应用密度梯度离心法分离扩增兔骨髓基质细胞的体会

The isolation and proliferation of bone marrow stromal cells of rabbits in vitro by density gradient separation method

目的 探讨密度梯度离心法分离培养兔骨髓基质干细胞 (BMSc)的条件 ,为组织工程学选择合适的种子细胞奠定基础。方法 采用密度梯度离心法将兔BMSc自小量骨髓中分离并培养 ,观察其增殖和生长特性 ,绘制生长曲线 ,测定分裂指数和贴壁率。结果 兔BMSc在第 7代前性状稳定 ,生长曲线相似 ;第 4天分裂指数最高为 15 % ;传代后 10h贴壁率高达 90 %。结论 密度梯度离心法是分离兔BMSc的理想方法 ,BMSc能为未来组织工程提供足量种子细胞。

Objective To investigate the isolation and culturing condition of the bone marrow stromal cell(BMSc)of rabbits in vitro by density gradient separation method and select the suitable seed-cells for bone tissue engineering.Methods BMSc of the bone marrow of the rabbits was isolated by density gradient separation method and cultured to observe the proliferation and growth characteristics in primary and passage cultures,draw the growth curve,measure the split index and the adhesion rate.Results The living characteristic of the cultured cells is quite stable before the seventh generation,and the growth curve is similar.The division index of the 4th day reaches the highest(15%). About 90% subcultured cells adhered to the wall in 10 hours.Conclusion The density gradient separation method is the ideal way for the isolation of BMSc.BMSc can provide enough seed-cells for the bone tissue engineering in the future.