人外周血内皮祖细胞的分离培养及鉴定

Isolation,culture and identification of human endothelial progenitor cells from peripheral blood

目的:从人外周血中分离培养内皮祖细胞(EPCs),并探讨其体外诱导培养的条件。方法:密度梯度离心法分离正常人外周血单个核细胞,置于鼠尾胶原包被的培养瓶中进行体外培养,流式细胞术和免疫荧光法检测分化细胞表面特异性抗原标记物的表达。结果:人外周血单个核细胞在鼠尾胶原包被的培养瓶中培养后呈梭形,并表达内皮细胞的特异性抗原CD34和KDR,和干/祖细胞抗原CD133。提示这些培养细胞既具有内皮细胞的表面标志和功能,又具有祖细胞特性。结论:成功从外周血中培养出EPCs。鼠尾胶原可取代EPCs培养中常用的纤维连接蛋白,是体外分离培养外周血EPCs的更节省的一种方法。

Objective：To investigate the methods of isolating and culturing endothelial progenitor cells （EPCs） from human peripheral blood. Method：Human peripheral blood mononuclear cells （PBMC） were isolated from normal persons and put into the culture plate previously coated by rat tail collagen. Flow cytometry was used to detect the expression of CD34, CD133 and KDR of the cultured cells, and immunofluorescence was performed to display the character of endocytosing UEA I and acLDL. Result： Most of the cultured ceils displayed a fibroblast-like morphology adhering to the culture plate. They expressed CD34, CD133 and KDR. They could show endocytose of UEA I and acLDL, too. Conelusion：EPCs were successfully cultured from PBMC. Rat tail collagen is much cheaper than fibronectin and can be used in culturing EPCs.