Print Email Facebook Twitter A sandwich-cultured rat hepatocyte system with increased metabolic competence evaluated by gene expression profiling Title A sandwich-cultured rat hepatocyte system with increased metabolic competence evaluated by gene expression profiling Author Kienhuis, A.S. Wortelboer, H.M. Maas, W.J. van Herwijnen, M. Kleinjans, J.C.S. van Delft, J.H.M. Stierum, R.H. TNO Kwaliteit van Leven Publication year 2007 Abstract A rapid decline of cytochrome P450 (CYP450) enzyme activities remains a drawback of rat hepatocyte-based in vitro cultures. Consequently, judgment of the toxic potential of compounds that need bioactivation by CYP450s may not be adequate using this model. In the present study, an improved hepatocyte-based in vitro system was developed with special focus on metabolic competence. Therefore, a mixture of CYP450 inducers, phenobarbital, dexamethasone and β-naphthoflavone, was added to culture medium of sandwich-cultured rat hepatocytes. The resulting modified model was evaluated by comparing its genome-wide expression profiles with liver and a standard model without the inducer mixture. Metabolic capacity for CYP450 enzymes showed that the modified model resembled more closely the in vivo situation. Gene expression results revealed large differences between in vivo and both in vitro models. The slight differences between the two sandwich models were predominantly represented by gene expression changes in CYP450s. Importantly, in the modified model, expression ratios of the phase I and the majority of phase II genes more closely resembled liver in vivo. The CYP450 enzyme activities corresponded with gene expression data. In conclusion, for toxicological applications using sandwich-cultured hepatocytes, the modified model may be preferred. © 2007 Elsevier Ltd. All rights reserved. Subject BiologyBiomedical ResearchGene expression profilingIn vitro modelsMetabolic competencePrimary rat hepatocytesSandwich-culturebeta naphthoflavonecytochrome P450dexamethasonephenobarbitalanimal cellarticlecell cultureculture mediumenzyme activitygene expressiongene expression profilingin vitro studyin vivo studyliverliver cellmalemetabolic capacitymodelnonhumannucleotide sequenceratActinsAnimalsBiotransformationCytochrome P-450 Enzyme SystemCytological TechniquesData Interpretation, StatisticalGene Expression ProfilingHepatocytesHydroxylationMaleNucleic Acid HybridizationOligonucleotide Array Sequence AnalysisRatsRats, WistarReverse Transcriptase Polymerase Chain ReactionRNATestosteroneRattus To reference this document use: http://resolver.tudelft.nl/uuid:b9dc3c02-d1bb-48a7-866b-b65afde5ee58 DOI https://doi.org/10.1016/j.tiv.2007.01.010 TNO identifier 240122 ISSN 0887-2333 Source Toxicology in Vitro, 21 (5), 892-901 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.