The study was designed to investigate the effects of phenobarbital (PB), 3-methylcholanthrene (3-MC), and oltipraz (OPZ), a synthetic derivative of 1,2-dithiole-3-thione, on the levels of cytochrome P450 1A1/2 and gluthathione transferase (GST) mRNAs in both fresh and cryopreserved human, monkey, and dog hepatocytes in primary culture. GST alpha mRNAs were demonstrated in liver parenchymal cells from the three species: after 4 days of culture, their basal levels were decreased, but were strongly higher in PB- and OPZ-treated cells from the three species. In contrast 3-MC was mostly effective on human hepatocytes. The increased levels of GST alpha mRNAs in the presence of PB or OPZ were not observed in all cell populations. GST mu mRNAs, which were detected in both dog and monkey hepatocytes, were induced only in the presence of OPZ. GST pi mRNAs were expressed in dog hepatocytes but did not respond to any of the inducers. In all cases, similar effects were observed in fresh and thawed hepatocytes. Similarly, CYP1A1/2 transcripts were induced by 3-MC in both fresh and cryopreserved cells from the three species but also after OPZ treatment for monkey hepatocytes. These findings demonstrate that enzymes which play a major role in bioactivation/detoxication of xenobiotics remain expressed and inducible in hepatocytes from various species after cryopreservation and thawing.