Abstract
Over the last decade HepaRG cells have emerged as a promising alternative to primary human hepatocytes (PHH) and have been featured in over 300 research publications. Most of these reports employed freshly differentiated HepaRG cells that require time-consuming culture (∼28 days) for full differentiation. Recently, a cryopreserved, predifferentiated format of HepaRG cells (termed here “cryo-HepaRG”) has emerged as a new model that improves global availability and experimental flexibility; however, it is largely unknown whether HepaRG cells in this format fully retain their hepatic characteristics. Therefore, we systematically investigated the hepatocyte functionality of cryo-HepaRG cultures in context with the range of interindividual variation observed with PHH in both sandwich-culture and suspension formats. These evaluations uncovered a novel adaptation period for the cryo-HepaRG format and demonstrated the impact of extracellular matrix on cryo-HepaRG functionality. Pharmacologically important drug-metabolizing alleles were genotyped in HepaRG cells and poor metabolizer alleles for CYP2D6, CYP2C9, and CYP3A5 were identified and consistent with higher frequency alleles found in individuals of Caucasian decent. We observed liver enzyme inducibility with aryl hydrocarbon receptor, constitutive androstane receptor (CAR), and pregnane X receptor activators comparable to that of sandwich-cultured PHH. Finally, we show for the first time that cryo-HepaRG supports proper CAR cytosolic sequestration and translocation to hepatocyte nuclei in response to phenobarbital treatment. Taken together, these data reveal important considerations for the use of this cell model and demonstrate that cryo-HepaRG are suitable for metabolism and toxicology screening.
Footnotes
- Received February 1, 2016.
- Accepted June 22, 2016.
↵1 Current affiliation: J.P.J. currently with Qualyst Transporter Solutions (QTS).
↵2 Current affiliation: E.D.C. currently with Quest Pharmaceutical Services (QPS).
↵3 Current affiliation: S.S.F. currently with the National Toxicology Program Division, National Institute of Environmental Health Sciences (NIEHS).
The laboratory research was funded by Life Technologies (Thermo Fisher) research and development. Resources for data analysis and manuscript preparation were supported by the National Institutes of Health, National Institute of Environmental Health Sciences, the National Toxicology Program Division, and by Qualyst Transporter Solutions, Inc.
Some of the data presented were part of a poster abstract: Jackson JP, Edwards M, Chamberlain E, and Ferguson, SS (2011) Cryopreserved HepaRG cells: an alternative in vitro screening tool for human hepatic drug metabolism, induction of metabolism, and safety applications, at the 2011 ISSX meeting; Atlanta, GA. International Society for the Study of Xenobiotics, Atlanta GA.
↵This article has supplemental material available at dmd.aspetjournals.org.
- U.S. Government work not protected by U.S. copyright
DMD articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|