Intraarterial injection of muscle-derived CD34(+)Sca-1(+) stem cells restores dystrophin in mdx mice.
Duchenne muscular dystrophy is a lethal recessive disease characterized by widespread muscle damage throughout the body. This increases the difficulty of cell or gene therapy based on direct injections into muscles. One way to circumvent this obstacle would be to use circulating cells capable of homing to the sites of lesions. Here, we showed that stem cell antigen 1 (Sca-1), CD34 double-positive cells purified from the muscle tissues of newborn mice are multipotent in vitro and can undergo both myogenic and multimyeloid differentiation. These muscle-derived stem cells were isolated from newborn mice expressing the LacZ gene under the control of the muscle-specific desmin or troponin I promoter and injected into arterial circulation of the hindlimb of mdx mice. The ability of these cells to interact and firmly adhere to endothelium in mdx muscles microcirculation was demonstrated by intravital microscopy after an intraarterial injection. Donor Sca-1, CD34 muscle-derived stem cells were able to migrate from the circulation into host muscle tissues. Histochemical analysis showed colocalization of LacZ and dystrophin expression in all muscles of the injected hindlimb in all of five out of five 8-wk-old treated mdx mice. Their participation in the formation of muscle fibers was significantly increased by muscle damage done 48 h after their intraarterial injection, as indicated by the presence of 12% beta-galactosidase-positive fibers in muscle cross sections. Normal dystrophin transcripts detected enzymes in the muscles of the hind limb injected intraarterially by the mdx reverse transcription polymerase chain reaction method, which differentiates between normal and mdx message. Our results showed that the muscle-derived stem cells first attach to the capillaries of the muscles and then participate in regeneration after muscle damage.
Torrente
Y
Y
Instituto de Ricovero e Cura a Carattere Scientifico (IRCCS) Ospedale Maggiore Policlinico, 20122 Milan, Italy.
Tremblay
J P
JP
Pisati
F
F
Belicchi
M
M
Rossi
B
B
Sironi
M
M
Fortunato
F
F
El Fahime
M
M
D'Angelo
M G
MG
Caron
N J
NJ
Constantin
G
G
Paulin
D
D
Scarlato
G
G
Bresolin
N
N
D000199:Actins / Q000032:analysis; D000818:Animals; D000831:Animals, Newborn; D018952:Antigens, CD34 / Q000032:analysis; D000950:Antigens, Ly / Q000032:analysis; D002448:Cell Adhesion; D002454:Cell Differentiation; D002460:Cell Line; D017690:Cell Transplantation / Q000379:methods*; D016189:Dystrophin / Q000032:analysis / Q000235:genetics*; D004730:Endothelium, Vascular / Q000502:physiology; D015316:Genetic Therapy; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000502:physiology*; D006614:Hindlimb; D016130:Immunophenotyping; D007269:Injections, Intra-Arterial; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D018101:Mice, Inbred mdx; D008822:Mice, Transgenic; D008833:Microcirculation / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000502:physiology; D018482:Muscle, Skeletal / Q000166:cytology*; D009137:Muscular Dystrophy, Animal / Q000628:therapy*; D009218:Myosins / Q000032:analysis; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D000199:Actins; D018952:Antigens, CD34; D000950:Antigens, Ly; D016189:Dystrophin; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D001616:beta-Galactosidase; D009218:Myosins