| Objective the rabbit bone marrow mesenchymal stem cells were observed the biological characteristics and the differentiation potential, which to provide experimental basis for selection in the seed cells for tissue engineering.Methods The rabbit bone marrow-derived MSCs were isolated and cultured from rabbit bone marrow by the whole bone marrow adherent cultivation,Morphology of MSCs was examined by phase contrast microscopy,surface antigen CD29,CD44 and CD45 from 3 passage BMSCs were detected by flow cytometry.The BMSCs were cryopreserved by Mr.Frosty Cryo 1℃Freezing Container.3 months later,obverved BMSCs after recovery by microscope, Survival were confirmed by the Trypan blue dye exclusion.The BMSCs were treated with adipose inductor (10%FBS+H-DMEM,4×10-6g/L Dexamethasone,10mg/L IBMX,10mg/L Insulin),osteogenetic inductor(10%FBS+H-DMEM,4×10-6g/L Dexamethasone,0.5g/L Vitamin-C,1g/L sodiumβ-glycerophosphate) and neuronal inducter(DMEM/F12,10ng/mL b-FGF,10ng/mL EGF,10ng/mL PDGF,100μg/mL transferrin,0.03μM Sodium Selenite)to differentiated into adipocytes,osteoblast and neuron-like cells in vitro.And the differentiated cells were identified by oil red O staining ,alizarin bordeaux staining, Vankossa silver staining and alkaline phosphatase staining.The BMSCs that treated with osteogenetic inductor were obtained on days 3,5,7,9,12,15,18,21,then type I procollagen were investigated by ELISA. Specific surface marker from neuron-like cells were detected by Immunohistochemistry.Result the BMSCs were isolated by the whole bone marrow adherent cultivation in vitro. through flow cytometry analyzed, surface antigen CD29(95.36%)CD44(87.92%)were positive,CD45(2.51%)was negative.the biological characteristics of 3 passage BMSCs were consistent, which were induced into adipocytes,osteoblast and neuron-like cells. The BMSCs survival after recovery was 85%. After passage,the cells restore the original state. After adipose induction, Oil red O staining showed that red lipid droplet existed in adipose cells.After osteogenetic induction, mineralized nodules were detected by alizarin Bordeaux staining and Vankossa silver staining, the controlled cells were weak positive to alkaline phosphates staining.The induced-cells were strong positive. type I procollagen Gradually increased from 3d to 21d after Osteogenic.two weeks later, Cell morphology changed after neuronal inducted.NSE was weak expressed at the control group,Nestin was weak expressed and NSE was strong expressed at the factor group .Conclution the whole bone marrow adherent cultivation is a simple and feasible way, which isolate and culture MSCs from rabbit bone marrow. The MSCs are superior seed cells of tissue engineering, which have rich sources and multiplex differentiation potential.
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