Characterization and induction of xenobiotic metabolizing enzyme activities in a primary culture of rainbow trout hepatocytes

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Abstract
1. A primary cell culture from rainbow trout (Oncorhynchus mykiss) liver was prepared and evaluated for biotransformation of xenobiotics. 2. The hepatocytes maintained cytochrome P-450 content, as well as their cytochrome P-450-dependent activities, stable for 5-6 days in serum-free medium. Protein and glutathione levels, as well as other enzyme activities important for biotransformation, were close to their fresh cell levels throughout the culture period. 3. The cells were also responsive to cytochrome P-450 inducers. Both beta-naphthoflavone (BNF) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) caused an increase in ethoxyresorufin O-deethylase (EROD) activity, which was dose-dependent over the concentration ranges of 3.6-360 nM and 2.5-100 pM, respectively. The induced activities in BNF exposed cells returned to basal levels within 48 h after replacing the medium with a BNF-free medium. Exposure of cells to TCDD (100 pM) for 48 h induced EROD activity which, in contrast to response of BNF-exposed cells, continued to increase after the medium had been replaced with TCDD-free medium. 4. The results show that trout hepatocytes in primary culture afford a reliable in vitro method for studying the regulation and functions of xenobiotic biotransformation enzymes, and for defining toxic effects of aquatic pollutants in cells.

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