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Cytochrome P450 expression profile of the PICM-19H pig liver cell line: potential application to rapid liver toxicity assays

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Abstract

Liver in vitro models are needed to replace animal models for rapid assessment of drug biotransformation and toxicity. The PICM-19 pig liver stem cell line may fulfill this need since these cells have activities associated with xenobiotic phase I and II metabolism lacking in other liver cell lines. The objective of this study was to characterize phase I and II metabolic functions of a PICM-19 derivative cell line, PICM-19H, compared to the tumor-derived human HepG2 C3A cell line and primary cultures of adult porcine hepatocytes. Following exposure of PICM-19H cells to either 3-methylcholanthrene, rifampicin or phenobarbital, the induced activities of cytochrome P450 (CYP450) isozymes CYP-1A, -2, and-3A were assessed. Relative to adult porcine hepatocytes, PICM-19H cells exhibited 30% and 43%, respectively, of CYP1A and 3A activities, while HepG2 C3A cells exhibited 7% and 0% of those activities. Fluorescent metabolites were extensively conjugated, i.e., 52% and 96% of CYP450-1A and-3A metabolites were released from medium samples following treatment with β-glucuronidase/arylsulfatase. Rifampicin induction of CYP450 isozyme activities was confirmed by conversion of testosterone to 6β-OH-, 2α-OH- and 2β-OH-testosterone, as determined by mass spectrometry. Susceptibility of PICM-19H cells to acetaminophen toxicity was determined; CD50 was calculated to be 14.9 ± 0.9 mM. Toxicity and bioactivation of aflatoxin B1 was determined in 3-methylcholanthrene-treated cultures and untreated controls; CD50 were 1.59 μM and 31 μM, respectively. These results demonstrate the potential use of PICM-19H cells in drug biotransformation and toxicity testing and further support their use in extracorporeal artificial liver device technology.

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Acknowledgments

We thank Dr. John McMurtry and Dr. Wes Garrett for reading the manuscript and for offering helpful editorial and scientific comments in its final preparation. We also thank Ms. Amy Shannon for assistance with enzyme assays and Grant Harrington for assistance with LC MS/MS method development.

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Authors and Affiliations

  1. USDA, ARS, Animal and Natural Resources Institute, Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, Bldg. 200, Rm. 13, BARC-East, Beltsville, MD, 20705, USA

    Ryan R. Willard, Neil C. Talbot & Thomas J. Caperna

  2. USDA-ARS, Animal Metabolism Unit, Biosciences Research Laboratory, Fargo, ND, 58105, USA

    Nancy W. Shappell

  3. Hepalife Technologies, Inc., 60 State Street, Suite 700, Boston, MA, 02109, USA

    John H. Meekin

Authors
  1. Ryan R. Willard
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  2. Nancy W. Shappell
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  3. John H. Meekin
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  4. Neil C. Talbot
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  5. Thomas J. Caperna
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Correspondence to Neil C. Talbot.

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Editor: J. Denry Sato

Mention of trade names or commercial products in this publication is solely for the purposes of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture

The study was supported in part by Hepalife Technologies, Inc., 60 State Street, Suite 700, Boston, MA 02109 under USDA Cooperative Research and Development Agreement No. 58-3K95-8-1238.

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Willard, R.R., Shappell, N.W., Meekin, J.H. et al. Cytochrome P450 expression profile of the PICM-19H pig liver cell line: potential application to rapid liver toxicity assays. In Vitro Cell.Dev.Biol.-Animal 46, 11–19 (2010). https://doi.org/10.1007/s11626-009-9244-z

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  • DOI: https://doi.org/10.1007/s11626-009-9244-z

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