Isolation and characterization of myogenic satellite cells from the muscular dystrophic hamster

Myogenic satellite cells were isolated from control and dystrophic hamster diaphragms to examine cellular mechanisms involved in the physiology of mus cular dystrophy, The Bio 14.6 dystrophic hamster, which possesses a defect in the delta-sarcoglycan gene, develops biochemical and physical symptoms o f Duchenne-like and limb girdle muscular dystrophies. Because primary cultu res of the control and dystrophic satellite cells became extensively contam inated with non-myogenic cells during proliferation, cell clones were devel oped to provide pure cultures for study. Cell culture conditions were optim ized with the use of Ham's F-12K medium containing 10% fetal bovine serum 5% horse serum + 10 ng/mL basic fibroblast growth factor + 50 mu g/mL porci ne gelatin. Proliferation rates of the two clonal cultures were similar bet ween the two lines. Satellite cell-derived myotubes from both primary cultu res and clones differed between control and dystrophic animals. Dystrophic myotubes tended to be long and narrow, while the control-derived myotubes w ere broader. Measurement of muscle-specific creatine kinase during differen tiation revealed that the dystrophic myotubes possessed higher creatine kin ase levels than control myotubes (up to 146-fold at 168 h). The results dem onstrate that satellite cells can be isolated from the hamster and may prov ide a useful tool to study muscular dystrophies associated with defects in the sarcoglycan complex and the involvement of sarcoglycans in normal skele tal muscle growth and development. (C) 2000 Harcourt Publishers Ltd.