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

Methods Mol Biol. 2017;1668:93-103. doi: 10.1007/978-1-4939-7283-8_7.

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.

Keywords: FAPs; Fibro-adipogenic progenitors; Fibrosis; MSCs; Mesenchymal progenitors; Myogenesis; Satellite cells; Skeletal muscle; Skeletal muscle regeneration.

MeSH terms

  • Adipogenesis
  • Animals
  • Antigens, Surface / analysis
  • Cell Differentiation*
  • Cell Separation / methods*
  • Flow Cytometry / methods*
  • Mice
  • Multipotent Stem Cells / cytology
  • Multipotent Stem Cells / physiology*
  • Muscle Development
  • Osteogenesis
  • Primary Cell Culture
  • Regeneration
  • Satellite Cells, Skeletal Muscle / cytology
  • Satellite Cells, Skeletal Muscle / physiology*

Substances

  • Antigens, Surface