Infectious Disease Laboratory, China Medical University, Shengjing.
Intervirology. 2011;54(5):276-81. doi: 10.1159/000324528. Epub 2011 Apr 1.
To build a hepatitis B virus (HBV)-infected human trophoblast cell model in vitro and determine the mechanism of intrauterine HBV infection.
Serum from hepatitis B-infected patients containing HBV DNA >10(9) was drawn, subsequently inoculated into human trophoblast cells in vitro (JEG3) and passage-cultured. The supernatants and intracellular HBV viral load of inoculated cells were tested by real-time PCR, and HBV DNA was determined by Southern blot.
From inoculation of the 1st passage JEG3 cells, the supernatant viral load of the 1st passage was seen increasing over time, which peaked at 120 h, whereas the HBV viral load was seen decreasing gradually in subsequent passages, and tested negative after the 6th passage. In addition, infected cells of HBV DNA were tested by Southern blot, and showed continual expression in the subsequent cell passages 1-5 while passage 6 was negative. HBsAg was tested as positive from different passages 1-5, and its concentration was also seen decreasing with each subsequent passage cultured until the 6th passage when it tested negative.
HBV could infect human trophoblast cells (JEG3) in vitro, and it showed continual expression in subsequent cell passages 1-5.
建立体外乙型肝炎病毒（HBV）感染人滋养层细胞模型，探讨宫内HBV 感染的机制。
采集 HBV 感染者血清，HBV-DNA>10(9)拷贝/ml，接种体外培养的人滋养层细胞（JEG3），传代培养。实时荧光定量 PCR 法检测接种细胞上清及细胞内 HBV 病毒载量，Southern blot 法检测 HBV-DNA。
第 1 代 JEG3 细胞接种后，上清液 HBV 病毒载量逐渐升高，120 h 达峰值，随后逐渐下降，至第 6 代检测不到；Southern blot 检测到感染细胞持续表达 HBV-DNA，至第 6 代检测不到；HBsAg 阳性表达从第 1 代至第 5 代逐渐下降，至第 6 代检测不到。
HBV 可体外感染人滋养层细胞（JEG3），并在随后的细胞传代中持续表达。
