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Human Hepatic Hepa RG Cells Maintain an Organotypic Phenotype with High Intrinsic CYP 450 Activity/Metabolism and Significantly Outperform Standard HepG2/C3A Cells for Pharmaceutical and Therapeutic Applications
Author(s) -
Nelson Leonard J.,
Morgan Katie,
Treskes Philipp,
Samuel Kay,
Henderson Catherine J.,
LeBled Claire,
Homer Natalie,
Grant M. Helen,
Hayes Peter C.,
Plevris John N.
Publication year - 2017
Publication title -
basic and clinical pharmacology and toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.805
H-Index - 90
eISSN - 1742-7843
pISSN - 1742-7835
DOI - 10.1111/bcpt.12631
Subject(s) - drug metabolism , cyp3a4 , phenotype , biology , phenotypic screening , cell culture , phenacetin , transcriptome , cell growth , metabolite , microbiology and biotechnology , in vitro , pharmacology , metabolism , cytochrome p450 , drug , gene expression , biochemistry , gene , genetics
Conventional in vitro human hepatic models for drug testing are based on the use of standard cell lines derived from hepatomas or primary human hepatocytes ( PHH s). Limited availability, interdonor functional variability and early phenotypic alterations in PHH s restrict their use, whilst standard cell lines such as HepG2 lack a substantial and variable set of liver‐specific functions such as CYP 450 activity. Alternatives include the HepG2‐derivative C3A cells selected as a more differentiated and metabolically active hepatic phenotype. Human Hepa RG cells are an alternative organotypic co‐culture model of hepatocytes and cholangiocytes reported to maintain in vivo ‐like liver‐specific functions, including intact Phase I–III drug metabolism. In this study, we compared C3A and human Hepa RG cells using phenotypic profiling, CYP 450 activity and drug metabolism parameters to assess their value as hepatic models for pre‐clinical drug testing or therapeutics. Compared with C3As, Hepa RG co‐cultures exhibit a more organotypic phenotype, including evidence of hepatic polarity with the strong expression of CYP 3A4, the major isoform involved in the metabolism of over 60% of marketed drugs. Significantly greater CYP 450 activity and expression of CYP 1A2, CYP 2E1 and CYP 3A4 genes in Hepa RG cells (comparable with that of human liver tissue) was demonstrated. Moreover, Hepa RG cells also preferentially expressed the hepatic integrin α 5 β 1 – an important modulator of cell behaviour including growth and survival, differentiation and polarity. Drug metabolite profiling of phenacetin ( CYP 1A2) and testosterone ( CYP 3A4) using LC ‐ MS / MS and HPLC , respectively, revealed that Hepa RG s had more intact (Phase I–II) metabolism profile. Thus, Hepa RG cells significantly outperform C3A cells for the potential pharmaceutical and therapeutic applications.