兔骨髓间充质干细胞的体外培养及多向诱导分化

髓间充质干细胞的分离获取及培养鉴定一直以来仍无统一标准和定论。 目的：体外获取兔骨髓间充质干细胞、纯化、扩增，同时研究其生物学特性及多向分化潜能。 方法：采用贴壁筛选法分离培养兔骨髓间充质干细胞，观察其增殖和生长特性，绘制生长曲线；倒置相差显微镜、苏木精-伊红染色及碱性磷酸酶染色观察细胞形态；并向成骨细胞、软骨细胞和血管内皮细胞多向诱导分化，分别采用钙结节茜素红染色、甲苯胺蓝染色和荆豆凝集素染色鉴定。 结果与结论：经贴壁筛选法得到的骨髓间充质干细胞性状稳定，为纺锤状，呈克隆样生长，碱性磷酸酶染色弱阳性；经成骨、成软骨、成血管内皮细胞诱导培养的骨髓间充质干细胞，表现出相应细胞的形态特征和生物学特征。提示，全骨髓培养法取材方便，骨髓间充质干细胞增殖能力强，在适宜条件下能够多向分化。

In vitro cultivation and multi-inducing differentiation of rabbit bone marrow mesenchymal stem cells

There is still no unified standard and final conclusion of isolation, culture and evaluation of bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To observe the biological characteristics and multi-differentiation potential of BMSCs through in vitro cultivation, purification, and amplification. METHODS: BMSCs were isolated and cultured by adherence screening method, followed by observation of proliferation and growth characteristics, for drawing growth curves; inverted phase contrast microscope, hemotoxylun and eosin staining, and alkaline phosphatase staining were used to observe the cell morphology; BMSCs were induced to differentiate into osteoblasts, chondrocytes and vascular endothelial cells, which were identified by calcified nodules-Alizarin red, toluidine blue and Ules europaeus agglutinin (UEA) staining respectively. RESULTS AND CONCLUSION: By the method of adherence screening, BMSCs were found in stable characteristic, appeared morphologically in spindle and colony-shape, expressed weakly positive of Alkaline phosphatase staining. BMSCs presented with the corresponding cytomorphology and cytobiology characteristics after inducing cultivation of osteoblasts, chondrocytes and vascular endothelial cells. It revealed that adherence screening method is relatively simple and feasible. Furthermore, BMSCs have a good ability of proliferation and could multi-differenrate under suitable condition.