The invention relates to the in vitro viral cell culture of hepatitis non-A, non-B hepatitis (NANBH). Were isolated and cultured primary hepatocytes from a chimpanzee during the acute phase of an experimental infection with NANBH virus. The culture of hepatocytes differentiated cells was maintained in serum-free medium comprising a Williams medium E, a hepatocyte proliférogène, transferrin, serum albumin, a corticosteroid, prolactin, a factor of release of thyrotropin, a cholera toxin and ethanolamine. The cultured hepatocytes were positive for expression of a cytoplasmic antigen associated with NANBH. The presence of the cytoplasmic marker suggested persistence of infection in vitro. The production of infectious virus in vitro was confirmed by inoculating a tissue culture infected with NANBH virus and documenting subsequent development of NANBH in chimpanzees.
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