| Project/Area Number |
13555222
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
生物・生体工学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SAKAI Yasuyuki Institute of Industrial Science, University of Tokyo, Associate Professor, 生産技術研究所, 助教授 (00235128)
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| Co-Investigator(Kenkyū-buntansha) |
TANIGUTCHI Yoshitaka Department of Biological Sciences, Toray Research Center, Researcher, 生物科学研究部, 研究員
SAKODA Akiyoshi Institute of Industrial Science, University of Tokyo, Professor, 生産技術研究所, 教授 (30170658)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥13,200,000 (Direct Cost: ¥13,200,000)
Fiscal Year 2003: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2001: ¥8,500,000 (Direct Cost: ¥8,500,000)
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| Keywords | Perfusion cocultivation / absorption and biotransformation / shim ulator / Benzolalovrene / PBPK simulation model / umu test / Microtox / 小腸膜 / 肝組織 / チトクロームP450 / 生理学的薬物・毒物動力学モデル / 極性輸送 / 代謝活性化 / バイオハイブリッド / 人体代謝シュミレータ / 小腸 / 肝臓 / 灌流培養 / 臓器間相互作用 / 薬物代謝酵素 / ベンゾ[a]ピレン |
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| Research Abstract |
Conventional cytotoxicity tests cannot usually include various metabolic processes in humans. We therefore developed a physiologically-based multi-compartment perfusion coculture system (bio-hybrid simulator) using a Caco-2 cell monolayer on a semipermeable membrane and microcarrier-based three-dimensional culture of Hep G2 cells to mimic the absorption across the small intestine and biotransformation of the small intestine and the liver. Stable operations enabled to maintain various activities of the both cells for at least four days.
Cocultivation improved the growth of Hep G2 cells and enhanced the cytochrome P450 lAl/2 capacities of both cell lines. When benzo[a]pyrene (BaP) was, loaded to the apical side of the Caco-2 cell layer, the enhanced P450 capacities produced a larger amount of BaP-7,8-hydrodiol, an immediate precursor to the highly-reactive ultimate toxicant of BaP, BaP-7,8-dihydrodiol-9,l0-epoxide (BPDE). This led to initially retarded and later stronger expression of BaP toxicity in the coculture system than in pure culture, which agreed well with the largest time integral of the concentration (area under curve, AUC) of BaP-7,8-hydrodiol in the Hep G2 cells compartment of the coculture system.
Because this kind of system can reproduce such complicated phenomena including those derived from Organ-to-organ interactions, it is useful as a new in vitro experimental system when we understand unknown mechanisms involved in final toxicity in humans and develop physiologically-based pharmacokinetic (PBPK) numerical simulation models.
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