Abstract
Cytochrome P450 enzymes are a diverse group of catalytic enzymes in the liver that are mainly responsible for the biotransformation of organic substances. Cytochrome P450 activity as well as both its induction and inhibition are key factors in drug biotransformation and can be involved in deactivation, activation, detoxification and toxification processes. Thus, the modulation of cytochrome P450 activity is an important parameter when evaluating the potential toxicity of chemical compounds using an in vitro system. The cytochrome P450 3A subfamily proteins are among the most important drug-metabolizing enzymes in human liver and are responsible for about half of all cytochrome P450-dependent drug oxidations. In vitro, these enzymes are active not only in primary human hepatocyte cultures, but also in differentiated human hepatoma HepaRG cells. The present protocol describes the culture of cryopreserved differentiated HepaRG cells and the evaluation of its cytochrome P450 activity upon exposure to a chemical compound using a commercially available luminogenic cytochrome P450 assay. This in vitro model can be used to monitor the induction and inhibition of cytochrome P450 3A following exposure to a particular test compound.
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Acknowledgements
This work has received funding from grants of the Fund for Scientific Research in Flanders (FWO-Vlaanderen, GO16312N), the European Community’s Seventh Framework Programme (FP7/2007–2013) under grant agreement no. 266838 (DETECTIVE) and no. 266777 (HEMIBIO) and from the Brussels research fund INNOVIRIS (Brustem).
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Rodrigues, R.M., De Kock, J., Doktorova, T.Y., Rogiers, V., Vanhaecke, T. (2015). Measurement of Cytochrome P450 Enzyme Induction and Inhibition in Human Hepatoma Cells. In: Vinken, M., Rogiers, V. (eds) Protocols in In Vitro Hepatocyte Research. Methods in Molecular Biology, vol 1250. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2074-7_20
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DOI: https://doi.org/10.1007/978-1-4939-2074-7_20
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Online ISBN: 978-1-4939-2074-7
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