Synergistic induction of drug-metabolizing enzymes in co-cultures of bovine hepatocytic and sinusoidal cell lines

  • Miyako YoshiokaEmail author
  • Takato Takenouchi
  • Hiroshi Kitani
  • Keerthi S. Guruge
  • Noriko Yamanaka


Hepatocyte-derived cell lines provide useful experimental systems for studying liver metabolism. Unlike human and rodents, few hepatocyte-derived cell lines have been generated from cattle. Here, we established two immortalized bovine hepatocyte-derived cell lines (BH4 and BH5) via transfection with a SV40 large T-antigen construct. Morphological and immunocytochemical analyses revealed that BH4 and BH5 originated from hepatocytes and biliary-epithelial cells, respectively. A potent carcinogen, 3-methylcholanthrene (3-MC), upregulated gene expression of cytochrome P450 (CYP)1A1, CYP1A2, and CYP1B1 in BH4 and BH5, but only to levels less than one-fifteenth of those in primary cultured bovine hepatocytes. Phenobarbital (PB) also increased expression levels of CYP2B6, CYP2C18, and CYP3A4 in BH4 and BH, but at a lower level than 3-MC. By contrast, when BH4 or BH5 was co-cultured with previously established bovine liver sinusoidal cell lines and treated with 3-MC, the gene expression levels of CYP1A1, CYP1A2, and CYP1B1 increased by 38~290%, compared with those in BH4 or BH5 cells cultured alone. PB-treated co-cultures of BH4 or BH5 cells and liver sinusoidal cell lines also showed synergistic increases in CYP2B6 and CYP2C18 expression. Together, the results suggest that these co-cultures could provide an in vitro model for investigations into pharmacological and toxicological properties of drugs in cattle liver.


Hepatocytes Liver sinusoidal cells Bovine liver cell lines Cytochrome P450 mRNA expression 


Funding information

This work was supported by JSPS KAKENHI Grant No. 19K06431.


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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  1. 1.Pathology and Pathophysiology Research DivisionNational Institute of Animal Health, NAROIbarakiJapan
  2. 2.Division of Animal SciencesInstitute of Agrobiological Sciences, NAROIbarakiJapan

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