Human Skeletal Muscle-Derived Mesenchymal Stem/Stromal Cell Isolation and Growth Kinetics Analysis

: The most studied sources of mesenchymal stem/stromal cells (MSCs) are bone marrow and adipose tissue. However skeletal muscle represents an interesting source of diverse subpopulations of MSCs, such as paired box 7 (Pax-7)-positive satellite cells, fibro-/adipogenic progenitors, PW1-positive interstitial cells and others. The specific properties of some of these muscle-derived cells have encouraged the development of cell therapies for muscle regeneration. However, the identity and multilineage potential of the diverse muscle-resident cells should first be evaluated in vitro, followed by in vivo clinical trials to predict their regenerative capacity. Here, we present protocols for the isolation of MSCs from skeletal muscle using enzymatic digestion and mechanical trituration. We also provide a method to determine their specific growth rate, a feature that is of particular interest when designing cell therapies.