原代人脐静脉内皮细胞的分离培养及其对人巨细胞病毒的易感性研究  

Primary Culture of Human Umbilical Vein Endothelial Cells and Their Susceptibility to Human Cytomegalovirus

作　　者：董丽群[1] 周淑[1] 关林波[1] 王巍[1] 张立[2] 于萍[1] 

机构地区：[1]四川大学华西第二医院儿科,成都610041 [2]四川大学华西医院移植免疫实验室,成都610041

出　　处：《四川大学学报（医学版）》2010年第3期424-427,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30600677及30500483)资助

摘　　要：目的通过检测原代人脐静脉内皮细胞(HUVEC)对人巨细胞病毒(HCMV)的易感性,探讨胎儿宫内感染HCMV的可能途径。方法无菌条件下取30例健康产妇正常娩出的胎盘端游离脐带,向脐静脉灌注0.1%胶原酶Ⅱ37℃消化15 min,通过对细胞的形态学观察和免疫荧光染色方法对细胞进行纯度鉴定;用HCMV病毒株体外感染HUVEC,通过对HCMV基因及蛋白检测确定HUVEC对HCMV的易感性。结果 0.1%胶原酶Ⅱ、15 min的消化方法对HUVEC有较好的分离效果,免疫荧光染色结果表明HUVEC细胞内有Ⅷ因子相关抗原表达;HCMV体外感染HUVEC 72 h后形态学有明显改变,病毒基因及蛋白均有阳性表达。结论原代HUEVC对HCMV易感,通过脐静脉的血液传播可能是胎儿宫内感染HCMV的重要途径之一。Objective To detect whether the primary culture of human umbilical vein endothelial cells（HUVEC） is susceptible to human cytomegalovirus（HCMV） infection so as to explain how virus from mothers traverses the placenta to fetus.Methods Thirty umbilical cords of healthy parturient were used in this study.The HUVEC were harvested with 0.1% collagenase for 15 minutes.The cells were identified by morphology and immunofluorescence.Then the HUVEC were infected with HCMV in vitro.The DNA of cytomegalovirus and protein of infected cells were detected 72 hours later.Results The HUVEC with adequate quality were harvested, which expressed Ⅷ related antigen.Positive expressions of CMV gB gene and virus proteins were identified in all of the HUVEC infected with HCMV.Conclusion HUVEC is susceptible to HCMV infection,which suggests a possible route of HCMV infection through the fetal-maternal interface.

关 键 词：人原代脐静脉内皮细胞 HCMV 

分 类 号：R363[医药卫生—病理学]