Macs skeletal muscle dissociation kit for mouse and rat

Six-week old enhanced GFP transgenic C57BL/6 mice were anesthetized with isoflurane and sacrificed by exsanguination from the heart. Connective tissue, blood vessels, and fat were removed from skeletal muscle collected from the lower limbs. The collected muscle tissue was placed in Hanks' balanced salt solution (HBSS-, FUJIFILM Wako Pure Chemical Corporation) containing 1% PS and gently shaken to avoid contamination. Myoblast-containing cell suspensions were prepared using the MACS skeletal muscle dissociation kit for mouse and rat (Miltenyi Biotec, North Rhine, Germany). Then, mouse myoblasts were isolated by MACS with a purity of 98.5%, using the satellite cell isolation kit (Miltenyi Biotec), according to the manufacturers’ protocols [45 (link)]. Obtained cells were expanded on cell culture dishes coated with type I collagen solution (FUJIFILM Wako Pure Chemical Corporation) with myoblast culture medium. Since Motohashi et al. reported that these myoblasts engrafted to muscle when intramuscularly injected, it is assumed that the cells in this study, obtained by the same method, would also be able to engraft if they migrated into the muscle [45 (link)].