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Objective: To observe the rabbit bone marrow mesenchymal stem cells (BMSCs) growth characteristics and potential differentiation potential to provide an experimental basis to select seed cells for tissue engineering. Methods: whole bone marrow adherent were isolated cultured rabbit BMSCs by phase contrast microscopy to observe the growth characteristics, Flow cytometry for third-generation cell surface antigen CD29, CD44 and CD45 identified. The programmed cooling box cryopreservation of BMSCs cells under a microscope to observe the proliferation ability of recovery after three months, the trypan blue staining its survival. Adipogenic (1O? S H-DMEM was 4 × 10 -5 g / L dexamethasone, 10mg / L IBMX, 10mg / L, insulin), osteoblasts (10? S H-DMEM was 4 × 10 -6 g / L dexamethasone, 0.5g / L Vitamin-c, 1g/Lβ- phosphorylated glycerol sodium), and into the nerve (DMEM/F12, 10ng/mL b-FGF 10ng/mL EGF and 10ng/mL of PDGF 100μg/mL transferrin, 0.03μM sodium selenite) induce BMSCs induced liquid to the fat cells, osteoblasts, neuron-like cell differentiation. Induced cells two weeks after oil red O staining, alizarin red staining, Vankossa silver staining staining and alkaline phosphatase staining. Osteogenic induction 3d, 5d, 7d, 9d, 12d, 15d, 18d, 21d cell rabbit type Ⅰ procollagen (procol Ⅰ) enzyme-linked immunoassay. Immunocytochemical methods to detect specific surface markers induced into neuron-like cells. Results: The whole bone marrow adherence method available of BMSCs rabbit the BMSCs surface antigen by flow cytometry, CD44 expression was 87.92%, CD29 expression was 95.36%, CD45 expression was 2.51%, the third generation BMSCs biometric basic consistent. After cryopreservation of cell viability was 85%, and passaged cells to restore the original proliferative ability. Under certain conditions, can induce differentiation into adipocytes, osteoblasts and neuron-like cells, red lipid droplets in the oil red O staining cells after adipogenic, osteogenic induction Alizarin Red staining, Vankossa silver staining can be observed to mineralized nodules. Alkaline phosphatase activity staining control group was weakly positive induction group was strongly positive. The osteogenic induction 3d to 21d, procol Ⅰ showed an increasing trend. Into neural induction two weeks after cell morphology changes, the control group BMSCs by immunohistochemical staining weak expression of neuron-specific enolase (NSE); factor group cells strong expression of neuron-specific enolase enzyme (NSE), weakly expressed in nest protein (nstin). Conclusion: The whole bone marrow adherence method isolated and cultured rabbit BMSCs simple and feasible method. A rich source of mesenchymal stem cells in the bone marrow, easily isolated and cultured in vitro, and a large number of amplification, with multiple differentiation potential, excellent seed cells for tissue engineering.
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