Xiao Juan, Zhang Dandan, Chen Juanjuan, Yin Zongzhi, Liu Tao, Ai Jihui, Chen Suhua
Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Feb;30(1):94-7. doi: 10.1007/s11596-010-0117-1. Epub 2010 Feb 14.
Human trophoblast cells were isolated and cultured in vitro in order to investigate possible pathogenesis of intrauterine infection caused by HCMV. Trophoblast cells were obtained by compound enzymes digestion and discontinuous percoll gradient. Cells and purity were identified by using immunocytochemistry assay with anti-CK7, Vim and beta-hCG antibodies. HCMV AD169 strain replication in isolated trophoblast cells and cell apoptosis were detected at different time points post infection (p.i.). The results showed that highly purified trophoblast cells were obtained. Specific virus replication was increased dramatically at the 24th h p.i., and then increased slowly during 48 h and 72 h. Apoptosis rate of trophoblast cells infected with HCMV was (34.68+/-3.14)% at 24th h p.i., while that in control group was (15.32+/-2.34)% (P<0.05). It was suggested that highly purified trophoblast cells can be isolated by the simplified cell purification method. HCMV can infect human trophoblast cells, and be quickly replicated, resulting in the accelerated apoptosis of human trophoblast cells during early time.
为了研究人巨细胞病毒(HCMV)引起的宫内感染的可能发病机制,分离并体外培养了人滋养层细胞。通过复合酶消化和不连续 Percoll 梯度法获得滋养层细胞。使用抗 CK7、波形蛋白(Vim)和β-人绒毛膜促性腺激素(β-hCG)抗体的免疫细胞化学分析法鉴定细胞及其纯度。在感染后(p.i.)的不同时间点检测 HCMV AD169 株在分离的滋养层细胞中的复制情况以及细胞凋亡情况。结果显示获得了高度纯化的滋养层细胞。特异性病毒复制在感染后第 24 小时急剧增加,然后在 48 小时和 72 小时期间缓慢增加。感染 HCMV 的滋养层细胞在感染后第 24 小时的凋亡率为(34.68±3.14)%,而对照组为(15.32±2.34)%(P<0.05)。提示通过简化的细胞纯化方法可分离出高度纯化的滋养层细胞。HCMV 可感染人滋养层细胞,并快速复制,导致人滋养层细胞在早期加速凋亡。