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Isolation of Muscle Stem Cells from Mouse Skeletal Muscle

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Muscle Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1556))

Abstract

Isolation of muscle stem cells from skeletal muscle is a critical step for the study of skeletal myogenesis and regeneration. Although stem cell isolation has been performed for decades, the emergence of flow cytometry with defined cell surface markers, or transgenic mouse models, has allowed the efficient isolation of highly enriched stem cell populations. Here, we describe the isolation of mouse muscle stem cells using two different combinations of enzyme treatments allowing the release of mononucleated muscle stem cells from their niche. Mouse muscle stem cells can be further isolated as a highly enriched population by flow cytometry using fluorescent reporters or cell surface markers. We will present advantages and drawbacks of these different approaches.

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Abbreviations

FACS:

Fluorescent-activated cell sorting

TA:

Tibialis anterior

GFP:

Green fluorescent protein

FSC:

Forward scatter

SSC:

Side scatter

C/T:

Collagenase D/Trypsin

C/D:

Collagenase A/Dispase II

FBS:

Fetal bovine serum

CD:

Cluster of differentiation

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Acknowledgments

We acknowledge the funding support from the Institut Pasteur, Centre National pour la Recherche Scientifique, Association Française contre les Myopathies, Agence Nationale de la Recherche (Laboratoire d’Excellence Revive, Investissement d’Avenir; ANR-10-LABX-73), Association pour la Recherche sur le Cancer, EU Advanced ERC grant, and Fondation pour la Recherche Médicale. H. Sakai is funded by the ERC and F. Pala by the LabEx Revive/Pasteur PPU program.

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Authors and Affiliations

  1. Department of Developmental and Stem Cell Biology, Stem Cells and Development, CNRS URA 2578, Institut Pasteur, 25 rue du Dr. Roux, Paris, 75015, France

    Barbara Gayraud-Morel, Francesca Pala, Hiroshi Sakai & Shahragim Tajbakhsh

Authors
  1. Barbara Gayraud-Morel
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  2. Francesca Pala
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  3. Hiroshi Sakai
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  4. Shahragim Tajbakhsh
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Corresponding author

Correspondence to Barbara Gayraud-Morel .

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Editors and Affiliations

  1. Cell Biology Group, Department of Experimental and Health Sciences, University Pompeu Fabra (UPF), Barcelona, Spain

    Eusebio Perdiguero

  2. Division of Biological Sciences and Christopher S. Bond Life Sciences Center, University of Missouri, Columbia, MO, USA

    DDW Cornelison

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Gayraud-Morel, B., Pala, F., Sakai, H., Tajbakhsh, S. (2017). Isolation of Muscle Stem Cells from Mouse Skeletal Muscle. In: Perdiguero, E., Cornelison, D. (eds) Muscle Stem Cells. Methods in Molecular Biology, vol 1556. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-6771-1_2

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  • DOI: https://doi.org/10.1007/978-1-4939-6771-1_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-6769-8

  • Online ISBN: 978-1-4939-6771-1

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