Replication of hepatitis E virus in three-dimensional cell culture

Area of Science:

• Virology
• Hepatology
• Public Health

Background:

• Hepatitis E virus (HEV) causes epidemics in developing regions and sporadic cases in developed areas.
• Pigs are identified as reservoirs for zoonotic HEV transmission, with undercooked pork consumption a suspected route.
• Challenges in in vitro HEV propagation hinder research into zoonotic transmission and food safety.

Purpose of the Study:

• To develop a 3D cell culture system for efficient Hepatitis E virus propagation.
• To utilize the Rotating Wall Vessel (RVW) for culturing HEV in a 3D environment.
• To facilitate studies on HEV viability in contaminated food products, particularly pig tissues.

Main Methods:

• Utilized the Rotating Wall Vessel (RVW) to establish a 3D cell culture system.
• Employed human hepatoblastoma PLC/PRF/5 cells for HEV culture.
• Assessed HEV replication and virion presence using real-time RT-PCR and electron microscopy.

Main Results:

• Demonstrated efficient HEV replication in 3D cultures for up to 5 months.
• Confirmed HEV replication through detection of HEV RNA and complete virions.
• Showcased the production of infectious HEV progeny capable of infecting new 3D cultures.

Conclusions:

• The RVW 3D cell culture system effectively supports HEV replication and production of infectious virions.
• This model system offers a valuable tool for investigating HEV contamination in food sources.
• Advances understanding of HEV zoonotic potential and aids in developing food safety strategies.