HepaRG Cells as an in Vitro Model for Evaluation of Cytochrome P450 Induction in Humans

Kanebratt, Kajsa P.; Andersson, Tommy

doi:10.1124/dmd.107.017418

Cited by 183 publications

(140 citation statements)

References 44 publications

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“…A major difference between HepaRG cells and primary hepatocytes was the continuous expression of CYP1A1 in the former; this activity could be related to their transformed state rather than to the presence of biliary cells. RIF is another well established inducer, and our data (Table 1) reproduced well the previous findings in primary hepatocytes (Morel et al, 1990;Lahoz et al, 2008b) and HepaRG cells (Aninat et al, 2006;Kanebratt and Andersson, 2008b). Therefore, HepaRG cells appear to be representative of a large fraction of human hepatocyte populations.…”

Section: Discussionsupporting

confidence: 79%

“…Differentiated HepaRG cells express various liver functions, including P450s, phase II enzymes, transporters, and nuclear receptors at levels comparable with those found in primary hepatocytes and are responsive to prototypical inducers, suggesting that they could represent a surrogate to the latter in drug metabolism and toxicity studies (Aninat et al, 2006;Le Vee et al, 2006;Guillouzo et al, 2007;Josse et al, 2008;Kanebratt and Andersson, 2008b;Turpeinen et al, 2009). Moreover, some evidence has been provided that LD-seeded HepaRG cells can retain relatively stable expression and activities of P450s for several weeks at confluence (Josse et al, 2008;Kanebratt and Andersson, 2008a).…”

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confidence: 99%

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Stable Expression, Activity, and Inducibility of Cytochromes P450 in Differentiated HepaRG Cells

Anthérieu

Chesné²,

Li³

et al. 2009

Drug Metab Dispos

234

143

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ABSTRACT:HepaRG cells possess the unique property to differentiate in vitro and to express various functions of mature hepatocytes, including the major cytochromes P450 (P450s). In the present study, we carefully analyzed mRNA expression and activity of the major P450s and their responsiveness to three prototypical inducers, phenobarbital, rifampicin, and omeprazole, in differentiated HepaRG cell cultures over a 4-week period after low and high seeding. Only minor differences were observed in P450 activities when measured by two cocktails of probe substrates, probably related to the choice and/or concentration of substrates. Similar results were obtained from the two cell seeding conditions. Expression and activities of several P450s were dimethyl sulfoxidedependent. However, basal P450 expression and activities as well as their responsiveness to the prototypical inducers were well maintained over the 4-week period, and a good correlation was observed between transcript levels and corresponding activities. Thus, CYP1A2, CYP2B6, and CYP3A4 were found to accurately respond to their respective prototypical inducers, i.e., omeprazole, phenobarbital, and rifampicin. Likewise, basal expression of several phase II enzymes, transporters, and nuclear receptors, and response to inducers were also well preserved. More genes were found to be induced in HepaRG cells than in primary human hepatocytes, and no marked variation was noticed between the different passages. Taken together, these data support the conclusion that HepaRG cells represent a promising surrogate to primary human hepatocytes for xenobiotic metabolism and toxicity studies.

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Section: Discussionsupporting

confidence: 79%

mentioning

confidence: 99%

Stable Expression, Activity, and Inducibility of Cytochromes P450 in Differentiated HepaRG Cells

Anthérieu

Chesné²,

Li³

et al. 2009

Drug Metab Dispos

234

143

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“…In particular, they are able to express numerous P450 cytochromes, allowing the performance of many normal metabolic liver functions such as the production of phase I and II enzymes and transmembrane transport proteins, unlike other hepatocyte cell lines such as HepG2 (Guillouzo et al 2007;Kanebratt and Andersson 2008;Turpeinen et al 2009;Jennen et al 2010). …”

Section: Discussionmentioning

confidence: 99%

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

et al. 2017

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Magnetic mesoporous silica nanoparticles (M-MSNs) are a promising class of nanoparticles for drug delivery. However, a deep understanding of the toxicological mechanisms of action of these nanocarriers is essential, especially in the liver. The potential toxicity on HepaRG cells of pristine, pegylated (PEG), and lipid (DMPC) M-MSNs were compared. Based on MTT assay and real-time cell impedance, none of these NPs presented an extensive toxicity on hepatic cells. However, we observed by transmission electron microscopy (TEM) that the DMPC and pristine M-MSNs were greatly internalized. In comparison, PEG M-MSNs showed a slower cellular uptake. Whole gene expression profiling revealed the M-MSNs molecular modes of action in a time-and dose-dependent manner. The lowest dose tested (1.6 mg/cm 2 ) induced no molecular effect and was defined as 'No Observed Transcriptional Effect level.' The dose 16 mg/cm 2 revealed nascent but transient effects. At the highest dose (80 mg/cm 2 ), adverse effects have clearly arisen and increased over time. The limit of biocompatibility for HepaRG cells could be set at 16 mg/cm 2 for these NPs. Thanks to a comparative pathway-driven analysis, we highlighted the sequence of events that leads to the disruption of hepatobiliary system, elicited by the three types of MMSNs, at the highest dose. The Adverse Outcome Pathway of hepatic cholestasis was implicated. Toxicogenomics applied to cell cultures is an effective tool to characterize and compare the modes of action of many substances. We propose this strategy as an asset for upstream selection of the safest nanocarriers in the framework of regulation for nanobiosafety. ARTICLE HISTORY

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“…Among these alternatives, the human HepaRG cell line is one of the most suitable human hepatic cell lines due to the retention of key liver functionality (Kanebratt and Andersson, 2008;Turpeinen et al, 2009;Templeton et al 2011). This model is considered useful for the evaluation of DDIs as most of the common CYP isoform activities have been measured in this cell line and shown to be both selectively inhibited and induced by prototypical CYP-selective inhibitors and inducers at comparable levels to those of primary cultures of human hepatocytes (Turpeinen et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Normalization Methods To Predict CYP3A4 Induction in Six Fully Characterized Cryopreserved Human Hepatocyte Preparations and HepaRG Cells

Vermet

Raoust

Ngo

et al. 2015

Drug Metabolism and Disposition

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Prediction of drug-drug interactions due to cytochrome P450 isoform 3A4 (CYP3A4) overexpression is important because this CYP isoform is involved in the metabolism of about 30% of clinically used drugs from almost all therapeutic categories. Therefore, it is mandatory to attempt to predict the potential of a new compound to induce CYP3A4. Among several in vitro-in vivo extrapolation methods recently proposed in the literature, an approach using a scaling factor, called a d factor, for a given hepatocyte batch to provide extrapolation between in vitro induction data and clinical outcome has been adopted by leading health authorities. We challenged the relevance of the calibration factor determined using a set of 15 well-known clinical CYP3A4 inducers or the potent CYP3A4 inducer rifampicin only. These investigations were conducted using six batches of human hepatocytes and an established HepaRG cell line. Our findings show that use of a calibration factor is preferable for clinical predictions, as shown previously by other investigators. Moreover, the present results also suggest that the accuracy of prediction through calculation of this factor is sufficient when rifampicin is considered alone, and the use of a larger set of fully characterized CYP3A4 clinical inducers is not required. For the established HepaRG cell line, the findings obtained in three experiments using a single batch of cells show a good prediction accuracy with or without the d factor. Additional investigations with different batches of HepaRG cell lines are needed to confirm these results.

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HepaRG Cells as an in Vitro Model for Evaluation of Cytochrome P450 Induction in Humans

Abstract: ABSTRACT:HepaRG is a highly differentiated cell line that displays several hepatocyte-like functions, including drug-metabolizing enzymes. In this study, the HepaRG cells were characterized and evaluated as an in vitro model to predict cytochrome P450 (

Cited by 183 publications

References 44 publications

Stable Expression, Activity, and Inducibility of Cytochromes P450 in Differentiated HepaRG Cells

Stable Expression, Activity, and Inducibility of Cytochromes P450 in Differentiated HepaRG Cells

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

Evaluation of Normalization Methods To Predict CYP3A4 Induction in Six Fully Characterized Cryopreserved Human Hepatocyte Preparations and HepaRG Cells

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