HepG2-NTCP Subclones Exhibiting High Susceptibility to Hepatitis B Virus Infection

Area of Science:

• Hepatology
• Virology
• Cell Biology

Background:

• HepG2 cells engineered with sodium taurocholate co-transporting polypeptide (NTCP) are crucial for in vitro Hepatitis B virus (HBV) studies.
• Variability in NTCP expression can impact HBV infection efficiency and data reliability.

Purpose of the Study:

• To isolate and characterize HepG2-NTCP-A3 cell subclones with improved and stable HBV infection efficiency.
• To evaluate the potential of these subclones for developing a more robust in vitro HBV infection model.

Main Methods:

• Single cell-cloning of HepG2-NTCP-A3 cells using limiting dilution.
• Assessment of HBV infection levels and viral antigen expression (HBcAg) via immunofluorescence.
• Evaluation of cell permissiveness under standard and polyethylene glycol (PEG)-free conditions.

Main Results:

• Multiple subclones with increased HBV permissiveness were obtained, with one (HepG2-NTCP-A3/C2) showing >4-fold higher infection than parental cells.
• Subclone C2 demonstrated relatively higher HBV infectivity in the absence of PEG, indicating potential for PEG-free models.
• Increased viral production correlated with elevated intracellular HBcAg expression.
• Isolated subclones exhibited varying NTCP expression levels without significant differences in morphology or growth.

Conclusions:

• Isolated HepG2-NTCP-A3 subclones support significantly more efficient HBV production.
• These subclones represent an improved in vitro model for studying HBV replication and infection dynamics.