外周血来源的内皮祖细胞的体外培养与分化  

Culture and differentiation of endothelial progenitor cells from peripheral blood in vitro

作　　者：吴少峰[1] 黄惠民[2] 

机构地区：[1]上海第二医科大学新华医院,上海200127 [2]上海儿童医学中心心胸外科,上海200127

出　　处：《上海第二医科大学学报》2004年第3期159-161,共3页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：上海市自然科学基金(99ZB14018)

摘　　要：目的 研究内皮祖细胞(EPC)的分离、培养、分化及鉴定。方法 Ficoll密度梯度离心分离外周血获取单核细胞,在加有EGM-2-MV-SingleQuots的EBM-2培养液中培养和扩增,免疫组化和免疫荧光测定分化细胞表面特异性抗原的表达。结果 培养后10d细胞融合,呈克隆样生长,20d后细胞呈现典型的鹅卵石样内皮细胞形态;免疫组化和免疫荧光显示,CD31、vWF、FLKI/KDR/VEGFR-2和VE-cadherin均呈阳性。结论 外周血来源的单核细胞含有内皮祖细胞,能在一定的培养条件下分化成为血管内皮细胞。Objective To investigate the methods of isolating, culturing, differentiating and evaluating the endothelial progenitor cell (EPC ) . Methods The mononuclear cells were obtained by Ficoll density-gradient centrifu-gation. The cells were suspended in endothelial basal medium (EBM-2) supplemented with EGM-2-MV-SingleQuots for culturing, differentiating and proliferating. The expressions of specific antigens on cell surface were analysed by immunohistochemistry and immunofluorescence. Results The EPCs exhibited the clonal morphology and classical cobblestone morphology after 10 and 20 days culture respectively. The endothelial phenotype was confirmed by positive immunostaining with endothelial cell-specific markers, such as CD31, vWF, FLK-1/KDR/VEGFR-2 , and VE-cadherin. Conclusion EPC can be obtained from mononuclear cells in peripheral blood and be differentiated into endothelial cell in vitro.

关 键 词：内皮祖细胞 体外培养 细胞分化 外周血 细胞表面特异性抗原 血管内皮细胞 组织工程 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]