JoVE Encyclopedia of Experiments
Microbiology
0 views • 2:52 min • August 31st, 2026
Begin with a culture of human liver cancer cells transfected with hepatitis E virus (HEV) RNA to produce viral particles.
Some HEVs acquire a host-derived endosomal membrane and exit the cell, while others remain within the cell as non-enveloped HEVs.
To harvest extracellular HEVs, collect and filter the culture supernatant to remove cellular debris.
Store at an ultra-low temperature to preserve infectivity.
To harvest intracellular HEVs, wash the cells with a buffer.
Add trypsin-EDTA to detach the cells, then add a medium to stop enzymatic activity.
Transfer the suspension into a tube and centrifuge.
Discard the supernatant containing residual trypsin. Resuspend the pellet and transfer it to a tube.
Freeze the cells in liquid nitrogen and thaw them on ice.
Repeat the freeze-thaw cycles to lyse the cells, releasing the viruses.
Centrifuge, collect the supernatant containing the viruses, and store at an ultra-low temperature.
On day six, filter the supernatant through a 0.45 micrometer mesh to remove any cell debris and store the harvested extracellular cell culture-derived hepatitis E virus at four degrees Celsius. For intracellular cell culture-derived hepatitis E virus particle harvest, was
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