Flow cytometry and transplantation-based quantitative assays for satellite cell self-renewal and differentiation

Robert W Arpke, Michael Kyba

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages163-179
Number of pages17
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1460
ISSN (Print)1064-3745

Bibliographical note

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

Publisher Copyright:
© Springer Science+Business Media New York 2016.

Keywords

  • Myogenesis
  • Pax7
  • Satellite cells
  • Transplantation

Fingerprint

Dive into the research topics of 'Flow cytometry and transplantation-based quantitative assays for satellite cell self-renewal and differentiation'. Together they form a unique fingerprint.

Cite this