Superior survival and proliferation after transplantation of myoblasts obtained from adult mice compared with neonatal mice.
BACKGROUND
Myoblast transfer therapy (MTT) is a strategy designed to compensate for the defective gene in myopathies such as Duchenne muscular dystrophy (DMD). Experimental MTT in the mdx mouse (an animal model of DMD) has used donor myoblasts derived from mice of various ages; however, to date, there has been no direct quantitative comparison between the efficacy of MTT using myoblasts isolated from adult and neonate donor muscle.


METHODS
Donor normal male myoblasts were injected into Tibialis Anterior muscles of dystrophic female host mice and the survival and proliferation of male myoblasts quantitated using Y-chromosome specific real-time quantitative polymerase chain reaction. The survival of late preplate (PP6) myoblasts derived from neonatal (3-5 days old) or adult (6-8 weeks old) donor mice after MTT were compared. The influence of the number of tissue culture passages, on survival post-MTT, was also evaluated for both types of myoblasts.


RESULTS
Surprisingly, superior transplantation efficiency was observed for adult-derived compared with neonatal myoblasts (both early and late passage). Extended expansion (>17 passages) in tissue culture resulted in inferior survival and proliferation of both adult and neonatal myoblasts; however, proliferation of early passage myoblasts (both adult and neonate) was evident between 3 weeks and 3 months.


CONCLUSIONS
Myoblasts derived from neonatal mice were inferior for transplantation, and early passage donor myoblasts from adult mice are recommended for MTT in this model.
Lee-Pullen
Tracey F
TF
Discipline of Microbiology, School of Biomedical and Chemical Sciences, The University of Western Australia, QEII Medical Centre, Nedlands, Perth, WA 6009, Australia.
Bennett
Alayne L
AL
Beilharz
Manfred W
MW
Grounds
Miranda D
MD
Sammels
Leanne M
LM
D000375:Aging; D000818:Animals; D000831:Animals, Newborn; D002452:Cell Count; D002455:Cell Division; D002470:Cell Survival; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D005260:Female; D007273:Injections, Intramuscular; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D018482:Muscle, Skeletal / Q000601:surgery; D009137:Muscular Dystrophy, Animal / Q000601:surgery*; D032446:Myoblasts / Q000473:pathology / Q000637:transplantation*; D013997:Time Factors; D014019:Tissue Donors