[32] Hepatocyte culture in study of P450 regulation

This chapter discusses the hepatocyte culture in study of P450 regulation. Primary cultures of adult rat hepatocytes provide a unique system for the study of many differentiated liver functions including induction of the cytochromes P450. These nonproliferating hepatocytes can be maintained in a chemically defined, serum-free medium for weeks while retaining the capabilities of expressing a wide array of liver-specific functions. This chapter describes the essential elements in isolating hepatocytes with the use of proteolytic enzymes and culturing the viable cells. Adult male Sprague–Dawley rats weighing 200–250 g or females weighing 185–210 g can be purchased from commercial animal breeding sources. Rats are maintained in pairs in wire-bottomed cages with free access to chow diet and kept under regulated lighting.