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Methods Mol Biol. 2018;1732:539-549. doi: 10.1007/978-1-4939-7598-3_34.

Analysis of Muscle Stem Cell Fate Through Modulation of AMPK Activity.

Author information

1
The Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada.
2
Faculty of Medicine, The University of British Columbia, Vancouver, BC, Canada.
3
Institut NeuroMyogène, CNRS UMR 5310, INSERM U1217, Université Claude Bernard Lyon 1, Villeurbanne, France.
4
Institut NeuroMyogène, CNRS UMR 5310, INSERM U1217, Université Claude Bernard Lyon 1, Villeurbanne, France. remi.mounier@univ-lyon1.fr.

Abstract

In this chapter, we describe the methods to isolate and culture muscle stem cells (MuSCs) from murine skeletal muscle in order to decipher the intrinsic effect of AMP-activated kinase activity on MuSC fate. Culture of MuSCs is a powerful model to recapitulate every step of stem cell behavior observed in vivo: activation, proliferation, differentiation, fusion and also self-renewal. We provide the detailed procedures to isolate pure MuSCs by a flow cytometry-based method using the selection of a combination of specific markers and to characterize MuSC fate (quiescence, activation, and differentiation) in response to AMPK activity modulation by assessing of the expression of stem cell (e.g., Pax7) and myogenic marker (e.g., MyoD).

KEYWORDS:

Differentiation; Flow cytometry; Metabolism; Muscle; Myogenesis; Pax7; Quiescence; Stem cell behavior

PMID:
29480498
DOI:
10.1007/978-1-4939-7598-3_34
[Indexed for MEDLINE]

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