Isolation of Mouse, Rat, and Human Muscle Stem Cells by Fluorescence-Activated Cell Sorting Based on α7-Integrin

Muscle stem cells (MuSC) are necessary for skeletal muscle tissue homeostasis and repair following injury. These cells are therefore extensively studied and are considered promising candidates for cell-based therapies, rendering their purification a recurring technical requirement. For this purpose, adhesion molecules expressed on MuSC surface represent ideal positive selection markers, for magnetic-activated cell sorting (MACS) or fluorescence-activated cell sorting (FACS). Here, we describe a method for efficiently isolating MuSC from postnatal and adult muscles of mice, rats, and humans by FACS, using α7-integrin (ITGA7) as common selection marker. We provide a detailed protocol for the immunostaining of mononucleated cells derived from digested skeletal muscle and the gating strategies to obtain highly pure MuSC from the muscles of these three species.