Isolation，cultivation and differentiation of rabbit bone marrow mesenchymal stem cells in vitro

Abstract:

Objective：To investigate the isolation and cultivation of rabbit bone marrow mesenchymal stem cells（rBMSCs） in vitro，to observe their biological characteristics and to discuss the paths of inducing the rBMSCs into osteoblasts and lipoblasts in order to pro－vide experimental basis for selecting seed cells. Methods：The rabbit was put to death by air embolism to the ear marginal vein with the anesthesia of urethane. The femur of the rabbits was got under the sterilized condition. rBMSCs were separated by percoll den－sity gradient centrifugation method and the whole bone marrow adherence culture method in vitro. The morphology of cells was ob－served by inverted microscope. At the third passage，rBMSCs were chosen to be induced into osteoblasts and lipoblasts. After 14 d of incubation，alkaline phosphatase detection was used in osteoblast-induced group and oil red O staining was performed in lipoblast-induced group. Results：The rBMSCs with good reproductive activities were obtained successfully by percoll density gradient lentrifu－gation and the whole bone marrow adherence culture method. The biological characteristics of the cells in the third generation were i－dentical. In osteoblast-induced group，alkaline phosphatase test showed strong positive expression. After oil red O staining，there was a large number of intracytoplasmic lipid droplets stained red in lipoblast-induced group. Conclusions：rBMSCs can be isolat－ed and proliferated effectively by percoll density gradient cen－trifugation method and the whole bone marrow adherence culture method.The cultured rBMSCs grow up and reproduce stably and could be induced and differentiated into osteoblasts and lipoblasts. So rBMSCs are excellent seed cells in tissue engineering.