• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development of a multi-compartment cell culture system for quantitative evaluation of chemical impacts to humans

Research Project

Project/Area Number 11450310
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field 生物・生体工学
Research InstitutionUniversity of Tokyo

Principal Investigator

SAKAI Yasuyuki  Institute of Industrial Science, University of Tokyo, Lecturer, 生産技術研究所, 講師 (00235128)

Co-Investigator(Kenkyū-buntansha) SAKODA Akiyoshi  Institute of Industrial Science, University of Tokyo, Professor, 生産技術研究所, 教授 (30170658)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 2000: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1999: ¥6,200,000 (Direct Cost: ¥6,200,000)
Keywordsmodel small intestine / model liver tissue / multi-compartment cell culture system / bioactivation / active transport / static culture / perfusion culture / oral intake / 毒性発現 / 小腸上皮 / 肝細胞 / 吸収 / 代謝 / アセトアミノフェン
Research Abstract

The final goal of this research is to develop a human body simulator that mimics various responses to added chemicals by combining compartments havingorgan-derived cells in a physiologically-relevant manner.
First, we developed a simple double-layered culture system consisting of a model small intestine (cultured Caco-2 cell layer) and a model target organs (cultured human diploid fibroblasts, TIG-1 cells). All ED50 values for four model chemicals in the double-layered system changed into higher concentration ranges compared those obtained in a single-layered culture system (without the Caco-2 cell layer), according to the in vitro absorbability of the model chemicals. This resulted in enhancement of predictivity of in vivo toxicity because in vivo absorbability is considered and affects the final toxicity in the double-layered culture system. An interesting phenomenon was that detoxification and active transport mechanisms were postulated for some chemicals in the Caco-2 cell layer (AA … More TEX, published).
Second, in the similar double-layered culture system using human hepatoma, Hep G2 cells, instead of the TIG-1 cells, we focused on involvement of detoxification and active transport of the Caco-2 cell layer in the toxicity expression mechanisms of benzo[a]pyrene used as a model chemical that exhibits strong toxicity through bioactivation by cytochrome P450 enzymes in humans. Quantification of benzo[a]pyrene metabolites (some of them are procarcinogens) showed that only less than one-tenth amount could permeate the Caco-2 membrane. P450 1A1/2 was strongly induced not only in the Hep G2 cells but also in the Caco-2 cells. These observations indicate that such a double-layered culture system is advantageous over conventional single-population-based cytotoxicity tests because it can closely mimic very complex toxicity expression mechanism occurring in in vivo humans.
Although such simple culture systems are effective in screening or ranking of chemical toxicities in vivo, they are not suitable for quantitative or kinetic analyses of toxicity expressions in humans, because of their very low cell density. Therefore, we developed a perfusion culture system consisting of Caco-2 cells and Hep G2 cells combined with a physiologically-relevant circuit. In a series of experiments using acetaminophen as a model chemical that is well absorbed across the small intestine, biologically-activated in the liver, and expresses specific toxicity in the liver. Unexpectedly, observed toxicity was higher in the Caco-2 cell-containing system than in the Caco-2-cell-free system. Measurement of cytochrome P450 3A that is responsible for acetaminophen toxicity in humans showed the very high enzymatic activity in the Caco-2 cells. We therefore concluded that acetaminophen was transformed into more toxic metabolites when it permeates across the Caco-2 cell layer.
To improve duration of culture system, we are developing a new perfusion culture system in which cells are continuously shaken to meet oxygen consumption of the cells in the system. Less

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Y.Sakai,T.Arai,A.Sakoda and M.Suzuki: "Development of a simple double-layered cell culture system using Caco-2 and TIG-1 cells as a new cytotoxicity test"AATEX. 7(2-3). 47-58 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 酒井康行: "複合動物細胞培養,動物細胞工学ハンドブック,動物細胞工学会 編"朝倉書店(東京). 216-217 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Sakai, T.Arai, A.Sakoda, and M.Suzuki: "Development of a simple double-layered culture system using Caco-2 and TIG-1 cells as a new cytotoxicity test"AATEX. 7(2-3). 47-58 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Sakai: "Multi-compartment mammalian cell culture (Japanese)"Handbook for animal cell culture engineering Etd. by Japanese Association of Animal Cell Culture. 216-217 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Sakai,T.Arai,A.Sakoda and M.Suzuki: "Development of a simple double-layered cell culture system using Caco-2 and TIG-1 cells as a new cytotoxicity test"AATEX. 7(2-3). 47-58 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 酒井康行: "複合動物細胞培養,動物細胞工学ハンドブック,動物細胞工学会編"朝倉書店(東京). pp.216-217 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 酒井康行: "複合動物細胞培養"動物細胞工学ハンドブック. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] 酒井康行: "細胞から組織へ"化学工学. 63・4. 214-215 (1999)

    • Related Report
      1999 Annual Research Report

URL: 

Published: 1999-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi