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Flow Cytometry and Transplantation-Based Quantitative Assays for Satellite Cell Self-Renewal and Differentiation

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Book cover Skeletal Muscle Regeneration in the Mouse

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1460))

Abstract

In response to muscle damage, satellite cells proliferate and undertake both differentiation and self-renewal, generating new functional muscle tissue and repopulating this new muscle with stem cells for future injury responses. For many questions relating to the physiological regulation of satellite cells, quantitative readouts of self-renewal and differentiation can be very useful. There is a particular need for a quantitative assay for satellite cell self-renewal that does not rely solely upon sectioning, staining and counting cells in sections. In this chapter, we provide detailed methods for quantifying the self-renewal and differentiation potential of a given population of satellite cells using an assay involving transplantation into injured, regenerating muscle together with specific markers for donor cell identity and state of differentiation. In particular, using the Pax7-ZsGreen transgene as a marker of satellite cell state, self-renewal can be quantified by FACS on transplanted muscle to actually count the total number of resident satellite cells at time points following transplantation.

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Acknowledgments

This work was supported by grants from the NIH (R01 AR055685) and the Muscular Dystrophy Association (MDA351022).

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Authors and Affiliations

  1. Lillehei Heart Institute and Department of Pediatrics, University of Minnesota, Cancer and Cardiovascular Research Building 4-127, 2231 6th St. S.E., Minneapolis, MN, 55455, USA

    Robert W. Arpke & Michael Kyba Ph.D.

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  1. Robert W. Arpke
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  2. Michael Kyba Ph.D.
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Corresponding author

Correspondence to Michael Kyba Ph.D. .

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Editors and Affiliations

  1. Lillehei Heart Institute, University of Minnesota, Minneapolis, Minnesota, USA

    Michael Kyba

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Arpke, R.W., Kyba, M. (2016). Flow Cytometry and Transplantation-Based Quantitative Assays for Satellite Cell Self-Renewal and Differentiation. In: Kyba, M. (eds) Skeletal Muscle Regeneration in the Mouse. Methods in Molecular Biology, vol 1460. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-3810-0_12

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  • DOI: https://doi.org/10.1007/978-1-4939-3810-0_12

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-3808-7

  • Online ISBN: 978-1-4939-3810-0

  • eBook Packages: Springer Protocols

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