大鼠骨髓间充质干细胞的优化获取及生物学鉴定

目的 优化骨髓间充质干细胞的体外获取方法并鉴定。方法 取幼年SD大鼠骨髓，分离骨髓间充质干细胞，观察细胞形态特征、生长状况及表面抗原表达。结果 分离培养的细胞有两种不同的形态，一种为小梭形细胞，一种为大扁平细胞。第9代以前生长性状稳定，增殖能力强，细胞倍增时间为48．2h；超微结构显示为早期幼稚细胞形态；免疫细胞化学检测细胞表达CD44、CD54、CD71，但不表达CD34、CD68、层粘连蛋白；在有限的传代数内，抗原表达无改变；与传统方法相比，细胞纯度高。结论 分离培养的细胞成分单一，具有干细胞特性，适用于干细胞生物学和组织工程学的研究。

Optimal Isolation and Identification of Rat Autogenous Bone Marrow-derived Mesenchymal Stem Cells

Objective To optimize the accumulation method of rat autogenous bone marrow derived mesenchymal stem cells (MSCs) and to identify them. Methods MSCs were isolated from the bone marrow of the young SD rats. Light and electron microscopies were used to study the morphologic features and growth status. Immunocytochemistry technique was employed to examine the cell surface antigens. Results The cultured cells contained two morphologically distinct kinds of cells: small spindle shaped cells and large flattened cells. It was found that both grewed stably and proliferated rapidly before the ninth generation. Their population doubling time was 48 2 h. The cells showed the characteristics of stem cells under electron microscopy. The postive expression of CD44, CD54 and CD71 was detectable, while CD34, CD68 and Laminin not detectable. There was no difference in the cell surface antigens between the 1st generation and the 7th one. The purity of the obtained cells by the present method was higher than that by traditional methods. Conclusion The cultured cells were only composed of undifferentiated MSCs, being of the property of stem cells. The characterization of MSCs would facilitate future studies of stem cell biology and tissue engineering.