title: Stable integration of an mdx skeletal muscle cell line into dystrophic (mdx) skeletal muscle: evidence for stem cell status.
abstract: We have previously described a method for the derivation of long term cultures of undifferentiated myoblasts from the skeletal muscle of dystrophic (mdx) mice (J. Smith and P. N. Schofield, Exp. Cell Res., 210: 86-93, 1994). We now show that a clonal mdx-derived skeletal muscle cell line labeled with a retrovirus conferring beta-galactosidase activity and G418 resistance (PD50A) is capable of incorporation into mdx skeletal muscle myofibers for up to 14 months with no incidence of tumor formation. After a lag period of 5 days, injected PD50A cells disperse throughout the injected tibialis anterior muscle and take up satellite cell positions on the perimeter of myofiber bundles. PD50A cells begin to incorporate into fused muscle syncitium as early as 8 weeks after injection and persist for at least 14 months. We have rederived myoblasts expressing beta-galactosidase from PD50A-injected muscles 12 months after injection, demonstrating that a reserve of mononuclear proliferation-competent PD50A cells are present in host muscle up to a year after their original introduction. These data support the contention that myoblasts derived by this culture method are functionally representative of a class of skeletal muscle "stem cells" and thus have potential both as agents for cellular therapy of intransigent diseases such as Duchenne muscular dystrophy as well as being a useful tool for the further investigation of normal muscle development.
authors.affiliation: Department of Anatomy, University of Cambridge, United Kingdom. JS10046@cam.ac.uk
authors.forename: J
authors.initials: J
authors.lastname: Smith
authors.forename: P N
authors.initials: PN
authors.lastname: Schofield
mesh_terms: D000818:Animals; D002460:Cell Line; D017690:Cell Transplantation / Q000379:methods*; D005260:Female; D018014:Gene Transfer Techniques; D007621:Karyotyping; D008297:Male; D051379:Mice; D018101:Mice, Inbred mdx; D018485:Muscle Fibers, Skeletal / Q000166:cytology*; D018482:Muscle, Skeletal / Q000166:cytology*; D012190:Retroviridae / Q000235:genetics; D013234:Stem Cells / Q000166:cytology*; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D001616:beta-Galactosidase