Protocol for cell proliferation and cell death analysis of primary muscle stem cell culture using flow cytometry

Skeletal muscle is critically dependent on the function of muscle stem cells (MuSCs) for effective muscle repair following injury. Here, we detail a protocol for the isolation of primary muscle cells and subsequent analysis of proliferation capacity in vitro using EdU (5-ethynyl-2'-deoxyuridine) on fixed cells. We also describe a cell death analysis on living cells with the identification of early- and late-apoptotic cells, as well as necrotic cells, through the incorporation of propidium iodide and YO-PRO-1 staining. For complete details on the use and execution of this protocol, please refer to Garcia et al.<sup>1</sup>.

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Publication Details

Journal
STAR Protocols
Published
2024-10-23
DOI
https://doi.org/10.1016/j.xpro.2024.103411
Primary Topic
Muscle Physiology and Disorders
Type
article
Field-Weighted Citation Impact
0.00

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article

Protocol for cell proliferation and cell death analysis of primary muscle stem cell culture using flow cytometry

Fabien Le Grand, Jade Ravent, Pauline Garcia, Orane Mercier
STAR Protocols
Muscle Physiology and Disorders
article

Protocol for cell proliferation and cell death analysis of primary muscle stem cell culture using flow cytometry

Fabien Le Grand, Jade Ravent, Pauline Garcia, Orane Mercier
article en

Abstract

Skeletal muscle is critically dependent on the function of muscle stem cells (MuSCs) for effective muscle repair following injury. Here, we detail a protocol for the isolation of primary muscle cells and subsequent analysis of proliferation capacity in vitro using EdU (5-ethynyl-2'-deoxyuridine) on fixed cells. We also describe a cell death analysis on living cells with the identification of early- and late-apoptotic cells, as well as necrotic cells, through the incorporation of propidium iodide and YO-PRO-1 staining. For complete details on the use and execution of this protocol, please refer to Garcia et al.<sup>1</sup>.

STAR ProtocolsVol. 5(4)
Université Claude Bernard Lyon 1 (FR), Centre National de la Recherche Scientifique (FR), Inserm (FR), Institut NeuroMyoGène (FR)
Association Française contre les Myopathies, Agence Nationale de la Recherche, Institut National de la Santé et de la Recherche Médicale, Fondation pour la Recherche Médicale, Centre National de la Recherche Scientifique, AFM-Téléthon
Good health and well-being
Openalex Percentile: Top 83%
Muscle Physiology and Disorders
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Protocol for cell proliferation and cell death analysis of primary muscle stem cell culture using flow cytometry — Fabien Le Grand, Jade Ravent, et al. · STAR Protocols (2024) | TGRS Research Map | TGRS