Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system

Nakamura, Kazuaki; Mizutani, Reiko; Sanbe, Atsushi; Enosawa, Shin; Kasahara, Mureo; Nakagawa, Atsuko; Ejiri, Yoko; Murayama, Norie; Miyamoto, Yuki; Torii, Tomohiro; Kusakawa, Shinji; Yamauchi, Junji; Fukuda, Motohiro; Yamazaki, Hiroshi; Tanoue, Akito

doi:10.1016/j.jbiosc.2010.08.008

Cited by 61 publications

(48 citation statements)

References 33 publications

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“…Several examples confirm the relevance of 3D culture models to improve the structure and the prediction rate, not only for toxicity but also in screening for pharmacological assays (Dash et al, 2009;Lan and Starly, 2011;Meng, 2010;Nakamura et al, 2011;Toh et al, 2009). Similarly, co-culture methods will give a better idea of the relevance of interaction and crosstalk between the different cell types.…”

Section: Culture Methodsmentioning

confidence: 84%

A roadmap for the development of alternative (non-animal) methods for systemic toxicity testing

Basketter

Clewell

Kimber

et al. 2012

ALTEX

203

165

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Section: Culture Methodsmentioning

confidence: 84%

A roadmap for the development of alternative (non-animal) methods for systemic toxicity testing

Basketter

Clewell

Kimber

et al. 2012

ALTEX

203

165

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“…Recently, Nakamura et al (16) demonstrated that the 3D-culture method enhances hepatocyte-specific functions of hepatocytes, including drug-metabolizing enzyme activities in HepG2 cells. They also identified higher sensitivity to acetaminophen cytotoxicity in 3D-cultured HepG2 cells compared with conventional monolayer cultured HepG2 cells and suggested that this 3D-cultured HepG2 cell system may be useful as a human model of APAPinduced hepatotoxicity.…”

Section: Discussionmentioning

confidence: 99%

“…Recent reports have demonstrated that the micro-space cell culture plate method, a three-dimensional (3D) culture system, can induce hepatocyte-specific functions, including CYP2E1 expression, in HepG2 cells, which results in an increase in cytotoxicity by acetaminophen compared with a conventional two-dimensional (2D) culture system (16). These results suggest that 3D-cultured HepG2 cells may be a useful tool as an in vitro human model of APAP-induced hepatotoxicity.…”

mentioning

confidence: 96%

Evaluation of Three-Dimensional Cultured HepG2 Cells in a Nano Culture Plate System: an In Vitro Human Model of Acetaminophen Hepatotoxicity

et al. 2014

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“…HepG2 cells were maintained at 37 C under an atmosphere of 5% CO 2 in complete RPMI 1640 medium containing 10% fetal bovine serum and cultured in 10 5 cells/well. 16 APAP (5 mM), BOC-1 (10 À5 M; MP Biomedicals LLC, Solon, OH), and DF2156a (10 À6 M; Dompe Pharmaceutical, Milan, Italy) were dissolved in dimethyl sulfoxide, then in culture medium, and then incubated throughout the experiments. Cell viability was assessed by tetrazolium (Sigma-Aldrich) metabolism assay or by the trypan blue exclusion test (in neutrophil viability tests).…”

Section: Methodsmentioning

confidence: 99%

Chemokines and mitochondrial products activate neutrophils to amplify organ injury during mouse acute liver failure

Marques¹,

Amaral²,

Pires³

et al. 2012

Hepatology

285

269

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Acetaminophen (APAP) is a safe analgesic and antipyretic drug. However, APAP overdose leads to massive hepatocyte death. Cell death during APAP toxicity occurs by oncotic necrosis, in which the release of intracellular contents can elicit a reactive inflammatory response. We have previously demonstrated that an intravascular gradient of chemokines and mitochondria-derived formyl peptides collaborate to guide neutrophils to sites of liver necrosis by CXC chemokine receptor 2 (CXCR2) and formyl peptide receptor 1 (FPR1), respectively. Here, we investigated the role of CXCR2 chemokines and mitochondrial products during APAP-induced liver injury and in liver neutrophil influx and hepatotoxicity. During APAP overdose, neutrophils accumulated into the liver, and blockage of neutrophil infiltration by anti-granulocyte receptor 1 depletion or combined CXCR2-FPR1 antagonism significantly prevented hepatotoxicity. In agreement with our in vivo data, isolated human neutrophils were cytotoxic to HepG2 cells when cocultured, and the mechanism of neutrophil killing was dependent on direct contact with HepG2 cells and the CXCR2-FPR1-signaling pathway. Also, in mice and humans, serum levels of both mitochondrial DNA (mitDNA) and CXCR2 chemokines were higher during acute liver injury, suggesting that necrosis products may reach remote organs through the circulation, leading to a systemic inflammatory response. Accordingly, APAP-treated mice exhibited marked systemic inflammation and lung injury, which was prevented by CXCR2-FPR1 blockage and Toll-like receptor 9 (TLR9) absence (TLR9 2/2 mice). Conclusion: Chemokines and mitochondrial products (e.g., formyl peptides and mitDNA) collaborate in neutrophil-mediated injury and systemic inflammation during acute liver failure. Hepatocyte death is amplified by liver neutrophil infiltration, and the release of necrotic products into the circulation may trigger a systemic inflammatory response and remote lung injury.

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Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system

Cited by 61 publications

References 33 publications

A roadmap for the development of alternative (non-animal) methods for systemic toxicity testing

A roadmap for the development of alternative (non-animal) methods for systemic toxicity testing

Evaluation of Three-Dimensional Cultured HepG2 Cells in a Nano Culture Plate System: an In Vitro Human Model of Acetaminophen Hepatotoxicity

Chemokines and mitochondrial products activate neutrophils to amplify organ injury during mouse acute liver failure

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