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Generation of fluorescent cell-derived-matrix to study 3D cell migration

Amélie Luise Godeau, Hélène Delanoë-Ayari, Daniel Riveline
doi: https://doi.org/10.1101/746941
Amélie Luise Godeau
Laboratory of Cell Physics ISIS/IGBMC, CNRS and University of Strasbourg, Strasbourg, FranceInstitut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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Hélène Delanoë-Ayari
Université Claude Bernard Lyon 1, CNRS UMR 5306 10 rue Ada Byron, 69622 Villeurbanne Cedex, France
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Daniel Riveline
Laboratory of Cell Physics ISIS/IGBMC, CNRS and University of Strasbourg, Strasbourg, FranceInstitut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, FranceCentre National de la Recherche Scientifique, UMR7104, Illkirch, FranceInstitut National de la Santé et de la Recherche Médicale, U964, Illkirch, FranceUniversité de Strasbourg, Illkirch, France
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  • For correspondence: riveline@unistra.fr
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Abstract

Cell migration is involved in key phenomena in biology, ranging from development to cancer. Fibroblasts move between organs in 3D polymeric networks. So far, motile cells were mainly tracked in vitro on Petri dishes or on coverslips, i.e. 2D flat surfaces, which made the extrapolation to 3D physiological environments difficult. We therefore prepared 3D Cell Derived Matrix (CDM) with specific characteristics with the goal of extracting the main readouts required to measure and characterise cell motion: cell specific matrix deformation through the tracking of fluorescent fibronectin within CDM, focal contacts as the cell anchor and acto-myosin cytoskeleton which applies cellular forces. We report our method for generating this assay of physiological-like gel with relevant readouts together with its potential impact in explaining cell motility in vivo.

  • List of abbreviations

    CDM
    Cell Derived Matrix
    CS
    CoverSlip
    ECM
    ExtraCellular Matrix
    FN
    FibroNectin
    KLT
    Kanade-Lucas-Tomasi
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    The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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    Posted September 01, 2019.
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    Generation of fluorescent cell-derived-matrix to study 3D cell migration
    Amélie Luise Godeau, Hélène Delanoë-Ayari, Daniel Riveline
    bioRxiv 746941; doi: https://doi.org/10.1101/746941
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    Generation of fluorescent cell-derived-matrix to study 3D cell migration
    Amélie Luise Godeau, Hélène Delanoë-Ayari, Daniel Riveline
    bioRxiv 746941; doi: https://doi.org/10.1101/746941

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