摘要
目的探讨体外分离、培养成人外周血单个核细胞,并将其定向诱导分化为内皮祖细胞及成熟血管内皮细胞的方法.方法密度梯度离心法提取人外周血单个核细胞,用含有生长因子的内皮培养基将其体外培养、定向诱导分化为内皮祖细胞及成熟血管内皮细胞;分别用流式细胞技术及RT-PCR通过对内皮祖细胞及内皮细胞表面特异性抗原的检测进行细胞鉴定.结果分离获得的单个核细胞培养7 d后形成梭状的内皮样细胞,部分细胞积聚成团形成克隆集落,流式细胞仪鉴定该细胞表达内皮祖细胞特异性抗原CD34、CD133及VEGFR.继续培养4周后细胞形成典型铺路石样改变,RT-PCR检测有成熟血管内皮细胞特异性基因vWF、eNOS表达.结论可成功分离人外周血单个核细胞,并可将其定向诱导分化为内皮祖细胞及成熟内皮细胞.
Objective To investigate the methods of isolation and culture of human peripheral blood mononuclear cells （PBMC） for differentiating into endothelial progenitor cells （EPCs） and endothelial cell （ECs）. Method Peripheral blood mononuclear cells were isolated by density-gradient centrifugation and incubated in endothelial medium in the presence of vascular endothelial growth factor. Peripheral blood mononuclear cells were differentiated into endothelial progenitor cells and endothelial cell. Flow cytometry and PT-PCR were used to identify the endothelial progenitor cells and endothelial cell. Results After 7 days, most of the cultured cells displayed a fibroblast-like morphology adhering to the culture plate. They expressed CD34, CD133 and VEGFR. After 4 weeks, most of the cultured cells expressed vWF and eNOS. Conclusion The endothelial progenitor cells and endothelial cell were successfully cultured from peripheral blood mononuclear cells
出处
《昆明医学院学报》
2012年第1期33-36,53,共5页
Journal of Kunming Medical College
基金
云南省科技计划基金资助项目(2011FB150)
关键词
内皮祖细胞
内皮细胞
细胞培养
Endothelial progenitor cells
Endothelial cells
Cell culture
