INDUCTION OF CYTOCHROME-P-450-DEPENDENT ENZYME-ACTIVITIES IN CULTURED RAT-LIVER SLICES

Precision-cut liver slices were prepared from male Sprague-Dawley rats with a Krumdieck tissue slicer and cultured in RPMI 1640 medium for up to 72 hr. After 48 hr, cytochrome P-450 content in the slices declined to 36% of levels present in freshly cut rat liver slices. The addition of either beta-naphthoflavone (BNF) or Aroclor 1254 (ARO) partially prevented the loss of cytochrome P-450. Culture of liver slices with phenobarbitone (PB), BNF and ARO resulted in the induction of 7-ethoxy-coumarin O-deethylase, 7-benzoxyresorufin O-debenzylase and 7-ethoxyresorufin O-deethlylase activities. Generally, the induction of mixed-function oxidase enzymes was greater in 72- than in 48-hr cultured slices, and at the concentrations examined ARO produced a greater stimulation of enzyme activities than did either PB or BNF. These results demonstrate that rat liver slices may be maintained in culture for up to 72 hr, and that they respond in a similar manner to rat primary hepatocyte cultures to some inducers of xenobiotic metabolism. Precision-cut liver slices may therefore be a useful alternative in vitro system to hepatocyte cultures for screening compounds for effects on mixed-function oxidases and for assessing species differences in response.