To improve our understanding of the mechanisms of hepatitis E virus (HEV) replication, a novel mouse model that is humanized using primary human hepatocytes has been established. The mice were infected with two HEV genotypes (1 and 3), and infection and spreading parameters were monitored in various tissues and cell types. These techniques revealed that HEV virions in faeces were more infective than those from serum, and that HEV genotype 1 spread with faster kinetics. Infected mice were also treated with ribavirin, after which a strong reduction in viraemia was seen, demonstrating the suitability of this model for evaluating preclinical antiviral agents.