Experimental Study Of Myoblast And Mesenchymal Stem Cells Transplantation To Treat Progressive Musclar Dystrophy

Objective: To verificate culture,amplification,induction anddifferentiation of myoblast and Mesenchymal stem cells (MSCs) of themouse in vitro; to transplant the myoblast and MSCs to mdx mice, themodel animal of DMD, by vena caudalis; To observe the growth andfusion of myoblast and MSCs in the host after transplantation. Investigatethe method and feasibility of cell and gene therapy for DMD. Methods:Take the gastrocnemius muscle from C57/BL10 mouse of 5～7 days old, anduse the methods of steps enzyme digestion and differential adhesion toobtain sufficient myoblasts in vitro, marking the cells with BrdU,whileinject the 4th generation cells into mdx mouse via vena caudalisintravenous, then observe the growth of myoblast in the mdx mouse after4 weeks' breeding; Take out the tibia of 4 weeks old C57/BL10 mice inasepsis condition,and rinse the marrow with L-DMEM to culture the MSCs with the method of differential adhesion and density gradient.centrifugation, marking with BrdU after the third generation and inducingwith 5-azacytidine, then inject the inducted differenciation cells into themdx mice via vena caudalis, while observe the growth of MSCs in hostafter 4 weeks' breeding. Inject the control group with 1ml DF. Result:The primary culture of myoblastcan be generated after 3～4 days with themethods of steps enzyme digestion and differential adhesion so as to getstable and enough cells to satisfy the need of experiment. The primaryculture of mesenchymal stem cells (MSCs) can be generated after 7～10days with the methods of differential adhesion and density gradientcentrifugation, and the induced mesenchymal stem cells can differenciateto myoblast expressing desmin, the motor activity of the injecton group isobviously enhanced than the control group in exhausted swimmingexperiment; Execute the mice after 4 weeks and test the expression ofBrdU,dsmin and dystrophin of the muscle while observe fusion oftransplanted myoblast with local myofiber. Conclusion: Myoblast that isabundent and easy to get can fuse with the local myoblast to formpolynuclemyoblast and live in myofiber after transplantation.Mesenchymal stem cells (MSCs) are one of promising vector cells in celland gene projection for their extensive resource, easy to culture, and potential ability to differenciate into multiple cell. Transplanted cellsthrough venous can partly fuse with cells of the host and celltransplantation can play a role in DMD animal model.