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Development of novel co-culture system of parenchymal cells and nonparenchymal cells on thermoresponsive polymer-grafted dish surfaces

Research Project

Project/Area Number 13308055
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Biomedical engineering/Biological material science
Research InstitutionTokyo Women's Medical University

Principal Investigator

OKANO Teruo  Tokyo Women's Medical University, Department of Medicine, Professor, 医学部, 教授 (00130237)

Co-Investigator(Kenkyū-buntansha) YAMATO Masayuki  Tokyo Women's Medical University, Department of Medicine, Associate Professor, 医学部, 助教授 (40267117)
KIKUCHI Akihiko  Tokyo Women's Medical University, Department of Medicine, Associate Professor, 医学部, 助教授 (40266820)
YOKOYAMA Masayuki  Tokyo Women's Medical University, Department of Medicine, Associate Professor, 医学部, 助教授 (20220577)
AKIYAMA Yoshikatsu  Tokyo Women's Medical University, Department of Medicine, Research Assistant Professor, 医学部, 助手 (20349640)
SHIMIZU Tatsuya  Tokyo Women's Medical University, Department of Medicine, Assistant Professor, 医学部, 講師 (40318100)
KUSHIDA Ai  Tokyo Women's Medical University, Department of Medicine, Research Assistant Professor (10338981)
青柳 隆夫  東京女子医科大学, 医学部, 助教授 (40277132)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥53,040,000 (Direct Cost: ¥40,800,000、Indirect Cost: ¥12,240,000)
Fiscal Year 2003: ¥12,350,000 (Direct Cost: ¥9,500,000、Indirect Cost: ¥2,850,000)
Fiscal Year 2002: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Fiscal Year 2001: ¥27,300,000 (Direct Cost: ¥21,000,000、Indirect Cost: ¥6,300,000)
Keywordspoly(N-isopropylacrylamide) / patterned thermoresponsive surfaces / pattern co-culture / parenchymal cells / endothelial cells / albumin production / urea synthesis / patterned domain sizes / 温度応答性表面 / 細胞シートのマニピュレーション / 相転移挙動
Research Abstract

This research focused on the design and characterization of novel patterned dual thermoresponsive surfaces for possible patterned co-culture and recovery of co-cultured cell monolayers by temperature modulation. Thermoresponsive polymers of poly(N-isopropylacrylamide) and its hydrophobic derivatives containing n-butyl methacrylate (BMA) as hydrophobic units were pattern grafted using electron beam irradiated polymerization. In this fiscal year, several types of the patterned dual thermoresponsive surfaces were prepared by applying limited electron beam irradiated polymerization of BMA onto PIPAAm-grafted surfaces through aluminum masks having different sizes of circular holes. In case of the 100μmφ-patterned masks, irradiated electron beam was diffracted with the mask and meniscus of the monomer solution, resulting in the formation of hydrophobic patterns with >100μmφ in size, and we did not obtain patterned dual thermoresponsive surfaces reproducibly. Using metal masks with larger pat … More tern sizes (0.5mmφ and 1.0mmφ), we successfully prepared patterned, dual thermoresponsive surfaces. Patterned co-culture of hepatocytes (HCs) and endothelial cells (ECs) were then performed by modulation of surface hydrophilic/hydrophobic property alteration with temperature. HCs were seeded and cultured at 27℃ to culture only onto relatively hydrophobic domains, followed by ECs culture at 37℃ to realize patterned co-culture of these two cell types. By lowering culture temperature to 20℃ at which entire surfaces become hydrophilic, co-cultured cells spontaneously detach from the culture surfaces in a single cell sheet maintaining patterned structure. We then investigate the effect of the pattern sizes on hepatic function changes in patterned co-culture systems, in terms of the albumin production and urea syntheses by ammonium metabolism. Both physiological functions increased with decreasing patterned domain sizes at constant HCs/ECs ratio than homotypic culture of HCs. More functional augmentation was observed with smaller patterned domains with longer culture periods. These results are consistent with the previously obtained results, in which HC monolayers were covered with EC monolayers obtained by using PIPAAm-grafted dishes. Thus, the control of the distance between HCs and ECs has preferential influences to co-cultured cell functionalities including cellular viabilities. Less

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] OH Kwon, A Kikuchi, M Yamato, T Kano: "Accerelated cell sheet recovery by co-grafting of PEG with PIPAAm onto porous cell culture membranes"Biomaterials. 24(7). 1223-1232 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M Ebara, M Yamato, M Hirose, T Aoyagi, A Kikuchi, K Sakai, T Okano: "Copolymerization of 2-carbozyisoppopylacrylamide with N-isopropylacrylamide accerelates cell detachment from grafted surfaces by reducing temperature"Biomacromolecules. 4. 334-349 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y Shiroyanagi, M Yamato, Y Yamazaki, H Toma, T Okano: "Transplantable urothelial cell sheets harvested noninvasively from temperature-responsive culture surfaces by reducing temperature"Tissue Engineering. 9(5). 1005-1012 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M Yamato, C Konno, S Koike, Y Isoi, T Shimizu, A Kikuchi, K Makino, T Okano: "Nanofabrication for microoatterned cell arrays by combining electron beam irradiated polymer-grafting and localized laser ablations"Journal of Biomedical Materials Research. 67A. 1065-1071 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M Harimoto, M Yamato, A Kikuchi, T Okano: "Cell sheet engineering : Intelligent polymer patterned surfaces for tissue engineered liver"Macromolecular Symposia. 195. 231-235 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y Tsuda, A Kikuchi, M Yamato, Y Sakurai, M Umezu, T Okano: "Control of cell adhesion and detachment using temperature and thermoresponsive copolymer grafted culture surfaces"Journal of Biomedical Materials Research. 69A. 70-78 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] OH Kwon, A.Kikuchi, M.Yamato, T.Okano: "Accelerated cell sheet recovery by co-grafting of PEG with PIPAAm onto porous cell culture membranes"Biomaterials. 24(7). 1223-1232 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Ebara, M.Yamato, M.Hirose, T.Aoyagi, A.Kikuchi, K.Sakai, T.Okano: "Copolymerization of 2-carboxyisopropylacrylamide with N-isopropylacrylamide accelerate cell detachment from grafted surfaces by reducing temperature"Biomacromolecules. 4. 334-349 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Shiroyanagi, M.Yamato, Y.Yamazaki, H, Toma, Teruo Okano: "Transplantable urothelial cell sheets harvested noninvasively from temperature-responsive culture surfaces by reducing temperature"Tissue Engineering. 9(5). 1005-1012 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Yamato, C.Konno, S.Koike, Y.Isoi, T.Shimizu, A.Kikuchi, K.Makino, Teruo Okano: "Nanofabrication for micropatterned cell arrays by combining electron beam irradiated polymer-grafting and localized laser ablations"Journal of Biomedical Materials Research. 67A. 1065-1071 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Harimoto, M.Yamato, A.Kikuchi, Teruo Okano: "Cell sheet engineering : Intelligent polymer patterned surfaces for tissue engineered liver"Macromolecular Symposia. 195. 231-235 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Tsuda, A.Kikuchi, M.Yamato, Y.Sakurai, M.Umezu, T.Okano: "Control of cell adhesion and detachment using temperature and thermoresponsive copolymer grafted culture surfaces"Journal of Biomedical Materials Research. 69A. 70-78 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] OH Kwon, A Kikuchi, M Yamato, T Okano: "Accelerated cell sheet recovery by co-grafting of PEG with PIPAAm onto porous cell culture membranes"Biomaterials. 24(7). 1223-1232 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M Ebara, M Yamato, M Hirose, T Aoyagi, A Kikuchi, K Sakai, T Okano: "Copolymerization of 2-carbozyisopropylacrylamide with N-isopropylaclrylamide accelerates cell detachment from grafted surfaces by reducing temperature"Biomacromolecules. 4. 334-349 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Y Shiroyanagi, M Yamato, Y Yamazaki, H Toma, T Okano: "Transplantable urothelial cell sheets harvested noninvasively from temperature-responsive culture surfaces by reducing temperature"Tissue Engineering. 9(5). 1005-1012 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M Yamato, C Konno, S Koike, Y Isoi, T Shimizu, A Kikuchi, K Makino, T Okano: "Nanofabrication for micropatterned cell arrays by combining electron beam irradiated polymer-grafting and localized laser ablations"Journal of Biomedical Materials Research. 67A. 1065-1071 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] M Harimoto, M Yamato, A Kikuchi, T Okano: "Cell sheet engineering : Intelligent polymer patterned surfaces for tissue engineered liver"Macromolecular Symposia. 195. 231-235 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Y Tsuda, A Kikuchi, M Yamato, Y Sakurai, M Umezu, T Okano: "Control of cell adhesion and detachment using temperature and thermoresponsive copolymer grafted cultures surfaces"Journal of Biomedical Materials Research. (In Press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.Harimoto, M.Yamato, M.Hirose, C.Takahashi, Y.Isoi, A.Kikuchi, T.Okano: "Novel approach for achieving double-layered cell sheets co-culture : overlaying endothelial cell sheets on to monolayer hepatocytes utilizing temperature-responsive culture dishes"J. Biomed. Mater. Res.. 62(3). 464-470 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Yamato, C.Konno, M.Utsumi, A.Kikuchi, T.Okano: "Thermally responsive polymer-grafted surfaces facilitate patterned cell seeding and co-culture"Biomaterials. 23. 561-567 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.A.Nandkumar, M.Yamato, A.Kushida, C.Konno, M.Hirose, A.Kikuchi, T.Okano: "Two-dimensional cell sheet manipulation of heterotypical co-cultured lung cells utilizing temperature-responsive culture dishes results in long-term maintenance of differentiated epithelial cell function"Biomaterials. 23. 1121-1130 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Shimizu, M.Yamato, Y.Isoi, T.Akutsu, T.Setomaru, K.Abe, A.Kikuchi, M.Umezu, T.Okano: "Fabrication of pulsatile cardiac tissue grafts using a novel 3-dimensional cell sheet manipulation technique and temperature-responsive cell culture surfaces"Circ. Res.. 90. e40-e48 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.O.Collier, J.M.Anderson, A.Kikuchi, T.Okano: "Adhesion behavior of monocyte, macrophages and foreign body giant cells on poly (N-isopropylacrylamide) temperature-responsive surfaces"J. Biomed. Mater. Res.. 59. 136-143 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Shimizu, M.Yamato, T.Akutsu, T.Shibata, Y.Isoi, A.Kikuchi, M.Umezu, T.Okano: "Electrically communicating three-dimensional cardiac tissue mimic fabricated by layered cultured cardiomyocyte sheets"J. Biomed. Mater. Res.. 60. 110-117 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Harimoto, M.Yamato, A.Kikuchi, T.Okano: "Tissue Engineering for Therapeutic Use 6, Y.Ikada, Y.Umakoshi, T.Hotta, Eds."Double layerd co-culture of hepatocytes and endothelial cell sheets by cell sheet engineering. 4 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yamato M: "Thermally responsive polymer-grafted surfaces facilitate patterned cell seeding and co-culture"Biomaterials. 23. 561-567 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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