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1. Large deviations of a velocity jump process with a Hamilton-Jacobi approach

Author

Caillerie, Nils, Modélisation mathématique, calcul scientifique (MMCS), Institut Camille Jordan [Villeurbanne] (ICJ), École Centrale de Lyon (ECL), Université de Lyon-Université de Lyon-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université Jean Monnet [Saint-Étienne] (UJM)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Lyon (ECL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 639638)., and European Project: European Research Council

Subjects
Kinetic Equations, PDMP, [MATH.MATH-AP]Mathematics [math]/Analysis of PDEs [math.AP], Hamilton Jacobi equations, hyperbolic limit, perturbed test function methos
Abstract

We study a random process on R n moving in straight lines and changing randomly its velocity at random exponential times. We focus more precisely on the Kolmogorov equation in the hyperbolic scale (t, x, v) → t ε , x ε , v , with ε > 0, before proceeding to a Hopf-Cole transform, which gives a kinetic equation on a potential. We show convergence as ε → 0 of the potential towards the viscosity solution of a Hamilton-Jacobi equation ∂tϕ + H (∇xϕ) = 0 where the hamiltonian may lack C 1 regularity, which is quite unseen in this type of studies. Résumé

Published
2016

2. Architecture and Connectivity Govern Actin Network Contractility

Author

Manuel Théry, Jean-Louis Martiel, François Nédélec, Wenxiang Cao, Christophe Guérin, Enrique M. De La Cruz, Hajer Ennomani, Gaëlle Letort, Laurent Blanchoin, Laboratoire de physiologie cellulaire végétale (LPCV), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Department of Molecular Biophysics and Biochemistry, Yale University [New Haven], Cell Biology and Cell Biophysics Unit, European Molecular Biology Laboratory, European Molecular Biology Laboratory [Grenoble] (EMBL), Grants from the Human Frontier Science Program (RGP0004/2011)- NIH (GM097348)- Burroughs Wellcome Fund (BWF) 2014 Collaborative Research Travel Grant CRTG- Projet International de Cooperation Scientifique (PICS) award from CNRS, ANR-12-BSV5-0014,Contract,Nouveaux systèmes biomimétiques pour étudier la contractilité acto-myosine cellulaire(2012), European Project: European Research Council, Physiologie cellulaire et végétale (LPCV), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
0301 basic medicine, macromolecular substances, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Myosins, Microfilament, General Biochemistry, Genetics and Molecular Biology, Article, Contractility, 03 medical and health sciences, 0302 clinical medicine, Actin filament branching, Cell polarity, Myosin, Animals, Micropatterning, Actin filament dynamics, Actin network contractility, Cytoskeleton, Actin, ComputingMilieux_MISCELLANEOUS, Actomyosin contractility, Agricultural and Biological Sciences(all), Biochemistry, Genetics and Molecular Biology(all), Network architecture, Actin remodeling, Actomyosin, Actins, Cell biology, 030104 developmental biology, Rabbits, General Agricultural and Biological Sciences, 030217 neurology & neurosurgery, Muscle Contraction
Abstract

International audience; Actomyosin contractility plays a central role in a wide range of cellular processes, including the establishment of cell polarity, cell migration, tissue integrity, and morphogenesis during development. The contractile response is variable and depends on actomyosin network architecture and biochemical composition. To determine how this coupling regulates actomyosin-driven contraction, we used a micropatterning method that enables the spatial control of actin assembly. We generated a variety of actin templates and measured how defined actin structures respond to myosin-induced forces. We found that the same actin filament crosslinkers either enhance or inhibit the contractility of a network, depending on the organization of actin within the network. Numerical simulations unified the roles of actin filament branching and crosslinking during actomyosin contraction. Specifically, we introduce the concept of "network connectivity'' and show that the contractions of distinct actin architectures are described by the same master curve when considering their degree of connectivity. This makes it possible to predict the dynamic response of defined actin structures to transient changes in connectivity. We propose that, depending on the connectivity and the architecture, network contraction is dominated by either sarcomeric-like or buckling mechanisms. More generally, this study reveals how actin network contractility depends on its architecture under a defined set of biochemical conditions.

Published
2016
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3. The centrosome is an actin-organizing centre

Author

James Sillibourne, Laurent Blanchoin, Manuel Théry, Christophe Guérin, Jérémie Gaillard, Francesca Farina, Yohann Couté, Physiologie cellulaire et végétale (LPCV), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Etude de la dynamique des protéomes (EDyP), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA), Alloimmunité-Autoimmunité-Transplantation (A2T), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Universitaire d'Hématologie (IUH), Université Paris Diderot - Paris 7 (UPD7)-Université Paris Diderot - Paris 7 (UPD7), Grenoble Alliance for Integrated Structural Cell Biology, ANR-12-BSV5-0004,MITOTUBES,Disséquer la Dynamique des Microtubules en Mitose par Reconstitution de Sous-Modules du Fuseau(2012), ANR-10-INBS-08-01/10-INBS-0008,ProFI,Infrastructure Française de Protéomique(2010), European Project: European Research Council, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Institut Universitaire d'Hématologie (IUH), Université Paris Diderot - Paris 7 (UPD7)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Etude de la dynamique des protéomes (EDyP ), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), ANR-10-INBS-0008,ProFI,Infrastructure Française de Protéomique(2010), Physico-Chimie-Curie (PCC), Centre National de la Recherche Scientifique (CNRS)-Institut Curie-Université Pierre et Marie Curie - Paris 6 (UPMC), Laboratoire de physiologie cellulaire végétale (LPCV), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP)-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG)

Subjects
0301 basic medicine, [SDV]Life Sciences [q-bio], Arp2/3 complex, Cell architecture, Centrosome cycle, macromolecular substances, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Microtubules, Article, Actin-Related Protein 2-3 Complex, 03 medical and health sciences, Actin remodeling of neurons, Cell growth, PCM1, WASH, Actin filament assembly, Humans, Cytoskeleton, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, Pericentriolar material, Centrosome, Actin remodeling, Cell Polarity, Cell Biology, Actin binding protein, Actins, Cell biology, Actin Cytoskeleton, 030104 developmental biology, Actin filament nucleation, Actin network organization, biology.protein, Arp2/3
Abstract

International audience; Microtubules and actin filaments are the two main cytoskeleton networks supporting intracellular architecture and cell polarity. The centrosome nucleates and anchors microtubules and is therefore considered to be the main microtubule-organizing centre. However, recurring, yet unexplained, observations have pointed towards a connection between the centrosome and actin filaments. Here we have used isolated centrosomes to demonstrate that the centrosome can directly promote actin-filament assembly. A cloud of centrosome-associated actin filaments could be identified in living cells as well. Actin-filament nucleation at the centrosome was mediated by the nucleation-promoting factor WASH in combination with the Arp2/3 complex. Pericentriolar material 1 (PCM1) seemed to modulate the centrosomal actin network by regulating Arp2/3 complex and WASH recruitment to the centrosome. Hence, our results reveal an additional facet of the centrosome as an intracellular organizer and provide mechanistic insights into how the centrosome can function as an actin-filament-organizing centre.

Published
2016
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4. Large deviations of a velocity jump process with a Hamilton-Jacobi approach

Author

Caillerie, Nils, Modélisation mathématique, calcul scientifique (MMCS), Institut Camille Jordan [Villeurbanne] (ICJ), École Centrale de Lyon (ECL), Université de Lyon-Université de Lyon-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université Jean Monnet [Saint-Étienne] (UJM)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Lyon (ECL), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 639638)., European Project: European Research Council, Institut Camille Jordan (ICJ), Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Jean Monnet - Saint-Étienne (UJM)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Lyon (ECL), and Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Jean Monnet - Saint-Étienne (UJM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Mathematics - Analysis of PDEs, Kinetic Equations, PDMP, FOS: Mathematics, [MATH.MATH-AP]Mathematics [math]/Analysis of PDEs [math.AP], Hamilton Jacobi equations, hyperbolic limit, perturbed test function methos, Analysis of PDEs (math.AP)
Abstract

We study a random process on R n moving in straight lines and changing randomly its velocity at random exponential times. We focus more precisely on the Kolmogorov equation in the hyperbolic scale (t, x, v) $\to$ t $\epsilon$, x $\epsilon$, v, with $\epsilon$ \textgreater{} 0, before proceeding to a Hopf-Cole transform, which gives a kinetic equation on a potential. We show convergence as $\epsilon$ $\to$ 0 of the potential towards the viscosity solution of a Hamilton-Jacobi equation $\partial$t\"I + H ($\nabla$x\"I) = 0 where the hamiltonian may lack C 1 regularity, which is quite unseen in this type of studies. R{\'e}sum{\'e}

Published
2016
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5. Microtubules self-repair in response to mechanical stress

Author

Manuel Théry, Karin John, Laurent Blanchoin, Jérémie Gaillard, Laura Schaedel, Maxence V. Nachury, Laboratoire de physiologie cellulaire végétale (LPCV), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), LIPHY-DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Stanford University [Stanford], HFSP funding (RGY0088/2012), European Project: European Research Council, Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM-LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Stanford University, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Recherche Agronomique (INRA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)

Subjects
Materials science, Mechanical stress, Nanotechnology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Flagellum, Plasticity, Microtubules, Article, Softening, Microtubule, Tubulin, Lab-On-A-Chip Devices, medicine, Animals, Humans, General Materials Science, Cytoskeleton, Microtubule nucleation, biology, Mechanical Engineering, Cilium, Stiffness, General Chemistry, Condensed Matter Physics, Self repairing, Mechanics of Materials, Biophysics, biology.protein, Stress, Mechanical, medicine.symptom
Abstract

Supplementary information : Supplementary figure S1 – S8 Supplementary video 1 – 6 legends; International audience; Microtubules--which define the shape of axons, cilia and flagella, and provide tracks for intracellular transport--can be highly bent by intracellular forces, and microtubule structure and stiffness are thought to be affected by physical constraints. Yet how microtubules tolerate the vast forces exerted on them remains unknown. Here, by using a microfluidic device, we show that microtubule stiffness decreases incrementally with each cycle of bending and release. Similar to other cases of material fatigue, the concentration of mechanical stresses on pre-existing defects in the microtubule lattice is responsible for the generation of more extensive damage, which further decreases microtubule stiffness. Strikingly, damaged microtubules were able to incorporate new tubulin dimers into their lattice and recover their initial stiffness. Our findings demonstrate that microtubules are ductile materials with self-healing properties, that their dynamics does not exclusively occur at their ends, and that their lattice plasticity enables the microtubules' adaptation to mechanical stresses.

Published
2015
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6. Design of a 2D no-flow chamber to monitor hematopoietic stem cells

Author

Valérie Vanneaux, Laurent Blanchoin, Manuel Théry, Jean Berthier, Théo Cambier, Jérôme Larghero, Thibault Honegger, David Peyrade, Laboratoire de physiologie cellulaire végétale (LPCV), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Recherche Agronomique (INRA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Laboratoire des technologies de la microélectronique (LTM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), CIC - Biotherapie - Saint Louis ((CIC-BT 301)), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Agence National pour la Recherche (ANR-05PRIB01103, ANR-10-IBHU-0002), Université de Grenoble, ANR-05-PRIB-0011,MYOSOTIS,MyoSoTIS : Microtechnologies et modélisation mathématique appliquées à la thérapie cellulaire(2005), ANR-10-IBHU-0002,SLI,Institut Saint-Louis(2010), European Project: European Research Council, Martin-Laffon, Jacqueline, Biotechnologies - MyoSoTIS : Microtechnologies et modélisation mathématique appliquées à la thérapie cellulaire - - MYOSOTIS2005 - ANR-05-PRIB-0011 - RIB - VALID, Instituts Hospitalo-Universitaires B - Institut Saint-Louis - - SLI2010 - ANR-10-IBHU-0002 - IBHU - VALID, ERC-310472 - European Research Council - INCOMING, Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-Commissariat à l'Énergie Atomique et aux Énergies Alternatives (CEA) - Grenoble-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Laboratoire d'Electronique et des Technologies de l'Information (CEA-LETI), Université Grenoble Alpes (UGA)-Direction de Recherche Technologique (CEA) (DRT (CEA)), ANR: ANR-05PRIB01103, ANR: ANR-10-IBHU-0002, and Théry, Manuel

Subjects
Cell type, [SDV]Life Sciences [q-bio], Cytological Techniques, Large array, Biomedical Engineering, Bioengineering, Sciences du Vivant, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, 01 natural sciences, Biochemistry, 03 medical and health sciences, blood, stem cells, Chamber design, Human Umbilical Vein Endothelial Cells, Humans, human, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Cells, Cultured, Fluorescent Dyes, 030304 developmental biology, 0303 health sciences, 010401 analytical chemistry, Life Sciences, Cell Differentiation, Equipment Design, General Chemistry, Microfluidic Analytical Techniques, Hematopoietic Stem Cells, 0104 chemical sciences, Cell biology, Technique, no flow chamber design, [SDV] Life Sciences [q-bio], Haematopoiesis, Cell tracking, Stem cell, Biomedical engineering
Abstract

International audience; Hematopoietic stem cells (HSCs) are the most commonly used cell type in cell-based therapy. However, the investigation of their behavior in vitro has been limited by the difficulty of monitoring these non-adherent cells under classical culture conditions. Indeed, fluid flow moves cells away from the video-recording position and prevents single cell tracking over long periods of time. Here we describe a large array of 2D no-flow chambers allowing the monitoring of single HSCs for several days. The chamber design has been optimized to facilitate manufacturing and routine use. The chip contains a single inlet and 800 chambers. The chamber medium can be renewed by diffusion within a few minutes. This allowed us to stain live human HSCs with fluorescent primary antibodies in order to reveal their stage in the hematopoiesis differentiation pathway. Thus we were able to correlate human HSCs' growth rate, polarization and migration to their differentiation stage.

Published
2015
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