Cultured human adult hepatocytes: a new model for drug metabolism studies

Bégué, Jean-Marc; Bigot, J. F. Le; Guguen‐Guillouzo, Christiane; Kiechel, J. R.; Guillouzo, André

doi:10.1016/0006-2952(83)90341-6

Cited by 66 publications

(10 citation statements)

References 12 publications

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“…These observations strongly support the view that the sodium-dependent bile acid uptake system present in normal hepatocytes was maintained in the co-culture system. This appears to be yet another liver-specific function maintained in this culture system, as previously demonstrated for albumin synthesis and secretion (5), drug metabolism (17), and cytochrome P-450 activity (18).…”

Section: Nacl (Mm)mentioning

confidence: 69%

Long-Term maintenance of taurocholate uptake by adult rat hepatocytes co-cultured with a liver epithelial cell line

et al. 1985

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Taurocholate (TC) uptake by adult rat hepatocytes co-cultured with other rat liver epithelial cells (RLEC) was studied comparatively to hepatocytes in primary culture. Cells were cultured on Petri dishes for desired times prior to measuring their ability to transport TC. TC uptake was linear for 150 sec in both culture conditions. In hepatocytes cultured alone, the initial rate of TC uptake at an extracellular concentration of 100 microM was 0.19 +/- 0.02 nmole per min per 10(6) cells after 48 hr of culture and decreased by 75% after 4 to 6 days. In hepatocytes co-cultured with RLEC, the rate of uptake at 48 hr (0.31 +/- 0.01 nmole per min per 10(6) cells) was significantly higher than in hepatocytes cultured alone (p less than 0.01); in addition, TC uptake remained stable at an average rate of 0.17 +/- 0.01 nmole per min per 10(6) cells for up to 56 days. No detectable uptake was found in RLEC cultured alone. TC uptake exhibited both saturable (Vmax = 0.30 +/- 0.03 nmole per min per 10(6) cells and Km = 42.6 +/- 4.4 microM) and nonsaturable components. These kinetic parameters were similar to those previously reported in isolated hepatocytes and in short-term cultured hepatocytes. TC uptake exhibited sodium dependence and was significantly reduced when extracellular sodium was replaced by lithium and sucrose, or in the presence of 1 mM ouabain. After 18 days of co-culture, TC uptake had qualitatively the same characteristics as at 48 hr, with a saturable and a nonsaturable component.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Nacl (Mm)mentioning

confidence: 69%

Long-Term maintenance of taurocholate uptake by adult rat hepatocytes co-cultured with a liver epithelial cell line

et al. 1985

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“…However, with a few compounds major differences can be observed. Thus, cycloheximide inhibits protein synthesis but not urea synthesis, whereas norvaline does the opposite (192 (127). In a recent study we found that minor metabolites were detected only after two or three daily exposures to the compounds (203).…”

Section: However Survival and Metabolic Compe-mentioning

confidence: 85%

“…A considerable step forward in long-term maintenance of differentiated hepatocytes was made by adding another liver epithelial cell type (126). When cocultured with rat liver epithelial cells probably derived from primitive biliary cells, hepatocytes from various species including humans survive for several weeks and retain various liverspecific functions; these include production of plasma proteins, expression of both phase I and phase II drug metabolizing enzymes, and taurocholate uptake (52,71,(127)(128)(129)(130)(131)(132)(133)(134)(135) (71,126,128) and with preservation of communication through gap junctions (136). These two features could be related because in pure hepatocyte cultures the addition of proteoglycans improves gap junction activity (137).…”

Section: However Survival and Metabolic Compe-mentioning

confidence: 99%

Liver cell models in in vitro toxicology.

Guillouzo

1998

Environ Health Perspect

297

125

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In vitro liver preparations are increasingly used for the study of hepatotoxicity of chemicals. In recent years their actual advantages and limitations have been better defined. The cell models, slices, and mainly primary hepatocyte cultures, appear to be the most powerful in vitro systems, as liver-specific functions and responsiveness to inducers are retained either for a few days or several weeks depending on culture conditions. Maintenance of phase and phase 11 xenobiotic metabolizing enzyme activities allows various chemical investigations to be performed, including determination of kinetic parameters, metabolic profile, interspecies comparison, inhibition and induction effects, and drug-drug interactions. In vitro liver cell models also have various applications in toxicology: screening of cytotoxic and genotoxic compounds, evaluation of chemoprotective agents, and determination of characteristic liver lesions and associated biochemical mechanisms induced by toxic compounds. Extrapolation of the results to the in vivo situation remains a matter of debate. Presently, the most convincing applications of liver cell models are the studies on different aspects of metabolism and mechanisms of toxicity. For the future, there is a need for better culture conditions and differentiated hepatocyte cell lines to overcome the limited availability of human liver tissues. In addition, strategies for in vitro analysis of potentially toxic chemicals must be better defined. Environ Health Perspect 106(Suppl 2):511-532 (1998). http.//ehpnetl.niehs.nih.gov/docs/1998/Suppl-2/51 1-532guillouzo/abstract.html

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“…Third, and certainly the most original feature, when passaged at low density, they recover characteristic features of progenitor cells able to differentiate in both hepatocytes and biliary epithelial cells (Parent et al, 2004) and able to form a coculture system that strongly resembles that which we have, a long time ago, described by associating primary normal hepatocytes with rat liver epithelial cells and resulting in long-term maintenance of liver-specific functions at high levels . Indeed, several studies have demonstrated that various functions, including phase 1 and phase 2 xenobiotic enzyme activities, were better and longer preserved in cocultures versus pure cultures of human and animal hepatocytes (Begue et al, 1983;Fraslin et al, 1985;Niemann et al, 1991). Such a model should represent a unique tool to distinguish the liver epithelial cell-type target of hepatotoxicants.…”

Section: Aninat Et Almentioning

confidence: 99%

EXPRESSION OF CYTOCHROMES P450, CONJUGATING ENZYMES AND NUCLEAR RECEPTORS IN HUMAN HEPATOMA HepaRG CELLS

Aninat¹,

Piton²,

Glaise³

et al. 2005

Drug Metab Dispos

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584

453

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ABSTRACT:Most human hepatocyte cell lines lack a substantial set of liverspecific functions, especially major cytochrome P450 (P450)-related enzyme activities, making them unrepresentative of in vivo hepatocytes. We have used the HepaRG cells, derived from a human hepatocellular carcinoma, which exhibit a high differentiation pattern after 2 weeks at confluency to determine whether they could mimic human hepatocytes for drug metabolism and toxicity studies. We show that when passaged at low density, these cells reversed to an undifferentiated morphology, actively divided, and, after having reached confluency, formed typical hepatocyte-like colonies surrounded by biliary epithelial-like cells. By contrast, when seeded at high density, hepatocyte-like clusters retained their typical differentiated morphology. Transcripts of various nuclear receptors (aryl hydrocarbon receptor, pregnane X receptor, constitutive androstane receptor, peroxisome proliferator-activated receptor ␣), P450s (CYP1A2, 2C9, 2D6, 2E1, 3A4), phase 2 enzymes (UGT1A1, GSTA1, GSTA4, GSTM1), and other liver-specific functions were estimated by reverse transcriptase-quantitative polymerase chain reaction and were found to be expressed, for most of them, at comparable levels in both confluent differentiated and high-density differentiated HepaRG cells and in cultured primary human hepatocytes. For several transcripts, the levels were strongly increased in the presence of 2% dimethyl sulfoxide. Measurement of basal activities of several P450s and their response to prototypical inducers as well as analysis of metabolic profiles and cytotoxicity of several compounds confirmed the functional resemblance of HepaRG cells to primary cultured human hepatocytes. In conclusion, HepaRG cells constitute the first human hepatoma cell line expressing high levels of the major P450s involved in xenobiotic metabolism and represent a reliable surrogate to human hepatocytes for drug metabolism and toxicity studies.

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Cultured human adult hepatocytes: a new model for drug metabolism studies

Cited by 66 publications

References 12 publications

Long-Term maintenance of taurocholate uptake by adult rat hepatocytes co-cultured with a liver epithelial cell line

Long-Term maintenance of taurocholate uptake by adult rat hepatocytes co-cultured with a liver epithelial cell line

Liver cell models in in vitro toxicology.

EXPRESSION OF CYTOCHROMES P450, CONJUGATING ENZYMES AND NUCLEAR RECEPTORS IN HUMAN HEPATOMA HepaRG CELLS

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