人脐血中血管内皮祖细胞体外扩增和鉴定的实验研究

目的：探讨从人脐血中分离、培养、体外扩增血管内皮祖细胞(EPC)的方法及体外进行EPC的鉴定。方法：采用密度梯度离心法从脐血中分离EPC，体外分别培养在包被有FN和不包被FN的培养皿中，采用免疫组织化学技术(SABC法)鉴定培养贴壁细胞表面标志CD31、CD34及KDR的表达；流式细胞仪检测细胞表面标志CD133及表达CD133细胞比例。结果：包被FN的培养皿中细胞贴壁及增殖均比未包被的多，免疫组化染色CD31、CD34及KDR均呈阳性，培养第4天流式细胞仪检测CD133^ 占21．3％。结论：密度梯度离心法可用于体外分离脐血中EPC进行实验研究，FN对于体外培养EPC有非常重要的作用。

Study of cultivate and identification of endothelial progenitor cells from human umbilical cord blood in vitro

Objective To study the isolation?cultivate?proliferation and identification of endothelial progenitor cells from human umbilical cord blood in vitro. Methods EPC separation was performed by density gradient centrifugation,isolated cells were cultured in dishes coated with or without FN,the expression of cell markers CD31?CD34 and KDR was assessed using immunohistochemistry(SABC method), the expression of CD133 and the percentage of CD133 cells in adherent cells were assessed using fluorescence-activated cell sorting(FACS). Results Compared with the cells cultured in dish without FN,number of Adherent Cell and proliferation of cells cultured in dish with FN were increased. Immunostaining of cell markers CD31?CD34 and KDR of adherent cells were positive. The percentage of CD133+ cells of adherent cells was 21.3% at 4d. Conclusion Density gradient centrifugation is a good method of EPC separation from human umbilical cord blood in vitro,and FN is very useful for culturing EPC in vitro.