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AMPK pp 539-549 | Cite as

Analysis of Muscle Stem Cell Fate Through Modulation of AMPK Activity

  • Marine Theret
  • Linda Gsaier
  • Sabrina Ben Larbi
  • Michèle Weiss-Gayet
  • Rémi MounierEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1732)

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).

Key words

Metabolism Stem cell behavior Myogenesis Muscle Quiescence Differentiation Pax7 Flow cytometry 

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Marine Theret
    • 1
    • 2
    • 3
  • Linda Gsaier
    • 3
  • Sabrina Ben Larbi
    • 3
  • Michèle Weiss-Gayet
    • 3
  • Rémi Mounier
    • 3
    Email author
  1. 1.The Biomedical Research CentreUniversity of British ColumbiaVancouverCanada
  2. 2.Faculty of MedicineThe University of British ColumbiaVancouverCanada
  3. 3.Institut NeuroMyogène, CNRS UMR 5310, INSERM U1217Université Claude Bernard Lyon 1VilleurbanneFrance

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