A cultivation method for highly differentiated primary chimpanzee hepatocytes permissive for hepatitis C virus replication.

The liver performs a wide array of functions, a few of which include the synthesis and secretion of most of the plasma proteins, including the lipoproteins, cholesterol, and bile acid metabolism, and detoxification of the blood. In vitro analysis of most liver functions has been hampered by the difficulties encountered in isolating and maintaining functional cultures of primary hepatocytes. Although in vivo the liver has an amazing capacity for regeneration, hepatocytes in culture have limited proliferation capacity and are normally short-lived. We have developed methods for the isolation and cultivation of highly differentiated primate hepatocyte cultures that can be maintained for over 100 d without significant loss of differentiated function. This system has been used in our lab for the analysis of lipoprotein synthesis and hepatotropic virus replication. This chapter is designed to provide a detailed methodology of our approach. Numerous alternative hepatocyte cultivation systems have been described, but owing to space limitations, these systems will not be described here. A number of excellent reviews are dedicated to this subject, one of which is in a previous volume of this series (1), and another that is an entire book dedicated to the subject (2).