兔骨髓基质细胞的体外成骨定向诱导培养

目的:体外分离培养兔骨髓基质细胞(bonemarrowstromalcells,BMSCs),并向成骨定向诱导培养,为骨组织工程提供适宜的种子细胞。方法:抽取新西兰白兔的骨髓,体外分离纯化得到BMSCs,并利用条件培养液将其向成骨定向诱导培养、扩增。采用倒置相差显微镜观察细胞增殖分化情况;使用碱性磷酸酶(ALP)和VonKossa染色的方法,鉴定其成骨性能;用MTT法描绘标准细胞浓度-OD值曲线。结果:BMSCs在培养皿中贴壁生长、增殖;经ALP和VonKossa染色证实其有成骨潜能;标准BMSCs细胞浓度-OD值曲线显示细胞浓度和OD值呈线性正相关。结论:可以通过在体外分离纯化骨髓并诱导培养,获得足够数量、有成骨潜能的BMSCs作为骨组织工程的种子细胞。

Differentiation and Proliferation of Rabbit Bone Marrow Stromal Cells into Osteoblasts in Vitro

Objective: To prepare seed cells for bone tissue engineering,differentiate and proliferate bone marrow stromal cells(BMSCs) from rabbit's marrow into osteoblasts in vitro.Methods: Some bone marrow of a rabbit was drew out and cultured by differentiation culture medium to harvest BMSCs.The proliferation of BMSCs was observed by light microscope.Von Kossa staining and alkaline phosphatase(ALP) activity test were employed to assess BMSCs' osteoblastic differentiation and the generation of calcified extracellular matrix.MTT Assay was used to draw the standard cell amount-OD curve.Results: BMSCs were cultivated,differentiated and proliferated well in the differentiation culture medium with active function,which were confirmed by observation of light microscope,Von Kossa staining and ALP activity test.A standard cell amount-OD curve was drawn by the results of MTT Assay.Conclusion: BMSC could be cultured,differentiated and proliferated with active osteogenic function by differentiation culture medium in vitro,so it could be suitable seed cell for bone tissue engineering.