Differentiation of endothelial progenitor cells from human umbilical cord blood CD 34^+ cells in vitro  

人脐血CD34^+内皮祖细胞的体外分化(英文)

作　　者：范春玲[1] 李燕[1] 高平进[1] 刘建军[1] 张学军[2] 朱鼎良 

机构地区：[1]上海第二医科大学附属瑞金医院上海市高血压研究所医学遗传学国家重点实验室,上海200025 [2]中国科学院上海生命科学研究院生物化学与细胞生物学研究所分子细胞生物学重点实验室

出　　处：《Acta Pharmacologica Sinica》2003年第3期212-218,共7页中国药理学报（英文版）

基　　金：Project supported by the grants from the National Key Program of Basic Research (№ G2000056904), the Shanghai Science and Technology Committee, and the Health Science Center,Shanghai Second Medical University and Chinese Academy of Sciences.

摘　　要：AIM: To study the time course of the expression of stem cell marker and endothelial cell markers on human cord blood CD34^+ cells during in vitro differentiation process of endothelial progenitor cells (EPC). METHODS: CD34^+ cells were selected and enriched from human cord blood by magnetically activated cell sorting (MACS), and cultured in dishes coated with or without fibronectin (Fn). Endothelial cells were identified by staining the cells with anti Flk-1 and vWF antibodies. The percentage of AC133^+ cells in adherent CD34^+ cell population was analyzed by fluorescence-activated cell sorting (FACS). RESULTS: The expression of Flk-1 and vWF on adherent CD34^+ cells increased during the culture time, with 27.0％positive for Flk-1 and negative for vWF at d 3, and 100％positive for both Flk-1 and vWF at d 7. When cells were cultured in Fn-treated dishes, the percentages of Flk-1 and vWF positive cells increased to 34％and 47％, respectively at d 3, and 100％at d 7. In contrast, the percentages of AC 133^+ cells among the adherent cell population decreased rapidly, and similar changes occurred in cells cultured in the presence of Fn. CONCLUSION: The gradual appearance of endothelial cell markers and the disappearance of stem cell marker characterized the in vitro differentiation of endothelial progenitor cells. Fibronectin accelerated the differentiation process of EPC.目的:研究人脐血CD34^+细胞群体中内皮祖细胞在体外分化为内皮细胞的过程中,干细胞标志以及内皮细胞表型随时间的变化。方法:将免疫磁珠细胞分选法(MACS)得到的CD34^+细胞体外培养于纤连蛋白和无纤连蛋白处理的培养皿中,以免疫细胞化学鉴定贴壁细胞的内皮标志Flk-1和vWF,并以流式细胞仪分析其干细胞标志AC133。结果:贴壁细胞的内皮标志Flk-1和vWF是逐步出现的,d3时有27.0％贴壁细胞表达Flk-1,vWF不表达,d7时已100％表达vWF和Flk-1。纤连蛋白促进贴壁细胞内皮标志Flk-1和vWF的表达,d3时的表达百分率分别为34.0％和47.0％,d7时Flk-1和vWF的表达均为100％。在培养过程中,AC133阳性细胞的比例迅速下降,但纤连蛋白对AC133的表达无显著影响。结论:在内皮祖细胞分化的过程中,干细胞标志迅速消失,向内皮细胞分化,内皮细胞的表型是逐步出现的。纤连蛋白促进内皮祖细胞的分化。

关 键 词：vascular endothelium cell differentiation CD34^+ antigens pregency genetic markers hematopoietic stem cells FIBRONECTINS 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]