In situ fixation redefines quiescence and early activation of skeletal muscle stem cells
Open accessAbstract
State of the art techniques have been developed to isolate and analyze cells from various tissues, aiming to capture their in vivo state. However, the majority of cell isolation protocols involve lengthy mechanical and enzymatic dissociation steps followed by flow cytometry, exposing cells to stress and disrupting their physiological niche. Focusing on adult skeletal muscle stem cells, we have developed a protocol that circumvents the impact of isolation procedures and captures cells in their native quiescent state. We show that current isolation protocols induce major transcriptional changes accompanied by specific histone modifications while having negligible effects on DNA methylation. In addition to proposing a protocol to avoid isolation-induced artifacts, our study reveals previously undetected quiescence and early activation genes of potential biological interest.
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FACS-isolated cells used here express some markers of early MuSC activation, even immediately after isolation.
“Recent studies have reported that the FACS-isolated cells used here express some markers of early MuSC activation, even immediately after isolation. Therefore, our ‘quiescent’ cell populations probably experience some early activation stimulus during isolation (Machado et al., 2017; Velthoven et al., 2017).”
From Aging induces aberrant state transition kinetics in murine muscle stem cells (2020) · cites this paper as Machado et al., 2017
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References 40 · Cited by 328
- Authors
- Machado, L., Esteves de Lima, J., Fabre, O., Proux, C., Legendre, R., Szegedi, A., Varet, H., Ingerslev, L. R., Barrès, R., Relaix, F., & Mourikis, P.
- Affiliations
- Inserm (FR)
- University of Copenhagen (DK)
- Institut Pasteur (FR)
- Centre National de la Recherche Scientifique (FR)
- Corresponding author
- Relaix, Frédéric
- ORCID
- Venue
- Cell Reports
- Publisher
- Elsevier BV
- Published
- 2017-11
- Volume
- 21
- Issue
- 7
- Pages
- 1982-1993
- ISSN
- 2211-1247 (ISSN-L)
- OpenAlex
- W2769209008
- Cited by
- 328
- References
- 40
- Citations by year
- 2026: 35 · 2025: 28 · 2024: 37 · 2023: 44 · 2022: 42 · 2021: 63 · 2020: 40 · 2019: 24 · 2018: 12 · 2017: 2
- FWCI
- 8.83
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Cell Reports (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by-nc-nd)
- HAL (submittedVersion, cc-by-nc-nd)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Fondation pour la Recherche Médicale (DEQ20130326526, FDT20130928236)
- Novo Nordisk Fonden
- Association Française contre les Myopathies (19507, ANR-10-LABX-73)
- Danish Diabetes Academy (1077471001)
- Agence Nationale de la Recherche (ANR 11 BSV2 017 02, ANR-13-BSV1-0011-02)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- skeletal muscle stem cells, satellite cells, in situ fixation, quiescence, early activation, histone modifications, DNA methylation, gene expression changes
- MeSH terms
- Animals, Cells, Cultured, Female, Tissue Fixation, DNA Methylation, Myoblasts, Skeletal, Histone Code, Mice, Transcriptome, Primary Cell Culture
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