, Volume 71, Issue 1, pp 231–244 | Cite as

Evaluation of the hepatocyte-derived cell line BFH12 as an in vitro model for bovine biotransformation

  • Alexander Gleich
  • Bastian Kaiser
  • Walther Honscha
  • Herbert Fuhrmann
  • Axel SchoenigerEmail author
Original Article


The knowledge of drug metabolising enzymes (DMEs) in cattle is rather limited. The capability of the bovine foetal hepatocyte-derived cell line BFH12 to serve as model for biotransformation was evaluated. Gene expression analysis of DMEs was performed by reverse transcription PCR (RT-PCR). The presence of efflux transporters was visualised by immunocytochemistry, and functional induction of cytochrome P450 (CYP) 1A was assessed by the ethoxyresorufin-O-deethylase (EROD) assay. The production of bile acids was measured by liquid chromatography-tandem mass spectrometry (LC–MS/MS). RT-PCR revealed the expression of cytochromes 1A1, 1A2, 3A4 and phase II enzymes UGT1A1, UGT1A6 and GSTM1. Immunofluorescence demonstrated efflux transporters ABCG2 and ABCC1. The EROD assay revealed a dose-dependent CYP1A induction after treatment with benzo[a]pyrene (BP). LC–MS/MS analysis of cell culture supernatants showed the production of bile acids including taurocholic acid, tauro-chenodeoxycholic acid, taurodeoxycholic acid and taurolithocholic acid. The results strongly suggest the applicability of the cell line BFH12 for subsequent experiments in the emerging field of bovine biotransformation.


Biotransformation Cytochrome P450 Bovine hepatocyte Cattle In vitro model Bile acid 



ATP-binding cassette sub-family C member 1


ATP-binding cassette sub-family G member 2




aryl hydrocarbon receptor


Analysis of variance


AhR nuclear translocator




complementary DNA






Cytochrome P450


Cytochrome P450 1A1


Cytochrome P450 1A2


Cytochrome P450 2B6


Cytochrome P450 2C19


Cytochrome P450 2C9


DAPI, 4′,6-diamidino-2-phenylindole


drug metabolising enzymes


Ethylenediaminetetraacetic acid




Foetal bovine serum


Glucose transporter 2


Glutathione S-Transferase M1


Liquid chromatography tandem mass spectroscopy


Maximum residue level


3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide


Sodium-taurocholate cotransporting polypeptide


Organic anion transporter


Organic anion-transporting polypeptide


Organic anion-transporting polypeptide 1B3


Organic cation transporter 1




Polycyclic aromatic hydrocarbons


Polymerase chain reaction


Reverse transcription polymerase chain reaction


Quantitative reverse transcription PCR


Standard deviation


Sodium dodecyl sulfate


Solute Carrier Family 22 Member 1


Solute carrier organic anion transporter family member 1B3


SV40 large-T antigen


Taurocholic acid


Taurochenodeoxycholic acid


Taurodeoxycholic acid


Taurolithocholic acid


UDP glucuronosyltransferase 1 family polypeptide A1


UDP glucuronosyltransferase 1 family polypeptide A6



We gratefully thank the group of Professor Michaela Schulz-Siegmund of the Institute of Pharmaceutical Technology (University of Leipzig) for discussions and feedback. Adult liver tissue samples were obtained from the group of Professor Alexander Starke of the Medizinische Tierklinik (University of Leipzig). We also thank the University Hospital Regensburg for conducting LC/MS–MS measurements of bile acids. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10616_2018_279_MOESM1_ESM.docx (814 kb)
Supplementary material 1 (DOCX 814 kb)


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© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Institute of BiochemistryUniversity of LeipzigLeipzigGermany
  2. 2.Institute of Veterinary PhysiologyUniversity of LeipzigLeipzigGermany
  3. 3.Institute of Veterinary Pharmacology and ToxicologyUniversity of LeipzigLeipzigGermany

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