Isolation of Skeletal Muscle Stem Cells for Phenotypic Screens for Modulators of Proliferation.

Adult skeletal muscle contains a population of resident stem cells known as muscle stem cells (MuSC) or satellite cells. This population of cells is required for regeneration of functional myofibers after damage. Aging reduces the proliferative response of satellite cells post-injury. This deficient response is thought to contribute to slowed recovery of muscle function after damage in the elderly and may also contribute to age-related loss of muscle function (sarcopenia). Numerous techniques are now available for the isolation of highly purified satellite cells from mice and humans (Sherwood, et al. Cell 119:543-554, 2004; Cerletti, et al. Cell 134:37-47; 2008; Conboy, et al. Methods Mol Biol 621:165-173, 2010; Bareja, et al. PLoS One 9:e90398; 2014; Castiglioni et al. Stem Cell Rep 2:92-106, 2014; Charville, et al. Stem Cell Rep 5:621-632, 2015; Liu et al. Nat Protoc 10:1612-1624, 2015; Sincennes et al. Methods Mol Biol 1556:41-50, 2017), thus opening an opportunity to use satellite cells in phenotypic screens for regulators of satellite cell proliferation and differentiation. In this chapter, we describe a technique for the prospective isolation of mouse satellite cells that we have recently used in a phenotypic screen of a focused set of small molecules.