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01.03.2018 | Molecular Toxicology

Monitoring cytochrome P450 activity in living hepatocytes by chromogenic substrates in response to drug treatment or during cell maturation

verfasst von: Jannick Theobald, Xinlai Cheng, Ali Ghanem, Haristi Gaitantzi, Guangqi Song, Edda Klipp, Judith Wodke, Holger Becker, Ralf Mrowka, Katja Breitkopf-Heinlein, Steven Dooley, Stefan Wölfl

Erschienen in: Archives of Toxicology | Ausgabe 3/2018

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Abstract

The metabolic activity of hepatocytes is a central prerequisite for drug activity and a key element in drug–drug interaction. This central role in metabolism largely depends on the activity of the cytochrome P450 (CYP450) enzyme family, which is not only dependent on liver cell maturation but is also controlled in response to drug and chemical exposure. Here, we report the use of VividDye fluorogenic CYP450 substrates to directly measure and continuously monitor metabolic activity in living hepatocytes. We observed time- and dose-dependent correlation in response to established and putative CYP450 inducers acting through the aryl hydrocarbon receptor and drug combinations. Using repetitive addition of VividDye fluorogenic substrate on a daily basis, we demonstrated the new application of VividDye for monitoring the maturation and dedifferentiation of hepatic cells. Despite a lack of high specificity for individual CYP450 isoenzymes, our approach enables continuous monitoring of metabolic activity in living cells with no need to disrupt cultivation. Our assay can be integrated in in vitro liver-mimetic models for on-line monitoring and thus should enhance the reliability of these tissue model systems.

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Titel: Monitoring cytochrome P450 activity in living hepatocytes by chromogenic substrates in response to drug treatment or during cell maturation
verfasst von: Jannick Theobald
Xinlai Cheng
Ali Ghanem
Haristi Gaitantzi
Guangqi Song
Edda Klipp
Judith Wodke
Holger Becker
Ralf Mrowka
Katja Breitkopf-Heinlein
Steven Dooley
Stefan Wölfl
Publikationsdatum: 01.03.2018
Verlag: Springer Berlin Heidelberg
Erschienen in: Archives of Toxicology / Ausgabe 3/2018
Print ISSN: 0340-5761
Elektronische ISSN: 1432-0738
DOI: https://doi.org/10.1007/s00204-017-2128-1

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