Study On Rattit Mesenchymal Stem Cell Induced Into Cardiomyocytes In Vitro

Nowadays, MSCs induced into other type cells is one focus of biology research .Because impaired cardiomyocytes is irreversible , the necrotic cardiomyocytes can only be replaced by scar tissue. The application of transplant of heart is limited because of lacking of donator, moreover, the ordinary therapeutic method such as medication and coronary artery bypass can't reverse necrotic cardiac myocytes .The bone marrow-derived mesenchymal stem cell have some advantages,such as potential of hyperplasia and differentiation,more easily be got,adherence and be isolated easily, stable quantity, be suitable for auto grafting, be avoided dispute of ethic. Therefore, cellular transplantation is being explored as a newly developed strategy for patients with end-stage heart disease, and bone marrow mesenchymal stem cells (BMSCs)have been proposed as candidate to repair the necrotic myocardial tissue.The present study is to culture rabbit bone marrow mesenchymal stem cells, study part of their biologic characteristics, observe the effect of 5-Azachtine to Macs. To establish an effective method for inducing rabbit marrow mesenchymal stem cells to cardiomyocytes. It might provide a basis data's for farther studies on MSCs differentiating into cardiomyocytes. The experimental results are as fellows:1. The result shows that MSCs were adherent cells with a similar fibroblastic spindle-shaped morphology during different passages. MSCs show fibroblast-like morphology from first passage to passage 8 before treatment, the growth curve of different generation of MSCs indicated that the activity of MSCs declined with the passage of cells. Rabbit MSCs can be isolated and cultured in vitro successfully. It put the foundation for further studies.2. From the comparative study of MSCs of different age rabbit, we found that the MSCs from baby rabbit showed a stronger capability of proliferation compared with the MSCs from adult and aged rabbit(P<0.05). The OD of rabbit MSCs of different ages was compared, the result showed that the discrepancy was evident (P<0.05) at 3rd, 4th, 5th day. The adherence rate of rabbit MSCs of different ages was compared, the result showed that the adherence rate of baby rabbit, adult rabbit and old rabbit MSCs was 80.67%, 75.33%, 56.33%, respectively (P<0.05) at 6th day. Baby rabbits MSCs had better abilities of adaptation, proliferation, cell viability and refraction by comparison with adult and aged rabbits.3. In experimental group, after induction, some MSCs cells gradually increased in size and formed a stick-like appearance. One month after treatment, most of the long stick-like cells connected with adjoining cells and began to form my tube-like structures, the direction of the cells arraying was similar gradually. Cardiac cell-specific proteinα-actin and cTnT expressed positively, the expression rate of cardiac cell-specific protein was different after MSCs was induced by different concentration 5-Aza, of which 10μmol/L 5-Aza was the optimal concentration. MSCs are multipotential stem cells and they can be induced into cardiomyocytes by 5-Aza in vitro which have the characteristics of cardicmyocytes in structure. The optimal concentration of 5-Aza induced MSCs into cardicmyocytes is 10μmol/L.