大鼠骨髓间充质干细胞的分离与培养

Cultivation and Isolation of the Rat Bone Marrow Mesenchymal Stem Cells

目的 观察体外培养的大鼠骨髓间充质干细胞（MSCs）的生物学特性。方法 通过贴壁法培养、鉴定大鼠骨髓MSCs，观察其生长规律及光镜结构特点。结果 大鼠MSCs 7代以前增殖速度较快，细胞接种后1～2d为滞留期，3d后达对数生长期，第6天进入平台期。免疫组织化学染色显示MSCs表达CI）44，不表达CD45；光镜下可见MSCs呈梭形、纺锤形和多角形，核居中，有1～2个核仁。结论 培养的细胞为骨髓MSCs；在体外培养条件下细胞生长性状稳定，可作为组织工程的种子细胞。

Objective To investigate the biological characteristics of bone marrow mesenchymal stem cell （MSCs） from rat. Methods The rat MSCs from bone marrow were culture by adhering to the culture plastic and identified using immunocytochemical methods. Their growth axiom, light microscopic features were observed. Results MSCs proliferate fast before the seventh generation. The cells present stagnatio, after inoculation 1 ～ 2 days and after 3 days the cells present logarithm growth period. At the sixth day MSCs are into platform period. The MSCs are, in the present of CD44 and in the absent of CD45 by immunocytochemical techniques. Under the light microscope MSCs present fusiform, spindle and polygon shape. The nuclei seat in the middle of the cells and have one or two nucleoli. Conclusion The eultured cells are identified the bone marrow MSCs. The growth traits of MSCs are stable in vitro and they can be used as the seed-cells in the study of tissue engineering.