Print Email Facebook Twitter Comparison of coumarin-induced toxicity between sandwich-cultured primary rat hepatocytes and rats in vivo: A toxicogenomics approach Title Comparison of coumarin-induced toxicity between sandwich-cultured primary rat hepatocytes and rats in vivo: A toxicogenomics approach Author Kienhuis, A.S. Wortelboer, H.M. Hoflack, J.C. Moonen, E.J. Kleinjans, J.C.S. van Ommen, B. van Delft, J.H.M. Stierum, R.H. TNO Kwaliteit van Leven Publication year 2006 Abstract Sandwich-cultured primary rat hepatocytes are often used as an in vitro model in toxicology and pharmacology. However, loss of liver-specific functions, in particular, the decline of cytochrome P450 (P450) enzyme activity, limits the value of this model for prediction of in vivo toxicity. In this study, we investigated whether a hepatic in vitro system with improved metabolic competence enhances the predictability for coumarin-induced in vivo toxicity by using a toxicogenomics approach. Therefore, primary rat hepatocytes were cultured in sandwich configuration in medium containing a mixture of low concentrations of P450 inducers, phenobarbital, dexamethasone, and β-naphthoflavone. The toxicogenomics approach used enabled comparison of similar mechanistic endpoints at the molecular level between in vitro and in vivo conditions, namely, compound-induced changes in multiple genes and signaling pathways. Toxicant-induced cytotoxic effects and gene expression profiles observed in hepatocytes cultured in modified medium and hepatocytes cultured in standard medium (without inducers) were compared with results from a rat in vivo study. Coumarin was used as a model compound because its toxicity depends on bioactivation by P450 enzymes. Metabolism of coumarin toward active metabolites, coumarin-induced cytotoxicity, and gene expression modulation were more pronounced in hepatocytes cultured in modified medium compared with hepatocytes cultured in standard medium. In addition, more genes and biological pathways were similarly affected by coumarin in hepatocytes cultured in modified medium and in vivo. In conclusion, these experiments showed that for coumarin-induced toxicity, sandwich-cultured hepatocytes maintained in modified medium better represent the situation in vivo compared with hepatocytes cultured in standard medium. Copyright © 2006 by The American Society for Pharmacology and Experimental Therapeutics. Subject BiologyBiomedical Researchbeta naphthoflavonecoumarincytochrome P450dexamethasonedrug metaboliteenzyme inducing agentphenobarbitalanimal cellanimal experimentanimal modelanimal tissuearticlecontrolled studyculture mediumcytotoxicitydrug activationdrug metabolismenzyme activityexperimental ratgene expressionhistopathologyin vitro studyin vivo studyliver cellliver toxicitymalenonhumannucleotide sequencepredictionprincipal component analysispriority journalratsignal transductiontoxicogeneticsAlanine TransaminaseAnimalsAspartate AminotransferasesCell Culture TechniquesCells, CulturedCholesterolCoumarinsgamma-GlutamyltransferaseGene Expression ProfilingHepatocytesLiverMaleOligonucleotide Array Sequence AnalysisRatsRats, WistarToxicogenetics To reference this document use: http://resolver.tudelft.nl/uuid:3eea1fec-79cc-4614-a6b5-0d42049fbc97 DOI https://doi.org/10.1124/dmd.106.011262 TNO identifier 239699 ISSN 0090-9556 Source Drug Metabolism and Disposition, 34 (12), 2083-2090 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.