Isolation and Culture of Primary Myoblasts from Humans and Mice

Primary myoblasts are generated from committed stem cells, responsible for repairing skeletal muscle fibers upon injury. They are a valuable resource for scientific research in muscle disease and drug screening and hold promise for cell-based therapies. However, skeletal muscle biopsies contain a plethora of non-myogenic cells, exacerbated in diseased muscle tissues. Thus, purification methods of the myogenic cell population are necessary. This protocol describes techniques for dissociating cells from human and mouse skeletal muscle biopsies and enrichment for a highly myogenic population using magnetic isolation for mouse muscles and by fluorescence-activated cell sorting (FACS) using cell surface antibodies for humans. We also describe methods to expand myoblasts in culture and assess myogenicity.