应用DiI标记犬脐血内皮祖细胞的实验研究  

Labeling Endothelial Progenitor Cells from Dog’s Core Blood with DiI

作　　者：赵宏光[1] 单根法[1] 钟竑[1] 许勤[2] 李小波[1] 申通[3] 

机构地区：[1]上海第二医科大学附属新华医院胸心外科,上海200092 [2]上海第二医科大学附属新华医院科研中心,上海200092 [3]上海第二医科大学附属新华医院麻醉科,上海200092

出　　处：《中国微循环》2006年第2期126-129,i0003,共5页Journal of Chinese Microcirculation

基　　金：上海市科委重点基础研究项目(02JC14012)

摘　　要：目的 应用DiI标记犬脐血内皮祖细胞，为内皮祖细胞的研究提供细胞标记方法。方法 取犬的脐带血，用添加有EGM-2MV的EBM-2在纤连蛋白包被的六孔板上培养。检测细胞表面抗CD31、CD34、vWF及FLK-1的表达。用DiI标记内皮祖细胞，用荧光显微镜观察内皮祖细胞的荧光情况。结果 4-6d有细胞贴壁生长，7-14d可见贴壁细胞呈克隆型生长，呈典型的“铺路石”样外观。传代后，细胞生长良好。细胞表面抗体检测显示为：CD31（＋），CD34（＋），vW因子（＋），FLK-1（＋）。荧光显微镜下见内皮祖细胞的细胞浆内有红色荧光。阳性率（68．8±1．6）％。结论 应用DiI标记犬脐血内皮祖细胞．方法简单，染色效果好，是内皮祖细胞示踪的好方法。Objective To label endothelial progenitor cells from dog＇s core blood with 1, 1＇ -dioetadeeyl-3, 3, 3＇, 3＇ -tetramethylindoearboeyanine perehlorate（DiI）, to provide method of cells labeling for research of EPCs. Methods Got core blood from dogs and cultured the cells with EBM-2 adding EGM-2 MV in 6 well eel culture clusters coating with fibroneetin. Determined the expression of CD31, CD34, vW factor and FLK- 1 with immunohistoehemieal method. To label endothelial progenitor cells from dog＇ s core blood with DiI, and observe the fluorescence in endothelial progenitor cells with fluorescence microscope. Results Cells could be seen to adhere to the well in 4 to 6 days, they grew up in clones in 7 to 14 days. They grew in a cobble appearance, and grew well after passage. Cell antigen showed CD31（ ＋ ）, CD34（ ＋ ）, vW factor（ ＋ ）, FLK-I（ ＋ ）. Red fluorescence could be seen in endothelial progenitor cells＇ plasma. The positive rate was 68.8 ± 1.6%. Conclusion The method of labeling endothelial progenitor cells with DiI was simple and the results were ideal. It was a good method to label endothelial progenitor cells.

关 键 词：内皮祖细胞 脐血 标记 荧光 

分 类 号：Q26[生物学—细胞生物学]