HepaRG-cell
  • Biologically relevant alternative to primary human hepatocytes
  • High batch-to-batch reproducibility
  • Complete media systems to support the growth and use of your HepaRG cells

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Biologically relevant in vitro tool for ADME/Tox studies

Gibco HepaRG cells are terminally differentiated hepatic cells derived from a human hepatic progenitor cell line that retain many characteristics of primary human hepatocytes. HepaRG cells comprise a biologically relevant alternative model that exhibits many characteristics of primary human hepatocytes, including morphology and expression of key metabolic enzymes (Phase I and II), nuclear receptors (CAR, PXR, and AhR), and drug transporters. Unlike HepG2 and Fa2N-4 cells, HepaRG cells have high P450 activity and complete expression of all nuclear receptors. For scientists who need reproducible metabolism data, HepaRG cells are an in vitro tool designed to provide reproducible results in a metabolically complete and scalable system.

Consistent performance

HepaRG cells lack donor variability and lot sizes are not limited by donor tissue availability, helping to ensure an indefinite and consistent supply of cells with high batch-to-batch reproducibility. This enables users to obtain physiologically relevant results for metabolism-based drug-drug interactions without the concern of donor variability and lot sizes. 

Gibco HepaRG cells offered through Thermo Fisher Scientific are terminally differentiated and provided in a convenient cryopreserved format of 107cells/vial (enough for an entire 96-well plate).

Complete media systems to support your research needs

To simplify the growth and use of HepaRG cells, Thermo Fisher Scientific also offers HepaRG media supplements. Simply add the HepaRG media supplement to basal media (William’s E and GlutaMAX supplement) to create a complete media system for your research needs.

Use HepaRG Thaw, Plate & General Purpose supplement (HPRG670/HPRG770) in your media to thaw and seed HepaRG cells.

Continue to grow your HepaRG cells with the appropriate HepaRG supplements for your applications. (all formulations below are used in conjunction with Williams E Media and GlutaMAX supplements.

Incubate the cells in monolayer with the test substrates according to your protocol.

Reagents and application notes are available to help complete your HepaRG workflow:

Use HepaRG Thaw, Plate & General Purpose supplement (HPRG670/HPRG770) in your media to thaw and seed HepaRG cells.

Continue to grow your HepaRG cells with the appropriate HepaRG supplements for your applications. (all formulations below are used in conjunction with Williams E Media and GlutaMAX supplements.

Incubate the cells in monolayer with the test substrates according to your protocol.

Reagents and application notes are available to help complete your HepaRG workflow:

Comparative data: PHH vs HepaRG cells for various applications

The following section contains comparative data generated from Primary Human Hepatocytes vs HepaRG model for multiple applications.

Induction screening

A unique in vitro system for induction screening

P450 enzymes can be induced as a result of drug exposure, which may cause increased formation of toxic metabolites and/or decreased systemic levels of a co-administered drug potentially resulting in drug toxicity or decreased drug efficacy. The use of primary human hepatocytes in screening applications is limited by tissue availability, donor variability, cost, and a relative short culture lifespan. The use of HepaRG cells solves these limitations without sacrificing critical hepatocyte traits such as drug metabolizing enzyme expression, functional transport proteins, and expression of key nuclear receptor pathways.

HepaRG cells respond to prototypical P450 inducers such as omeprazole (OMP), phenobarbital (PB), and rifampicin (RIF) demonstrating the utility of this cell system in the evaluation of in vitro enzyme induction (Figure 1).

Graph of fold-change in enzyme activity for 3 enzymes following induction in PHH and HepaRG cells
Graph of fold-change in mRNA expression for 3 enzymes following induction in PHH and HepaRG cells

Figure 1. Results of induction screening assay. Induction of (A) CYP1A2, CYP2B6, and CYP3A4 enzyme activity and (B) mRNA expression in HepaRG cells or primary human hepatocytes (PHH) treated with omeprazole (OMP), phenobarbital (PB), or rifampicin (RIF) for 72 hr in culture. Box and whisker plots (box = 25th to 75th percentile, line within box = median, whiskers = extreme values observed) were generated using data from multiple PHH preparations, illustrating the large donor-to-donor variability observed in PHH. HepaRG data (activity and mRNA) for CYP1A2, CYP2B6, and CYP3A4 are denoted as red diamonds and were generated from three separate vials.

Metabolism

Assess metabolic stability

Estimates of in vivo metabolic drug clearance can be determined from in vitro metabolism kinetic data. Metabolic stability studies are typically performed to estimate a drug candidate’s metabolic half-life and intrinsic clearance rates, which are major determinants of in vivo drug efficacy. Compounds with short half-lives may require multiple doses to maintain effective plasma levels, whereas compounds with slower elimination kinetics require fewer doses.

Unlike other cell lines (e.g., HepG2 and Fa2N-4) HepaRG cells have expression levels of multiple functional Phase 1 and 2 drug metabolizing enzymes (DME) and nuclear receptors consistent with levels observed in primary human hepatocytes, and therefore, are more suitable to assess the metabolic stability of candidate compounds (Figure 2).

Graph of clearance rates for 11 drugs in PHH and HepaRG cells
Figure 2. Results from metabolic clearance assay.Intrinsic clearance of the reference drugs aminodarone, carbamezepine, clozapine, diclofenac, dextromethorphan, lovastatin, methotrexate, rifampicine, tacrine, troglitazone, and verapamil in cultures of primary human hepatocytes (PHH, n=6) and HepaRG cells (n = 2). Results are shown as means + SD [2].

Toxicity

Investigate acute and chronic toxicity

The liver plays a central role in metabolizing and eliminating xenobiotics and as a result is susceptible to injury from drug toxicity. Liver toxicity has led to withdrawal or severe use limitations of marketed drugs and is a major problem in drug development.

HepaRG cells are a metabolically competent system and tolerant of long culture periods (i.e., ≥22 days).  In addition, they are well suited for in vitro determinations of acute and chronic toxicity resulting from intrinsic and/or metabolism-based mechanisms (Figures 3 and 4).

Bar chart of HepaRG cell viability following treatment with toxicants at 2 concentrations

Figure 3. Results from viability assay. HepaRG cell viability after 24 hr and 72 hr treatment with metabolism-dependent toxicants [3].

Bar chart of HepG2 and HepaRG cell viability following treatment with aflatoxin B1 at 3 concentrations

Figure 4. Results from cytotoxicity assay. Comparative cytotoxicity of aflatoxin B1 in HepG2 and HepaRG cells after a 3-day treatment. Cell viability was estimated using a standard MTT test. The values were normalized to untreated cells and expressed as means ± SD (n = 3 cultures) [4].

Transporters

Assess potential transporter-mediated drug interactions

Transporters often work together with drug-metabolizing enzymes in drug absorption and elimination, resulting in altered drug efficacy and adverse drug effects. HepaRG cells have superior expression levels of key uptake and efflux transporters compared to other cell lines and expression levels closely resembling those of human hepatocytes (Figures 5 and 6). They also form tight junctions and bile canaliculi, making them ideal for uptake and biliary secretion studies.

Bar chart of mRNA amounts from 6 uptake transporter genes in HepaRG and HepG2 cells
Figure 5. Results from uptake transporter gene expression assay. Uptake transporter gene expression, HepaRG and HepG2 cells vs. PHH [5].
Bar chart of mRNA amounts from 4 efflux transporter genes in HepaRG and HepG2 cells
Figure 6. Results from efflux transporter gene expression assay. Efflux transporter gene expression, HepaRG and HepG2 cells vs. PHH [5].

References

  1. Aninat C et al. (2008) Crit Care Med 36:848–854.
  2. Lübberstedt M et al. (2011) J Pharmacol Toxicol Methods 63:59–68.
  3. Aninat C et al. (2006) Drug Metab Dispos 34:75–83.
  4. Guillouzo A et al. (2007) Chem Biol Interact 168:66–73.
  5. Le Vee M et al. (2006) Eur J Pharm Sci 28:109–117.

HepaRG™ is a trademark of BioPredic International.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.

Limited Use License
HepaRG™ cells are patented and their use is strictly limited; consider the cells as a single-use, disposable product that must be destroyed upon conclusion of a study or experiment. Propagating, reproducing, cloning, subcloning or any other use of the cells following the conclusion of a study is prohibited. Use of the cells to produce or manufacture commercial products for general sale or for use in the manufacture of products intended for general sale is prohibited. Transfer of the cells to anyone not employed within the same organization, whether for financial benefit or not, is also prohibited.

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