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Human and Murine Skeletal Muscle Reserve Cells

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1035)

Abstract

Study of stem cell phenotype and functions requires their proper isolation. Stem cells isolated from skeletal muscle are a useful tool to explore molecular pathways involved in the regulation of myogenesis. Among progenitor cells, a subset of cells, called reserve cells, has been identified, in vitro, in myogenic cell cultures. This subset of cells remains undifferentiated while the main population of progenitor cells commits to terminal myogenic differentiation. When replated, these reserve cells grow as new colonies of progenitors. At the time of differentiation, they reform both differentiated myotubes and undifferentiated reserve cells. Here, we present a protocol to obtain and further isolate reserve cells from both human and murine myogenic cell cultures, together with techniques to analyze their cell cycle status.

Key words

Skeletal muscle Stem cells Satellite cells Myogenic progenitor cells Myogenesis Human Mouse Cell cycle Nocodazole synchronization Reserve cells 

Notes

Acknowledgments

We thank Jyotsna Dhawan for her advices concerning the synchronization of myogenic cell cultures. We thank Marie-Claude Gendron for the setting up of the flow cytometry analysis of Pyronin/Hoechst staining.

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.INSERM U781, Hôpital Necker-Enfants MaladesParisFrance
  2. 2.Institut Cochin, INSERM U1016ParisFrance

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