四川大学学报（医学版）2012，Vol.43Issue(3)：344-347,377,5.
原代人绒毛滋养层细胞分离培养及其对巨细胞病毒的易感性研究
Primary Culture of Human Trophoblastic Cells and Their Susceptibility to Huma Cytomegalovirus
摘要
Abstract
Objective to determine the susceptibility of primary human trophoblastic cells (HTCs) to human cytomegalovirus (HCMV) infection. Methods 30 villus of healthy pregnant women were digested with a mixture of 0. 25% trypsin and 0.2% DNase. The digestion was then filtered by a 200 mesh stainless steel sieve. HTCs were purified by Percoll gradient centrifugation and repeat purification with trypsin during cell passage. HTCs were identified by cell morphology and immunofluorescence. PCR, Western blot and immunofluorescent staining were performed to detect HCMV gene and protein 72 hours post infection of HCMV. Results The combination of 0.25% trypsin and 0. 2% DNase with Percoll gradient centrifugation isolated and purified primary HTCs. More than 90 percent cells expressed CK-7 related antigen. The results of PCR, western blot and immunofluorescence showed positive expressions of CMV gB and virus proteins. Conclusion HTCs is susceptible to HCMV infection, which suggests a possible route of HCMV infection at the fetal-maternal interface.关键词
人绒毛滋养层细胞/人巨细胞病毒Key words
Human trophoblastic cell/ HCMV引用本文复制引用
周淑,董丽群,关林波,于萍..原代人绒毛滋养层细胞分离培养及其对巨细胞病毒的易感性研究[J].四川大学学报（医学版）,2012,43(3):344-347,377,5.基金项目
国家自然科学基金(批准号81070499)、四川省科技厅应用基础项目(2009JY0048)、早期发育与损伤的基础与临床嵌合研究教育部创新团队(PCSIRT0935)和四川省科技厅科技支撑项目(2009SZ0198)资助 （批准号81070499）
