人脐血和外周血内皮祖细胞的分离培养和鉴定

背景：国内外有关内皮祖细胞的分离培养和鉴定方面的文章很多,但是人脐血和外周血内皮祖细胞的鉴别方面文章不多。目的：从人脐血和外周血中分离出内皮祖细胞,并对其进行培养和鉴定。方法：选取密度梯度离心法从脐血和外周血中分离获得单个核细胞,按照1× 10⁶/cm²的浓度种植于预先铺有纤维连接蛋白的培养板中,用含血管内皮生长因子的M199培养基进行诱导培养。 结果与结论：人脐血和外周血中存在内皮祖细胞,浓度梯度离心联合贴壁筛选获得的单个核细胞在血管内皮生长因子的诱导培养下可分化成内皮祖细胞,内皮祖细胞表达CD34、CD133、CD105、KDR和CD31,能吞噬乙酰化低密度脂蛋白,结合荆豆凝集素1,它们可作为体外分选内皮祖细胞的标志。

Endothelial progenitor cells from human umbilical cord blood and peripheral blood: Isolation,culture and identification

BACKGROUND:There are many articles on the isolation, culture and identification of endothelial progenitor cells at home and abroad, while the articles regarding the endothelial progenitor cells from the umbilical cord blood and peripheral blood are rare. OBJECTIVE:To isolate, culture and identify the endothelial progenitor cells from umbilical cord blood and peripheral blood. METHODS:Mononuclear cells were isolated from umbilical cord blood and peripheral blood with density gradient centrifugation method, and then the cells were implanted on the culture plate pre-paved with fibronectin with the concentration of 1×10⁶/cm². After that, the cells were induced and cultured with M199 medium containing vascular endothelial growth factor. RESULTS AND CONCLUSION:The endothelial progenitor cells could be isolated and obtained from peripheral blood and umbilical cord blood, and the mononuclear cells obtained with density gradient centrifugation and adherence screening method could be differentiated into endothelial progenitor cells after induced with vascular endothelial growth factor. The endothelial progenitor cells could express CD34, CD133, CD105, KDR and CD13, could swallow acetylated low density lipoprotein and be combined with with ulex europaeus agglutinin 1, which could be considered as the sign of in vitro sorting of endothelial progenitor cells.