Anti sca 1 pe

TA muscle was harvested, and satellite cell-derived primary myoblasts were isolated as described previously (Motohashi et al., 2014 ). Briefly, TA muscles of MyoD^−/− mice or their wild-type littermates were dissected and minced and then digested in type I collagenase/dispase B mixture (Roche Applied Science). The digestions were filtered from debris for magnetic-activated cell sorting (MACS). Dissociated muscle cells were incubated with Anti-CD45-PE, anti-Sca-1-PE and anti-CD31-PE antibodies (all from eBiosciences) followed by anti-PE beads (Miltenyi Biotec). LD column (Miltenyi Biotec) was used for negative selection to eliminate hematopoietic cells, endothelial cells and other non-muscle cells from the muscle preparation. Flow through fraction was incubated with integrin α-biotin antibody (Miltenyi Biotec) followed by anti-biotin beads (Miltenyi Biotec). MS column (Miltenyi Biotec) was used for primary myoblast isolation as a positive fraction. Isolated primary myoblasts were cultured in growth media (F-10 Ham's media supplemented with 17% FBS, 4 ng/ml basic fibroblast growth factor and 1% penicillin-streptomycin) on collagen-coated dishes, and the growth medium was changed every two days. Primary myoblasts were cultured in normal humidified tissue culture incubators with 5% CO₂.