Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of a Cell Line HFH-T2, Producing Viral Particles, from Primary Human Fetal Hepatocytes Infected with Hepatitis B Virus

  • Shim, Jae-Kyoung (Department of Biochemistry, Molecular Biology and Pathology, College of Oriental Medicine, Dongguk University) ;
  • Kim, Dong-Wook (Department of Biochemistry, Molecular Biology and Pathology, College of Oriental Medicine, Dongguk University) ;
  • Chung, Tae-Ho (Institute of Liver Research, College of Medicine, Kyungpook National University) ;
  • Kim, June-Ki (Department of Biochemistry, Molecular Biology and Pathology, College of Oriental Medicine, Dongguk University) ;
  • Suh, Jeong-Ill (Department of Internal Medicine, College of Medicine, Dongguk University) ;
  • Park, Chun (Department of Biochemistry, Molecular Biology and Pathology, College of Oriental Medicine, Dongguk University) ;
  • Lee, Young-Choon (Faculty of Life Sciences and Bioresources, Dong-A University) ;
  • Chung, Tae-Wha (Korea Research Institute of Bioscience an Biotechnology, and KOBIAS Co.) ;
  • Song, Eun-Young (Korea Research Institute of Bioscience an Biotechnology, and KOBIAS Co.) ;
  • Kim, Cheorl-Ho (Department of Biochemistry, Molecular Biology and Pathology, College of Oriental Medicine, Dongguk University)
  • Published : 2001.04.01
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Abstract

A primary culture of human fetal hepatocytes was obtained through a therapeutic abortion process at 26 weeks of gestation period. More than $10^8$ cells were seeded on a plastic plate. These hepatocytes were infected with hepatitis B virus (HBV). The HBV was purified from serum of one chronic HBV carrier. Transformed hepatocytes were subcultured in a 10% FBS-supplemented medium. The morphology of the transformed cell was epithelial-like. The cells from the first pass showed signs of early proliferation and had a latent period of more than 3 months after 6-7 passages. After the rest period, the transformed cell proliferated actively and they were subcultured every three days. Transformed hepatocytes were characterized by detection of the HBV transcript by RT-PCR. The secretion of virions from transformed cells was investigated by PCR with the cell medium. Two types of virions secreted into the culture medium were examined by using the transmission electron microscope. Another approach to study the secretion of virions in to culture medium was carried out with HBV antibody. HBsAg was detected in the culture medium of transformed cells using ELISA and Western blot analyses. These data suggested that the human fetal hepatocyte cell line has been established by infection of HBV, in which this cell line secreted viral particles into the culture medium.

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