兔骨髓基质干细胞的分离培养及鉴定

目的探讨兔骨髓基质干细胞（MSC）体外分离培养及鉴定。方法自兔髂骨抽取骨髓,采用密度梯度离心法分离纯化出MSC,并增殖。观察MSC的生长情况及形态学特点,流式细胞仪检测第3代MSC表面抗原的表达情况。结果体外培养的兔MSC贴壁生长,呈长梭形,可增殖形成克隆;MSC阳性表达CD29,CD90,但CD34,CD45呈阴性。结论利用密度梯度离心法获取的MSC具有大量增殖的能力,表达CD29,CD90,不表达CD34,CD45。

Isolation, Culture and Identification of the Rabbit Marrow Stromal Cells in Vitro

Objective To investigate the methods of isolation,culture and identification of the rabbit marrow stromal cells（MSC） in vitro.Methods MSC were aspirated from rabbit iliac and purified by density gradient centrifugation.MSC were cultured and expanded in flasks.The MSC morphologic features and growth status were observed,and the cell surface molecules on MSC at passage 3 was investigated by flowcytometry.Results The rabbit MSC attached to the surface of plates,exhibiting a fibroblast-like spindle shape.In vitro culture,MSC proliferated quickly to form colonies.These expanded attached MSC were positive for CD29,CD90. In contrast,the cells were negative for CD34,CD45.Conclusion Using density gradient centrifugation,MSC have great proliferation ability,positively express CD29,CD90,but absence of CD34,CD45.