应用粒细胞集落刺激因子动员的外周血体外扩增内皮祖细胞

Ex vivo expansion of endothelial progenitor cells from granulocyte colony stimulating factor-mobilized peripheral blood

目的探讨从正常人经粒细胞集落刺激因子(granulocyte colony simulating factor,G-CSF)动员的外周血中分离扩增内皮祖细胞(endothelial progenitor cells,EPCs)。方法经G-CSF动员后,富集的外周血单个核细胞(peripheralblood mononuclear cells,PBMNC)贴壁培养法定向扩增EPCs,免疫组织化学染色和荧光标记鉴定其内皮细胞特性。流式细胞仪检测细胞表面标记物变化。结果获得的贴壁细胞数为(8.825±2.958)个/100个PBMNC,具有多种内皮细胞特征,血管性假血友病因子阳性,同时结合荆豆凝集素I并内吞乙酰化低密度脂蛋白呈红绿荧光双染色。培养后血细胞标记物减少,内皮细胞标记物增加(P<0.01)。结论从G-CSF动员的外周血中可获得较多数量EPCs,并具有多种内皮细胞特征。

Objective To isolate and expand endothelial progenitor cells （EPCs） from granulocyte colony simulating factor（G-CSF）-mobilized peripheral blood of healthy donors. Methods EPCs were obtained from G-CSF-mobilized peripheral blood mononuclear cells （PBMNC） of healthy donors by directed culture ex vivo . The endothelial characteristics of EPCs were identified by immunohistochemical staining and fluorescent labeling. Flow cytometry（FACS） was used to determine the cell phenotype changes. Results There were 8. 825 ± 2.958 adherent cells/100 PBMNC. The cells had many endothelial characteristics and were vWF positive. They could bind to UEA-Ⅰ and endocytose acLDL. The phenotype of hematopeietic cells reduced while the endothelial markers increased after culture （ P 〈 0.01）. Conclusion Many EPCs can be obtained by directed culture from G-CSF-mobilized peripheral blood. EPCs have many characteristics of endothelial cells.