Human Hepatic HepaRG Cells Maintain an Organotypic Phenotype with High Intrinsic CYP450 Activity/Metabolism and Significantly Outperform Standard HepG2/C3A Cells for Pharmaceutical and Therapeutic Applications

Abstract

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 (PHHs). Limited availability, interdonor functional variability and early phenotypic alterations in PHHs restrict their use, whilst standard cell lines such as HepG2 lack a substantial and variable set of liver‐specific functions such as CYP450 activity. Alternatives include the HepG2‐derivative C3A cells selected as a more differentiated and metabolically active hepatic phenotype. Human HepaRG 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 HepaRG cells using phenotypic profiling, CYP450 activity and drug metabolism parameters to assess their value as hepatic models for pre‐clinical drug testing or therapeutics. Compared with C3As, HepaRG co‐cultures exhibit a more organotypic phenotype, including evidence of hepatic polarity with the strong expression of CYP3A4, the major isoform involved in the metabolism of over 60% of marketed drugs. Significantly greater CYP450 activity and expression of CYP1A2, CYP2E1 and CYP3A4 genes in HepaRG cells (comparable with that of human liver tissue) was demonstrated. Moreover, HepaRG cells also preferentially expressed the hepatic integrin α5β1 – an important modulator of cell behaviour including growth and survival, differentiation and polarity. Drug metabolite profiling of phenacetin (CYP1A2) and testosterone (CYP3A4) using LC‐MS/MS and HPLC, respectively, revealed that HepaRGs had more intact (Phase I–II) metabolism profile. Thus, HepaRG cells significantly outperform C3A cells for the potential pharmaceutical and therapeutic applications.

Nondestructive Optical Toxicity Assays of 3D Liver Spheroids with Optical Coherence Tomography

“…When plotting the same information, i.e., speckle variance as calculated by equation, binned on the X axis of a histogram with pixel count in the Y axis, we found striking differences in the distribution in a dose‐dependent manner ( Figure 4 ). This step was also necessary to validate the choice of the mean speckle variance (MSV) over the spheroids as a valid descriptive statistic parameters of the distribution, onto which further statistical tests could be conducted.…”

“…Moreover, given the central role of the liver in drug metabolism and detoxification, strong incentives exist to create new physiologically relevant, human‐based, in vitro liver models which would offer better prediction of drug‐induced liver injury in humans. Human hepatic HepaRG cells offer a spontaneous coculture model of hepatocytes and cholangiocytes, and maintain in vivo liver‐specific functions, including phase I–III metabolism in 2D and 3D culture . HepaRG cells are considered a sustainable surrogate to primary human hepatocytes and a good cell model for pharmaceutical purposes, and therefore were the chosen cell type for this study.…”

“…Human hepatic HepaRG cells offer a spontaneous coculture model of hepatocytes and cholangiocytes, and maintain in vivo liver‐specific functions, including phase I–III metabolism in 2D and 3D culture . HepaRG cells are considered a sustainable surrogate to primary human hepatocytes and a good cell model for pharmaceutical purposes, and therefore were the chosen cell type for this study.…”

Letter to the editor: ‘Human based systems: Mechanistic NASH modelling just around the corner?’

“…(iii) HepaRG cells display a stable genotype and maintain phenotypically-differentiated functions in culture for at least four weeks, thus making them ideal cells for studying the pathophysiology of NAFLD [4,5].…”

β2-adrenergic receptor-mediated in vitro regulation of human hepatic drug transporter expression by epinephrine

“…Human hepatoma HepaRG cells were also used in the study, owing to the limited, scarce and unpredictable availability of human hepatocytes. Indeed, differentiated HepaRG cells are now recognized as surrogates for human hepatocytes in drug metabolism and transport studies (Bachour-El Azzi et al, 2015;Lubberstedt et al, 2011;Nelson et al, 2017), even if differences between HepaRG cells and human hepatocytes for expression of some transporters such as OATP1B3 (SLCO1B3) exist and have likely to be kept in mind (Le Vee et al, 2013b). HepaRG cells were cultured as previously reported (Le Vee et al, 2013b).…”