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

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Abstract

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.

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Funding

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

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Authors and Affiliations

  1. Pathology and Pathophysiology Research Division, National Institute of Animal Health, NARO, Kannondai 3-1-5, Tsukuba, Ibaraki, 305-0856, Japan

    Miyako Yoshioka, Keerthi S. Guruge & Noriko Yamanaka

  2. Division of Animal Sciences, Institute of Agrobiological Sciences, NARO, 1-2 Ohwashi, Tsukuba, Ibaraki, 305-8634, Japan

    Takato Takenouchi & Hiroshi Kitani

Authors
  1. Miyako Yoshioka
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  2. Takato Takenouchi
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  3. Hiroshi Kitani
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  4. Keerthi S. Guruge
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  5. Noriko Yamanaka
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Correspondence to Miyako Yoshioka.

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Editor: Tetsuji Okamoto

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Yoshioka, M., Takenouchi, T., Kitani, H. et al. Synergistic induction of drug-metabolizing enzymes in co-cultures of bovine hepatocytic and sinusoidal cell lines. In Vitro Cell.Dev.Biol.-Animal 56, 2–9 (2020). https://doi.org/10.1007/s11626-019-00408-6

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  • DOI: https://doi.org/10.1007/s11626-019-00408-6

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