Study of multiple genetic variations caused by persistent hepatitis C virus replication in long-term cell culture

Nobuyuki Kato, Youki Ueda, Hiroe Sejima, Weilin Gu, Shinya Satoh, Hiromichi Dansako, Masanori Ikeda, Kunitada Shimotohno

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


The most characteristic feature of the hepatitis C virus (HCV) genome in patients with chronic hepatitis C is its remarkable variability and diversity. To better understand this feature, we performed genetic analysis of HCV replicons recovered from two human hepatoma HuH-7-derived cell lines after 1, 3, 5, 7, and 9 years in culture: The cell lines 50-1 and sO harbored HCV 1B-1 and O strain-derived HCV replicons established in 2002 and 2003, respectively. The results revealed that genetic variations in both replicons accumulated in a time-dependent manner at a constant rate despite the maintenance of moderate diversity (less than 1.8% difference) between the clones and that the mutation rate in the 50-1 and sO replicons was 2.5 and 2.9 × 10-3 base substitutions/site/year, respectively. We found that the genetic distance of both replicons increased from 7.9% to 10.5% after 9 years in culture. In addition, we observed that the guanine + cytosine (GC) content of both replicon RNAs increased in a time-dependent manner, as observed in our previous studies. Finally, we demonstrated that the high sensitivity of both replicons to direct-acting antivirals was maintained even after 9 years in culture. Our results suggest that long-term cultured HCV replicon-harboring cells are a useful model for understanding the variability and diversity of the HCV genome and the drug sensitivity of HCV in patients with chronic hepatitis C.

Original languageEnglish
Pages (from-to)331-343
Number of pages13
JournalArchives of Virology
Issue number2
Publication statusPublished - Feb 2020
Externally publishedYes


  • Carcinoma, Hepatocellular/virology
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Genes, Reporter/genetics
  • Genetic Variation/genetics
  • Genome, Viral/genetics
  • Genotype
  • Hepacivirus/genetics
  • Hepatitis C, Chronic/virology
  • Humans
  • Liver Neoplasms/virology
  • RNA, Viral/genetics
  • Replicon/genetics
  • Virus Replication/genetics

ASJC Scopus subject areas

  • Virology


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