In-vitro toxicity testing applications: Hepatotoxicity (reactive metabolites, lipidosis, choleastasis etc), Virology (HBV, HCV…), Infection with hepatitis and other viruses for replication, ...
Liver assist devices/ BioArtificial Livers
Transgenic liver humanized animals with HepaRG™ cells
…. an ever-expanding range of applications!
HeparRG™ cells are more economical, convenient, and predictable than fresh or cryopreserved primary human hepatocytes. It's available in proliferative state to be expanded and differentiatedin-house, or as cryopreserved, fully differentiated and ready-to-use hepatic cells.
Full array of functions, responses, and regulatory pathways of primary human hepatocytes including:Phase I and II, and transporter activities consistent with those found within a population of primary human hepatocytes + Intact response elements, PXR, CAR and PPARa
Form bile canaculi
Has the potential to express major properties of stem cells
High plasticity & complete transdifferentiation capacity
Abstract : Non-alcoholic Fatty Liver Disease (NAFLD) is a frequently encountered Drug-Induced Liver Injury (DILI). Although this stage of the disease is reversible, it can lead to irreversible damage provoked by non-alcoholic steatohepatitis (NASH), fibrosis and cirrhosis. Therefore, the assessment of NAFLD is a paramount objective in toxicological screenings of new drug candidates. In this study, a metabolomic fingerprint of NAFLD induced in HepaRG® cells at four dosing schemes by a reference toxicant, sodium valproate (NaVPA), was obtained using liquid-liquid extraction followed by liquid chromatography and accurate mass-mass spectrometry (LC-AM/MS). The combination of a strict design of experiment with a robust detection method, applied on sodium valproate, validated the possibilities of untargeted metabolomics in hepatic toxicological research. Distinctive patterns between exposed and control cells were consistently observed, multivariate analyses selected up to 200 features of interest, revealing hallmark NAFLD-biomarkers, such as diacylglycerol and triglyceride accumulation and carnitine deficiency. Initial toxic responses show increased levels of S-adenosylmethionine and mono-acetylspermidine in combination with only a moderate increase in triglycerides. New specific markers of toxicity have been observed, such as spermidines, creatine, and acetylcholine. The described design of experiment provides a valuable metabolomics platform for mechanistic research of toxicological hazards and identified new markers for steatotic progression.
Abstract : BACKGROUND:
The HepaRG cells have key drug metabolism functionalities comparable to those of primary human hepatocytes. Many studies have reported that this cell line can be used as a reliable in vitro model for human drug metabolism studies, including the assessment of cytochrome P450 (CYP) induction.
The objective of this study is to determine whether CYP mRNA level measurement is superior to the CYP enzyme activity measurement as a convenient high-throughput method for evaluating CYP induction potential using HepaRG cells.
QuantiGene Plex 2.0 Assay and LC/MS/MS. mRNA expression levels and enzyme activities of CYP1A2, CYP2B6, and CYP3A in HepaRG cells treated with prototypical inducers of each CYP isoform [omeprazole (OME) for CYP1A2, phenobarbital (PB) for CYP2B6, and rifampicin (RIF) for CYP3A] were evaluated.
Although the activities of CYP2B6 and CYP3A was were induced by treatment with PB and RIF, we found that the activity of phenacetin O-deethylase (PHOD), which is known as a marker of the activity of CYP1A2, was also enhanced by treatment with these non-CYP1A2 inducers in HepaRG cells. Based on previously published reports, we hypothesized that the expression ratio of CYP3A to CYP1A2 is much higher in HepaRG cells than in human hepatocytes; this may result in a nonnegligible contribution of CYP3A to the PHOD reaction in HepaRG cells. Studies using CYP3A inhibitor and pregnane X receptor-knockout HepaRG cells supported this hypothesis.
The measurement of mRNA serves as a higher reliable indicator for the evaluation of CYP induction potential when using HepaRG cells.
Time lapse (Total recording time: 2 hours). Chlormoprazine superior to 40µM constricts the bile lumen and provokes cholestasis. Constriction of canaliculus lumen has been observed in presence of CPZ after 2 hours.