Secretion of Albumin and Induction of CYP1A2 and CYP3A4 in Novel Three-dimensional Culture System for Human Hepatocytes using Micro-space Plate

Nishimura, Masuhiro; Hagi, Mieko; Ejiri, Yoko; Kishimoto, Sanae; Horie, Tetsuhiro; Narimatsu, Shizuo; Naito, Shinsaku

doi:10.2133/dmpk.25.236

Cited by 19 publications

(21 citation statements)

References 21 publications

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“…In the present study, 0.1% DMSO was used to dissolve the inducers, and hepatocytes were exposed to the inducers for 24 hr; it had previously been reported that 0.1% DMSO had no effect on induction (Nishimura et al, 2010). In addition, the induction of drug-metabolizing enzyme mRNAs could be evaluated after 24 hr of exposure to these inducers (Nishimura et al, 2010).…”

Section: Induction Of Cyps Mrna Expression In Hepg2 Cells By Rifampicmentioning

confidence: 97%

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Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

Nakamura¹,

Kato²,

Aizawa³

et al. 2011

J. Toxicol. Sci.

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-The Nanoculture plate (NCP) is a recently developed plate which essentially consists of a textured surface with specific characteristics that induce spheroid formation: microfabrications with a micro-square pattern on the culture surface. The NCP can be used to generate uniform adhesive spheroids of cancer cell lines using conventional techniques without the need of any animal compounds. In this study, we assessed the performance of human hepatoma cell line HepG2 cells cultured with an NCP to evaluate the effects of the NCP on their hepatocyte-specific functions. The NCP facilitated the formation of three-dimensional (3D) HepG2 cell architecture. HepG2 cells cultured with an NCP exhibited enhanced mRNA expression levels of albumin and cytochrome P450 (CYP) enzymes compared to those cultured with a two-dimensional (2D) conventional plate. The expression levels of two specific liver-enriched transcription factors, hepatocyte nuclear factor 4α (HNF4α) and CCAAT/enhancer binding protein α (C/EBPα), were higher in HepG2 cells grown with the NCP than those in HepG2 cells grown with conventional plates before albumin and CYP enzymes expression levels were increased. The inducibility of CYP1A2 and CYP3A4 mRNA following exposure to inducers in HepG2 cells cultured with an NCP was comparable to that in HepG2 cells cultured with conventional plates, while the expression levels of CYP1A2 and CYP3A4 mRNA following exposure to inducers were higher when using an NCP than when using conventional plates. These results suggest that the use of an NCP enhances the hepatocyte-specific functions of HepG2 cells, such as drug-metabolizing enzyme expression, making the NCP/ HepG2 system a useful tool for evaluating drug metabolism in vitro.

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Section: Induction Of Cyps Mrna Expression In Hepg2 Cells By Rifampicmentioning

confidence: 97%

“…In addition, the induction of drug-metabolizing enzyme mRNAs could be evaluated after 24 hr of exposure to these inducers (Nishimura et al, 2010). The changes in CYP1A2 mRNA levels induced by exposure to Ome in HepG2 cultured with monolayer and NCP systems are shown in Fig.…”

Section: Induction Of Cyps Mrna Expression In Hepg2 Cells By Rifampicmentioning

confidence: 99%

Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

Nakamura¹,

Kato²,

Aizawa³

et al. 2011

J. Toxicol. Sci.

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“…In the three-dimensional culture of hepatocytes, freshly isolated hepatocytes in co-culture with activated stellate cells rapidly aggregate to form well-defined viable spheroids; the spheroids have a complex extracellular matrix support and hepatic ultrastructure including bile canaliculi, tight junctions, desmosomes, and lipid storage [7,8]. Therefore, the metabolism by this system is expected to be relatively similar to that by the intact liver in living mammals.…”

Section: Discussionmentioning

confidence: 99%

“…In recent years, the techniques for three-dimensional cell culture have been used in various fields. According to the published data, much higher mRNA expression for albumin and CYP450 enzymes could be obtained by three-dimensional hepatocyte culture than by the traditional monolayer hepatocyte culture, and thus much increased inducibility of CYP functions can be expected [7,8], suggesting that drug metabolite profiles can be precisely investigated by the three-dimensional hepatocyte culture. TESTLIVER TM (Toyobo, Osaka, Japan) [9] is a bundle of hollow fibers filled with hepatocytes that enables simple three-dimensional hepatocyte culture (Fig.…”

Section: Introductionmentioning

confidence: 99%

A model system for prediction of the in vivo metabolism of designer drugs using three-dimensional culture of rat and human hepatocytes

et al. 2011

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The in vitro metabolisms of 4-bromo-2,5-dimethoxyphenethylamine (2C-B), 5-methoxy-N,N-diisopropyltryptamine (5-MeO-DIPT), and 2,5-dimethoxy-4-propylthiophenethylamine (2C-T-7) were studied using TESTLIVER TM -rat and TESTLIVER TM -human, which are new three-dimensional rat and human hepatocyte culture systems, respectively. The metabolites produced in the incubation media were measured by liquid chromatographytandem mass spectrometry or gas chromatography-mass spectrometry. The data obtained with the in vitro system of rat origin were carefully compared with those obtained by rat in vivo experiments; most of the metabolites found in the in vivo experiments could be reproduced in the present threedimensional in vitro experiments, although quantitative metabolite distribution patterns obtained with the threedimensional system was somewhat different from those of in vivo experiments. Because human in vivo experiments are not possible, especially for dubious designer drugs, the in vitro experiments using the three-dimensional human hepatocyte culture system seem very useful for studying human metabolism of new designer drugs as an alternative to human experiments.Keywords Three-dimensional culture Á Hepatocyte Á Metabolism Á 2C-B Á 5-MeO-DIPT Á 2C-T-7

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“…132,133 Parallelization and miniaturization of cell-based assays is now common practice in two dimensions but is also becoming more feasible in 3D. Indeed, microspace plates have been used to establish three-dimensional cultures of primary hepatocytes, 134 agarose-coated 384-well plates have been used to perform high-throughput compound screens on spheroids from primary colon cancer cells, 135 and even a perfusion-based microfluidic chip has been conceived for parallelized 3D culture of primary chondrocytes. 136 Compatibility with LSFM will require alternative well plate geometries as well as a convenient interface for rapid exchange of samples.…”

Section: Future Space Microscopymentioning

confidence: 99%

Invited Review Article: Advanced light microscopy for biological space research

Vos

Beghuin²,

Schwarz

et al. 2014

Review of Scientific Instruments

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As commercial space flights have become feasible and long-term extraterrestrial missions are planned, it is imperative that the impact of space travel and the space environment on human physiology be thoroughly characterized. Scrutinizing the effects of potentially detrimental factors such as ionizing radiation and microgravity at the cellular and tissue level demands adequate visualization technology. Advanced light microscopy (ALM) is the leading tool for non-destructive structural and functional investigation of static as well as dynamic biological systems. In recent years, technological developments and advances in photochemistry and genetic engineering have boosted all aspects of resolution, readout and throughput, rendering ALM ideally suited for biological space research. While various microscopy-based studies have addressed cellular response to space-related environmental stressors, biological endpoints have typically been determined only after the mission, leaving an experimental gap that is prone to bias results. An on-board, real-time microscopical monitoring device can bridge this gap. Breadboards and even fully operational microscope setups have been conceived, but they need to be rendered more compact and versatile. Most importantly, they must allow addressing the impact of gravity, or the lack thereof, on physiologically relevant biological systems in space and in ground-based simulations. In order to delineate the essential functionalities for such a system, we have reviewed the pending questions in space science, the relevant biological model systems, and the state-of-the art in ALM. Based on a rigorous trade-off, in which we recognize the relevance of multi-cellular systems and the cellular microenvironment, we propose a compact, but flexible concept for space-related cell biological research that is based on light sheet microscopy.

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Secretion of Albumin and Induction of CYP1A2 and CYP3A4 in Novel Three-dimensional Culture System for Human Hepatocytes using Micro-space Plate

Cited by 19 publications

References 21 publications

Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

Expression of albumin and cytochrome P450 enzymes in HepG2 cells cultured with a nanotechnology-based culture plate with microfabricated scaffold

A model system for prediction of the in vivo metabolism of designer drugs using three-dimensional culture of rat and human hepatocytes

Invited Review Article: Advanced light microscopy for biological space research

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