Adult Canine Skeletal Myoblast Autologous Transplantation Treatment Of Acute Myocardial Infarction In Experimental Research

Modified culture, identification and characteritics of adult cannine skeletal myoblastsObjective: To establish a thrifty, convenient procedure for culture, identification and biological characteristics of adult cannine skeletal myoblasts.Methods: Adult cannine myoblasts were obtained through two-step digestion method(inclduing collagenase IA and trypsin) combined with mechanical mincing, and were subjected to primary as well as secondary culture in vitro. Morphorlogical characteristics and growth curve were observed to evaluate the replicative and differential capacity of adult cannin skeletal myoblasts. The myoblasts were identified by cellular immunochemical stain. To reevaluate the necessity of using different attatchment capacity as a purificative way through calculation of the positive rate of anti- α -sarcometric-actin stained cell obtaind with or without purificative procedure respectively.Results: Adult cannine myoblats obtaind through two-step digestion method combined with mechanical mincing, can replicate very fast in proliferative medium, and form myotubes in differential medium which can be observed clearly by fluorescent microscopy after labbled by DAPI. Though the myoblasts were stained weakly positively by cellular immunochemical stain with anti-α-sarcometric actin and anti-myosin antibody, formed myotubes were stained strongly positively at the same conditions. Anti-myogenin immuochemical stain result of myoblasts showed strongly positive. The positive rate of anti-sarcometric actin-stained cell obtaind with or without purificative procedure has no significant differnece.Conclusion: The two-step digestion combined with mechanical mincing can be used as a thrifty and convenient alterative to get adult cannine myoblasts. The purificative procedure utilizing the different attatchment...