人脐血内皮祖细胞的体外培养

目的从人脐带血中分离内皮祖细胞（endothelial progenitor cells,EPCs）,建立人脐血内皮祖细胞体外培养的方法,为实现内皮祖细胞的移植及实验研究提供足量的细胞来源。方法采用密度梯度离心法分离人脐带血内皮祖细胞,在EGM-2培养基中培养,采用流式细胞仪、免疫组化和免疫荧光鉴定EPCs。结果脐带血单个核细胞在经EGM-2培养过程中出现梭形贴壁和铺路石样等形态;1周后既分化成EPCs,细胞免疫荧光CD133染色率在培养第7天为（67.2±2.12）%,免疫组化CD34染色率为（89.67±2.05）%,流式细胞仪鉴定该种细胞CD133与KDR的双阳性率达87.8%。可确定该细胞为EPSc。结论采用本培养方法可获得良好的内皮祖细胞用于实验研究。

Cultivation in vitro of people umbilical cord blood endothelial progenitor cells

Objective To establish a method of endothelial progenitor cells isolated from human umbilical cord blood in vitro,and to provide a sufficient amount of source for endothelial progenitor cell transplantation and experimental study.Methods Isolate from human umbilical cord blood endothelial progenitor cells with density gradient centrifugation,and culture in EGM-2 medium.EPCs were identificatd by flow cytometry and immunofluorescence.Results The cells cultured in EGM-2 were displayed spindle-shaped and cobble-stone morphology,and differentiated into EPCs a week later.The rate of CD133 immunofluorescence staining was(67.2±2.12)% on seven days.The rate of CD34 Immunohistochemical staining was(89.67±2.05)% on seven days.The double positive rate of CD133 and KDR was 87.8% in these cells.It could determine that the cell was EPSc.Conclusion We can obtain available endothelial progenitor cells using this culture method for experimental research.