Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins

Abstract

In the pharmaceutical industry, improving the early detection of drug-induced hepatotoxicity is essential as it is one of the most important reasons for attrition of candidate drugs during the later stages of drug development. The first objective of this study was to better characterize different cellular models (i.e., HepG2, HepaRG cells, and fresh primary human hepatocytes) at the gene expression level and analyze their metabolic cytochrome P450 capabilities. The cellular models were exposed to three different CYP450 inducers; beta-naphthoflavone (BNF), phenobarbital (PB), and rifampicin (RIF). HepG2 cells responded very weakly to the different inducers at the gene expression level, and this translated generally into low CYP450 activities in the induced cells compared with the control cells. On the contrary, HepaRG cells and the three human donors were inducible after exposure to BNF, PB, and RIF according to gene expression responses and CYP450 activities. Consequently, HepaRG cells could be used in screening as a substitute and/or in complement to primary hepatocytes for CYP induction studies. The second objective was to investigate the predictivity of the different cellular models to detect hepatotoxins (16 hepatotoxic and 5 nonhepatotoxic compounds). Specificity was 100% with the different cellular models tested. Cryopreserved human hepatocytes gave the highest sensitivity, ranging from 31% to 44% (depending on the donor), followed by lower sensitivity (13%) for HepaRG and HepG2 cells (6.3%). Overall, none of the models under study gave desirable sensitivities (80–100%). Consequently, a high metabolic capacity and CYP inducibility in cell lines does not necessarily correlate with a high sensitivity for the detection of hepatotoxic drugs. Further investigations are necessary to compare different cellular models and determine those that are best suited for the detection of hepatotoxic compounds.

Liver X receptor α (LXRα/NR1H3) regulates differentiation of hepatocyte-like cells via reciprocal regulation of HNF4α

“…Since the HepaRG cell line was first described in 2002, many reports [33,34] have appeared that have described its unique features. These include the ability to differentiate towards both hepatocyte-like and biliary-like cells and the ability to stably express a range of important hepatic functions.…”

Contextualizing Hepatocyte Functionality of Cryopreserved HepaRG Cell Cultures

“…To reach phenotypic maturity, HepaRG cells grow to confluence and differentiate over 4 weeks (from progenitor cells) into cocultures of hepatocyte-like and cholangiocyte-like cells (Gripon et al, 2002). Since this model was discovered, many studies have shown that freshly differentiated HepaRG cultures exhibit cellular interactions, drug metabolism/transport, and drug induction responsiveness comparable to PHH cultures (Grime et al, 2010;McGill et al, 2011;Gerets et al, 2012;Le Vee et al, 2013;Szabo et al, 2013). A cryopreserved format of differentiated HepaRG cells (cryo-HepaRG) has recently become available, improving the global availability and experimental flexibility of this model.…”

Utility of human hepatocyte spheroids without feeder cells for evaluation of hepatotoxicity

“…Several in vitro studies using primary-cultured human hepatocytes or human immortalized cells have been reported for evaluating DILI, using flutamide (Gerets et al, 2012), diclofenac (Bort et al, 1999), isoniazid (Wang et al, 2002) or chlorpromazine (Gerets et al, 2012) as model drugs. These studies all utilized the IC 50 value of albumin secretion as a toxicity parameter, and all of them yielded values similar to or greater than our IC 50 value.…”