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Isolation, Culture and Immunostaining of Skeletal Muscle Fibres to Study Myogenic Progression in Satellite Cells

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Stem Cells and Tissue Repair

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1210))

Abstract

Satellite cells are the resident stem cells of skeletal muscle, located on the surface of a myofibre, beneath the surrounding basal lamina. Satellite cells are responsible for the homeostasis, hypertrophy and repair of skeletal muscle fibres, being activated to enter proliferation and generate myoblasts that either fuse to existing myofibres, or fuse together for de novo myofibre formation. Isolating muscle fibres allows the associated satellite cells to be obtained while remaining in their anatomical niche beneath the basal lamina, free of interstitial and vascular tissue. Myofibres can then be immunostained to examine gene expression in quiescent satellite cells, or cultured to activate satellite cells before immunostaining to investigate gene expression dynamics during myogenic progression and self-renewal. Here, we describe methods for the isolation, culture and immunostaining of muscle fibres for examining satellite cell biology.

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Acknowledgements

We would like to thank Farah Patell for the confocal image of a satellite cell (Fig. 2e). Louise Moyle is supported by a Muscular Dystrophy Campaign PhD studentship (RA4/817). The laboratory of Pete Zammit is currently also supported by the Medical Research Council (G1100193), and Association Française Contre les Myopathies (SB/CP/2012-0218/15814 and SB/CF/2012-0910), together with OPTISTEM (223098) and BIODESIGN (262948-2) from the European Commission 7th Framework Programme.

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Authors and Affiliations

  1. Randall Division of Cell and Molecular Biophysics, New Hunt’s House, King’s College London, Guy’s Campus, London, SE1 1UL, UK

    Louise A. Moyle & Peter S. Zammit

Authors
  1. Louise A. Moyle
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  2. Peter S. Zammit
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Corresponding author

Correspondence to Peter S. Zammit .

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Editors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, Oregon, USA

    Chrissa Kioussi

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Moyle, L.A., Zammit, P.S. (2014). Isolation, Culture and Immunostaining of Skeletal Muscle Fibres to Study Myogenic Progression in Satellite Cells. In: Kioussi, C. (eds) Stem Cells and Tissue Repair. Methods in Molecular Biology, vol 1210. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1435-7_6

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  • DOI: https://doi.org/10.1007/978-1-4939-1435-7_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1434-0

  • Online ISBN: 978-1-4939-1435-7

  • eBook Packages: Springer Protocols

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