title: Isolation and in vitro propagation of human skeletal muscle progenitor cells from fetal muscle.
abstract: Skeletal muscle progenitor cells (SMPCs) are considered one of the most valuable cells for cell-based therapy targeting skeletal muscle. However, an efficient protocol for isolating and maintaining human myogenic progenitors in vitro has not been fully established. In this study, we demonstrate that human myogenic progenitors can be expanded and proliferated from human fetal muscles. Human SMPCs were prepared from fetal hind limb muscles and induced to proliferate as free-floating spheres termed myospheres in the medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Both myogenic progenitors and myoblast populations from human fetal muscles were effectively propagated in myospheres and passaged by a mechanical chopping. After expanding these spheres in culture, we tested whether myogenic progenitor cells can differentiate into multinucleated myotubes. The myospheres were dissociated, plated down on coverslips and cultured in the medium for terminal differentiation. We could confirm that the plated cells formed well-developed, multinucleated myotubes. This culture method using myospheres is an effective protocol to isolate and maintain SMPCs from human fetal skeletal muscles in culture.
authors: Department of Comparative Biosciences, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors: Tohru
authors: T
authors: Hosoyama
authors: Michael G
authors: MG
authors: Meyer
authors: Dan
authors: D
authors: Krakora
authors: Masatoshi
authors: M
authors: Suzuki
mesh_terms: D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000378:metabolism; D005333:Fetus / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D006801:Humans; D018482:Muscle, Skeletal / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor