|HepaRG Cells Bibliography|
|HepaRG cells and their application to diverse endpoints|
|Applications of HepaRG Cells|
|Cryopreserved HepaRG cells|
HepaRG™ cells are terminally differentiated human hepatocellular carcinoma cells that reproducibly express the activities of transporters and drug metabolizing enzymes. Functionally equivalent to primary human hepatocytes, they facilitate studies of uptake, metabolism, and disposition of drug candidates. EMD Millipore is the exclusive supplier of Biopredic’s International’s original, widely cited HepaRG™ cells, manufactured by Biopredic using their proprietary process.
Advantages: Quality, Convenience and Value
- Respond and function like primary human hepatocytes across a range of applications
- Do not require daily medium changes, are reproducible and can be stored until required
- Can be used for diverse studies at a lower overall cost than human primary hepatocytes
Single-use format and simple workflow
- For the resources listed below, click on the Supporting Documentation tab above:
- HepaRG™ White Paper: a detailed characterization of HepaRG and comparison to other cell models
- HepaRG™ Manual: thawing, culture, and use of cells
- HepaRG™ Top Applications List
- HepaRG™ Full Bibliography
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Dependable alternative to primary human hepatocytes
The supply of primary human hepatocytes (PHHs) is limited and sporadic. PHHs have little growth capacity, have a short lifespan, and lose their differentiated functions when cultured even for a few days. PHHs also suffer from large donor variations in initial and longer-term functions and enzyme activities, especially CYPs.
By contrast, HepaRG cells are a hepatic model that exhibit long-term stability of differentiated activities and lack donor variability. Together with the advantage of an abundant supply and a single use format, HepaRG cells represent a powerful alternative to PHHs.
Origin of HepaRG cells
The HepaRG cell line was established from a tumor of a female patient suffering from chronic hepatitis C infection and hepatocarcinoma. The cells are likely to have originated from ductular structures –rather than mature hepatocytes or bile ducts – associated with long-term HCV infection. HepaRG cells do not contain any part of the HCV genome or express any HCV protein. When passaged at low density, HepaRG cells can recover and differentiate into both hepatocytes and biliary epithelial cells and are thus considered to be progenitor cells.
Differentiated HepaRG cells are hepatocyte-like in nature, with morphology close to that of PHHs. Once differentiated, HepaRG cells stop proliferating and retain their hepatocyte-like features. Genes up-regulated during differentiation are those relating to cell cycle inhibition, increased susceptibility to apoptosis, innate immunity and liver-enriched transcripts involved in lipid homeostasis and drug metabolism. The expression of different hepatic nuclear factors (HNFs) involved in hepatic-specific gene expression changes as the cells grow and differentiate. The cells surrounding the hepatocyte-like cells are biliary epithelial cells.
HepaRG cells do not produce urea (due to poor or disturbed nitrogen elimination via the urea cycle), but they do regulate carbohydrate metabolism (glycogenolysis and/or gluconeogenesis), produce lactate (a product of anaerobic metabolism) and albumin, and eliminate galactose and sorbitol at comparable rates to PHHs.
High levels of drug metabolizing enzymes (DMEs) and transporters in HepaRG cells allow them to be used for metabolism studies. The mRNA content of CYPs in HepaRG cells is similar to that in PHHs, suggesting that HepaRG cells are not overabundant in a single CYP isoform. Intrinsic clearance of a number of compounds in differentiated HepaRG has been compared with that in PHHs.
HepaRG cells can be used for CYP inhibition studies as they have sufficient levels of DMEs. Studies by Turpeinen et al. showed a good correlation between IC50 values of inhibitors of CYPs in HepaRG cells and PHHs.
High expression levels of CAR, AhR, PXR, and PPAR – the transcription factors involved in regulation of DME and transporters – make HepaRG a strong alternative to PHHs for hepatic testing.
Drug-induced liver injury
The stable metabolic activity of HepaRG cells makes them suitable for long-term and repeated dose toxicity studies in which the toxic effects are metabolism-dependent and/or only evident after days or weeks. The presence of biliary cells and hepatocytes provides information as to whether toxicity is specific to one cell type or whether both are affected.
Steosis – the accumulation triglycerides – caused by drugs has been demonstrated in vitro in HepaRG cells treated with a number of polyunsaturated fatty acids and derivatives.
Phospholipidosis – accumulation of phospholipids and formation of lamellar bodies – has also been shown to occur in HepaRG cells after treatment with amiodarone. The HepaRG cells were able to discriminate between amiodarone which causes steatosis (after 24 h) and phospholipidosis (evident after 2 weeks) and tetracycline, which causes steatosis only.
Cultured HepaRG cells are suitable for biliary secretion studies due to the presence and function of efflux and uptake transporters, with formation of tight junctions and correct location of canaliculi.
HepaRG cells are responsive to lipopolysaccharides and show the classical response of PHHs to inflammatory stimulation. They are able to differentiate between β-receptor specific modulations of inflammatory responses and provide information on which catecholamine to use for treatment of sepsis.
The permissiveness of HepaRG cells to HBV and HCV infection allow use of HepaRG cells as a model for viral hepatitis research.