Long‐term maintenance of hepatocyte functional activity in co‐culture: Requirements for sinusoidal endothelial cells and dexamethasone

Morin, O.; Normand, Claire

doi:10.1002/jcp.1041290115

Cited by 131 publications

(68 citation statements)

References 20 publications

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“…These two features could be related because in pure hepatocyte cultures the addition of proteoglycans improves gap junction activity (137). The enhanced survival and function of hepatocytes cocultured with rat liver epithelial cells have been confirmed by many investigators (134,(138)(139)(140)(141)(142)(143); the induction of CYP2B1 and CYP2B2 by phenobarbital has also been demonstrated (137,144). Moreover, other cells have been effective, including both liver endothelial cells and nonhepatic cells (145)(146)(147).…”

Section: However Survival and Metabolic Compe-mentioning

confidence: 89%

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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Section: However Survival and Metabolic Compe-mentioning

confidence: 89%

Liver cell models in in vitro toxicology.

Guillouzo

1998

Environ Health Perspect

297

125

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“…8. Hepatocytes cultured in a collagen double-gel have been shown to exhibit native cell-cell contacts such as E-cadherin and bile canaliculi, 34 but do not express sinusoidal receptors such as EGF-R 34 and LDL-R. Hepatocytes cocultured with 3T3 fibroblasts exhibit hepatic cell-cell contacts such as connexin-32 44 and bile canaliculi, 24 but also do not express the EGF-R and LDL-R. On the other hand, hepatocytes cocultured with LSEC show both traditional polarity markers, 45,46 and express a high level of sinusoidal receptors (EGF-R, LDL-R) at the interface between the hepatocytes and the LSEC. At least part of the interaction between hepatocytes and LSECs has been shown to be mediated by growth factors.…”

Section: Discussionmentioning

confidence: 99%

Liver endothelial cells promote LDL-R expression and the uptake of HCV-like particles in primary rat and human hepatocytes

et al. 2006

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Low-density lipoprotein (LDL) is an important carrier of plasma cholesterol and triglycerides whose concentration is regulated by the liver parenchymal cells. Abnormal LDL regulation is thought to cause atherosclerosis, while viral binding to LDL has been suggested to facilitate hepatitis C infection. Primary hepatocytes quickly lose the ability to clear LDL during in vitro culture. Here we show that the coculture of hepatocytes with liver sinusoidal endothelial cells (LSEC) significantly increases the ability of hepatocytes to uptake LDL in vitro. LDL uptake does not increase when hepatocytes are cocultured with other cell types such as fibroblasts or umbilical vein endothelial cells. We find that LSECs induce the hepatic expression of the LDL receptor and the epidermal growth factor receptor. In addition, while hepatocytes in single culture did not take up hepatitis C virus (HCV)-like particles, the hepatocytes cocultured with LSECs showed a high level of HCV-like particle uptake. We suggest that coculture with LSECs induces the emergence of a sinusoidal surface in primary hepatocytes conducive to the uptake of HCV-like particles. In conclusion, our findings describe a novel model of polarized hepatocytes in vitro that can be used for the study of LDL metabolism and hepatitis C infection. L ow-density lipoprotein (LDL) is a plasma-carried particle whose lipid component includes cholesterol and triglycerides. 1 LDL originates from very low-density lipoprotein (vLDL) synthesized by the liver with apoprotein B-100. The vLDL is converted to LDL by an endothelial lipase, which releases free fatty acids, increasing the density of the particle to form LDL. Excess LDL is then taken up by hepatocytes through LDL receptor (LDL-R)-mediated endocytosis. 1 Improper hepatic clearance of LDL results in elevated plasma levels of LDL which is a serious risk factor for the development of atherosclerosis. 2 One of the first steps in atherosclerosis is the passage of LDL into the vascular wall. Once trapped in the vessel wall, LDL can undergo oxidation. 3 Oxidized LDL is no longer a ligand for the LDL receptor. 4 Accumulation of oxidized LDL and LDL aggregates in the vessel wall stimulates an inflammatory response causing endothelial injury, macrophage recruitment, foam cell formation, and smooth muscle cell proliferation. 3 Current therapeutic intervention includes administration of statins, which block cholesterol production and increase expression of the LDL-R by the liver parenchymal cells, 2 removing LDL from circulation. 5 In addition, liver sinusoidal endothelial cells (LSECs) remove oxidized LDL from the circulation via the scavenger cell receptor. 5 LDL-and LDL-R-mediated pathways have also been shown to play a role in hepatitis C virus (HCV) infection. 6 The HCV surface receptors, glycoproteins E1 and E2, have been shown to associate with LDL and vLDL. 7 The virus particle itself was shown to associate with the

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“…However, the co-culture of mature hepatocytes with non-parenchymal (stromal) cells from the liver itself has been shown to be more effective in modulating cell differentiation (Bader et al 1996;Bhatia et al 1999;Ries et al 2000). Various two and three dimensional co-culture models have demonstrated their ability to mimic the liver microenvironment, suggesting that each hepatic non-parenchymal cell population (sinusoidal endothelium, Kupffer cells and hepatic stellate cells (HSCs)) may contribute to the overall beneficial effect on hepatocytes (Morin and Normand 1986;Okamoto et al 1998;Riccalton-Banks et al 2003; Thomas et al 2005;Zinchenko et al 2006a).…”

Section: Introductionmentioning

confidence: 99%

Maintaining hepatocyte differentiation in vitro through co-culture with hepatic stellate cells

Krause

Saghatolislam

Koenig

et al. 2009

In Vitro Cell.Dev.Biol.-Animal

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Primary hepatocytes lose their differentiated functions rapidly when in culture. Our aim was to maintain the differentiated status of hepatocytes in vitro by means of vital hepatic stellate cells (HSCs), their soluble and particulate factors and lipid extracts. Hepatocytes were placed into collagen-coated culture dishes in the presence of HSCs at different ages of pre-culture, with or without direct cell to cell contacts, at different cell ratios and in monoculture with cellular HSC components in place of vital cells. Changes in morphology and enhancement of phosphoenolpyruvate carboxykinase (PCK) activity by glucagon were used to determine the differentiated status of hepatocytes in 2d-short-term culture. HSCs proved able to maintain the differentiated function of hepatocytes in coculture either by direct cell contacts or via factors derived from HSC-conditioned medium. In comparison, however, without cellular contact to hepatocytes five to ten times as many HSCs were necessary to increase the PCK activity to the same degree as in the presence of intercellular contacts. Whereas stimulation in the presence of HSC/hepatocyte contacts was independent of HSC culture age only quiescent, resting HSCs (precultured for 1-2 d) were able to stimulate hepatocytes significantly via soluble factors. Culturing of hepatocytes with a lipid extract or a particulate fraction from HSCs clearly displayed a very strong beneficial effect on enzyme activity and morphology. HSCs maintain hepatocyte function and structure through preferentially cell-bound signalling and transfer of lipids.

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Long‐term maintenance of hepatocyte functional activity in co‐culture: Requirements for sinusoidal endothelial cells and dexamethasone

Cited by 131 publications

References 20 publications

Liver cell models in in vitro toxicology.

Liver cell models in in vitro toxicology.

Liver endothelial cells promote LDL-R expression and the uptake of HCV-like particles in primary rat and human hepatocytes

Maintaining hepatocyte differentiation in vitro through co-culture with hepatic stellate cells

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