Isolation, Culture, and Differentiation of Fibro/Adipogenic Progenitors (FAPs) from Skeletal Muscle

  • Robert N. Judson
  • Marcela Low
  • Christine Eisner
  • Fabio M. Rossi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1668)

Abstract

Fibro/Adipogenic Progenitors (FAPs) are a multipotent progenitor population resident in skeletal muscle. During development and regeneration, FAPs provide trophic support to myogenic progenitors that is required for muscle fiber maturation and specification. FAPs also represent a major cellular source of fibrosis in degenerative disease states, highlighting them as a potential cellular target for anti-fibrotic muscle therapies. Effective and reproducible methods to isolate and culture highly purified FAP populations are therefore critical to further understand their biology. Here, we describe a fluorescent activated cell sorting (FACS) based protocol to isolate CD31−/CD45−/Integrin-α7−/Sca1+ FAPs from murine skeletal muscle including details of tissue collection and enzymatic muscle digestion. We also incorporate optimized methods of expanding and differentiated FAPs in vitro. Together, this protocol provides a complete workflow to study skeletal muscle derived FAPs and compliments downstream analytical, drug screening, and disease modeling applications.

Key words

FAPs Fibro-adipogenic progenitors Skeletal muscle MSCs Mesenchymal progenitors Myogenesis Skeletal muscle regeneration Satellite cells Fibrosis 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Robert N. Judson
    • 1
    • 2
  • Marcela Low
    • 1
  • Christine Eisner
    • 1
  • Fabio M. Rossi
    • 1
  1. 1.Biomedical Research CentreUniversity of British ColumbiaVancouverCanada
  2. 2.STEMCELL Technologies Inc.VancouverCanada

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