Quantification of Muscle Satellite Stem Cell Divisions by High-Content Analysis

William Chen; Theodore J Perkins; Michael A Rudnicki

doi:10.1007/978-1-0716-2772-3_29

Quantification of Muscle Satellite Stem Cell Divisions by High-Content Analysis

Methods Mol Biol. 2023:2587:537-553. doi: 10.1007/978-1-0716-2772-3_29.

Authors

William Chen^{1

2}, Theodore J Perkins^{1

2

3}, Michael A Rudnicki^{4

5

6}

Affiliations

¹ Sprott Center for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada.
² Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada.
³ Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada.
⁴ Sprott Center for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada. [email protected].
⁵ Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada. [email protected].
⁶ Department of Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada. [email protected].

PMID: 36401049
DOI: 10.1007/978-1-0716-2772-3_29

Abstract

High-content screening is commonly performed on 2D cultured cells, which is high throughput but has low biological relevance. In contrast, single myofiber culture assay preserves the satellite cell niche between the basal lamina and sarcolemma and consequently has high biological relevance but is low throughput. We describe here a high-content screening method that utilizes single myofiber culture that addresses the caveats of both techniques. Our method utilizes the transgenic reporter allele Myf5-Cre:R26R-eYFP to differentiate stem and committed cells within a dividing couplet that can be quantified by high-content throughput immunodetection and bioinformatic analysis.

Keywords: Asymmetric self-renewal; High-content screening; Muscle satellite stem cell; Myofiber culture; Niche; Symmetric division.

Quantification of Muscle Satellite Stem Cell Divisions by High-Content Analysis

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Abstract

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