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>.
Authors
- Fabien Le Grand (ORCID: https://orcid.org/0000-0002-7843-3899)
- Jade Ravent
- Pauline Garcia (ORCID: https://orcid.org/0000-0002-4673-8583)
- Orane Mercier
Institutions
- Université Claude Bernard Lyon 1 (FR)
- Centre National de la Recherche Scientifique (FR)
- Inserm (FR)
- Institut NeuroMyoGène (FR)
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
Funders
- 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