• Hepatitis B Virus (HBV) Replication in HepG2 Cell Line
  • Reihaneh Kazemi,1 Mojtaba Hamidifard,2,* Elham Siasi,3 Mohammad Reza Aghasadeghi,4 Pooneh Rahimi,5
    1. Department of Genetics, NT.C., Islamic Azad University, Tehran, Iran
    2. Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran
    3. Department of Genetics, NT.C., Islamic Azad University, Tehran, Iran
    4. Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran
    5. Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran


  • Introduction: Hepatitis B virus (HBV) is a leading cause of liver diseases, including chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). Understanding the mechanisms of HBV replication in vitro is crucial for developing effective therapeutic strategies; however, this has proven challenging due to the virus’s reliance on host liver cells and the complexity of its life cycle. The HepG2 cell line is commonly employed in HBV research, yet its capacity to support replication of wild-type (non-engineered) HBV directly from clinical samples remains underexplored. In this study, we investigated the replication of HBV in HepG2 cells following infection with sera obtained from HBV-positive patients.
  • Methods: Serum samples were collected from patients previously diagnosed with HBV infection, confirmed by positive ELISA results for HBV surface antigens (HBsAg). The HEPG2 cell line was cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, and maintained at 37°C in 5% CO₂. Cells were seeded in 6-well plates and infected with HBV-positive patient serum. After incubation periods ranging from 3 to 7 days, the culture supernatants and cell lysates were collected for analysis. The presence of HBV antigens in supernatants was evaluated using a commercial ELISA kit specific for HBsAg.
  • Results: Following infection with HBV-positive serum, HEPG2 cells demonstrated detectable levels of HBsAg in the culture supernatants as measured by ELISA. The amount of HBsAg increased over time, indicating active viral replication and protein secretion.
  • Conclusion: Our findings demonstrate that HBV from infected human serum can effectively infect and replicate in HepG2 cells under in vitro conditions. The increasing levels of HBsAg detected by ELISA indicate successful viral replication, supporting the utility of HepG2 cells as a model for studying HBV infection and testing antiviral compounds.
  • Keywords: HBV-HEPG2-ELISA