Precision-cut rat liver slices were prepared with a Krumdieck tissue slicer and cultured in three standard hepatocyte culture media. Rat liver slices cultured in either RPMI 1640 medium or Williams Medium E could be maintained in culture for up to 72 hr. In contrast, Leibovitz's L-15 medium was unsatisfactory in that slice viability, assessed either by morphological examination or by measurement of enzyme activities, could not be maintained for periods greater than 24 hr. As a measure of functional viability liver slices were cultured with some known rodent peroxisome proliferators, namely clofibric acid, nafenopin, ciprofibrate and Wy-14,643. The peroxisome proliferators induced both palmitoyl CoA oxidation and carnitine acetyltransferase activities in 48- and 72-hr slice cultures. Ultrastructural examination of liver slices cultured with either ciprofibrate or Wy-14,643 for 72 hr revealed an increase in the number of peroxisomes. 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 peroxisome proliferators. Precision-cut liver slices may therefore be a useful alternative in vitro system to hepatocyte cultures for screening compounds for effects on enzyme activities and for assessing species differences in response.