Primary chimpanzee hepatocytes were maintained in vitro utilizing a serum‐free medium. Hepatocyte functions were sustained throughout the culture period as demonstrated by the synthesis and secretion of liver‐specific plasma proteins characteristic for differentiated hepatocytes. Hepatocyte cultures established from a chimpanzee chronically infected with human hepatitis B virus exhibited the synthesis and secretion of hepatitis B virus proteins into the medium. In addition, the de novo replication of hepatitis B virus was documented by the recovery of virus, exhibiting an endogenous DNA polymerase activity, from the tissue culture medium. Therefore, both the long‐term maintenance of differentiated hepatocytes and the expression of hepatitis B virus from these primary cultures were sustained in the serum‐free medium.
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