Therapeutic isolation and expansion of XXXX skeletal muscle-derived stem cells for the use of muscle-nerve-blood vessel reconstitution.

@entity1 @entity8236 @entity263 @entity19 @entity27064 @entity35 @entity22416
Skeletal muscle makes up 40-50% of body mass, and is thus considered to be a good adult stem cell source for autologous therapy. Although, several stem/progenitor cells have been fractionated from @entity19 skeletal muscle showing a high potential for therapeutic use, it is unclear whether this is the case in @entity1 . Differentiation and therapeutic potential of @entity1 skeletal muscle-derived cells (Sk-Cs) was examined. Samples (5-10 g) were obtained from the abdominal and leg muscles of 36 @entity1 (age, 17-79 years) @entity263 treatment or leg amputation surgery. All @entity1 gave informed consent. Sk-Cs were isolated using conditioned collagenase solution, and were then sorted as @entity22416 (-)/ @entity8236 (-)/ @entity27064 (+) (Sk-DN/29(+)) and @entity22416 (+)/ @entity8236 (-) (Sk-34) cells, in a similar manner as for the previous @entity19 Sk-Cs. Both cell fractions were appropriately expanded using conditioned culture medium for about 2 weeks. Differentiation potentials were then examined during cell culture and in vivo transplantation into the severely damaged muscles of athymic @entity19 and @entity35 . Interestingly, these two cell fractions could be divided into highly myogenic (Sk-DN/29(+)) and multipotent stem cell (Sk-34) fractions, in contrast to @entity19 Sk-Cs, which showed comparable capacities in both cells. At 6 weeks after the separate transplantation of both cell fractions, the former showed an active contribution to muscle fiber regeneration, but the latter showed vigorous engraftment to the interstitium associated with differentiation into Schwann cells, perineurial/endoneurial cells, and vascular endothelial cells and pericytes, which corresponded to previous observations with @entity19 SK-Cs. Importantly, mixed cultures of both cells resulted the reduction of tissue reconstitution capacities in vivo, whereas co-transplantation after separate expansion showed favorable results. Therefore, @entity1 Sk-Cs are potentially applicable to therapeutic autografts and show multiple differentiation potential in vivo.
