人脐静脉血管内皮细胞饲养层的制备  

A method for culture of human umbilical vein endothelial cells as feeder layers

作　　者：王志华[1] 何志旭[2] 汪浩文[3] 米蔷[2] 

机构地区：[1]天津市南开医院儿科,天津300100 [2]贵阳医学院干细胞中心,贵州贵阳550004 [3]安徽省池州市人民医院儿科,安徽池州247000

出　　处：《临床儿科杂志》2008年第10期890-892,共3页Journal of Clinical Pediatrics

基　　金：教育部新世纪优秀人才支持计划项目(No.050820);科技部国际科技合作重点项目(No.2004DFA03400);贵州省省长资金项目(No.200614)

摘　　要：目的探讨使用人脐静脉内皮细胞制备饲养层的方法。方法使用健康产妇剖宫产后同意遗弃的脐带,无菌状态下胶原酶消化法收集内皮细胞,进行传代培养和增殖扩增,免疫组化法进行Ⅷ因子相关抗原鉴定,分别取 P2～P5代内皮细胞经丝裂霉素 C(10μg/ml)分别处理30、60、90、120 min 后再接种 E14.1胚胎干细胞。观察 E14.1胚胎干细胞在内皮细胞上的生长情况。结果经丝裂霉素 C 处理30～60 min 后的 P2～P3代内皮细胞可以很好的支持胚胎干细胞的生长。结论人脐静脉血管内皮细胞可作为一种来源丰富、取材方便的人源化饲养层细胞,为以后临床应用打下基础。 [临床儿科杂志,2008,26(10):890-892]Objectives To explore the method for human umbilical vein endothelial cells cultured as feeder layers, netheds The umbilical cords were obtained from consented healthy pregnant women with cesarean section. The endothelial ceils were cultured and amplified under asepsis condition, and their morphologic characteristics were observed under inverted microscope and the factor Ⅷ antigen were identified by immunochemical method. At the same time, the endothelial cells from the second passage to the fifth passage had been treated with mitomycin-C （10 μg/ml） for 30, 60, 90, 120 minutes respectively. Results Endothelial cells between the second to the third passages were clearly superior to those of other passages and endothelial cells treated with mitomycin-C （ 10 μg/ml） for 30 - 60 minutes could support E14.1 embryonic stem cells in maintaining their undifferentiated phenotype very well. Conclusions Culture-expanded endothelial cell from multiple donors are used as feeder cells to support growth of embryonic stem cells. They can become an abundant source of human feeder layers and thus found the base for future clinical application.

关 键 词：人脐静脉 内皮细胞 饲养层 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]