Modular bioreactor for primary human hepatocyte culture: Medium flow stimulates expression and activity of detoxification genes

Vinci, Bruna; Duret, Cédric; Klieber, Sylvie; Gerbal‐Chaloin, Sabine; Cunha, Antonio Sa; Laporte, Sylvain; Suc, B.; Maurel, Patrick; Ahluwalia, Arti; Daujat-Chavanieu, Martine

doi:10.1002/biot.201000326

Cited by 97 publications

(79 citation statements)

References 45 publications

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“…These velocities are consistent with those typically reported for micro-fluidic cell culture devices [42]. For the MCmB model, the v in was set to 3.82 × 10 −3 m/s (corresponding to 180 μL/min inflow), on the basis of previous experiments with this bioreactor [31,[62][63][64][65][66]. Table 4.…”

Section: Model Implementationsupporting

confidence: 64%

Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors

2014

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Abstract:In this paper, we discuss the basic design requirements for the development of physiologically meaningful in vitro systems comprising cells, scaffolds and bioreactors, through a bottom up approach. Very simple micro-and milli-fluidic geometries are first used to illustrate the concepts, followed by a real device case-study. At each step, the fluidic and mass transport parameters in biological tissue design are considered, starting from basic questions such as the minimum number of cells and cell density required to represent a physiological system and the conditions necessary to ensure an adequate nutrient supply to tissues. At the next level, we consider the use of three-dimensional scaffolds, which are employed both for regenerative medicine applications and for the study of cells in environments which better recapitulate the physiological milieu. Here, the driving need is the rate of oxygen supply which must be maintained at an appropriate level to ensure cell viability throughout the thickness of a scaffold. Scaffold and bioreactor design are both critical in defining the oxygen profile in a cell construct and are considered together. We also discuss the oxygen-shear stress trade-off by considering the levels of mechanical stress required for hepatocytes, which are the limiting cell type in a multi-organ model. Similar considerations are also made for glucose consumption in cell constructs. Finally, the allometric approach for generating multi-tissue systemic models using bioreactors is described.

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Section: Model Implementationsupporting

confidence: 64%

Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors

2014

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“…It is noteworthy that we performed the minimum modification necessary to archive the hepatic tissue-like structure. Many ingenious attempts have been made to mimic the in vivo liver architecture and developed improved systems for evaluating hepatotoxicity, e.g., micropatterning (El-Ali et al, 2006;Khetani and Bhatia, 2008), shear stress (Vinci et al, 2011), P450-inducers such as phenobarbital, and others. Incorporation of these technologies will contribute to the further improvement of our strategy.…”

Section: Discussionmentioning

confidence: 99%

Acetaminophen-Induced Hepatotoxicity in a Liver Tissue Model Consisting of Primary Hepatocytes Assembling around an Endothelial Cell Network

Toyoda¹,

Tamai²,

Kashikura³

et al. 2011

Drug Metab Dispos

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ABSTRACT:Primary hepatocytes have been used in drug development for the evaluation of hepatotoxicity of candidate compounds. However, the rapid depression of their hepatic characters in vitro must be improved to predict toxicity with higher accuracy. We have hypothesized that a well organized tissue construct that includes nonparenchymal cells and appropriate scaffold material(s) could overcome this difficulty by remediating the viability and physiological function of primary hepatocytes. In this study, we constructed an in vitro liver tissue model, consisting of mouse primary hepatocytes assembling around an endothelial cell network on Engelbreth-Holm-Swarm gel, and examined its response to acetaminophen treatment. The increase in lactate dehydrogenase release after the exposure to acetaminophen was induced earlier in the liver tissue model than in monolayer hepatocytes alone, suggesting that the tissue model was more sensitive to an acetaminopheninduced toxicity. On the basis of our results, we conclude that liver tissue models of this kind may enhance the responses of hepatocytes against xenobiotics via the maintenance of hepatic genes and functions such as cytochrome P450s. These findings will contribute to the development of more accurate systems for evaluating hepatotoxicity.

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“…CYP3A4 expression in PHHs might therefore be regulated through other mechanisms. Indeed, CYP3A4 mRNA expression is upregulated by the medium flow in PHHs (Vinci et al, 2011), and the oxygen tension in rat hepatocytes (Allen and Bhatia, 2003). Alternatively, Cyp3a localization could also be regulated in a RAS/MAPK/ERK Braeuning et al, 2007), Dicer- (Sekine et al, 2009) or morphogen-dependent manners (Gebhardt and Hovhannisyan, 2010).…”

Section: B-catenin Regulates Drug Metabolismmentioning

confidence: 99%

The WNT/β-Catenin Pathway Is a Transcriptional Regulator ofCYP2E1,CYP1A2,and Aryl Hydrocarbon Receptor Gene Expression in Primary Human Hepatocytes

et al. 2014

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The wingless-type MMTV integration site family (WNT)/b-catenin/ adenomatous polyposis coli (CTNNB1/APC) pathway has been identified as a regulator of drug-metabolizing enzymes in the rodent liver. Conversely, little is known about the role of this pathway in drug metabolism regulation in human liver. Primary human hepatocytes (PHHs), which are the most physiologically relevant culture system to study drug metabolism in vitro, were used to investigate this issue. This study assessed the link between cytochrome P450 expression and WNT/b-catenin pathway activity in PHHs by modulating its activity with recombinant mouse Wnt3a (the canonical activator), inhibitors of glycogen synthase kinase 3b, and small-interfering RNA to invalidate CTNNB1 or its repressor APC, used separately or in combination. We found that the WNT/ b-catenin pathway can be activated in PHHs, as assessed by universal b-catenin target gene expression, leucine-rich repeat containing G protein-coupled receptor 5. Moreover, WNT/ b-catenin pathway activation induces the expression of CYP2E1, CYP1A2, and aryl hydrocarbon receptor, but not of CYP3A4, hepatocyte nuclear factor-4a, or pregnane X receptor (PXR) in PHHs. Specifically, we show for the first time that CYP2E1 is transcriptionally regulated by the WNT/b-catenin pathway. Moreover, CYP2E1 induction was accompanied by an increase in its metabolic activity, as indicated by the increased production of 6-OH-chlorzoxazone and by glutathione depletion after incubation with high doses of acetaminophen. In conclusion, the WNT/b-catenin pathway is functional in PHHs, and its induction in PHHs represents a powerful tool to evaluate the hepatotoxicity of drugs that are metabolized by CYP2E1.

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Modular bioreactor for primary human hepatocyte culture: Medium flow stimulates expression and activity of detoxification genes

Cited by 97 publications

References 45 publications

Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors

Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors

Acetaminophen-Induced Hepatotoxicity in a Liver Tissue Model Consisting of Primary Hepatocytes Assembling around an Endothelial Cell Network

The WNT/β-Catenin Pathway Is a Transcriptional Regulator ofCYP2E1,CYP1A2,and Aryl Hydrocarbon Receptor Gene Expression in Primary Human Hepatocytes

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