兔骨髓间充质干细胞的分离培养及鉴定

Isolated culture and identification of the rabbit marrow mesenchymal stem cells

目的:探讨一种简便可行的兔骨髓间充质干细胞(bone marrow derived mesenchymal stem cells,BMSCs)体外分离培养方法。方法:通过全骨髓贴壁分离法体外分离、扩增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;倒置相差显微镜观察细胞形态;并向软骨细胞诱导分化,采用番红快绿染色和甲苯胺蓝染色鉴定。结果:经全骨髓贴壁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;成软骨诱导培养的骨髓间充质干细胞,表现出软骨细胞的形态特征和生物学特征。结论:采用全骨髓贴壁分离法操作简便,是培养兔骨髓间充质干细胞的理想方案,在适当的条件下BMSCs能多向分化。

Objective：To establish the method of isolation and identification of the rabbit bone marrow mesenchymal stem cells in vitro. Methods： BMSCs were isolated and cultured by adherent screening method,followed by observation of proliferation and growth characteristics,for drawing growth curves. Inverted phase contrast microscope was used to observe the cell morphology. BMSCs were in- duced to differentiate into chondrocytes,which were identified by Safranin-O and fast green staining and toluidine blue staining respec- tively. Results： By the method of differentiation adherence, BMSCs were found in stable characteristic, appearing morphologically in spindle and colony-shape. BMSCs presented with the corresponding cytomorphology and cytobiology characteristics after cultivation of chondrocytes. Conclusion： The differentiation adherent method is relatively simple and feasible. Furthermore,BMSCs have a good a- bility of proliferation and can multi-differentiate under suitable condition.