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

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2023 Mar 30

PMID 36995586

Authors

Ahmed S Shams

Michael Kyba

Affiliations

Soon will be listed here.

Abstract

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.

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