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Research ArticleArticle

GENE EXPRESSION IN HUMAN HEPATOCYTES IN SUSPENSION AFTER ISOLATION IS SIMILAR TO THE LIVER OF ORIGIN, IS NOT AFFECTED BY HEPATOCYTE COLD STORAGE AND CRYOPRESERVATION, BUT IS STRONGLY CHANGED AFTER HEPATOCYTE PLATING

Lysiane Richert, Michael J. Liguori, Catherine Abadie, Bruno Heyd, Georges Mantion, Nermin Halkic and Jeffrey F. Waring
Drug Metabolism and Disposition May 2006, 34 (5) 870-879; DOI: https://doi.org/10.1124/dmd.105.007708
Lysiane Richert
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Michael J. Liguori
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Catherine Abadie
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Bruno Heyd
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Georges Mantion
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Nermin Halkic
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Jeffrey F. Waring
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Abstract

Isolated primary human hepatocytes are a well accepted system for evaluating pharmacological and toxicological effects in humans. However, questions remain regarding how culturing affects the liver-specific functions of the hepatocytes. In addition, cryopreservation could also potentially affect the differentiation state of the hepatocytes. The first aim of the present study was to compare gene expression in freshly isolated primary hepatocytes to that of the liver of origin and to evaluate the expression changes occurring after cryopreservation/thawing, both when maintained in suspension and after plating. The second aim of the present study was to evaluate gene expression in hepatocytes after cold storage of suspensions up to 24 h compared with freshly isolated hepatocytes in suspension. Our results show that the gene expression in freshly isolated human hepatocytes in suspension after isolation is similar to that of the liver of origin. Furthermore, gene expression in primary human hepatocytes in suspension is not affected by hepatocyte cold storage and cryopreservation. However, the gene expression is profoundly affected in monolayer cultures after plating. Specifically, gene expression changes were observed in cultured relative to suspensions of human hepatocytes that are involved in cellular processes such as phase I/II metabolism, basolateral and canalicular transport systems, fatty acid and lipid metabolism, apoptosis, and proteasomal protein recycling. An oxidative stress response may be partially involved in these changes in gene expression. Taken together, these results may aid in the interpretation of data collected from human hepatocyte experiments and suggest additional utility for cold storage and cryopreservation of hepatocytes.

Footnotes

  • L.R. and M.J.L. contributed equally to this work.

  • Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.

  • doi:10.1124/dmd.105.007708.

  • ABBREVIATIONS: P450, cytochrome P450; DMEM, Dulbecco's modified Eagle's medium; UW, University of Wisconsin solution; UGT, UDP-glucuronosyltransferase; CoA, coenzyme A.

  • ↵ Embedded Image The online version of this article (available at http://dmd.aspetjournals.org) contains supplemental material.

    • Received October 3, 2005.
    • Accepted February 3, 2006.
  • The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition: 34 (5)
Drug Metabolism and Disposition
Vol. 34, Issue 5
1 May 2006
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GENE EXPRESSION IN HUMAN HEPATOCYTES IN SUSPENSION AFTER ISOLATION IS SIMILAR TO THE LIVER OF ORIGIN, IS NOT AFFECTED BY HEPATOCYTE COLD STORAGE AND CRYOPRESERVATION, BUT IS STRONGLY CHANGED AFTER HEPATOCYTE PLATING
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Research ArticleArticle

GENE EXPRESSION IN HUMAN HEPATOCYTES IN SUSPENSION AFTER ISOLATION IS SIMILAR TO THE LIVER OF ORIGIN, IS NOT AFFECTED BY HEPATOCYTE COLD STORAGE AND CRYOPRESERVATION, BUT IS STRONGLY CHANGED AFTER HEPATOCYTE PLATING

Lysiane Richert, Michael J. Liguori, Catherine Abadie, Bruno Heyd, Georges Mantion, Nermin Halkic and Jeffrey F. Waring
Drug Metabolism and Disposition May 1, 2006, 34 (5) 870-879; DOI: https://doi.org/10.1124/dmd.105.007708

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Research ArticleArticle

GENE EXPRESSION IN HUMAN HEPATOCYTES IN SUSPENSION AFTER ISOLATION IS SIMILAR TO THE LIVER OF ORIGIN, IS NOT AFFECTED BY HEPATOCYTE COLD STORAGE AND CRYOPRESERVATION, BUT IS STRONGLY CHANGED AFTER HEPATOCYTE PLATING

Lysiane Richert, Michael J. Liguori, Catherine Abadie, Bruno Heyd, Georges Mantion, Nermin Halkic and Jeffrey F. Waring
Drug Metabolism and Disposition May 1, 2006, 34 (5) 870-879; DOI: https://doi.org/10.1124/dmd.105.007708
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