, Anna Kreshuk, Detlev Arendt, Can Aztekin, Joachim Lingner, Gioele La Manno, Martin Weigert<\/div>
Identifying spot-like structures in large and noisy microscopy images is a crucial step to produce high quality results in various life-science applications. Imaging-based spatial transcriptomics (iST) methods, in particular, critically depend on the precise detection of millions of transcripts in images with low signal-to-noise ratio. Despite advances in computer vision that have revolutionized many biological imaging tasks, currently adopted spot detection techniques are mostly still based on classical signal processing methods that often lack robustness to changing imaging conditions and thus require tedious manual tuning per dataset. In this work, we introduce Spotiflow, a deep learning method that achieves subpixel-accurate localizations by formulating the spot detection task as a multi-scale heatmap and stereographic flow regression problem. ...<\/div> Voir l'article<\/a><\/div>
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Soft Matter, 2024<\/div>

Global alignment and local curvature of microtubules in mouse fibroblasts are robust against perturbations of vimentin and actin<\/h2>\r\n
Anna Blob, David Ventzke, Ulrike R\u00f6lleke, Giacomo Nies, Axel Munk, Laura Schaedele and Sarah K\u00f6ster<\/div>
The eukaryotic cytoskeleton is an intricate network of three types of mechanically distinct biopolymers \u2013 actin filaments, microtubules and intermediate filaments (IFs). These filamentous networks determine essential cellular functions and properties. Among them, microtubules are important for intracellular transport and establishing cell polarity during migration. Despite their intrinsic stiffness, they exhibit characteristic bending and buckling in cells due to nonthermal forces acting on them. Interactions between cytoskeletal filaments have been found but are complex and diverse with respect to their effect on the mechanical behavior of the filaments and the architecture of networks. We systematically study how actin and vimentin IFs influence the network structure and local bending of microtubules by analyzing fluorescence microscopy images of mouse fibroblasts on protein micropatterns. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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ACS Applied Materials & Interfaces, 2024<\/div>

Cell Architecture and Dynamics of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) on Hydrogels with Spatially Patterned Laminin and N-Cadherin<\/h2>\r\n
Kerry V. Lane, Liam P. Dow, Erica A. Castilloa, R\u00e9mi Boros, Sam D. Feinstein, Gaspard Pardon, Beth L. Pruitt<\/div>
Controlling cellular shape with micropatterning extracellular matrix (ECM) proteins on hydrogels has been shown to improve the reproducibility of the cell structure, enhancing our ability to collect statistics on single-cell behaviors. (...) However, whether patterning single hiPSC-CMs on a protein associated with CM\u2013CM adhesion, like N-cadherin, can drive similar enhancement of the hiPSC-CM structure and function has not been tested. To address this, we developed a novel dual-protein patterning process featuring covalent binding of proteins at the hydrogel surface to ensure robust force transfer and force sensing. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature physics, 2025<\/div>

Single cell migration along and against confined haptotactic gradients<\/h2>\r\n
Isabela Corina Fortunato, David B. Br\u00fcckner, Steffen Grosser, Leone Rossetti, Miquel Bosch-Padr\u00f3s, Jonel Trebicka, Pere Roca-Cusachs, Raimon Sunyer, Edouard Hannezo and Xavier Trepat<\/div>
Haptotaxis is the process of directed cell migration along gradients of extracellular matrix density and is central to morphogenesis, immune responses and cancer invasion. It is commonly assumed that cells respond to these gradients by migrating directionally towards the regions of highest ligand density. In contrast with this view, here we show that cells exposed to micropatterned fibronectin gradients exhibit a wide range of complex trajectories, including directed haptotactic migration up the gradient but also linear oscillations and circles with extended periods of migration down the gradient.<\/div> Voir l'article<\/a><\/div>
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ChemRxiv, 2024<\/div>

Pressure-Controlled Nanopipette Sensing in the Asymmetric-Conductivity Configuration<\/h2>\r\n
Sebastian A. Skaanvik, Xinyu Zhang, Ian J. McPherson, Yuqing Wang, Anne-Kathrine K. Larsen, Steffan M. S\u00f8nderskov, Patrick R. Unwin,Tomaso Zambelli, Mingdong Dong<\/div>
Nanopipettes are important tools across diverse disciplines including biology, physics, materials science, and electrochemistry, and precisely adjusting their characteristics is vital for many applications. Recent progress in this endeavor has involved using the asymmetric-conductivity configuration with different electrolyte solutions inside and outside of the nanopipette, which can greatly improve nanopipette sensing. However, understanding such measurements is challenging due to the complex interplay between diffusion, electromigration, and electroosmosis. Herein, we studied the simplified case of the asymmetric-conductivity configuration where classical ion current rectification due to ion-selective migration is minimized, while the effect of electroosmotic flow is maximized ...<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Structural organization of p62 filaments and the cellular ultrastructure of calcium-rich p62-enwrapped lipid droplet cargo<\/h2>\r\n
Sabrina Berkamp, Lisa jungbluth, Alexandros Katranidis, Siavash Mostafavi, Olivera Korculanin, Peng-Han Lu, Lokesh Sharma, Lipi Thukral, J\u00f6rg Fitter, Rafal E. Dunin-Borkowski and Carsten Sachse<\/div>
The selective autophagy receptor p62\/SQSTM1 (from hereon p62) is known to form higher-order filaments in vitro and to undergo liquid-liquid phase separation when mixed with poly-ubiquitin. We determined the full-length cryo-EM structure of p62 and elucidated a structured double helical filament scaffold composed of the PB1-domain associated with the flexible C-terminal part residing in the lumen and the solvent-accessible major groove.<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Endogenous retrovirus-like proteins recruit UBQLN2 to stress granules and alter their functional properties<\/h2>\r\n
Harihar M. Mohan, Martin G. Fernandez, Camellia Huang, Rita Lin, Jaimie H. Ryou, Donald Seyfried, Nikolas Grotewold, Alexandra M. Whiteley, Sami J. Barmada, Venkatesha Basrur, Shyamal Mosalaganti, Henry L. Paulson, Lisa M. Sharkey<\/div>
The human genome is replete with sequences derived from foreign elements including endogenous retrovirus-like proteins of unknown function. Here we show that UBQLN2, a ubiquitin-proteasome shuttle factor implicated in neurodegenerative diseases, is regulated by the linked actions of two retrovirus-like proteins, RTL8 and PEG10. RTL8 confers on UBQLN2 the ability to complex with and regulate PEG10.<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Caging of membrane-to-cortex attachment proteins can trigger cellular symmetry breaking<\/h2>\r\n
Srishti Dar, Rub\u00e9n Tesoro Moreno, Ivan Palaia, Anusha B. Gopalan, Zachary Gao Sun, L\u00e9anne Strauss, Richard R. Sprenger, Julio M. Belmonte, Sarah K. Foster, Michael Murrell, Christer S. Ejsing, An\u0111ela \u0160ari\u0107, Maria Leptin and Alba Diz-Mu\u00f1oz<\/div>
To migrate, divide, and change shape, cells must regulate the mechanics of their periphery. The cell surface is a complex structure that consists of a thin, contractile cortical actin network tethered to the plasma membrane by specialized membrane-to-cortex attachment (MCA) proteins. This active and constantly fluctuating system maintains a delicate mechanochemical state which permits spontaneous polarization and shape change when needed. Combining\u00a0in silico<\/em>,\u00a0in vitro<\/em>, and\u00a0in vivo<\/em>\u00a0experiments we show how membrane viscosity and MCA protein length regulate cortical dynamics.<\/div> Voir l'article<\/a><\/div>
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Biophysical Journal, 2024<\/div>

Inferring cellular contractile forces and work using deep morphology traction microscopy<\/h2>\r\n
Yuanyuan Tao, Ajinkya Ghagre, Clayton W. Molter, Anna Clouvel, Jalal Al Rahbani, Claire M. Brown, Derek Nowrouzezahrai, Allen J. Ehrlicher<\/div>
Traction-force microscopy (TFM) has emerged as a widely used standard methodology to measure cell-generated traction forces and determine their role in regulating cell behavior. While TFM platforms have enabled many discoveries, their implementation remains limited (...). Here, we introduce deep morphology traction microscopy (DeepMorphoTM), a deep-learning alternative to conventional TFM approaches. DeepMorphoTM first infers cell-induced substrate displacement solely from a sequence of cell shapes and subsequently computes cellular traction forces, thus avoiding the requirement of a specialized fiduciarily marked deformable substrate or force-free reference image.<\/div> Voir l'article<\/a><\/div>
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Journal of Cell Science, 2024<\/div>

Nucleocytoplasmic transport senses mechanical forces independently of cell density in cell monolayers<\/h2>\r\n
Ignasi Granero-Moya, Valeria Venturini, Guillaume Belthier, Bart Groenen, Marc Molina-Jord\u00e1n, Miguel Gonz\u00e1lez-Mart\u00edn, Xavier Trepat, Jacco van Rheenen, Ion Andreu, Pere Roca-Cusachs<\/div>
Cells sense and respond to mechanical forces through mechanotransduction, which regulates processes in health and disease. In single adhesive cells, mechanotransduction involves the transmission of force from the extracellular matrix to the cell nucleus, where it affects nucleocytoplasmic transport (NCT) and the subsequent nuclear localization of transcriptional regulators, such as YAP (also known as YAP1). However, if and how NCT is mechanosensitive in multicellular systems is unclear. Here, we characterize and use a fluorescent sensor of nucleocytoplasmic transport (Sencyt) and demonstrate that NCT responds to mechanical forces but not cell density in cell monolayers. Using monolayers of both epithelial and mesenchymal phenotype, we show that NCT is altered in response both to osmotic shocks and to the inhibition of cell contractility.<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Balancing limited resources in actin networks competition<\/h2>\r\n
Christophe Gu\u00e9rin, Anne-Betty N\u2019Diaye, Laur\u00e8ne Gressin, Alex Mogilner, Manuel Th\u00e9ry, Laurent Blanchoin and Alexandra Colin<\/div>
In cells, multiple actin networks coexist in a dynamic manner. These networks compete for a common pool of actin monomers and actin-binding proteins. Interestingly, this competition does not result in the mere survival of the more consuming networks. Moreover, the co-existence of networks with various strengths is key to cell adaption to external changes. However, a comprehensive view of how these networks coexist in this competitive environment, where resources are limited, is still lacking. To address this question, we used a reconstituted system, in closed microwells, consisting of beads propelled by actin polymerization or micropatterns functionalized with lipids capable of initiating polymerization close to a membrane.<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Cell-mechanical parameter estimation from 1D cell trajectories using simulation-based inference<\/h2>\r\n
Johannes C. J. Heyn, Miguel Atienza Juanatey, Martin Falcke and Joachim O. R\u00e4dler<\/div>
Trajectories of motile cells represent a rich source of data that provide insights into the mechanisms of cell migration via mathematical modeling and statistical analysis. However, mechanistic models require cell type dependent parameter estimation, which in case of computational simulation is technically challenging due to the nonlinear and inherently stochastic nature of the models. Here, we employ simulation-based inference (SBI) to estimate cell specific model parameters from cell trajectories based on Bayesian inference. Using automated time-lapse image acquisition and image recognition large sets of 1D single cell trajectories are recorded from cells migrating on microfabricated lanes.<\/div> Voir l'article<\/a><\/div>
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ACS Applied Polymer Materials, 2024<\/div>

Advancing Semiconducting Polymer Patterning: Analysis and Predictive Modeling of Micropatterns Achieved via Photothermal Lithography<\/h2>\r\n
Meghna Jha, Joaquin Mogollon Santiana, Megan L. Hong, Emily Vong, Shiva Ahmadi, Harishankar Manikantan and Adam J. Moul\u00e9<\/div>
The industrial development of semiconducting polymers (SPs) faces a significant hurdle in the absence of an inexpensive, rapid, and viable patterning technology capable of producing submicron features. In this study, we explore photothermal patterning as a promising technique that leverages the solubility characteristics of SPs to address this challenge. We demonstrate the rapid adaptability of this technique using one of the commercially available direct-write photolithography apparatuses, the Alv\u00e9ole PRIMO that is commonly found in university clean rooms. (...) Put together, this method and the associated theoretical model set the stage for the development of a cost-effective and rapid photopatterning technology for SPs, opening up possibilities for industrial applications in microfabricating organic electronic devices.<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Spontaneous and Induced Oscillations in Confined Epithelia<\/h2>\r\n
Toshi Parmar, Liam P. Dow, Beth L. Pruitt and M. Cristina Marchetti<\/div>
The feedback between mechanical and chemical signals plays a key role in controlling many bisological processes and collective cell behavior. Here we focus on the emergence of spatiotemporal density waves in a one-dimensional \u201ccell train.\u201d Combining a minimal theoretical model with observations in an\u00a0in vitro<\/em>\u00a0experimental system of MDCK epithelial cells confined to a linear pattern, we examine the spontaneous oscillations driven by the feedback between myosin activation and mechanical deformations and their effect on the response of the tissue to externally applied deformations.<\/div> Voir l'article<\/a><\/div>
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PNAS Nexus, 2024<\/div>

Spatial regulation of substrate adhesion directs fibroblast morphotype and phenotype<\/h2>\r\n
Mirko D\u2019Urso, Ignasi Jorba, Atze van der Pol, Carlijn V C Bouten, Nicholas A Kurniawan<\/div>
The switching of the fibroblast phenotype to myofibroblast is a hallmark of a wide variety of tissue pathologies. This phenotypical switch is known to be influenced not only by humoral factors such as TGF-\u03b2, but also by mechanical and physical cues in the cellular environment, and is accompanied by distinctive changes in cell morphology. However, the causative link between these cues, the concomitant morphological changes, and the resulting phenotypic switch remain elusive. Here, we use protein micropatterning to spatially control dermal fibroblast adhesion without invoking exogenous mechanical changes and demonstrate that varying the spatial configuration of focal adhesions (FAs) is sufficient to direct fibroblast phenotype.<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Engineered microvasculature using maskless photolithography and on-chip hydrogel patterning: a facile approach<\/h2>\r\n
Dhanesh G. Kasi, Mees N. S. de Graaf, Dennis M. Nahon, Francijna E. van den Hil, Arn M. J. M. van den Maagdenberg, Christine L. Mummery and Valeria V. Orlova<\/div>
In vitro models of human microvasculature are increasingly used to understand blood vessel diseases and to support drug development. Most engineered models, however, are slow and labor-intensive to produce. Here, we used a single commercial digital micromirror device (DMD)-based setup for maskless photolithography to both fabricate microfluidic chips and pattern the inside of these chips with gelatin methacrylate (GelMA) hydrogels. ...<\/div> Voir l'article<\/a><\/div>
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ACS Nano, 2024<\/div>

Engineering the Cellular Microenvironment: Integrating Three-Dimensional Nontopographical and Two-Dimensional Biochemical Cues for Precise Control of Cellular Behavior<\/h2>\r\n
Einollah Sarikhani, Dhivya Pushpa Meganathan, Anne-Kathrine Kure Larsen, Keivan Rahmani, Ching-Ting Tsai, Chih-Hao Lu, Abel Marquez-Serrano, Leah Sadr, Xiao Li, Mingdong Dong, Francesca Santoro, Bianxiao Cui, Lasse Hyldgaard Klausen and Zeinab Jahed<\/div>
The development of biomaterials capable of regulating cellular processes and guiding cell fate decisions has broad implications in tissue engineering, regenerative medicine, and cell-based assays for drug development and disease modeling. Recent studies have shown that three-dimensional (3D) nanoscale physical cues such as nanotopography can modulate various cellular processes like adhesion and endocytosis by inducing nanoscale curvature on the plasma and nuclear membranes. Two-dimensional (2D) biochemical cues such as protein micropatterns can also regulate cell function and fate by controlling cellular geometries. Development of biomaterials with precise control over nanoscale physical and biochemical cues can significantly influence programming cell function and fate. In this study, we utilized a laser-assisted micropatterning technique to manipulate the 2D architectures of cells on 3D nanopillar platforms. ...<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Vimentin promotes collective cell migration through collagen networks via increased matrix remodeling and spheroid fluidity<\/h2>\r\n
Minh Tri Ho Thanh, Arun Poudel, Shabeeb Ameen, Bobby Carroll, M. Wu, Pranav Soman, Tao Zhang, J.M. Schwarz, Alison E. Patteson<\/div>
The intermediate filament (IF) protein vimentin is associated with many diseases with phenotypes of enhanced cellular migration and aggressive invasion through the extracellular matrix (ECM) of tissues, but vimentin\u2019s role in in-vivo cell migration is still largely unclear. Vimentin is important for proper cellular adhesion and force generation, which are critical to cell migration; yet the vimentin cytoskeleton also hinders the ability of cells to squeeze through small pores in ECM, resisting migration. To identify the role of vimentin in collective cell migration, we generate spheroids of wide-type and vimentin-null mouse embryonic fibroblasts (mEFs) and embed them in a 3D collagen matrix. ...<\/div> Voir l'article<\/a><\/div>
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Nature, 2024<\/div>

Endoplasmic reticulum\u2013plasma membrane contact gradients direct cell migration<\/h2>\r\n
Bo Gong, Jake D. Johnston, Alexander Thiemicke, Alex de Marco and Tobias Meyer<\/div>
Directed cell migration is driven by the front\u2013back polarization of intracellular signalling. Receptor tyrosine kinases and other inputs activate local signals that trigger membrane protrusions at the front. Equally important is a long-range inhibitory mechanism that suppresses signalling at the back to prevent the formation of multiple fronts. However, the identity of this mechanism is unknown. Here we report that endoplasmic reticulum\u2013plasma membrane (ER\u2013PM) contact sites are polarized in single and collectively migrating cells. ...<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Light-induced Extracellular Vesicle Adsorption<\/h2>\r\n
Colin L. Hisey, Xilal Y. Rima, Jacob Doon-Ralls, Chiranth K. Nagaraj, Sophia Mayone, Kim T. Nguyen, Sydney Wiggins, Kalpana D.P. Dorayappan, Karuppaiyah Selvendiran, David Wood, Chunyu Hu, Divya Patel, Andre Palmer, Derek Hansford and Eduardo Reategui<\/div>
The role of extracellular vesicles (EVs) in human health and disease has garnered considerable attention over the past two decades. However, while several types of EVs are known to interact dynamically with the extracellular matrix and there is great potential value in producing high-fidelity EV micropatterns, there are currently no label-free, high-resolution, and tunable platform technologies with this capability. We introduce Light-induced Extracellular Vesicle Adsorption (LEVA) as a powerful solution to rapidly advance the study of matrix- and surface-bound EVs and other particles. ...<\/div> Voir l'article<\/a><\/div>
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Biofabrication, 2024<\/div>

Steering cell orientation through light-based spatiotemporal modulation of the mechanical environment<\/h2>\r\n
Ignasi Jorba, Sil Gussenhoven, Atze van der Pol, Bart GW Groenen, Maarten van Zon, Marie Jos\u00e9 Goumans, Nicholas A Kurniawan, Tommaso Ristori and Carlijn VC Bouten<\/div>
The anisotropic organization of cells and the extracellular matrix (ECM) is essential for the physiological function of numerous biological tissues, including the myocardium. This organization changes gradually in space and time, during disease progression such as myocardial infarction. The role of mechanical stimuli has been demonstrated to be essential in obtaining, maintaining and de-railing this organization, but the underlying mechanisms are scarcely known. To enable the study of the mechanobiological mechanisms involved,\u00a0in vitro<\/i>\u00a0techniques able to spatiotemporally control the multiscale tissue mechanical environment are thus necessary. Here, by using light-sensitive materials combined with light-illumination techniques, we fabricated 2D and 3D\u00a0in vitro<\/i> model systems exposing cells to multiscale, spatiotemporally resolved stiffness anisotropies. ...<\/div> Voir l'article<\/a><\/div>
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Thesis, 2024<\/div>

Micropatterning subcellulaire pour \u00e9tudier la connectivit\u00e9 neuronale<\/h2>\r\n
Nathalie Piette<\/div>
Micropatterning was initially employed to replicate and understand the influence of the extracellular matrix on cells and some of their components. Over the past decade, subcellular printing has emerged, enabling the study of protein interactions and their role in signaling pathways as well as in the formation of synaptic, immunological, or neuronal pathways.The synaptic connection is mediated by synaptic adhesion proteins present on each side of the synapse. Due to the complexity of the synaptic environment and the lack of in vitro models to study synaptic connection in a biomimetic and controlled environment, the exact roles of these proteins in synaptogenesis remain uncertain. Subcellular protein printing presents a potential solution to address this gap. ...<\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2024<\/div>

Shape dynamics and migration of branched cells on complex networks<\/h2>\r\n
Jiayi Liu, Javier Boix-Campos, Jonathan E. Ron, Johan M. Kux, Nir S. Gov and Pablo J. S\u00e1ez<\/div>
Migratory and tissue resident cells exhibit highly branched morphologies to perform their function and to adapt to the microenvironment. Immune cells, for example, display transient branched shapes while exploring the surrounding tissues. In another example, to properly irrigate the tissues, blood vessels bifurcate thereby forcing the branching of cells moving on top or within the vessels. In both cases microenvironmental constraints force migrating cells to extend several highly dynamic protrusions. Here, we present a theoretical model for the shape dynamics and migration of cells that simultaneously span several junctions, which we validated by using micropatterns with an hexagonal array, and a neuronal network image analysis pipeline to monitor the macrophages and endothelial cell shapes and migration. ...<\/div> Voir l'article<\/a><\/div>
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PNAS, 2024<\/div>

Dual topologies of myotomal collagen XV and Tenascin C act in concert to guide and shape developing motor axons<\/h2>\r\n
Laurie Nemoz-Billet, Martial Balland, Laurent Gilquin, Benjamin Gillet, Isabelle St\u00e9vant, Emilie Guillon, Sandrine Hughes, Gilles Carpentier, Elisabeth Vaganay, Fr\u00e9d\u00e9ric Sohm, Vladimir Misiak, Mary-Julieth Gonzalez-Melo, Manuel Koch, Yad Ghavi-Helm, Sandrine Bretaud and Florence Ruggiero<\/div>
During development, motor axons are guided toward muscle target by various extrinsic cues including extracellular matrix (ECM) proteins whose identities and cellular source remain poorly characterized. Here, using single-cell RNAseq of sorted GFP+<\/sup>\u00a0cells from\u00a0smyhc1:gfp<\/i>-injected zebrafish embryos, we unravel the slow muscle progenitors (SMP) pseudotemporal trajectory at the single-cell level and show that differentiating SMPs are a major source of ECM proteins. (...)\u00a0Importantly, bioprinted micropatterns that mimic this in vivo ECM topology were sufficient to drive directional motor axon growth. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Plos Biology, 2024<\/div>

IntAct: A nondisruptive internal tagging strategy to study the organization and function of actin isoforms<\/h2>\r\n
Maxime C. van Zwam, Anubhav Dhar, Willem Bosman, Wendy van Straaten, Suzanne Weijers, Emiel Seta, Ben Joosten, Jeffrey van Haren, Saravanan Palani, Koen van den Dries <\/div>
Mammals have 6 highly conserved actin isoforms with nonredundant biological functions. The molecular basis of isoform specificity, however, remains elusive due to a lack of tools. Here, we describe the development of IntAct, an internal tagging strategy to study actin isoforms in fixed and living cells. ...<\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Dissecting the binding mechanisms of synaptic membrane adhesion complexes using a micropattern based cellular model<\/h2>\r\n
Nathalie Piette, Pierre-Olivier Strale, Matthieu Lagardere, Camille Saphy, Carsten Reissner, Matthieu Munier, Markus Missler, Ingrid Chamma, Matthieu Sainlos, olivier Thoumine, Vincent Studer<\/div>
The formation of adhesive cell-cell contacts is based on the intrinsic binding properties between specific transmembrane ligand-receptor pairs. In neurons, synaptic adhesion molecules provide a physical linkage between pre- and post-synaptic compartments, but the dynamics of these complexes in their actual membrane environments remain essentially unknown. To access such information, we developed a versatile assay to measure the affinity and binding kinetics of synaptic ligand-receptor interactions, based on the immobilization of Fc-tagged ligands on micropatterned substrates combined with live-cell imaging of fluorescently-tagged counter receptors in heterologous cells. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Doublecortin reinforces microtubules to promote growth cone advance in soft environments<\/h2>\r\n
Alessandro Dema, Rabab A. Charafeddine, Jeffrey van Haren, Shima Rahgozar, Giulia Viola, Kyle A. Jacobs, Matthew L. Kutys, Torsten Wittmann<\/div>
Doublecortin (DCX) is a microtubule-associated protein critical for brain development. Although most highly expressed in the developing central nervous system, the molecular function of DCX in neuron morphogenesis remains unknown and controversial. We demonstrate that DCX function is intimately linked to its microtubule-binding activity. (\u2026) Together with high resolution traction force microscopy data, we propose a model in which DCX-decorated, rigid growth cone microtubules provide intracellular mechanical resistance to actomyosin generated contractile forces in soft physiological environments in which weak and transient adhesion-mediated forces in the growth cone periphery may be insufficient for productive growth cone advance. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

Limiting Brownian Motion to Enhance Immunogold Phenotyping and Superimpose Optical and Non-Optical Single-EP Analyses<\/h2>\r\n
Kim Truc Nguyen, Xilal Y. Rima, Colin L. Hisey, Jacob Doon-Ralls, Chiranth K. Nagaraj, Eduardo Re\u00e1tegui<\/div>
Optical and non-optical techniques propelled the field of single extracellular particle (EP) research through phenotypic and morphological analyses, revealing the similarities, differences, and co-isolation of EP subpopulations. Overcoming the challenges of optical and non-optical techniques motivates the use of orthogonal techniques while analyzing extracellular particles (EPs), which require varying concentrations and preparations. Herein, we introduce the nano-positioning matrix (NPMx) technique capable of superimposing optical and non-optical modalities for a single-EP orthogonal analysis. The NPMx technique is realized by ultraviolet-mediated micropatterning to reduce the stochasticity of Brownian motion. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Mol Syst Biol, 2024<\/div>

The population context is a driver of the heterogeneous response of epithelial cells to interferons<\/h2>\r\n
Camila Metz-Zumaran, Zina M Uckeley, Patricio Doldan, Francesco Muraca, Yagmur Keser, Pascal Lukas, Benno Kuropka, Leonie K\u00fcchenhoff, Soheil Rastgou Talemi, Thomas H\u00f6fer, Christian Freund, Elisabetta Ada Cavalcanti-Adam, Frederik Graw, Megan Stanifer, Steeve Boulant<\/div>
Isogenic cells respond in a heterogeneous manner to interferon. Using a micropatterning approach combined with high-content imaging and spatial analyses, we characterized how the population context (position of a cell with respect to neighboring cells) of epithelial cells affects their response to interferons. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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\r\n
BioRxiv, 2024<\/div>

Optogenetic generation of leader cells reveals a force-velocity relation for collective cell migration<\/h2>\r\n
Leone Rossetti, Steffen Grosser, Juan Francisco Abenza, L\u00e9o Valon, Pere Roca-Cusachs, Ricard Alert, Xavier Trepat<\/div>
The front of migratory cellular clusters during development, wound healing and cancer invasion is typically populated with highly protrusive cells that are called leader cells. Leader cells are thought to physically pull and direct their cohort of followers, but how leaders and followers are mechanically organized to migrate collectively remains controversial. (\u2026) Here we show that the effectiveness of leader-follower organization is proportional to the asymmetry of traction and tension within the cellular cluster. By combining hydrogel micropatterning and optogenetic activation of Rac1, we locally generate highly protrusive leaders at the edge of minimal cell groups. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2024<\/div>

EasyGrid: a versatile platform for automated cryo-EM sample preparation and quality control<\/h2>\r\n
Olivier Gemin, Victor Armijo, Michael Hons, Caroline Bissardon, Romain Linares, Matthew W. Bowler, Georg Wolff, Kirill Kovalev, Anastasiia Babenko, Veijo T. Salo, Sarah Schneider, Christopher Rossi, L\u00e9a Lecomte, Thibault Deckers, K\u00e9vin Lauzier, Robert Janocha, Franck Felisaz, J\u00e9r\u00e9my Sinoir, Wojciech Galej, Julia Mahamid, Christoph W. M\u00fcller, Sebastian Eustermann, Simone Mattei, Florent Cipriani, Gergely Papp<\/div>
Imaging biological macromolecules in their native state with single-particle cryo-electron microscopy (cryo-EM) or\u00a0in situ<\/em>\u00a0cryo-electron tomography (cryo-ET) requires optimized approaches for the preparation and vitrification of biological samples. Here, we describe EasyGrid, a versatile technology enabling systematic, tailored and advanced sample preparation for cellular and structural biology. This automated, standalone platform combines in-line plasma treatment, microfluidic dispensing, blot-less sample spreading, jet-based vitrification and on-the-fly grid quality control using light interferometry to streamline cryo-EM sample optimization. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Sichuan Da Xue Xue Bao Yi Xue Ban, 2024<\/div>

Spatial Constraints of Rectangular Hydrogel Microgrooves Regulate the Morphology and Arrangement of Human Umbilical Vein Endothelial Cells<\/h2>\r\n
Wenli Jiang, Jian Zhong, Zhi Ouyang, Junyi Shen, Yan Qiu, Ye Zeng<\/div>
The objective was to construct microscale rectangular hydrogel grooves and to investigate the morphology and alignment of human umbilical vein endothelial cells (HUVECs) under spatial constraints. Vascular endothelial cell morphology and alignment are important factors in vascular development and the maintenance of homeostasis. Hydrogel microgrooves can regulate the morphology and orientation of HUVECs and mimic to a certain extent the\u00a0in vivo<\/i>\u00a0microenvironment of vascular endothelial cells, providing an experimental model that bears better resemblance to human physiology for the study of the unique physiological functions of vascular endothelial cells. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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\r\n
Sensors and Actuators, 2024<\/div>

Microfluidic integration of the single cell adhesion dot array (SCADA) technology for the real-time quantification of cell affinity<\/h2>\r\n
Alba Calatayud-Sanchez, Sara Caicedo de la Arada, Yara Alvarez-Bra\u00f1a, Fernando Benito-Lopez, Lourdes Basabe-Desmonts<\/div>
Understanding cell affinity to substrates and\u00a0biomolecules\u00a0is of great importance in disease research, drug development and general cell biology studies. Established techniques to measure cell affinity involve either expensive and cumbersome techniques that quantify the binding between cells and proteins in suspension (e.g., flow cytometry), or indirect methods that quantify the amount of cells on a surface (e.g., impedance sensors). Novel approaches exploit\u00a0microtechnologies\u00a0to reduce the number of cells needed and reach single cell resolution. However, the examples so far fail to provide a simple device to measure cell affinity with single cell resolution that can be adapted to several purposes and cell biology laboratories. Herein, we describe a tool for the real-time optical monitoring of cell affinity. It is based in the integration of the cell-based biosensing platform Single Cell Adhesion Dot Arrays (SCADA) into a\u00a0microfluidic device. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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\r\n
Molecular Oncology, 2025<\/div>

Unravelling the metastasis-preventing effect of miR-200c in vitro and in vivo<\/h2>\r\n
Bianca K\u00f6hler, Emily Brieger, Tom Brandst\u00e4tter, Elisa H\u00f6rterer, Ulrich Wilk, Jana P\u00f6hmerer, Anna J\u00f6tten, Philipp Paulitschke, Chase P Broedersz, Stefan Zahler, Joachim O R\u00e4dler, Ernst Wagner, Andreas Roidl<\/div>
Advanced breast cancer, as well as ineffective treatments leading to surviving cancer cells, can result in the dissemination of these malignant cells from the primary tumor to distant organs. Recent research has shown that microRNA 200c (miR-200c) can hamper certain steps of the invasion\u2013metastasis cascade. However, it is still unclear whether miR-200c expression alone is sufficient to prevent breast cancer cells from metastasis formation. Hence, we performed a xenograft mouse experiment with inducible miR-200c expression in MDA-MB 231 cells. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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\r\n
Bio-protocol, 2023<\/div>

Correlative Light and Electron Cryo-Microscopy Workflow Combining Micropatterning, Ice Shield, and an In-Chamber Fluorescence Light Microscope<\/h2>\r\n
Sabrina Berkamp, Deniz Daviran, Marit Smeets, Alexane Caignard, Riddhi A. Jani, Pia Sundermeyer, Caspar Jonker, Sven Gerlach, Bernd Hoffmann, Katherine Lau, Carsten Sachse<\/div>
In situ cryo-electron tomography (cryo-ET) is the most current, state-of-the-art technique to study cell machinery in its hydrated near-native state. The method provides ultrastructural details at sub-nanometer resolution for many components within the cellular context. Making use of recent advances in sample preparation techniques and combining this method with correlative light and electron microscopy (CLEM) approaches have enabled targeted molecular visualization. Nevertheless, the implementation has also added to the complexity of the workflow and introduced new obstacles in the way of streamlining and achieving high throughput, sample yield, and sample quality. Here, we report a detailed protocol by combining multiple newly available technologies to establish an integrated, high-throughput, optimized, and streamlined cryo-CLEM workflow for improved sample yield. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2023<\/div>

1D confinement mimicking microvessel geometry controls pericyte shape and motility<\/h2>\r\n
Aude Sagnimorte, Marie R. Adler, Gaspard de Tournemire, Pablo J. S\u00e1ez, David Gonzalez-Rodriguez, Claire A. Dessalles, Avin Babataheri<\/div>
Pericytes are mural cells of the microvasculature, characterised by their elongated distinct shape. Pericytes span along the axis of the vessels they adhere to, therefore they experience extreme lateral and longitudinal confinement. Pericyte shape is key for their function during vascular regulation and their spatial distribution is established by cell migration during the embryonic stage and maintained through controlled motility in the adult. However, how pericyte morphology is associated with migration and function remains unknown. We use micropatterns to mimic pericyte adhesion to vessels, and to reproduce in vitro the shapes adopted by pericytes in vivo.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Journal of Cell Science, 2023<\/div>

Microtubules under mechanical pressure can breach dense actin networks<\/h2>\r\n
Matthieu G\u00e9lin, Alexandre Schaeffer, J\u00e9r\u00e9mie Gaillard, Christophe Gu\u00e9rin, Benoit Vianay, Magali Orhant-Prioux, Marcus Braun, Christophe Leterrier, Laurent Blanchoin, Manuel Th\u00e9ry<\/div>
The crosstalk between actin network and microtubules is key to the establishment of cell polarity. It ensures that the asymmetry of actin architec ture along cell periphery directs the organization of microtubules in cell interior. In particular, the way the two networks are physically inter-twined regulates the spatial organization and the distribution of forces in the microtubule network. While their biochemical crosstalk is getting uncovered, their mechanical crosstalk is still poorly understood. Here we designed an in vitro reconstitution assay to study the physical interaction between dynamic microtubules with various structures made of actin filaments. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Journal of Extracellular Vesicles, 2023<\/div>

Engineering a tunable micropattern-array assay to sort single extracellular vesicles and particles to detect RNA and protein in situ<\/h2>\r\n
Jingjing Zhang, Xilal Y. Rima, Xinyu Wang, Luong T. H. Nguyen, Kristin Huntoon, Yifan Ma, Paola Loreto Palacio, Kim Truc Nguyen, Karunya Albert, Minh-Dao Duong-Thi, Nicole Walters, Kwang Joo Kwak, Min Jin Yoon, Hong Li, Jacob Doon-Ralls, Colin L. Hisey, Daeyong Lee, Yifan Wang, Jonghoon Ha, Kelsey Scherler, Shannon Fallen, Inyoul Lee, Andre F. Palmer, Wen Jiang, Setty M. Maga\u00f1a, Kai Wang, Betty Y. S. Kim, L. James Lee, Eduardo Re\u00e1tegui<\/div>
The molecular heterogeneity of extracellular vesicles (EVs) and the co-isolation of physically similar particles, such as lipoproteins (LPs), confounds and limits the sensitivity of EV bulk biomarker characterization. Herein, we present a single-EV and particle (siEVP) protein and RNA assay (siEVPPRA) to simultaneously detect mRNAs, miRNAs, and proteins in subpopulations of EVs and LPs. The\u00a0siEVPPRA immobilizes and sorts particles via positive immunoselection onto micropatterns and focuses biomolecular signals in situ. By detecting EVPs at a single-particle resolution, the\u00a0siEVPPRA outperformed the sensitivities of bulk-analysis benchmark assays for RNA and protein. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2023<\/div>

Nuclear deformation and dynamics of migrating cells in 3D confinement reveal adaptation of pulling and pushing forces<\/h2>\r\n
Stefan St\u00f6berl, Johannes Flommersfeld, Maximilian M. Kreft, Martin Benoit, Chase P. Broedersz, Joachim O. R\u00e4dler<\/div>
Eukaryotic cells show an astounding ability to migrate through pores and constrictions smaller than their nuclear diameter. However, the forces engaged in nuclear deformation and their effect on confined cell dynamics remain unclear. Here, we study the mechanics and dynamics of nuclei of mesenchymal cancer cells as they spontaneously and repeatedly transition through 3D compliant hydrogel channels. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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BioRxiv, 2023<\/div>

Comparative profiling of cellular gait on adhesive micropatterns defines statistical patterns of activity that underlie native and cancerous cell dynamics<\/h2>\r\n
John C. Ahn, Scott M. Coyle<\/div>
Cell dynamics are powered by patterns of activity, but it is not straightforward to quantify these patterns or compare them across different environmental conditions or cell-types. Here we digitize the long-term shape fluctuations of metazoan cells grown on micropatterned fibronectin islands to define and extract statistical features of cell dynamics without the need for genetic modification or fluorescence imaging. These shape fluctuations generate single-cell morphological signals that can be decomposed into two major components: a continuous, slow-timescale meandering of morphology about an average steady-state shape; and short-lived \u201cevents\u201d of rapid morphology change that sporadically occur throughout the timecourse.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Structural and Molecular Biology, 2023<\/div>

Molecular mechanism of glutaminase activation through filamentation and the role of filaments in mitophagy protection<\/h2>\r\n
Douglas Adamoski, Marilia Meira Dias, Jose Edwin Neciosup Ques\u00f1ay, Zhengyi Yang, Ievgeniia Zagoriy, Anna M. Steyer, Camila Tanimoto Rodrigues, Alliny Cristiny da Silva Bastos, Bianca Novaes da Silva, Renna Karoline Eloi Costa, Fl\u00e1via Mayumi Odahara de Abreu, Zeyaul Islam, Alexandre Cassago, Marin Gerard van Heel, S\u00edlvio Roberto Consonni, Simone Mattei, Julia Mahamid, Rodrigo Villares Portugal, Andre Luis Berteli Ambrosio, Sandra Martha Gomes Dias<\/div>
Glutaminase (GLS), which deaminates glutamine to form glutamate, is a mitochondrial tetrameric protein complex. Although inorganic phosphate (Pi) is known to promote GLS filamentation and activation, the molecular basis of this mechanism is unknown. Here we aimed to determine the molecular mechanism of Pi-induced mouse GLS filamentation and its impact on mitochondrial physiology. (\u2026) Human GLS filaments form inside tubulated mitochondria following glutamine withdrawal, as shown by in situ cryo-electron tomography of cells thinned by cryo-focused ion beam milling. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Advanced Healthcare Materials, 2023<\/div>

Mechanoresponse of Curved Epithelial Monolayers Lining Bowl-Shaped 3D Microwells<\/h2>\r\n
Marine Luciano, Marie Versaevel, Yohalie Kalukula, Sylvain Gabriele<\/div>
The optimal functioning of many organs relies on the curved architecture of their epithelial tissues. However, the mechanoresponse of epithelia to changes in curvature remains misunderstood. Here, bowl-shaped microwells in hydrogels are designed via photopolymerization to faithfully replicate the shape and dimensions of lobular structures. Leveraging these hydrogel-based microwells, curved epithelial monolayers are engineered, and how in-plane and Gaussian curvatures at the microwell entrance influence epithelial behavior is investigated. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Methods, 2023<\/div>

Correlative montage parallel array cryo-tomography for in situ structural cell biology<\/h2>\r\n
Jie E. Yang, Matthew R. Larson, Bryan S. Sibert, Joseph Y. Kim, Daniel Parrell, Juan C. Sanchez, Victoria Pappas, Anil Kumar, Kai Cai, Keith Thompson, Elizabeth Wright<\/div>
Imaging large fields of view while preserving high-resolution structural information remains a challenge in low-dose cryo-electron tomography. Here we present robust tools for montage parallel array cryo-tomography (MPACT) tailored for vitrified specimens. The combination of correlative cryo-fluorescence microscopy, focused-ion-beam milling, substrate micropatterning, and MPACT supports studies that contextually define the three-dimensional architecture of cells. To further extend the flexibility of MPACT, tilt series may be processed in their entirety or as individual tiles suitable for sub-tomogram averaging, enabling efficient data processing and analysis. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Materials, 2023<\/div>

The laminin-keratin link shields the nucleus from mechanical deformation and signalling<\/h2>\r\n
Zanetta Kechagia, Pablo S\u00e1ez, Manuel G\u00f3mez-Gonz\u00e1lez, Brenda Canales, Srivatsava Viswanadha, Mart\u00edn Zamarbide, Ion Andreu, Thijs Koorman, Amy E. M. Beedle, Alberto Elosegui-Artola, Patrick W. B. Derksen, Xavier Trepat, Marino Arroyo, Pere Roca-Cusachs<\/div>
The mechanical properties of the extracellular matrix (ECM) dictate tissue behaviour. In epithelial tissues, laminin is both a very abundant ECM component, and a key supporting element. Here we show that laminin hinders the mechanoresponses of breast epithelial cells by shielding the nucleus from mechanical deformation. Coating substrates with laminin-111, unlike fibronectin or collagen I, impairs cell response to substrate rigidity, and YAP nuclear localization. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Thesis, 2023<\/div>

Modelling physics with deep learning: An experimental case of cell contractility<\/h2>\r\n
Yuanyuan Tao<\/div>

Deep Learning (DL) algorithms have been used to model physical systems. However, the success heavily relies on how DL methodologies accommodate the properties of a system and data. (\u2026) Under this context, we study the case of Traction Force Microscopy (TFM), a class of experimental procedures and algorithms for measuring cell traction. (\u2026) We introduce Deep Morphology Traction Microscopy (DeepMorphoTM), a DL approach that infers cell traction from a shape sequence of a cell. By employing a deterministic framework, DeepMorphoTM effectively mitigates the biological variability in cell contractility for a given cell shape. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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BioRxiv, 2023<\/div>

A platform for dissecting force sensitivity and multivalency in actin networks<\/h2>\r\n
Joseph T. Levin, Ariel Pan, Michael T. Barrett, Gregory M. Alushin<\/div>

The physical structure and dynamics of cells are supported by micron-scale actin networks with diverse geometries, protein compositions, and mechanical properties. These networks are composed of actin filaments and numerous actin binding proteins (ABPs), many of which engage multiple filaments simultaneously to crosslink them into specific functional architectures. Mechanical force has been shown to modulate the interactions between several ABPs and individual actin filaments, but it is unclear how this phenomenon contributes to the emergent force-responsive functional dynamics of actin networks. Here, we engineer filament linker complexes and combine them with photo-micropatterning of myosin motor proteins to produce an\u00a0in vitro\u00a0reconstitution platform for examining how force impacts the behavior of ABPs within multi-filament assemblies.\u00a0\u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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iScience, 2023<\/div>

Na\u00efve T lymphocytes chemotax long distance to CCL21 but not to a source of bioactive S1P<\/h2>\r\n
Nicolas Garcia-Seyda, Solene Song, Valentine Seveau de Noray, Luc David-Broglio, Christoph Matti, Marc Artinger, Florian Dupuy, Martine Biarnes-Pelicot, Marie-Pierre Valignat, Daniel F. Legler, Marc Baj\u00e9noff, Olivier Theodoly<\/div>

Na\u00efve T lymphocytes traffic through the organism in search for antigen, alternating between blood and secondary lymphoid organs. Lymphocyte homing to lymph nodes relies on CCL21 chemokine sensing by CCR7 receptors, while exit into efferent lymphatics relies on sphingolipid S1P sensing by S1PR1 receptors. While both molecules are claimed chemotactic, a quantitative analysis of na\u00efve T lymphocyte migration along defined gradients is missing. Here, we used a reductionist approach to study the real-time single-cell response of na\u00efve T lymphocytes to CCL21 and serum rich in bioactive S1P. Using microfluidic and micropatterning ad hoc tools, we show that CCL21 triggers stable polarization and long-range chemotaxis of cells, whereas S1P-rich serum triggers a transient polarization only and no significant displacement, potentially representing a brief transmigration step through exit portals. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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Nature Communication, 2023<\/div>

Extracellular filaments revealed by affinity capture cryo-electron tomography of lymphocytes<\/h2>\r\n
Leeya Engel, Magda Zaoralova, Alexander R. Dunn, Stefan Oliver<\/div>
\r\n\r\nCryogenic-electron tomography (cryo-ET) has provided an unprecedented glimpse into the nanoscale architecture of cells by combining cryogenic preservation of biological structures with electron tomography. Micropatterning of extracellular matrix proteins is increasingly used as a method to prepare adherent cell types for cryo-ET as it promotes optimal positioning of cells and subcellular regions of interest for vitrification, cryo-focused ion beam (cryo-FIB) milling, and data acquisition. Here we demonstrate a micropatterning workflow for capturing minimally adherent cell types, human T-cells and Jurkat cells, for cryo-FIB and cryo-ET. \u2026\r\n\r\n<\/div><\/div> Voir l'article<\/a><\/div>
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Biophysical Journal, 2023<\/div>

Force generation in human blood platelets by filamentous actomyosin structures<\/h2>\r\n
Anna Zelena, Johannes Blumberg, Dimitri Probst, Ruta Gerasimait, Grazvydas Lukinavicius, Ulrich S. Schwarz, Sarah Koester<\/div>

Blood platelets are central elements of the blood clotting response after wounding. Upon vessel damage, they bind to the surrounding matrix and contract the forming thrombus, thus helping to restore normal blood circulation. The hemostatic function of platelets is directly connected to their mechanics and cytoskeletal organization. The reorganization of the platelet cytoskeleton during spreading occurs within minutes and leads to the formation of contractile actomyosin bundles, but it is not known if there is a direct correlation between the emerging actin structures and the force field that is exerted to the environment. In this study, we combine fluorescence imaging of the actin structures with simultaneous traction force measurements in a time-resolved manner. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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biorxiv, 2023<\/div>

Handling difficult cryo-ET samples: A study with primary neurons from Drosophila melanogaster<\/h2>\r\n
Joseph Y. Kim, Jie E. Yang, Josephine W. Mitchell, Lauren A. English, Sihui Z. Yang, Tanner Tenpas, Erik W. Dent, Jill Wildonger, Elizabeth R. Wright<\/div>

Cellular neurobiology has benefited from recent advances in the field of cryo-electron tomography (cryo-ET). Numerous structural and ultrastructural insights have been obtained from plunge-frozen primary neurons cultured on electron microscopy grids. With most primary neurons been derived from rodent sources, we sought to expand the breadth of sample availability by using primary neurons derived from 3rd\u00a0instar\u00a0Drosophila melanogaster\u00a0larval brains. Ultrastructural abnormalities were encountered while establishing this model system for cryo-ET, which were exemplified by excessive membrane blebbing and cellular fragmentation. To optimize neuronal samples, we integrated substrate selection, micropatterning, montage data collection, and chemical fixation. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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Nature Communications, 2023<\/div>

Mapping mechanical stress in curved epithelia of designed size and shape<\/h2>\r\n
Ariadna Mar\u00edn-Llaurad\u00f3, Sohan Kale, Adam Ouzeri, Tom Golde, Raimon Sunyer, Alejandro Torres-S\u00e1nchez, Ernest Latorre, Manuel G\u00f3mez-Gonz\u00e1lez, Pere Roca-Cusachs, Marino Arroyo, Xavier Trepat<\/div>
The function of organs such as lungs, kidneys and mammary glands relies on the three-dimensional geometry of their epithelium. To adopt shapes such as spheres, tubes and ellipsoids, epithelia generate mechanical stresses that are generally unknown. Here we engineered curved epithelial monolayers of controlled size and shape and mapped their state of stress. We designed pressurized epithelia with circular, rectangular and ellipsoidal footprints. (\u2026) Besides interrogating the fundamental mechanics of epithelia over a broad range of sizes and shapes, our approach will enable a systematic study of how geometry and stress influence epithelial fate and function in three-dimensions. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Journal of Cell Biology, 2023<\/div>

Mechanical control of the mammalian circadian clock via YAP\/TAZ and TEAD<\/h2>\r\n
Juan F. Abenza, Leone Rossetti, Mal\u00e8ke Mouelhi, Javier Burgu\u00e9s, Ion Andreu, Keith Kennedy, Pere Roca-Cusachs, Santiago Marco, Jordi Garc\u00eda-Ojalvo, Xavier Trepat<\/div>
Autonomous circadian clocks exist in nearly every mammalian cell type. These cellular clocks are subjected to a multilayered regulation sensitive to the mechanochemical cell microenvironment. Whereas the biochemical signaling that controls the cellular circadian clock is increasingly well understood, mechanisms underlying regulation by mechanical cues are largely unknown. Here we show that the fibroblast circadian clock is mechanically regulated through YAP\/TAZ nuclear levels. We use high-throughput analysis of single-cell circadian rhythms and apply controlled mechanical, biochemical, and genetic perturbations to study the expression of the clock gene\u00a0Rev-erb\u03b1. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Communications, 2023<\/div>

PPP2R1A regulates migration persistence through the NHSL1-containing WAVE Shell Complex<\/h2>\r\n
Yanan Wang, Giovanni Chiappetta, Rapha\u00ebl Gu\u00e9rois, Yijun Liu, St\u00e9phane Romero, Daniel J. Boesch, Matthias Krause, Claire A. Dessalles, Avin Babataheri, Abdul I. Barakat, Baoyu Chen, Joelle Vinh, Anna Polesskaya, Alexis M. Gautreau<\/div>

The RAC1-WAVE-Arp2\/3 signaling pathway generates branched actin networks that power lamellipodium protrusion of migrating cells. Feedback is thought to control protrusion lifetime and migration persistence, but its molecular circuitry remains elusive. Here, we identify PPP2R1A by proteomics as a protein differentially associated with the WAVE complex subunit ABI1 when RAC1 is activated and downstream generation of branched actin is blocked. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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Nature Communications, 2023<\/div>

A convolutional neural network STIFMap reveals associations between stromal stiffness and EMT in breast cancer<\/h2>\r\n
Connor Stashko, Mary-Kate Hayward, Jason J. Northey, Neil Pearson, Alastair J. Ironside, Johnathon N. Lakins, Roger Oria, Marie-Anne Goyette, Lakyn Mayo, Hege G. Russnes, E. Shelley Hwang, Matthew L. Kutys, Kornelia Polyak, Valerie M. Weaver<\/div>
Intratumor heterogeneity associates with poor patient outcome. Stromal stiffening also accompanies cancer. Whether cancers demonstrate stiffness heterogeneity, and if this is linked to tumor cell heterogeneity remains unclear. We developed a method to measure the stiffness heterogeneity in human breast tumors that quantifies the stromal stiffness each cell experiences and permits visual registration with biomarkers of tumor progression. We present Spatially\u00a0Transformed\u00a0Inferential\u00a0Force\u00a0Map\u00a0(STIFMap) which exploits computer vision to precisely automate atomic force microscopy (AFM) indentation combined with a trained convolutional neural network to predict stromal elasticity with micron-resolution using collagen morphological features and ground truth AFM data. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Thesis, 2023<\/div>

Spatiotemporal Analysis of Metazoan Cell Morphological Dynamics on Micropatterned Substrates<\/h2>\r\n
John C Ahn<\/div>

Spatial patterns of adhesive substrates can dictate cell motility trajectories.
Mammalian cells demonstrate idiosyncratic trajectories when subjected to barbell-shaped micropatterns with thin constrictions. However, the morphologies of these cells over time remained uncharacterized. We subjected 3T3 mouse fibroblast cells to an array of micropatterns, and compared their shape dynamics to a panel of triple-negative breast cancer cells. When exposed to a gradient of size, we found that the morphological dynamics of 3T3 cells display trends analogous to the potential energy curve of a molecule, where energy is high at small micropattern sizes, drops off at a minimum point, and rises again at large micropattern sizes. We also
found the breast cancer cell panel exhibited a gradient of morphological activity and could be classified using principal component analysis. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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biorxiv, 2023<\/div>

In vitro modelling of anterior primitive streak patterning with hESC reveals the dynamic of WNT and NODAL signalling required to specify notochord progenitors<\/h2>\r\n
M. Robles Garcia, C. Thimonier, K. Angoura, E. Ozga, H. MacPherson, G. Blin<\/div>

Notochord progenitors (NotoPs) are a rare, yet vital embryonic cell population that give rise to the cells that form and maintain intervertebral discs. An unlimited access to NotoPs would open new opportunities for basic biomedical research and regenerative medicine of the discs. However, the mechanisms responsible for the specification and the maintenance of NotoPs are not understood. This gap in understanding stems from the fact that NotoPs emerge during the gastrulation to axial elongation transition; an event that is ethically and technically challenging to investigate. Here, to circumvent this issue, we use micropatterning to guide the development of human ESCs into standardised patterns of anterior primitive streak cell fates.\u00a0\u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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biorxiv, 2023<\/div>

Engineering cell and nuclear morphology on nano topography by contact-free protein micropatterning<\/h2>\r\n
Einollah Sarikhani, Dhivya Pushpa Meganathan, Keivan Rahmani, Ching-Ting Tsai, Abel Marquez-Serrano, Xiao Li, Francesca Santoro, Bianxiao Cui, Lasse Hyldgaard Klausen, Zeinab Jahed<\/div>

Platforms with nanoscale topography have recently become powerful tools in cellular biophysics and bioengineering. Recent studies have shown that nanotopography affects various cellular processes like adhesion and endocytosis, as well as physical properties such as cell shape. To engineer nanopillars more effectively for biomedical applications, it is crucial to gain better control and understanding of how nanopillars affect cell and nuclear physical properties, such as shape and spreading area, and impact cellular processes like endocytosis and adhesion. In this study, we utilized a laser-assisted micropatterning technique to manipulate the 2D architectures of cells on 3D nanopillar platforms. We performed a comprehensive analysis of cellular and nuclear morphology and deformation on both nanopillar and flat substrates.\u00a0\u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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Advanced Science, 2023<\/div>

Downregulation of YAP Activity Restricts P53 Hyperactivation to Promote Cell Survival in Confinement<\/h2>\r\n
Farnaz Hemmati, Ayuba Akinpelu, Jiyeon Song, Farshad Amiri, Anya McDaniel, Collins McMurray, Alexandros Afthinos, Stelios T. Andreadis, Andrew V. Aitken, Vinicia C. Biancardi, Sharon Gerecht, Panagiotis Mistriotis<\/div>
Cell migration through confining three dimensional (3D) topographies can lead to loss of nuclear envelope integrity, DNA damage, and genomic instability. Despite these detrimental phenomena, cells transiently exposed to confinement do not usually die. Whether this is also true for cells subjected to long-term confinement remains unclear at present. To investigate this, photopatterning and microfluidics are employed to fabricate a high-throughput device that circumvents limitations of previous cell confinement models and enables prolonged culture of single cells in microchannels with physiologically relevant length scales. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2023<\/div>

Population context drives cell-to-cell variability in interferon response in epithelial cells<\/h2>\r\n
Camila Metz-Zumaran, Patricio Doldan, Francesco Muraca, Yagmur Keser, Pascal Lukas, Benno Kuropka, Leonie K\u00fcchenhoff, Soheil Rastgou Talemi, Thomas H\u00f6fer, Christian Freund, Elisabetta Ada Cavalcanti-Adam, Frederik Graw, Megan Stanifer, Steeve Boulant<\/div>
Isogenic cells respond in a heterogeneous manner to interferon. Using a micropatterning approach combined with high-content imaging and spatial analyses, we characterized how the population context (position of a cell with respect to the neighboring cells) of human intestinal epithelial cells affects single cell response to interferons. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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biorxiv, 2023<\/div>

Identification of the intracellular protein targets of a bio-active clickable half-sandwich iridium complex by chemical proteomics<\/h2>\r\n
Robin Ramos, Anthi Karaiskou, Candice Botuha, Micha\u00ebl Trichet, Florent Dingli, J\u00e9r\u00e9my Fort\u00e9, France Lam, Alexis Canette, Chlo\u00e9 Chaumeton, Murielle Salome, Thomas Chenuel, C\u00e9line Bergonzi, Philippe Meyer, Sylvain Bohic, Damarys Loew, Mich\u00e8le Salmain and Jo\u00eblle Sobczak-Th\u00e9pot<\/div>
Identification of intracellular targets of anticancer drug candidates provides key information on their mechanism of action. Exploiting the ability of the anticancer (C^N)-chelated half-sandwich iridium(III) complexes to covalently bind proteins, click chemistry with a bioorthogonal azido probe was used to localize a phenyloxazoline-chelated iridium complex within cells and profile its interactome at the proteome-wide scale. Proteins involved in protein folding and actin cytoskeleton regulation were identified as high affinity targets. Upon iridium complex treatment, HSP90 folding activity was inhibited\u00a0in vitro\u00a0and major cytoskeleton disorganization was observed. We used a wide array of imaging and biochemical methods to validate selected targets and obtain a multiscale overview of the effects of this complex on live human cells. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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ACS Applied Nano Materials, 2023<\/div>

Micropatterning of Quantum Dots for Biofunctionalization and Nanoimaging<\/h2>\r\n
Paul Robineau, J\u00e9r\u00e9mie B\u00e9al, Thomas Pons, Rodolphe Jaffiol and Cyrille V\u00e9zy<\/div>
Micron-scale patterning of colloidal quantum dots (QDs) is extremely important for the fabrication of high-performance Quantum dot Light-Emitting Diode (QLED) displays, biosensing, and super-resolution imaging. Thus, several nondestructive methods have been recently proposed, such as spatial self-organization. However, none of them can be useful for biofunctionalization or nanoimaging. To address this limitation, we propose a method to create micropatterns of QDs of any shape and size. UV photolithography assisted by a digital micromirror device (DMD) and silanization allow creating an adhesive layer, on which QDs micropatterns can be assembled with a 2 \u03bcm resolution.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Thesis, 2023<\/div>

Engineered 3D-Vessels-on-Chip to study effects of dynamic fluid flow on human induced pluripotent stem cell derived endothelial cells<\/h2>\r\n
Mees N. S. de Graaf<\/div>

Realistic models of the human vasculature would benefit understanding of normal physiology and disease pathology in the blood circulatory systems. Here we used a photo patterning system to form near vertical hydrogel walls inside a microfluidic device, generating a perfusable network. The hydrogels had realistic (tissue-like) viscoelastic properties and were permeable to large molecules like 70kD dextran. Endothelial cells from human pluripotent stem cells (hiPSC-ECs) cultured in the hydrogels could be guided to form complex networks. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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Biorxiv, 2023<\/div>

Self-organization of Long-lasting Human Endothelial Capillary Networks guided by DLP Bioprinting<\/h2>\r\n
Elsa Mazari-Arrighi, Matthieu L\u00e9pine, Dmitry Ayollo, Lionel Faivre, J\u00e9r\u00f4me Larghero, Fran\u00e7ois Chatelain, Alexandra Fuchs<\/div>
Tissue engineering holds great promise for regenerative medicine, drug discovery and as an alternative to animal models. However, as soon as the dimensions of engineered tissue exceed the diffusion limit of oxygen and nutriments, a necrotic core forms leading to irreversible damage. To overcome this constraint, the establishment of a functional perfusion network is essential and is a major challenge to be met. In this work, we explore a promising Digital Light Processing (DLP) bioprinting approach to encapsulate endothelial progenitor cells (EPCs) in 3D photopolymerized hydrogel scaffolds to guide them towards vascular network formation. We observed that EPCs encapsulated in the appropriate photopolymerized hydrogel can proliferate and self-organize within a few days into branched tubular structures with predefined geometry, forming capillary-like vascular tubes or trees of various diameters (in the range of 10 to 100 \u03bcm). \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biophysical Journal, 2023<\/div>

Engineering cell morphology using maskless 2D protein micropatterning on 3D nanostructures<\/h2>\r\n
Einollah Sarikhani, Lasse Klausen, Dhivya Pushpa Meganathan, Abel Marquez Serrano, Ching-Ting Tsai, Bianxiao Cui, Zeinab Jahed<\/div>
Mechanical cues such as the 2D and 3D shape of cellular microenvironments affect several cellular processes including adhesion and proliferation. Recent studies provide controlled conditions to recapitulate the 2D and 3D microenvironments to understand the mechanisms of cellular response to these mechanical cues. (\u2026) In this study, we present a micropatterning technique on nanostructured surfaces based on a maskless laser-assisted technique to study the cellular response to 3D nano-topographies in a controlled 2D microenvironment. We used a two-step dry and wet etching technique to fabricate transparent (SiO2) 3D nanostructured surfaces. Next, we micropatterned extracellular matrix proteins directly on the fabricated nanostructures by maskless micropatterning (PRIMO, Alv\u00e9ole) system mounted on an inverted microscope. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Communications, 2023<\/div>

Plasma FIB milling for the determination of structures in situ<\/h2>\r\n
Casper Berger, Maud Dumoux, Thomas Glen, Neville B.-y. Yee, John M. Mitchels, Zuzana Pat\u00e1kov\u00e1, Michele C. Darrow, James H Naismith, Michael Grange<\/div>
Structural biology studies inside cells and tissues require methods to thin vitrified specimens to electron transparency. Until now, focused ion beams based on gallium have been used. However, ion implantation, changes to surface chemistry and an inability to access high currents limit gallium application. Here, we show that plasma-coupled ion sources can produce cryogenic lamellae of vitrified human cells in a robust and automated manner, with quality sufficient for pseudo-atomic structure determination. Lamellae were produced in a prototype microscope equipped for long cryogenic run times (>\u00a01\u2009week) and with multi-specimen support fully compatible with modern-day transmission electron microscopes. We demonstrate that plasma ion sources can be used for structural biology within cells, determining a structure in situ to 4.9\u2009\u00c5, and characterise the resolution dependence on particle distance from the lamella edge. We describe a workflow upon which different plasmas can be examined to further streamline lamella fabrication. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2023<\/div>

The distance between the plasma membrane and the actomyosin cortex acts as a nanogate to control cell surface mechanics<\/h2>\r\n
Sergio Lembo, L\u00e9anne Strauss, Dorothy Cheng, Joseph Vermeil, Marc Siggel, Mauricio Toro-Nahuelpan, Chii Jou Chan, Jan Kosinski, Matthieu Piel, Olivia Du Roure, Julien Heuvingh, Julia Mahamid and Alba Diz-Mu\u00f1oz<\/div>
Animal cell shape changes are controlled by the actomyosin cortex, a peripheral actin network tethered to the plasma membrane by membrane-to-cortex attachment (MCA) proteins. Previous studies have focused on how myosin motors or actin turnover can generate the local deformations required for morphogenesis. However, how the cell controls local actin nucleation remains poorly understood. By combining molecular engineering with biophysical approaches and\u00a0in situ cryo-electron tomography characterization of cortical actin network architecture, we show that membrane-to-cortex tethering determines the distance between the plasma membrane and the actomyosin cortex at the nanoscale of single actin nucleators. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2023<\/div>

Notch1 cortical signaling regulates epithelial architecture and cell-cell adhesion<\/h2>\r\n
Matthew J. White, Kyle A. Jacobs, Tania Singh, Matthew L. Kutys<\/div>
Notch receptors control tissue morphogenic processes that involve coordinated changes in cell architecture and gene expression, but how a single receptor can produce these diverse biological outputs is unclear. Here we employ an organotypic microfluidic platform capable of recapitulating and dissecting three-dimensional (3D) morphogenic features of a ductal epithelium, to reveal tissue morphogenic defects result from loss of Notch1, but not Notch1 transcriptional signaling. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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ACS Synth. Biol., 2023<\/div>

Extracellular Cues Govern Shape and Cytoskeletal Organization in Giant Unilamellar Lipid Vesicles<\/h2>\r\n
Andreas Fink, Charlotte R. Doll, Ana Yag\u00fce Relimpio, Yannik Dreher, Joachim P. Spatz, Kerstin G\u00f6pfrich, and Elisabetta Ada Cavalcanti-Adam<\/div>
Spontaneous and induced front-rear polarization and a subsequent asymmetric actin cytoskeleton is a crucial event leading to cell migration, a key process involved in a variety of physiological and pathological conditions such as tissue development, wound healing, and cancer. Migration of adherent cells relies on the balance between adhesion to the underlying matrix and cytoskeleton-driven front protrusion and rear retraction. A current challenge is to uncouple the effect of adhesion and shape from the contribution of the cytoskeleton in regulating the onset of front-rear polarization. Here, we present a minimal model system that introduces an asymmetric actin cytoskeleton in synthetic cells, which are resembled by giant unilamellar lipid vesicles (GUVs) adhering onto symmetric and asymmetric micropatterned surfaces. Surface micropatterning of streptavidin-coated regions with varying adhesion shape and area was achieved by maskless UV photopatterning. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biomaterials, 2023<\/div>

Bioengineering a miniaturized in vitro 3D myotube contraction monitoring chip to model muscular dystrophies<\/h2>\r\n
Nicolas Rose, Berenice Estrada Chavez, Surabhi Sonam, Thao Nguyen, Gianluca Grenci, Anne Bigot, Antoine Muchir, Beno\u00eet Ladoux, Bruno Cadot, Fabien Le Grand, L\u00e9a Trichet<\/div>
Quantification of skeletal muscle functional contraction is essential to assess the outcomes of therapeutic procedures for neuromuscular disorders. Muscle three-dimensional \u201cOrgan-on-chip\u201d models usually require a substantial amount of biological material, which rarely can be obtained from patient biopsies. Here, we developed a miniaturized 3D myotube culture chip with contraction monitoring capacity at the single cell level. Optimized micropatterned substrate design enabled to obtain high culture yields in tightly controlled microenvironments, with myotubes derived from primary human myoblasts displaying spontaneous contractions.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2022<\/div>

Friction patterns guide actin network contraction<\/h2>\r\n
Alexandra Colin, Magali Orhant-Prioux, Christophe Gu\u00e9rin, Mariya Savinov, Ilaria Scarfone, Aurelien Roux, Enrique M. De La Cruz, Alex Mogilner, Manuel Th\u00e9ry and Laurent Blanchoin<\/div>
The shape of cells is the outcome of the balance of inner forces produced by the actomyosin network and the resistive forces produced by cell adhesion to their environment. The specific contributions of contractile, anchoring and friction forces to network deformation rate and orientation are difficult to disentangle in living cells where they influence each other. Here, we reconstituted contractile acto-myosin networks\u00a0in vitro to study specifically the role of the friction forces between the network and its anchoring substrate. To modulate the magnitude and spatial distribution of friction forces, we micropatterned actin nucleation promoting factors on glass or on a lipid bilayer. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Communications, 2022<\/div>

Morphological control enables nanometer-scale dissection of cell-cell signaling complexes<\/h2>\r\n
Liam P. Dow, Guido Gaietta, Yair Kaufman, Mark F. Swift, Moara Lemos, Kerry Lane, Matthew Hopcroft, Armel Bezault, C\u00e9cile Sauvanet, Niels Volkmann, Beth L. Pruitt & Dorit Hanein<\/div>
Protein micropatterning enables robust control of cell positioning on electron-microscopy substrates for cryogenic electron tomography (cryo-ET). However, the combination of regulated cell boundaries and the underlying electron-microscopy substrate (EM-grids) provides a poorly understood microenvironment for cell biology. Because substrate stiffness and morphology affect cellular behavior, we devised protocols to characterize the nanometer-scale details of the protein micropatterns on EM-grids by combining cryo-ET, atomic force microscopy, and scanning electron microscopy. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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STAR Protocols, 2022<\/div>

Protocol for live-cell fluorescence-guided cryoFIB-milling and electron cryo-tomography of virus-infected cells<\/h2>\r\n
Linda E. Franken, Rene Rosch, Ulrike Laugks, Kay Gr\u00fcnewald<\/div>
Here, we present a protocol for assessing virus-infected cells using electron cryo-tomography (cryoET). It includes the basic workflows of seeding cells, plunge-freezing, clipping, cryo-focused ion beam milling (cryoFIB-milling), and cryoET, as well as two optional modules: micropatterning and live-cell fluorescence microscopy. We use an A549 human cell line and the virus HAdV5-pIX-mcherry in this protocol, but the comprehensive workflow can be easily transferred to other cell types and different types of virus infection or treatment. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Physics, 2022<\/div>

Mechanical stress driven by rigidity sensing governs epithelial stability<\/h2>\r\n
Surabhi Sonam, Lakshmi Balasubramaniam, Shao-Zhen Lin, Ying Ming Yow Ivan, Irina Pi-Jaum\u00e0, Cecile Jebane, Marc Karnat, Yusuke Toyama, Philippe Marcq, Jacques Prost, Ren\u00e9-Marc M\u00e8ge, Jean-Fran\u00e7ois Rupprecht & Beno\u00eet Ladoux<\/div>
Epithelia act as barriers against environmental stresses. They are continuously exposed to various mechanical stress and abrasion, which impact epithelial integrity. The impact of the environment on epithelial integrity remains elusive. By culturing epithelial cells on two-dimensional hydrogels, we observe a loss of epithelial monolayer integrity on soft substrates through spontaneous hole formation. These monolayer ruptures are associated with local cellular stretching and cell-division events. (\u2026) Our results thus show that substrate stiffness provides feedback on the mechanical state of epithelial monolayers with potential application towards a mechanistic understanding of compromised epithelial integrity during normal and pathological human ontogenesis. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Cell Reports, 2022<\/div>

A mechanical G2 checkpoint controls epithelial cell division through E-cadherin-mediated regulation of Wee1-Cdk1<\/h2>\r\n
Lisa Donker, Ronja Houtekamer, Marjolein J. Vliem, Fran\u00e7ois Sipieter, Helena Canever, Manuel G\u00f3mez-Gonz\u00e1lez, Miquel Bosch-Padr\u00f3s, Willem-Jan Pannekoek, Xavier Trepat, Nicolas Borghi, Martijn Gloerich<\/div>
Epithelial cell divisions are coordinated with cell loss to preserve epithelial integrity. However, how epithelia adapt their rate of cell division to changes in cell number, for instance during homeostatic turnover or wounding, is not well understood. Here, we show that epithelial cells sense local cell density through mechanosensitive E-cadherin adhesions to control G2\/M cell-cycle progression. Micropatterning is used for monolayer stress microscopy, to map those intercellular forces. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Current Opinion in Structural Biology, 2022<\/div>

Electron microscopy of cellular ultrastructure in three dimensions<\/h2>\r\n
Neta Varsano, Sharon Grayer Wolf<\/div>
Electron microscopy in three dimensions (3D) of cells and tissues can be essential for understanding the ultrastructural aspects of\u00a0biological processes. The quest for 3D information reveals challenges at many stages of the workflow, from sample preparation, to imaging, data analysis and segmentation. Here, we outline several available methods, including volume SEM imaging, cryo-TEM and cryo-STEM tomography, each one occupying a different domain in the basic tradeoff between field-of-view and resolution. We discuss the considerations for choosing a suitable method depending on research needs and highlight recent developments that are essential for making 3D volume imaging of cells and tissues a standard tool for cellular and structural biologists. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Thesis, 2022<\/div>

Spatial control of angiogenesis by engineered patterns of Notch ligands<\/h2>\r\n
Laura Tiemeijer<\/div>
In tissue engineering and regenerative medicine, proper vascularization of engineered tissues is imperative, as diffusion of nutrients, oxygen and waste is limited in constructs larger than a few cells thick. Therefore, tissue vascularization has been extensively researched. However, most approaches rely on preset structural support for the cells provided by scaffolds and microfluidic chips to instruct vascular organization, which limit the integration into engineered and native tissues. (\u2026) In this thesis, we have developed an in vitro method where we used spatial patterns of parallel lines of Notch signaling ligands to locally modulate endothelial tip\/stalk cell selection and thereby gained spatial control over endothelial sprouting.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Cell Reports, 2022<\/div>

Pressure and curvature control of the cell cycle in epithelia growing under spherical confinement<\/h2>\r\n
Ilaria Di Meglio, Anastasiya Trushko, Pau Guillamat, Carles Blanch-Mercader, Shada Abuhattum, Aur\u00e9lien Roux<\/div>
Morphogenesis requires spatiotemporal regulation of proliferation, both by biochemical and mechanical cues. In epithelia, this regulation is called contact inhibition of proliferation, but disentangling biochemical from mechanical cues remains challenging. Here, we show that epithelia growing under confinement accumulate pressure that inhibits proliferation above a threshold value. (\u2026)\u00a0Altogether, our results suggest that different mechanical cues resulting from pressure inhibition of proliferation are at play through different mechano-sensing pathways: the \u03b2-catenin pathway sustains cell division under high pressure, and the\u00a0YAP pathway senses local curvature. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Communications, 2022<\/div>

Intrinsic cell rheology drives junction maturation<\/h2>\r\n
K. Sri-Ranjan, J. L. Sanchez-Alonso, P. Swiatlowska, S. Rothery, P. Novak , S. Gerlach, D. Koeninger, B. Hoffmann, R. Merkel, M. Stevens, S. X. Sun, J. Gorelik, Vania Braga<\/div>
A fundamental property of higher eukaryotes that underpins their evolutionary success is stable cell-cell cohesion. Yet, how intrinsic cell rheology and stiffness contributes to junction stabilization and maturation is poorly understood. We demonstrate that localized modulation of cell rheology governs the transition of a slack, undulated cell-cell contact (weak adhesion) to a mature, straight junction (optimal adhesion). Cell pairs con\ufb01ned on different geometries have heterogeneous elasticity maps and control their own intrinsic rheology co-ordinately. (\u2026) Our data inform on the minimal intrinsic rheology to generate a mature junction and provide a springboard towards understanding elements governing tissue-level mechanics. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2022<\/div>

Cryo-electron tomography reveals enrichment and identifies microtubule lumenal particles in neuronal differentiation<\/h2>\r\n
Saikat Chakraborty, Antonio Martinez-Sanchez, Florian Beck, Mauricio Toro-Nahuelpan, In-Young Hwang, Kyung-Min Noh, Wolfgang Baumeister, Julia Mahamid<\/div>
Functional architecture of the neuronal microtubule (MT) cytoskeleton is maintained by various MT-associated proteins (MAPs), most of which bind to the MT outer surface. Yet, electron microscopy (EM) has long revealed hitherto unknown electron-dense particles inside the lumens of neuronal MTs. Here, we use cryogenic electron tomography (cryo-ET) to analyze the native three-dimensional (3D) structures and organization of MT lumenal particles inside vitrified rodent primary neurons, pluripotent P19 cells and human induced pluripotent stem cell-derived neurons. We obtain 3D maps of several lumenal particles at molecular resolution that periodically decorate neuronal MTs. We show that increased lumenal particle localization is concomitant with neuronal differentiation and correlates with higher MT curvatures. Lumenal particles binding topology in MTs, their structural resemblance to tubulin binding cofactors (TBCs), enrichment around MT lattice defects and at plus-ends indicated their potential role in tubulin proteostasis for the maintenance of neuronal MTs. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Trends in Cell Biology, 2022<\/div>

Mechanobiological approaches to synthetic morphogenesis: learning by building<\/h2>\r\n
Marija Matej\u010di\u0107, Xavier Trepat<\/div>
Tissue morphogenesis occurs in a complex physicochemical microenvironment with limited experimental accessibility. This often prevents a clear identification of the processes that govern the formation of a given functional shape. By applying state-of-the-art methods to minimal tissue systems, synthetic morphogenesis aims to engineer the discrete events that are necessary and sufficient to build specific tissue shapes. Here, we review recent advances in synthetic morphogenesis, highlighting how a combination of microfabrication and mechanobiology is fostering our understanding of how tissues are built. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Current Biology, 2022<\/div>

MARK2 regulates directed cell migration through modulation of myosin II contractility and focal adhesion organization<\/h2>\r\n
Ana M. Pasapera, Sarah M. Heissler, Masumi Eto, Yukako Nishimura, Robert S. Fischer, Hawa R. Thiam, Clare M. Waterman<\/div>
Cancer cell migration during metastasis is mediated by a highly polarized\u00a0cytoskeleton. MARK2 and its invertebrate homolog Par1B are kinases that regulate the microtubule cytoskeleton to mediate polarization of neurons in mammals and embryos in invertebrates. However, the role of MARK2 in cancer cell migration is unclear. Using\u00a0osteosarcoma\u00a0cells, we found that in addition to its known localizations on microtubules and the\u00a0plasma membrane, MARK2 also associates with the\u00a0actomyosin\u00a0cytoskeleton and\u00a0focal adhesions. (\u2026) Together, our results define MARK2 as a master regulator of the actomyosin and microtubule cytoskeletal systems and focal adhesions to mediate directional cancer cell migration. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Lab on a Chip, 2022<\/div>

Microfluidic harvesting of breast cancer tumor spheroid-derived extracellular vesicles from immobilized microgels for single-vesicle analysis<\/h2>\r\n
Xilal Y. Rima, Jingjing Zhang, Luong TH Nguyen, Aaron Rajasuriyar, Min Jin Yoon, Chi-Ling Chiang, Nicole Walters, Kwang Joo Kwak, L.James Lee, Eduardo Re\u00e1tegui<\/div>
Investigating cellular and vesicular heterogeneity in breast cancer remains a challenge, which encourages the development of controllable\u00a0in vitro\u00a0systems that mimic the tumor microenvironment. Although three-dimensional cell culture better recapitulates the heterogeneity observed in tumor growth and extracellular vesicle (EV) biogenesis, the physiological relevance is often contrasted with the control offered by two-dimensional cell culture. Therefore, to challenge this misconception we developed a novel microfluidic system harboring highly tunable three-dimensional EV microbioreactors (EV\u00b5BRs) to model micrometastatic EV release in breast cancer while capitalizing on the convenient, low-volume, and sterile interface provided by microfluidics. (\u2026)\u00a0To immobilize the\u00a0EV\u00b5BRs within a microchannel and facilitate EV extraction, oxygen inhibition in free-radical polymerization was repurposed to rapidly generate two-layer hydrodynamic traps\u00a0in situ using a digital-micromirror device (DMD)-based ultraviolet (UV) projection system. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Methods in Molecular Biology, 2022<\/div>

Controlling Cell Shape and Microtubule Organization by Extracellular Matrix Micropatterning<\/h2>\r\n
Alessandro Dema, Shima Rahgozar, Laurent Siquier, Jeffrey van Haren, Torsten Wittmann<\/div>
Micropatterning of extracellular matrix proteins enables defining cell position and shape in experiments investigating intracellular dynamics and organization. While such standardization is advantageous in automated and quantitative analysis of many cells, the original methods generating such patterns are cumbersome and inflexible. However, recent development of contact-less methods that allow photochemical generation of protein patterns robustly and rapidly is boosting the broader availability of micropatterning approaches. Here, we describe an optimized protocol to achieve large micropatterned areas with high fidelity using a commercially available microscope-mounted UV projection system. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Adv. Funct. Mater., 2022<\/div>

Printing and Erasing of DNA-Based Photoresists Inside Synthetic Cells<\/h2>\r\n
Tobias Walther, Kevin Jahnke, Tobias Abele, Kerstin G\u00f6pfrich<\/div>
In the pursuit of producing functioning synthetic cells from the bottom-up, DNA nanotechnology has proven to be a powerful tool. However, the crowded yet highly organized arrangement in living cells, bridging from the nano- to the micron-scale, remains challenging to recreate with DNA-based architectures. Here, laser microprinting is established to print and erase shape-controlled DNA hydrogels inside the confinement of water-in-oil droplets and giant unilamellar lipid vesicles (GUVs). (\u2026) Overall, DNA-based photoresists for laser printing in confinement allow to build up architectures on the interior of synthetic cells with light, which diversifies the toolbox of bottom-up synthetic biology. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biorxiv, 2022<\/div>

Conserved basal lamina proteins, laminin and nidogen, are repurposed to organize mechanosensory complexes responsible for touch sensation<\/h2>\r\n
Alakananda Das, Joy Franco, Lingxin Wang, Dail Chapman, Lucy Wang, Chandni Jaisinghani, Beth Pruitt, Miriam Goodman<\/div>
The sense of touch is conferred by the conjoint function of somatosensory neurons and skin cells. These cells meet across a gap filled by a basal lamina, an ancient structure found in all metazoans. Using\u00a0Caenorhabditis elegans nematodes, we show that mechanosensory complexes essential for touch sensation reside at this interface and contain laminin, nidogen, and the MEC-4 mechano-electrical transduction channel proteins. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Materials, 2022<\/div>

Integer topological defects organize stresses driving tissue morphogenesis<\/h2>\r\n
Pau Guillamat, Carles Blanch-Mercader, Guillaume Pernollet, Karsten Kruse, Aur\u00e9lien Roux<\/div>
Tissues acquire function and shape via differentiation and morphogenesis. Both processes are driven by coordinating cellular forces and shapes at the tissue scale, but general principles governing this interplay remain to be discovered. Here we report that self-organization of myoblasts around integer topological defects, namely spirals and asters, suffices to establish complex multicellular architectures. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Jove, 2022<\/div>

Control of Cell Adhesion using Hydrogel Patterning Techniques for Applications in Traction Force Microscopy<\/h2>\r\n
Joel Christian, Johannes W. Blumberg, Dimitri Probst, Cristina Lo Giudice, Sandra Sindt, Christine Selhuber-Unkel, Ulrich S. Schwarz, Elisabetta Ada Cavalcanti-Adam<\/div>
For 2D-TFM (Traction Force Microscopy) on polyacrylamide, the difficulty in achieving high throughput results mainly from the large variability of cell shapes and tractions, calling for standardization. We present a protocol to rapidly and efficiently fabricate micropatterned PA hydrogels for 2D-TFM studies. The micropatterns are first created by maskless photolithography on PA hydrogels of different elasticity, and their displacement is tracked by embedded fluorescent beads. To further achieve precise recording of cell forces, we describe the use of a controlled dose of patterned light to release cell tractions in defined regions for single cells or groups of cells. We call this method local UV illumination traction force microscopy (LUVI-TFM). \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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PNAS, 2022<\/div>

ATP allosterically stabilizes integrin-linked kinase for efficient force generation<\/h2>\r\n
Isabel M. Martin, Michele M. Nava, Sara A. Wickstr\u00f6m, Frauke Gr\u00e4ter<\/div>
Focal adhesions link the actomyosin cytoskeleton to the extracellular matrix regulating cell adhesion, shape, and migration. Adhesions are dynamically assembled and disassembled in response to extrinsic and intrinsic forces, but how the essential adhesion component integrin-linked kinase (ILK) dynamically responds to mechanical force and what role adenosine triphosphate (ATP) bound to this pseudokinase plays remain elusive. Here, we apply force\u2013probe molecular-dynamics simulations of human ILK:\u03b1-parvin coupled to traction force microscopy to explore ILK mechanotransducing functions. Our study proposes a role of ATP as an obligatory binding partner for structural and mechanical integrity of the pseudokinase ILK, ensuring efficient cellular force generation and migration. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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ACS Appl. Mater. Interfaces, 2022<\/div>

Facile and Versatile Method for Micropatterning Poly(acrylamide) Hydrogels Using Photocleavable Comonomers<\/h2>\r\n
Dimitris Missirlis, Miguel Ban\u0303os, Felix Lussier, Joachim Spatz<\/div>
We here present a micropatterning strategy to introduce small molecules and ligands on patterns of arbitrary shapes on the surface of poly(acrylamide)-based hydrogels. To achieve the above, a monomer containing a caged amine was co-polymerized in the hydrogel network; upon UV light illumination using a commercially available setup, primary amines were locally deprotected and served as reactive groups for further functionalization. Cell patterning on various cell adhesive ligands was demonstrated, with cells responding to a combination of pattern shape and substrate elasticity. The approach is compatible with standard traction force microscopy (TFM) experimentation and can further be extended to reference-free TFM. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Micromachines, 2021<\/div>

Rapid Prototyping of Organ-on-a-Chip Devices Using Maskless Photolithography<\/h2>\r\n
Dhanesh Kasi, Mees de Graaf, Paul Motreuil-Ragot, Jean-Phillipe Frimat, Michel Ferrari, Pasqualina Sarro, Massimo Mastrangeli, Arn M. J. M. van den Maagdenberg, Christine Mummery, Valeria Orlova<\/div>
Organ-on-a-chip (OoC) and microfluidic devices are conventionally produced using microfabrication procedures that require cleanrooms, silicon wafers, photomasks, and multiple iterations of design steps. Here, we describe a rapid and cleanroom-free microfabrication method using a commercial digital micromirror device-based setup. Using this approach: digital photomasks can be designed, projected, and quickly
adjusted if needed; and SU-8 molds can be fabricated without cleanroom availability, which in turn reduces microfabrication time and costs and expedites prototyping of new OoC devices. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biomaterials, 2021<\/div>

Construction of functional biliary epithelial branched networks with predefined geometry using digital light stereolithography<\/h2>\r\n
Elsa Mazari-Arrighi, Dmitry Ayollo, Wissam Farhat, Auriane Marret, Emilie Gontran, Pascale Dupuis-Williams, Jerome Larghero, Francois Chatelain, Alexandra Fuchs<\/div>
Cholangiocytes, biliary epithelial cells, are known to spontaneously self-organize into spherical cysts with a central lumen. In this work, we explore a promising biocompatible stereolithographic approach to encapsulate cholangiocytes into geometrically-controlled 3D hydrogel structures to guide them towards the formation of branched tubular networks. We demonstrate that within the appropriate mix of hydrogels, normal rat cholangiocytes can proliferate, migrate and organize into branched tubular structures, form walls consisting of a cell monolayer, transport fluorescent dyes into the luminal space and show markers of epithelial maturation such as primary cilia. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Journal of Cell Biology, 2021<\/div>

Hematopoietic progenitors polarize in contact with bone marrow stromal cells in response to SDF1<\/h2>\r\n
Thomas Bessy, Adrian Candelas, Benoit Souquet, Khansa Saadallah, Alexandre Schaeffer, Benoit Vianay, Damien Cuvelier, Samy Gobaa, Cecilia Nakid-Cordero, Julien Lion, Jean-Christophe Bories, Nuala Mooney, Thierry Jaffredo, Jerome Larghero, Laurent Blanchoin, Lionel Faivre, Stephane Brunet, Manuel Th\u00e9ry<\/div>
Hematopoietic stem and progenitor cells (HSPCs) are located in the bone marrow, where they regulate the permanent production and renewal of all blood-cell types. HSPC proliferation and differentiation is locally regulated by their interaction with cells forming specific microenvironments close to the bone matrix or close to blood vessels. However, the cellular mechanisms underlying HSPC\u2019s interaction with these cells and their potential impact on HSPC polarity is still poorly understood. Here we modeled the bone-marrow niche using microfluidic technologies in a bone-marrow on a chip device. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Journal of structural biology, 2021<\/div>

Lattice micropatterning for cryo-electron tomography studies of cell-cell contacts<\/h2>\r\n
Leeya Engel, Claudia G. Vasquez, Elizabeth A. Montabana, Belle M. Sow, Marcin P. Walkiewicz, William I. Weis, Alexander R. Dunn<\/div>
Cryo-electron tomography is the highest resolution tool available for structural analysis of macromolecular complexes within their native cellular environment. At present, data acquisition suffers from low throughput, in part due to the low probability of positioning a cell such that the subcellular structure of interest is on a region of the electron microscopy (EM) grid that is suitable for imaging. Here, we leverage photo-micropatterning of EM grids to optimally position endothelial cells to enable high-throughput imaging of cell-cell contacts. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Jove, 2021<\/div>

Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows<\/h2>\r\n
Bryan S. Sibert, Joseph Y. Kim, Jie E. Yang, Elizabeth R. Wright<\/div>
There are challenges associated with culturing and\/or adhering cells onto TEM grids in a manner that is suitable for tomography while retaining the cells in their physiological state. Here, a detailed step-by-step protocol is presented on the use of micropatterning to direct and promote eukaryotic cell growth on TEM grids. Flexibility in the choice of surface coating and pattern design makes micropatterning broadly applicable for a wide range of cell types. Micropatterning is useful for studies of structures within individual cells as well as more complex experimental systems such as host-pathogen interactions or differentiated multi-cellular communities. Micropatterning may also be integrated into many downstream whole-cell cryo-ET workflows, including correlative light and electron microscopy (cryo-CLEM) and focused-ion beam milling (cryo-FIB). \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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nano letters, 2021<\/div>

Nanoscale Surface Topography Reduces Focal Adhesions and Cell Stiffness by Enhancing Integrin Endocytosis<\/h2>\r\n
Xiao Li, Lasse H. Klausen, Wei Zhang, Zeinab Jahed, Ching-Ting Tsai, Thomas L. Li, and Bianxiao Cui<\/div>
Both substrate stiffness and surface topography regulate cell behavior through mechanotransduction signaling pathways. However, the mechanisms by which cells recognize topographical features are not fully understood. Here we demonstrate that the presence of nanotopography drastically alters cell behavior such that neurons and stem cells cultured on rigid glass substrates behave as if they were on soft hydrogels. We further show that rigid nanotopography resembles the effect of soft hydrogels in reducing cell stiffness and membrane tension as measured by atomic force microscopy. Finally, we demonstrate that nanotopography reduces focal adhesions and cell stiffness by enhancing the endocytosis and the subsequent removal of integrin receptors. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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ACS Appl. Mater. Interfaces, 2021<\/div>

Composite Elastomer-Enabled Rapid Photofabrication of Microfluidic Devices<\/h2>\r\n
Futianchun Zhu , Yu He, Zefan Lu, Hongliang Fan, Tao Zhang<\/div>
Recently, photocurable resins, as a huge class of materials, have attracted extensive interest. However, very few of them can now be used in device fabrication due to the challenge in machining these materials. In response, we herein propose a novel concept of composite elastomers, which can covalently link with and consequently offer a flexible support to photocured thin films. This effect would allow most photocurable resins to be used in microfluidic device fabrication, greatly enriching the material choices for diverse applications. Moreover, the whole fabrication process becomes very simple and rapid, with an impressive throughput of at least hundreds of replicas per day.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biology of the Cell, 2021<\/div>

Direct measurement of near-nano-Newton forces developed by self-organizing actomyosin fibers bound \u03b1-catenin<\/h2>\r\n
Surabhi Sonam, Cl\u00e9mence Vigouroux, Antoine J\u00e9gou, Guillaume Romet-Lemonne, Christophe Le Clainche, Benoit Ladoux, Ren\u00e9 Marc M\u00e8ge<\/div>

Actin cytoskeleton contractility plays a critical role in morphogenetic processes by generating forces that are then transmitted to cell\u2013cell and cell-ECM adhesion complexes. In turn, mechanical properties of the environment are sensed and transmitted to the cytoskeleton at cell adhesion sites, influencing cellular processes such as cell migration, differentiation and survival. (\u2026) A key issue is to be able to measure the forces generated by actomyosin and transmitted to the adhesion complexes. Here, we applied an intermediate approach allowing reconstruction of the actomyosin-\u03b1-catenin complex in acellular conditions to probe directly the transmitted forces. For this, we combined micropatterning of purified \u03b1-catenin and spontaneous actomyosin network assembly in the presence of G-actin and Myosin II with micro force sensor arrays used so far to measure cell-generated forces. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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biorxiv, 2021<\/div>

Whole-cell cryo-electron tomography of cultured and primary eukaryotic cells on micropatterned TEM grids<\/h2>\r\n
Bryan S Sibert, Joseph Y Kim, Jie E Yang, Elizabeth R Wright<\/div>
Culture or adhere cells on TEM grids in a manner that is suitable for tomography while preserving the physiological state of the cells remains a challenge in whole-cell cryo-electron tomography (cryo-ET). Here, we demonstrate the versatility of micropatterning to direct and promote growth of both cultured and primary eukaryotic cells on TEM grids, by studying host-pathogen interactions using respiratory syncytial virus infected BEAS-2B cells as an example. We demonstrate the ability to use whole-cell tomography of primary Drosophila neuronal cells to identify organelles and cytoskeletal stuctures in cellular axons and the potential for micropatterning to dramatically increase throughput for these studies. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Methods in Molecular Biology, 2021<\/div>

Manufacturing a Bone Marrow-On-A-Chip Using Maskless Photolithography<\/h2>\r\n
Benoit Souquet, Matthieu Opitz, Benoit Vianay, St\u00e9phane Brunet, Manuel Th\u00e9ry<\/div>
The bone marrow (BM) is a complex microenvironment in which hematopoietic stem and progenitor cells (HSPCs) interact with multiple cell types that regulate their quiescence, growth, and differentiation. These cells constitute local niches where HSPCs are confined and subjected to specific set of physical and biochemical cues. (\u2026) Here, we present a method to manufacture a pseudo BM-on-a-chip with separated compartments mimicking the vascular and the endosteal niches. Such a configuration with connected but distant compartments allowed the investigation of the specific contribution of each niche to the regulation of HSPC behavior in vitro. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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PNAS, 2021<\/div>

Contractility, focal adhesion orientation, and stress fiber orientation drive cancer cell polarity and migration along wavy ECM substrates<\/h2>\r\n
Robert Fischer, Xiaoyu Sun, Michelle Baird, Matt Hourwitz, Bo Ri Seo, Ana Pasapera, Shalin Mehta, Wolfgang Losert, Claudia Fischbach, John Fourkas, and Clare Waterman<\/div>
Contact guidance is a powerful topographical cue that induces persistent directional cell migration. Healthy tissue stroma is characterized by a meshwork of wavy extracellular matrix (ECM) fiber bundles, whereas metastasis-prone stroma exhibit less wavy, more linear fibers. The latter topography correlates with poor prognosis, whereas more wavy bundles correlate with benign tumors. We designed nanotopographic ECM-coated substrates that mimic collagen fibril waveforms seen in tumors and healthy tissues to determine how these nanotopographies may regulate cancer cell polarization and migration machineries. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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ACS Applied Materials & Interfaces, 2021<\/div>

Protein Micropatterning in 2.5D: An Approach to Investigate Cellular Responses in Multi-Cue Environments<\/h2>\r\n
Cas van der Putten, Antonetta Buskermolen, Maike Werner, Hannah Brouwer, Paul Bartels, Patricia Dankers, Carlijn Bouten, and Nicholas Kurniawan<\/div>
Here, we present a new approach to investigate cellular responses in multi-cue environments, by combining optics-based protein patterning and lithography-based substrate microfabrication. Using a contactless and maskless UV-projection system, we created patterns of extracellular proteins (resembling contact-guidance cues) on a two-and-a-half-dimensional (2.5D) cell culture chip containing a library of well-defined microstructures (resembling topographical cues). \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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ACS Applied Materials & Interfaces, 2021<\/div>

Pattern-Based Contractility Screening, a Reference-Free Alternative to Traction Force Microscopy Methodology<\/h2>\r\n
Ajinkya Ghagre, Ali Amini, Luv Kishore Srivastava, Pouria Tirgar, Adele Khavari, Newsha Koushki, Allen Ehrlicher<\/div>
The sensing and generation of cellular forces are essential aspects of life. Traction Force Microscopy (TFM) has emerged as a standard broadly applicable methodology to measure cell contractility and its role in cell behavior. While TFM platforms have enabled diverse discoveries, their implementation remains limited in part due to various constraints, such as time-consuming substrate fabrication techniques, the need to detach cells to measure null force images, followed by complex imaging and analysis, and the unavailability of cells for post-processing. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Scientific Reports, 2021<\/div>

Collective migration during a gap closure in a two-dimensional haptotactic model<\/h2>\r\n
Marie Versaevel, Laura Alaimo, Valentine Seveau, Marine Luciano, Danahe Mohammed, C\u00e9line Bruy\u00e8re, El\u00e9onore Vercruysse, Olivier Th\u00e9odoly, Sylvain Gabriele<\/div>
The ability of cells to respond to substrate-bound protein gradients is crucial for many physiological processes, such as immune response, neurogenesis and cancer cell migration. Here we use a photopatterning technique to create well-controlled circular, square and linear fibronectin (FN) gradients on two-dimensional (2D) culture substrates, to understand collective cell migration in response to haptotaxis. Our findings provide a better understanding of the wound healing process over protein gradients, which are reminiscent of haptotaxis. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Jove, 2021<\/div>

Co-culture of Glioblastoma Stem-like Cells on Patterned Neurons to Study Migration and Cellular Interactions<\/h2>\r\n
Joris Guyon, Pierre-Olivier Strale, Irati Romero-Garmendia, Andreas Bikfalvi, Vincent Studer, Thomas Daubon<\/div>
Here, we present an easy-to-use co-culture assay to analyze glioblastoma (GBM) migration on patterned neurons. We developed a macro in FiJi software for easy quantification of GBM cell migration on neurons, and observed that neurons modify GBM cell invasive capacity. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Communications, 2021<\/div>

Mechanochemical control of epidermal stem cell divisions by B-plexins<\/h2>\r\n
Chen Jiang, Ahsan Javed, Laura Kaiser, Michele M. Nava, Rui Xu, Dominique T. Brandt, Dandan Zhao, Benjamin Mayer, Javier Fern\u00e1ndez-Baldovinos, Luping Zhou, Carsten H\u00f6\u00df, Kovilen Sawmynaden, Arkadiusz Oleksy, David Matthews, Lee S. Weinstein, Heidi Hahn, Hermann-Josef Gr\u00f6ne, Peter L. Graumann, Carien M. Niessen, Stefan Offermanns, Sara A. Wickstr\u00f6m, Thomas Worzfeld<\/div>

Epithelial cell divisions lead to tissue crowding and local changes in force distribution, which in turn suppress the rate of cell divisions. Our data define a central role of B-plexins in mechanosensation to couple cell density and cell division in development and disease. We identify a critical requirement of B-plexin transmembrane receptors in the response to crowding-induced mechanical forces during embryonic skin development. We show that B-plexins mediate mechanoresponses to crowding through stabilization of adhesive cell junctions and lowering of cortical stiffness; and provide evidence that the B-plexin-dependent mechanochemical feedback is also pathophysiologically relevant to limit tumor growth in basal cell carcinoma, the most common type of skin cancer. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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Cell Press, 2021<\/div>

Acto-myosin network geometry defines centrosome position<\/h2>\r\n
Ana Joaquina Jimenez, Alexandre Schaeffer, Chiara De Pascalis, ..., Matthieu Piel, Laurent Blanchoin, Manuel Th\u00e9ry<\/div>

Jimenez et al. show that the centrosome, which has long been thought to sit at the geometric center of the cell, is actually positioned at the center of a subcellular zone defined by the absence of contractile acto-myosin bundles. Centrosome position is defined by dyneins exerting pulling forces on microtubules specifically in this zone. \u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

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Methods in Microbiology, 2021<\/div>

Micropatterning of cells on EM grids for efficient cryo-correlative light electron microscopy<\/h2>\r\n
Lea Swistak, Anna Sartori-Ruppc, Matthijn Vos, Jost Enninga<\/div>
Cryo-transmission electron microscopy (cryo-TEM) provides access to high resolution information of adherent cell ultrastructures in a close to native environment but only volumes of less than 500\u00a0nm can be imaged. Cryo-focused ion beam (FIB) milling overcomes this obstacle through the generation of thin lamella of less than 200\u00a0nm. These lamellas can be imaged by cryo-electron tomography (cryo-ET) giving access to ultrastructural data within the volume of the imaged cells. Nevertheless, a lack of control on the positioning of the samples on the electron microscopy (EM) grids drastically constrains its throughput. The use of custom-designed micropatterned EM grids bypasses these issues by accurately positioning cells in areas that allow FIB milling followed by cryo-ET. Combined with fluorescent light microscopy in correlative light electron microscopy (CLEM) pipelines to pinpoint specific events and automated FIB milling, micropatterning of cells on EM grids has the potential of dramatically accelerating the workflow of cryo-ET. Here, a detailed description is provided of the key steps necessary to implement photomicropatterning of EM grids for improved CLEM pipelines. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Phys. Rev. Lett., 2021<\/div>

Quantifying Material Properties of Cell Monolayers by Analyzing Integer Topological Defects<\/h2>\r\n
Carles Blanch-Mercader, Pau Guillamat, Aur\u00e9lien Roux, and Karsten Kruse<\/div>
In developing organisms, internal cellular processes generate mechanical stresses at the tissue scale. The resulting deformations depend on the material properties of the tissue, which can exhibit long-ranged orientational order and topological defects. It remains a challenge to determine these properties on the time scales relevant for developmental processes. (\u2026) We illustrate our approach by analyzing monolayers of C2C12 cells in small circular confinements, where they form a single topological defect with integer charge. We find that such monolayers exert compressive stresses at the defect centers, where localized cell differentiation and formation of three-dimensional shapes is observed. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Scientific Reports, 2020<\/div>

A minimalist model to measure interactions between proteins and synaptic vesicles<\/h2>\r\n
Eleonora Perego, Sofiia Reshetniak, Charlotta Lorenz, Christian Hoffmann, Dragomir Milovanovi\u0107, Silvio O. Rizzoli & Sarah K\u00f6ster<\/div>
Protein dynamics in the synaptic bouton are still not well understood, despite many quantitative studies of synaptic structure and function. The complexity of the synaptic environment makes investigations of presynaptic protein mobility challenging. Here, we present an in vitro approach to create a minimalist model of the synaptic environment by patterning synaptic vesicles (SVs) on glass coverslips. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Materials, 2020<\/div>

Stress fibres are embedded in a contractile cortical network<\/h2>\r\n
Timoth\u00e9e Vignaud, Calina Copos, Christophe Leterrier, Mauricio Toro-Nahuelpan, Qingzong Tseng, Julia Mahamid, Laurent Blanchoin, Alex Mogilner, Manuel Th\u00e9ry & Laetitia Kurzawa<\/div>
Contractile actomyosin networks are responsible for the production of intracellular forces. There is increasing evidence that bundles of actin filaments form interconnected and interconvertible structures with the rest of the network. In this study, we explored the mechanical impact of these interconnections on the production and distribution of traction forces throughout the cell. By using a combination of hydrogel micropatterning, traction force microscopy and laser photoablation, we measured the relaxation of traction forces in response to local photoablations. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biology of the Cell, 2020<\/div>

Human neutrophils swim and phagocytise bacteria<\/h2>\r\n
Nicolas Garcia\u2010Seyda, Valentine Seveau, Fabio Manca, Martine Biarnes\u2010Pelicot, Marie\u2010Pierre Valignat, Marc Baj\u00e9noff, Olivier Theodoly<\/div>
Leukocytes migrate in an amoeboid fashion while patrolling our organism in the search for infection or tissue damage. Their capacity to migrate has been proven integrin independent, however, non\u2010specific adhesion or confinement remain a requisite in current models of cell migration. This idea has been challenged twice within the last decade with human neutrophils and effector T lymphocytes, which were shown to migrate in free suspension, a phenomenon termed swimming. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Nature Communications, 2020<\/div>

T-Plastin reinforces membrane protrusions to bridge matrix gaps during cell migration<\/h2>\r\n
Damien Garbett, Anjali Bisaria, Changsong Yang, Dannielle G. McCarthy, Arnold Hayer, W. E. Moerner, Tatyana M. Svitkina & Tobias Meyer<\/div>
Migrating cells move across diverse assemblies of extracellular matrix (ECM) that can be separated by micron-scale gaps. For membranes to protrude and reattach across a gap, actin filaments, which are relatively weak as single filaments, must polymerize outward from adhesion sites to push membranes towards distant sites of new adhesion. Here, using micropatterned ECMs, we identify T-Plastin, one of the most ancient actin bundling proteins, as an actin stabilizer that promotes membrane protrusions and enables bridging of ECM gaps. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Biophysical Journal, 2020<\/div>

Amoeboid Swimming Is Propelled by Molecular Paddling in Lymphocytes<\/h2>\r\n
Laurene Aoun, Alexander Farutin, Nicolas Garcia-Seyda, Paulin N\u00e8gre, Mohd Suhail Rizvi, Sham Tlili, Solene Song, Xuan Luo, Martine Biarnes-Pelicot, R\u00e9mi Galland, Jean-Baptiste Sibarita, Alph\u00e9e Michelot, Claire Hivroz, Salima Rafai, Marie-Pierre Valignat, Chaouqi Misbah,Olivier Theodoly<\/div>
Mammalian cells developed two main migration modes. The slow mesenchymatous mode, like crawling of fibroblasts, relies on maturation of adhesion complexes and actin fiber traction, whereas the fast amoeboid mode, observed exclusively for leukocytes and cancer cells, is characterized by weak adhesion, highly dynamic cell shapes, and ubiquitous motility on two-dimensional and in three-dimensional solid matrix. In both cases, interactions with the substrate by adhesion or friction are widely accepted as a prerequisite for mammalian cell motility, which precludes swimming. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Journal of Cell Science, 2020<\/div>

Lymphocytes perform reverse adhesive haptotaxis mediated by LFA-1 integrins<\/h2>\r\n
Xuan Luo, Valentine Seveau de Noray, Laurene Aoun, Martine Biarnes-Pelicot, Pierre-Olivier Strale, Vincent Studer, Marie-Pierre Valignat, Olivier Theodoly<\/div>
Cell guidance by anchored molecules, or haptotaxis, is crucial in development, immunology and cancer. Adhesive haptotaxis, or guidance by adhesion molecules, is well established for mesenchymal cells such as fibroblasts, whereas its existence remains unreported for amoeboid cells that require less or no adhesion in order to migrate. We show that,\u00a0in vitro, amoeboid human T lymphocytes develop adhesive haptotaxis mediated by densities of integrin ligands expressed by high endothelial venules.\u00a0 \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Advanced Healthcare Matherials, 2020<\/div>

Tailoring Common Hydrogels into 3D Cell Culture Templates<\/h2>\r\n
Aur\u00e9lien Pasturel, Pierre\u2010Olivier Strale, Vincent Studer<\/div>
Physiologically relevant cell\u2010based models require engineered microenvironments which recapitulate the topographical, biochemical, and mechanical properties encountered in vivo. In this context, hydrogels are the materials of choice. Here a light\u2010based toolbox is able to craft such microniches out of common place materials. Extensive use of benzophenone photoinitiators and their interaction with oxygen achieves this. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
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Science Advances, 2020<\/div>

Intercellular communication controls agonist-induced calcium oscillations independently of gap junctions in smooth muscle cells<\/h2>\r\n
S E Stasiak, R R Jamieson, J Bouffard, E J Cram and H Parameswaran<\/div>
In this study, we report the existence of a communication system among human smooth muscle cells that uses mechanical forces to frequency modulate long-range calcium waves. An important consequence of this mechanical signaling is that changes in stiffness of the underlying extracellular matrix can interfere with the frequency modulation of Ca2+ waves, causing smooth muscle cells from healthy human donors to falsely perceive a much higher agonist dose than they actually received. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Biology of the Cell, 2020<\/div>

Mechanobiology of antigen\u2010induced T cell arrest<\/h2>\r\n
M\u00e9lanie Chabaud, No\u00e9mie Paillon, Katharina Gaus, Claire Hivroz<\/div>
To mount an immune response, T\u00a0cells must first find rare antigens present at the surface of antigen\u2010presenting cells (APCs). They achieve this by migrating rapidly through the crowded space of tissues and constantly sampling the surface of APCs. Upon antigen recognition, T\u00a0cells decelerate and polarise towards the APC, ultimately forming a specialised interface known as the immunological synapse. These conjugates form as the result of the interaction between pairs of receptors\/ligands that are under mechanical stress due to the continuously reorganising cell cytoskeleton. In this review, \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
ACS Nano, 2020<\/div>

Coupling Polar Adhesion with Traction, Spring and Torque Forces Allows High Speed Helical Migration of the Protozoan Parasite Toxoplasma<\/h2>\r\n
Georgios Pavlou , Bastien Touquet, Luis Vigetti, Patricia Renesto, Alexandre Bougdour, Delphine Debarre, Martial Balland, and Isabelle Tardieux<\/div>
Among the eukaryotic cells that navigate through fully developed metazoan tissues, protozoans from the Apicomplexa phylum have evolved motile developmental stages that move much faster than the fastest crawling cells owing to a peculiar substrate-dependent type of motility, known as gliding. Best-studied models are the\u00a0Plasmodium\u00a0sporozoite and the\u00a0Toxoplasma tachyzoite polarized cells for which motility is vital to achieve their developmental programs in the metazoan hosts. The gliding machinery is shared between the two stages and functionally \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Lab On A Chip, 2020<\/div>

Microfluidic dialysis using photo-patterned hydrogel membranes in PDMS chips<\/h2>\r\n
Hoang-Thanh Nguyen, Morgan Massino, Camille Keita and Jean-Baptiste Salmon<\/div>
We report the fabrication of permeable membranes for microfluidic dialysis applications in poly(dimethylsiloxane) (PDMS) channels. A maskless UV projection device was used to photo-pattern long hydrogel membranes (mm\u2013cm) with a spatial resolution of a few microns in PDMS chips integrating also micro-valves. We show in particular that multi-layer soft lithography allows one to deplete oxygen from the PDMS walls using a nitrogen gas flow and therefore makes possible\u00a0in situ\u00a0UV-induced polymerization of hydrogels.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Nature Materials, 2020<\/div>

Biomimetic niches reveal the minimal cues to trigger apical lumen formation in single hepatocytes<\/h2>\r\n
Yue Zhang, Richard De Mets, Cornelia Monzel, Vidhyalakshmi Acharya, Pearlyn Toh, Jasmine Fei Li Chin, No\u00e9mi Van Hul, Inn Chuan Ng, Hanry Yu, Soon Seng Ng, S. Tamir Rashid & Virgile Viasnoff<\/div>
The symmetry breaking of protein distribution and cytoskeleton organization is an essential aspect for the development of apicobasal polarity. In embryonic cells this process is largely cell autonomous, while differentiated epithelial cells collectively polarize during epithelium formation. Here, we demonstrate that the de novo polarization of mature hepatocytes does not require the synchronized development of apical poles on neighbouring cells. De novo polarization at the single-cell level by mere contact with the extracellular matrix and immobilized cadherin defining a polarizing axis. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
CELL, 2020<\/div>

Defining the Design Principles of Skin Epidermis Postnatal Growth<\/h2>\r\n
Sophie Dekoninck, Edouard Hannezo, Alejandro Sifrim, Yekaterina A. Miroshnikova, Mariaceleste Aragona, Milan Malfait, Souhir Gargouri, Charlotte de Neunheuser, Christine Dubois, Thierry Voet, Sara A. Wickstr\u00f6m, Benjamin D. Simons and C\u00e9dric Blanpain<\/div>

During embryonic and postnatal development, or- gans and tissues grow steadily to achieve their final size at the end of puberty. However, little is known about the cellular dynamics that mediate postnatal growth. By combining in vivo clonal lineage tracing, proliferation kinetics, single-cell transcriptomics, and in vitro micro-pattern experiments, we resolved the cellular dynamics taking place during postnatal skin epidermis expansion. Our data revealed that harmonious growth is engineered by a single popula- tion of developmental progenitors presenting\u2026<\/p><\/div><\/div> Voir l'article<\/a><\/div>

<\/div>\r\n
\r\n
Nanoscale, 2020<\/div>

Transport and programmed release of nanoscale cargo from cells by using NETosis<\/h2>\r\n
Daniel Meyer, Saba Telele, Anna Zelen\u00e1, Alice J. Gillen, Alessandra Antonucci, Elsa Neubert, Robert Ni\u00dfler, Florian A. Mann, Luise Erpenbeck, Ardemis A. Boghossian, Sarah K\u00f6ster, Sebastian Kruss<\/div>
Transport and delivery of nanoscale materials are crucial for many applications in biomedicine. However, controlled uptake, transport and triggered release of such cargo remains challenging. In this study, we use human immune cells (neutrophilic granulocytes, neutrophils) and program them to perform these tasks in vitro. For this purpose, we let neutrophils phagocytose a nanoscale cargo. As an example, we used DNA-functionalized single-walled carbon nanotubes (SWCNT) that fluoresce in the near infrared (980 nm) and serve as sensors for small molecules. Cells still migrate, follow chemical gradients \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Jove, 2019<\/div>

Light-Induced Molecular Adsorption of Proteins Using the PRIMO System for Micro-Patterning to Study Cell Responses to Extracellular Matrix Proteins<\/h2>\r\n
Cristina Melero*, Aljona Kolmogorova*, Paul Atherton, Brian Derby, Adam Reid, Karin Jansen, Christoph Ballestrem<\/div>
Cells sense a variety of extracellular cues, including the composition and geometry of the extracellular matrix, which is synthesized and remodeled by the cells themselves. Here, we present the method of Light-Induced Molecular Adsorption of Proteins (LIMAP) using the PRIMO system as a patterning technique to produce micro-patterned extracellular matrix (ECM) substrates using a single or combination of proteins. The method enables printing of ECM patterns in micron resolution with excellent reproducibility. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
SCIENTIFIC REPORTS, 2019<\/div>

Extracellular matrix stiffness regulates human airway smooth muscle contraction by altering the cell-cell coupling<\/h2>\r\n
Samuel R. Polio, Suzanne E. Stasiak, Ryan R. Jamieson, Jenna L. Balestrini, Ramaswamy Krishnan & Harikrishnan Parameswaran<\/div>
For an airway or a blood vessel to narrow, there must be a connected path that links the smooth muscle (SM) cells with each other, and transmits forces around the organ, causing it to constrict. Currently, we know very little about the mechanisms that regulate force transmission pathways in a multicellular SM ensemble. Here, we used extracellular matrix (ECM) micropatterning to study force transmission in a two-cell ensemble of SM cells. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Nature Methods, 2019<\/div>

Tailoring cryo-electron microscopy grids by photo-micropatterning for in-cell structural studies<\/h2>\r\n
Mauricio Toro-Nahuelpan, Levgeniia Zagoriy, Fabrice Senger, Laurent Blanchoin, Manuel Thery & Julia Mahamid<\/div>
Spatially-controlled cell adhesion on electron microscopy (EM) supports remains a bottleneck in specimen preparation for cellular cryo-electron tomography. Here, we describe contactless and mask-free photo-micropatterning of EM grids for site-specific deposition of extracellular matrix-related proteins. We attained refined cell positioning for micromachining by cryo-focused ion beam milling. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
NAT PHYS, 2019<\/div>

Substrate area confinement is a key determinant of cell velocity in collective migration<\/h2>\r\n
Danahe Mohammed, Guillaume Charras, El\u00e9onore Vercruysse, Marie Versaevel, Jos\u00e9phine Lantoine, Laura Alaimo, C\u00e9line Bruy\u00e8re, Marine Luciano, Karine Glinel, Geoffrey Delhaye, Olivier Th\u00e9odoly & Sylvain Gabriele<\/div>
Collective cell migration is fundamental throughout development, during wound healing and in many diseases. Although much effort has focused on cell\u2013cell junctions, a role for physical confinement in collective cell migration remains unclear. Here, we used adhesive microstripes of varying widths to mimic the spatial confinement experienced by follower cells within epithelial tissues. Our results reveal that the substrate area confinement is sufficient to modulate the three-dimensional cellular morphology without the need for intercellular adhesive cues. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
NAT MAT, 2019<\/div>

Traction forces at the cytokinetic ring regulate cell division and polyploidy in the migrating zebrafish epicardium<\/h2>\r\n
Marina Uroz, Anna Garcia-Puig, Isil Tekeli, Alberto Elosegui-Artola, Juan F. Abenza, Ariadna Mar\u00edn-Llaurad\u00f3, Silvia Pujals, Vito Conte, Lorenzo Albertazzi, Pere Roca-Cusachs, \u00c1ngel Raya & Xavier Trepat<\/div>
Epithelial repair and regeneration are driven by collective cell migration and division. Both cellular functions involve tightly controlled mechanical events, but how physical forces regulate cell division in migrating epithelia is largely unknown. Here we show that cells dividing in the migrating zebrafish epicardium exert large cell\u2013extracellular matrix (ECM) forces during cytokinesis. These forces point towards the division axis and are exerted through focal adhesions that connect the cytokinetic ring to the underlying ECM.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
J. Micromech. Microeng., 2019<\/div>

Extracellular matrix micropatterning technology for whole cell cryogenic electron microscopy studies<\/h2>\r\n
Leeya Engel, Guido Gaietta, Liam P Dow, Mark F Swift, Gaspard Pardon, Niels Volkmann, William I Weis, Dorit Hanein, Beth L Pruitt<\/div>
Cryogenic electron tomography is the highest resolution tool available for structural analysis of macromolecular organization inside cells. Micropatterning of extracellular matrix (ECM) proteins is an established in vitro cell culture technique used to control cell shape. Recent traction force microscopy studies have shown correlation between cell morphology and the regulation of force transmission. However, it remains unknown how cells sustain increased strain energy states and localized stresses at the supramolecular level. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
NATURE MATERIALS, 2019<\/div>

A mechano-signalling network linking microtubules, myosin IIA filaments and integrin-based adhesions<\/h2>\r\n
Nisha Bte Mohd Rafiq, Yukako Nishimura, Sergey V. Plotnikov, Visalatchi Thiagarajan, Zhen Zhang, Shidong Shi, Meenubharathi Natarajan, Virgile Viasnoff, Pakorn Kanchanawong, Gareth E. Jones & Alexander D. Bershadsky<\/div>
The interrelationship between microtubules and the actin cytoskeleton in mechanoregulation of integrin-mediated adhesions is poorly understood. Here, we show that the effects of microtubules on two major types of cell-matrix adhesion, focal adhesions and podosomes, are mediated by KANK family proteins connecting the adhesion protein talin with microtubule tips. Both total microtubule disruption and microtubule uncoupling from adhesions by manipulations with KANKs trigger a massive assembly of myosin IIA filaments, augmenting focal adhesions and disrupting podosomes.\u00a0\u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Phil. Trans. R. Soc. B, 2019<\/div>

Forces and constraints controlling podosome assembly and disassembly<\/h2>\r\n
Nisha Bte Mohd Rafiq, Gianluca Grenci, Cheng Kai Lim, Michael M Kozlov, Gareth E Jones, Virgile Viasnoff and Alexander D Bershadsky<\/div>
Podosomes are a singular category of integrin-mediated adhesions important in the processes of cell migration, matrix degradation, and cancer cell invasion. Despite a wealth of biochemical studies, the effects of mechanical forces on podosome integrity and dynamics are poorly understood. Here, we show that podosomes are highly sensitive to two groups of physical factors. First, we describe the process of podosome disassembly induced by activation of myosin- IIA filament assembly. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
BIORXIV, 2019<\/div>

Autonomous induction of hepatic polarity to construct single cell liver<\/h2>\r\n
Yue Zhang, Richard de Mets, Cornelia Monzel, Pearlyn Toh, Noemi Van Hul, Soon Seng Ng, S. Tamir Rashid, Virgile Viasnoff<\/div>
Symmetry breaking of protein distribution and cytoskeleton organization is an essential aspect for development of apico-basal polarity. In embryonic cells this process is largely cell autonomous, while differentiated epithelial cells collectively polarize during epithelium formation. We report here that the\u00a0de novo\u00a0polarization of mature hepatocytes is a cell autonomous process. Single hepatocytes developed\u00a0bona fide secretory hemi-apical lumens upon adhesion to finely tuned substrates bio-functionalized with cadherin and extra cellular matrix. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
ADVANCED SCIENCE, 2019<\/div>

Photoactivatable Hsp47: A Tool to Regulate Collagen Secretion and Assembly<\/h2>\r\n
Essak S. Khan, Shrikrishnan Sankaran, Julieta I. Paez, Christina Muth, Mitchell K. L. Han, Ar\u00e1nzazu del Campo<\/div>
Collagen is the most abundant structural protein in mammals and is crucial for the mechanical integrity of tissues. Hsp47, an endoplasmic reticulum resident collagen\u2010specific chaperone, is involved in collagen biosynthesis and plays a fundamental role in the folding, stability, and intracellular transport of procollagen triple helices. This work reports on a photoactivatable derivative of Hsp47 that allows regulation of collagen biosynthesis within mammalian cells using light. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
PLOS One, 2018<\/div>

Live nanoscopic to mesoscopic topography reconstruction with an optical microscope for chemical and biological samples<\/h2>\r\n
Olivier Theodoly, Nicolas Garcia-Seyda, Fr\u00e9deric Bedu, Xuan Luo, Sylvain Gabriele, T\u00e2m Mignot, Joanna Giermanska, Jean-Paul Chapel, M\u00e9linda M\u00e9tivier, Marie-Pierre Valignat<\/div>
Macroscopic properties of physical and biological processes like friction, wetting, and adhesion or cell migration are controlled by interfacial properties at the nanoscopic scale. In an attempt to bridge simultaneously investigations at different scales, we demonstrate here how optical microscopy in Wet-Surface Ellipsometric Enhanced Contrast (Wet-SEEC) mode offers imaging and measurement of thin films at solid\/liquid interfaces in the range 1\u2013500 nm with lateral optical resolution. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Nano Lett, 2018<\/div>

Optical magnetometry of single biocompatible micromagnets for quantitative magnetogenetic and magnetomechanical assays<\/h2>\r\n
Lo\u00efc Toraille, Koceila A\u00efzel, Elie Balloul, Chiara Vicario, Cornelia Monzel, Mathieu Coppey, Emilie Secret, Jean-Michel Siaugue, Joao Sampaio, Stanislas Rohart, Nicolas Vernier, Louise Bonnemay, Thierry Debuisschert, Lo\u00efc Rondin, Jean-Francois ROCH, and Maxime Dahan<\/div>
The mechanical manipulation of magnetic nanoparticles is a powerful approach to probe and actuate biological processes in living systems. Implementing this technique in high-throughput assays can be achieved using biocompatible micomagnet arrays. Yet, the magnetic properties of these arrays are usually indirectly inferred from simulations or Stokes drag measurements, leaving unresolved questions about the actual profile of the magnetic fields at the micrometer scale and the exact magnetic forces that are applied. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Adv Biosys, 2018<\/div>

A new approach to design artificial 3D micro-niches with combined chemical, topographical and rheological cues<\/h2>\r\n
Celine Stoecklin, Zhang Yue, Wilhelm W. Chen, Richard de Mets, Eileen Fong, Vincent Studer, Virgile Viasnoff<\/div>
The in vitro methods to recapitulate environmental cues around cells are usually optimized to test a specific property of the environment (biochemical nature or the stiffness of the extra cellular matrix (ECM), or nanotopography) for its capability to induce defined cell behaviors (lineage commitment, migration). Approaches that combine different environmental cues in 3D to assess the biological response of cells to the spatial organization of different biophysical and biochemical cues are growingly being developed. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Lab Chip, 2018<\/div>

In situ photopatterning of pressure-resistant hydrogel membranes with controlled permeabilities in PEGDA microfluidic channels.<\/h2>\r\n
J\u00e9r\u00e9my Decock, Mathias Schlenk and Jean-Baptiste Salmon<\/div>
We report the fabrication of highly permeable membranes in poly(ethylene glycol) diacrylate (PEGDA) channels, for investigating ultra- or micro-filtration, at the microfluidic scale. More precisely, we used a maskless UV projection setup to photopattern PEG-based hydrogel membranes on a large scale (mm\u2013cm), and with a spatial resolution of a few microns.\u00a0We show that these membranes can withstand\u00a0trans-membrane pressure drops of up to 7 bar without any leakage, thanks to the strong anchoring of the hydrogel to the channel walls. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
BioRxiv, 2018<\/div>

Collagen assembly and turnover imaged with a CRISPR-Cas9 engineered Dendra2 tag<\/h2>\r\n
Adam Pickard, Antony Adamson, Yinhui Lu, Joan Chang, Richa Garva, Nigel Hodson, Karl Kadler<\/div>
Electron microscopy has been the gold standard for studying collagen networks but dynamic information on how cells synthesise the networks has been lacking. Live imaging methods have been unable to distinguish newly-synthesised fibrils from pre-existing fibrils and intracellular collagen. Here, we tagged endogenous collagen-I using CRISPR-Cas9 with photoswitchable Dendra2 and demonstrate live cells synthesising, migrating on, and interacting with, collagen fibrils. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Hum Mol Genet. 2016<\/div>

Altered microtubule dynamics and vesicular transport in mouse and human MeCP2-deficient astrocytes.<\/h2>\r\n
Del\u00e9pine C, Meziane H, Nectoux J, Opitz M, Smith AB, Ballatore C, Saillour Y, Bennaceur-Griscelli A, Chang Q, Williams EC, Dahan M, Duboin A, Billuart P, Herault Y, Bienvenu T.<\/div>
Rett syndrome (RTT) is a rare X-linked neurodevelopmental disorder, characterized by normal post-natal development followed by a sudden deceleration in brain growth with progressive loss of acquired motor and language skills, stereotypic hand movements and severe cognitive impairment.\u00a0Mutations in the methyl-CpG-binding protein 2 (MECP2) cause more than 95% of classic cases. \u2026<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n
\r\n
Adv Mater. 2016<\/div>

Multiprotein Printing by Light-Induced Molecular Adsorption.<\/h2>\r\n
Strale PO, Azioune A, Bugnicourt G, Lecomte Y, Chahid M, Studer V.<\/div>
Light-induced molecular adsorption of proteins (LIMAP) allows for quantitative sub-micrometer-resolution printing of multiple biomolecules. Surface-bound gradients are patterned within minutes over an entire glass cover-slip. LIMAP is used to perform selective immuno-assays, to dynamically control the adhesion of individual cells, and to achieve hierarchical co-cultures instrumental for tissue engineering.<\/div><\/div> Voir l'article<\/a><\/div>
<\/div>\r\n <\/div>\r\n[\/vc_column][\/vc_row]\n<\/div>","protected":false},"excerpt":{"rendered":"

A travers les publications de nos utilisateurs, d\u00e9couvrez les champs d’application du syst\u00e8me de photopatterning PRIMO: contr\u00f4le de la polarit\u00e9 cellulaire, adh\u00e9rence cellulaire, cryo-ET, microfabrication, etc.<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":13,"menu_order":4,"comment_status":"closed","ping_status":"closed","template":"","meta":{"inline_featured_image":false,"footnotes":""},"class_list":["post-5635","page","type-page","status-publish","hentry","themes-applications-biocell","themes-applications-3d","themes-application-microfab"],"yoast_head":"\nTechnologie de photopatterning PRIMO : articles scientifiques des utilisateurs<\/title>\n<meta name=\"description\" content=\"Lisez les articles scientifiques de nos utilisateurs et d\u00e9couvrez comment le photopatterning PRIMO les a aid\u00e9s dans divers domaines : contr\u00f4le de la polarit\u00e9 cellulaire, adh\u00e9sion cellulaire, microfabrication\" \/>\n<meta name=\"robots\" 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