Induction of cytochrome P450 enzymes in primary equine hepatocyte culture

Stefanski, Anja; Mevissen, Meike; Möller, A.-M.; Kühni-Boghenbor, Kathrin; Schmitz, Andrea (2013). Induction of cytochrome P450 enzymes in primary equine hepatocyte culture. Toxicology in vitro, 27(7), pp. 2023-2030. Elsevier 10.1016/j.tiv.2013.07.009

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In this study, we established cell culture conditions for primary equine hepatocytes allowing cytochrome P450 enzyme (CYP) induction experiments. Hepatocytes were isolated after a modified method of Bakala et al. (2003) and cultivated on collagen I coated plates. Three different media were compared for their influence on morphology, viability and CYP activity of the hepatocytes. CYP activity was evaluated with the fluorescent substrate 7-benzyloxy-4-trifluoromethylcoumarin. Induction experiments were carried out with rifampicin, dexamethasone or phenobarbital. Concentration-response curves for induction with rifampicin were created. Williams' medium E showed the best results on morphology and viability of the hepatocytes and was therefore used for the following induction experiments. Cells cultured in Dulbecco's Modified Eagle Medium were not inducible. Incubation with rifampicin increased the CYP activity in two different hepatocyte preparations in a dose dependent manner (EC50=1.20 μM and 6.06 μM; Emax=4.1- and 3.4-fold induction). No increase in CYP activity was detected after incubation with dexamethasone or phenobarbital. The hepatocyte culture conditions established in this study proved to be valuable for investigation of the induction of equine CYPs. In further studies, other equine drugs can be evaluated for CYP induction with this in vitro system.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Pharmacology and Toxicology
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Anatomy

UniBE Contributor:

Stefanski, Anja; Mevissen, Meike; Kühni, Kathrin and Schmitz, Andrea

Subjects:

600 Technology > 630 Agriculture
500 Science > 570 Life sciences; biology

ISSN:

0887-2333

Publisher:

Elsevier

Language:

English

Submitter:

Susanne Portner

Date Deposited:

22 Aug 2014 14:05

Last Modified:

14 Oct 2015 11:17

Publisher DOI:

10.1016/j.tiv.2013.07.009

PubMed ID:

23916975

Uncontrolled Keywords:

Horse, Cytochrome P450, Hepatocyte, Induction, Rifampicin

BORIS DOI:

10.7892/boris.46117

URI:

https://boris.unibe.ch/id/eprint/46117

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