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Analysis of Muscle Stem Cell Fate Through Modulation of AMPK Activity

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AMPK

Part of the book series: Methods in Molecular Biology ((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).

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References

  1. Mounier R, Chretien F, Chazaud B (2011) Blood vessels and the satellite cell niche. Curr Top Dev Biol 96:121–138

    Article  CAS  PubMed  Google Scholar 

  2. Kuang S, Kuroda K, Le Grand F, Rudnicki MA (2007) Asymmetric self-renewal and commitment of satellite stem cells in muscle. Cell 129:999–1010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Yin H, Price F, Rudnicki MA (2013) Satellite cells and the muscle stem cell niche. Physiol Rev 93:23–67

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Theret M, Gsaier L, Schaffer B, Juban G, Ben Larbi S, Weiss-Gayet M, Bultot L, Caterina C, Foretz M, Desplanches D, Sanz P, Zang Z, Yang L, Vial G, Viollet B, Sakamoto K, Brunet A, Chazaud B, Mounier R (2017) AMPKalpha1-LDH pathway regulates muscle stem cell self-renewal by controlling metabolic homeostasis. EMBO J 36:1946

    Article  CAS  PubMed  Google Scholar 

  5. Joe AW, Yi L, Natarajan A, Le Grand F, So L, Wang J, Rudnicki MA, Rossi FM (2010) Muscle injury activates resident fibro/adipogenic progenitors that facilitate myogenesis. Nat Cell Biol 12:153–163

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Correspondence to Rémi Mounier .

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Theret, M., Gsaier, L., Ben Larbi, S., Weiss-Gayet, M., Mounier, R. (2018). Analysis of Muscle Stem Cell Fate Through Modulation of AMPK Activity. In: Neumann, D., Viollet, B. (eds) AMPK. Methods in Molecular Biology, vol 1732. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7598-3_34

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  • DOI: https://doi.org/10.1007/978-1-4939-7598-3_34

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7597-6

  • Online ISBN: 978-1-4939-7598-3

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