Flow Cytometer Analyses, Isolation, and Staining of Murine Muscle Satellite Cells

Fluorescence-activated cell sorting (FACS) is a powerful and requisite tool for the analysis and purification of adult stem cells. However, it is difficult to separate adult stem cells from solid organs than from immune-related tissues/organs. This is because of the presence of large amounts of debris, which increases noise in the FACS profiles. In particular, it is extremely difficult for unfamiliar researchers to identify muscle stem cell (also known as muscle satellite cell: MuSC) fraction because all myofibers, which are mainly composed of skeletal muscle tissues, become debris during cell preparation. This chapter describes our FACS protocol, which we have used for more than a decade, to identify and purify MuSCs.