The loss of cytochrome P450 (CYP)-dependent activity continues to be a problem in the use of cultured hepatocytes in xenobiotic toxicity studies. It has been reported that the inclusion of pyruvate and various hormones in the culture medium improves the maintenance of various hepatic functions, including that of CYP2C11 mRNA expression. We have studied this further, by investigating the effects of the addition of pyruvate and hormones on various CYP-dependent enzyme activities and on the CYP-dependent toxicity of precocene II in rat hepatocyte cultures. No beneficial effects of this medium supplementation could be demonstrated.
SkettP. (1994). Problems in using isolated and cultured hepatocytes for xenobiotic metabolism/metabolism-based toxicity testing solutions?Toxicology in Vitro8, 491–504.
2.
TomitaY., YuasaC., NiR., IshimuraK., & IchiharaA. (1995). Long-term maintenance of functional rat hepatocytes in primary culture by additions of pyruvate and various hormones. Biochemica et Biophysica Acta1243, 329–335.
3.
HammondA.H., & FryJ.R. (1991). The use of hepatocytes cultured from inducer-treated rats in the detection of cytochrome P450-mediated cytotoxicity. Toxicology in Vitro5, 133–137.
4.
FryJ.R., & BridgesJ.W. (1980). The metabolism of 7-ethoxycoumarin in primary maintenance cultures of adult rat hepatocytes. Naunyn-Schmiedeberg's Archives of Pharmacology311, 85–90.
5.
PohlR.J., & FoutsJ.R. (1980). A rapid method for assaying the metabolism of 7-ethoxyresorufin by microsomal subcellular fractions. Analytical Biochemistry107, 150–155.
6.
HammondA.H., FryJ.R., & GarleM.J. (1996). Toxicity of dichloropropanols in rat hepatocyte cultures. Environmental Toxicology and Pharmacology1, 39–43.
7.
WaxmanD.J. (1985). Regulation of rat hepatic cytochrome P450: age-dependent expression, hormonal imprinting, and xenobiotic inducibility of sex-specific isoenzymes. Biochemistry24, 4409–4417.
8.
SoucekP., & GutI. (1992). Cytochromes P450 in rats: structures, functions, properties and relevant human forms. Xenobiotica22, 83–103.
9.
BurkeM.D., ThomsonS., WeaverR.J., WolfC.R., & MayerR.T. (1994). Cytochrome P450 specificities of alkoxyresorufin O-dealkylation in human and rat liver. Biochemical Pharmacology48, 923–936.
10.
NakajimaT., ElovaaraE., ParkS.S., GelboinH.V., HietanenE., & VainioH. (1990). Monoclonal antibody-directed characterization of benzene, ethoxyresorufin and pentoxyresorufin metabolism in rat liver microsomes. Biochemical Pharmacology40, 1255–1261.
11.
HsiaM., GrossmanS., & SchrankelK. (1981). Hepatotoxicity of the anti-juvenile hormone pre-cocene II and the generation of dihydrodiol metabolites. Chemico-Biological Interactions37, 265–277.
12.
HammondA.H., & FryJ.R. (1994). Toxicity of precocene II in rat hepatocyte cultures: effects of serum and culture time. Toxicology Letters70, 337–342.
13.
HammondA.H., GarleM.J., & FryJ.R. (1995). Mechanism of toxicity of precocene II in rat hepatocyte cultures. Journal of Biochemical Toxicology10, 265–273.
14.
VanhaeckeT., VercruysseA., & RogiersV. (1997). Effect of pyruvate on glutathione S-transferase expression in primary cultures of rat hepatocytes. Toxicology in Vitro11, 435–441.
