Chapter title |
Isolation, Culture, and Differentiation of Fibro/Adipogenic Progenitors (FAPs) from Skeletal Muscle
|
---|---|
Chapter number | 7 |
Book title |
Skeletal Muscle Development
|
Published in |
Methods in molecular biology, August 2017
|
DOI | 10.1007/978-1-4939-7283-8_7 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7282-1, 978-1-4939-7283-8
|
Authors |
Judson, Robert N., Low, Marcela, Eisner, Christine, Rossi, Fabio M., Robert N. Judson, Marcela Low, Christine Eisner, Fabio M. Rossi |
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. |
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