The Satellite Cell Colony Forming Cell Assay as a Tool to Measure Self-Renewal and Differentiation Potential

The muscle satellite cell population is responsible for homeostatic maintenance of muscle fibers in response to muscle injury and normal wear and tear. This population is heterogeneous, and its capacity for self-renewal and differentiation can be altered either by mutation of genes that regulate the...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2640; p. 45
Main Authors Shams, Ahmed S, Kyba, Michael
Format Journal Article
LanguageEnglish
Published United States 2023
Subjects
Cell Differentiation - physiology
Cells, Cultured
Humans
Muscle Fibers, Skeletal
Muscle, Skeletal
Muscular Diseases
Satellite Cells, Skeletal Muscle
Colony
Self-renewal
Satellite cells
Differentiation
Skeletal muscle
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Summary:The muscle satellite cell population is responsible for homeostatic maintenance of muscle fibers in response to muscle injury and normal wear and tear. This population is heterogeneous, and its capacity for self-renewal and differentiation can be altered either by mutation of genes that regulate these processes or with natural processes such as aging. The satellite cell colony assay is a facile way to extract information about the proliferation and differentiation potential of individual cells. Here, we provide a detailed protocol for the isolation, single cell plating, culture, and evaluation of colonies derived from single satellite cells. The variables of cell survival (cloning efficiency), proliferative potential (nuclei per colony), and differentiation propensity (ratio of nuclei within myosin heavy chain-positive cytoplasm to total nuclei) can thus be obtained.
ISSN:1940-6029
DOI:10.1007/978-1-0716-3036-5_4