BMSCs' culture, differentiation towards osteoblasts and recombination with coral materials

Objectives: To study the state of culture, differentiation and osteogenic of bone marrow stromal cells (BMSCs) in vitro and BMSCs' growth in the coral materials. Methods: Obtained BMSCs from the rabbits' femur, then cultured BMSCs and induced them to differentiate toward osteoblasts with special DMEM (contained Glycerophosphate disodium, L-Ascorbic acid, Dexamethasone and 10% fetal cattle serum) in vitro. Observed the condition of both induced cells and coral under an inverted phase contrast microscope(IPCM), scanning electron microscope (SEM) and electron probe microscope analysis (EPMA). cell morphology by HE, activity of alkaline phosphatase(ALP) using improved Gomori method, and content of osteocalcin with ELISA kit. Results: (1) BMSCs differentiated to osteoblasts and had strong capacity of proliferation. And ability of formation of calcifying nodules was detected. (2) When subcultured at about 12th, whatever induced BMSCs or BMSCs in complete culture medium changed in shape. And cells were transformed entirely at 16th or so, its' shape became bigger and flatter than before, and adhere to bottom of bottle irregularly. Its' activity decreased and tended to decrepitude. (3) BMSCs could grow, differentiate and proliferate in and on the coral materials normally. Conclusion: (1) Obtaining the BMSCs from bone is convenient and have a strong activity of proliferation and osteogenic capacity. So BMSCs are apt to serve as seed cells . (2) BMSCs could grow, proliferate and form mineralization area on/in the coral . Therefore, Coral is a favorable bone tissue engineering scaffold.