人循环血管内皮祖细胞的分离培养和诱导分化

目的研究外周血来源的血管内皮祖细胞(EPCs)的分离、培养、分化及鉴定方法,探索EPCs体外扩增所需的条件。方法采集健康成人外周血,经Ficoll密度梯度离心法获取单个核细胞(MNC),接种在纤维连接蛋白包被的培养板上,在加有EGM-2-MV-SingleQuots的培养液中培养和扩增。于培养第7天选择免疫荧光(Dil-acLDL/FITC-UEA-1)鉴定培养所得的细胞是否具有EPCs的特征。结果MNC在培养的第2天开始出现成簇现象,第4天细胞由圆形转变为梭形贴壁细胞,呈条索状或管状分布;激光共聚焦显微镜显示双染色阳性的细胞为正在分化的EPCs。结论人外周血MNC在体外经适当条件的培养,完全可以分化成EPCs。

The Culture and Differentiation of Endothelial Progenitor Cells from Peripheral Blood in Vitro

Objective To investigate the isolation,culture,differentiation and identification of endothelial progenitor cells(EPCs) from human peripheral blood in vitro.Methods MNCs(mononuclear cells) were isolated from the human peripheral anticoagulant blood sample by Ficoll-density centrifugation,and then the cells were plated on fibronectin-coated culture dishes.The cells were suspended in endothelial basal medium supplemented with ECG-2-MV-SingleQuots.Cultured cells were subjected to immunofluorescence to detect dual binding of Ac-LDL and UEA-1 on day 7.Result Cluster attaching cells appeared in the second day.Cord-like or duct-like structure formed among much more spindle-shaped cells was observed on day 4.EPCs characterized as double positive for Dil-Ac-LDL and FITC-UEI-1.Conclusion EPC can be obtained from mononuclear cells in peripheral blood and be differentiated into endothelial cells at specific conditions in vitro.