ICR小鼠骨髓间充质干细胞分离及生物学特性的探讨

Investigation on the Isolation and Biological Characteristics of Bone Marrow Mesenchymal Stem Cells of ICR Mice

目的探讨分离、培养的ICR（Institute of Cancer Research，ICR）小鼠骨髓间充质干细胞（mouse bone marrow mesenchymal stem cells，mBM—MSCs）的细胞生物学特性。方法取6～8周龄ICR小鼠，利用全骨髓反复贴壁和有限稀释培养法分离纯化mBM—MSCs，观察形态特点，测定生长曲线和活力，利用流式细胞仪分析细胞的周期和鉴定表面抗原，诱导其向成骨、软骨及脂肪细胞分化，采用染色法鉴定。结果新分离的mBM—MSCs多呈小圆形，形态规整。培养传代后，细胞多变为梭形，大小较均匀，形态较一致。随着传代的次数增加，细胞的生长曲线、活力及周期呈现快速发育期、平台期和缓慢期；流式结果细胞CD44、CD73、SCA-1呈阳性反应，部分细胞CD90、CD105、STRO-1呈阳性反应，CD11b和CD45呈阴性反应；诱导分化为成骨细胞后其碱性磷酸酶、茜素红和YonKossa银染色均呈阳性反应，分化成脂肪细胞后油红O染色呈阳性反应，分化成软骨细胞后阿尔新蓝染色呈阳性反应。结论ICR小鼠骨髓中分离培养出的MSCs，生物学特点鲜明，适于做进一步研究。

Objective To investigate the cell biological properties of isolated and cultured mBM - MSCs of ICR. Methods Mouse bone marrow mesenchymal stem cells of 6 - 8 weeks old ICR mice were in vitro isolated and purified by whole bone marrow repeated adherent culture and limiting dilution culture method. Cell morphology was observed. Its growth curve and vitality were measured, and its cell cycle and surface antigen markers were analyzed by flow cytometry and differentiation ability was detected to identify whether they are the BM - MSCs. Results The newly isolated mBM - MSCs were small round, regular shape. After subculture, cells size was uniform, and their morphology was consistent and multi - spindle. Along with the increased number of cell passage, the growth curve, viability and cy- cle of cells were showed the first rapid development stage, platform stage and slow activity period. Flow cytometry results showed that cell＇ s surface maker CD44, CD73 and SCA - 1 were positive, part of the cells CD90, CD105 and STRO - 1 were positive, CDllb, CD45 were negative. After osteogenic, adipogenic and chondrogenic differentiation, alkaline phosphatase, alizarin red, Von Kossa silver, oil red O and alcian blue staining were positive respectively. Conclusion The biological characteristic of isolated and cultured mBM - MSCs of ICR was homogeneous and distinct. These cells were suitable for further research.