慢病毒载体介导SV40LT致人脐静脉内皮细胞永生化  

Identification and Immortalization of Human Umbilical Vein Endothelial Cells by SV40 Large T

作　　者：梁淑丽[1] 薛增福[1] 吕艳香[1] 刘洋 梁树辉[1] 殷继鹏[1] 窦维佳[1] 赵晓迪[1] 赵宏喜[3] 聂勇战[1] 吴开春[1] 

机构地区：[1]第四军医大学西京消化病医院,西安710032 [2]西安市第五医院,西安710082 [3]第四军医大学唐都医院妇产科,西安710038

出　　处：《科学技术与工程》2011年第11期2423-2427,共5页Science Technology and Engineering

基　　金：国家科技重大专项新药创制(2009ZX09103-667;2009ZX09301-009-RC06);863计划(2006AA02Z103;2006AA02A402);军队重点项目(06G087);国家自然科学基金重大项目(81090270;81090273);国家973项目(2010CB529302)资助

摘　　要：为成功分离与鉴定人原代脐静脉内皮细胞,用猿猴病毒40大T抗原(SV40LT)异位表达建立永生化人脐静脉内皮细胞系。将含SV40LT cDNA片段的慢病毒载体,转染人原代脐静脉内皮细胞,连续传代培养。通过形态、细胞免疫组织化学、RT-PCR及管状成形试验进行原代及转染后细胞形态学和功能学鉴定及检测SV40大T抗原表达。结果SV40LT转染后的人脐静脉内皮细胞为扁平多角形或短梭状,呈单层铺路石状镶嵌排列。特异性表达Ⅷ因子、KDR、SV40LT表达,并具有管状成型能力。说明成功分离与鉴定永生化的脐静脉内皮细胞系,为后续血管靶向治疗奠定了基础。To immortalize human umbilical vein endothelial cells（HUVECs）by ectopic expression of Simian Virus 40 Large T（SV40LT） antigen without malignant transformation,Lentivirus that contained SV40LT cDNA were transfected into the primary HUVECs.Light microscope and scanning electron microscope were used to observe the morphology and growth of the cells.The expression of factor Ⅷ,KDR and SV40LT were detected by immunofluorescence and RT-PCR.Tube formation assay was perfected to verify the functions of the primary HUVECs and the transfected cells.It is resulted that the immortalized cells were homogenous and displayed a characteristic ovoid nucleus.Immunofluorescence staining and RT-PCR showed the immortalized cells specifically expressedⅧ factor,KDR and SV40 LT.The transfected cells also keep the ability of tube formation.It is conclused that is identified and immortalized human umbilical vein endothelial cells by SV40 large T for future study.

关 键 词：内皮 血管 永生化 细胞 SV40LT 

分 类 号：R732.2[医药卫生—肿瘤]