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Fabrication and transplantation of canine corneal bio-sheet using in body tissue architecture (iBTA)

Research Project

Project/Area Number 17K08096
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Veterinary medical science
Research InstitutionThe University of Tokyo

Principal Investigator

Tsuzuki Keiko  東京大学, 大学院農学生命科学研究科(農学部), 特任助教 (30364251)

Co-Investigator(Kenkyū-buntansha) 中山 泰秀  大分大学, 医学部, 客員研究員 (50250262)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords犬 / 角膜損傷 / 生体内組織形成術 / 角膜移植 / 人工角膜 / 犬角膜内皮細胞 / ハイブリッド犬人工角膜 / 角膜 / 再生医療
Outline of Final Research Achievements

We tried to culture canine corneal endothelial cells using explant method, however, the cultured cells consisted of p75-positive immature cells and ZO-1 and Na+K+ATPase-positive mature endothelial cells. It was difficult to isolate mature cells. Therefore, we fabricate an artificial canine cornea only using a canine corneal epithelial cell line and bio-sheet fabricated using iBTA. Canine corneal epithelial cells were successfully cultured on a bio-sheet and 5 days of air exposure induced a sufficient stratified epithelium on the bio-sheet. We also established culture method of canine corneal stromal cells and successfully induced these cells into myofibroblasts. Using this induction system, we revealed that hepatocyte growth factor (HGF) can repress the differentiation from stromal cells into myofibroblasts. This result indicates that HGF can be a promising drug for opacity induced after corneal injury.

Academic Significance and Societal Importance of the Research Achievements

犬角膜内皮細胞の培養が可能であることが示された。今後、成熟細胞の誘導や純化を検討する必要があるが、犬の角膜内皮疾患の病態解明などへの応用が期待できる。また、iBTAを用いた角膜組織の作製には至らなかったが、犬角膜上皮細胞がバイオシートに定着し、重層化が誘導された。このことは、バイオシートを用いて体外で他の細胞と融合させた組織体の作製が可能であることを示し、iBTAを用いた組織再生療法において新たなアプローチを提案する。さらに、犬角膜実質細胞の分離培養と筋線維芽細胞分化の検討から、角膜混濁の病態を再現できた。今後、肝細胞成長因子の他、角膜混濁治療薬の開発につながることが期待される。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (7 results)

All 2019 2018 Other

All Journal Article (2 results) (of which Peer Reviewed: 2 results) Presentation (2 results) (of which Int'l Joint Research: 1 results) Remarks (3 results)

  • [Journal Article] Canine corneal epithelial cells possess a sustained proliferative capacity and generate a spontaneously derived cell line.2018

    • Author(s)
      Morita M, Fujita N, Abe M, Hayashimoto K, Nakagawa T, Nishimura R Tsuzuki K
    • Journal Title

      Experimental Eye Research

      Volume: 印刷中 Pages: 155-163

    • DOI

      10.1016/j.exer.2018.03.003

    • Related Report
      2018 Research-status Report 2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] A pilot study of transplantation of an autologous corneal epithelial cell sheet in a canine model of corneal injury2018

    • Author(s)
      Nam E, Fujita N, Morita M, Lin HY, Endo K, Nakagawa T, Nishimura R, Tsuzuki K.
    • Journal Title

      Jpn J Vet Res

      Volume: 80 Pages: 772-777

    • NAID

      120006469031

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] 犬角膜実質細胞の分離培養と筋線維芽細胞分化制御2019

    • Author(s)
      田中華鈴、藤田直己、林本考緒道、武田妙、陳君妍、羽生航、西村亮平、都築圭子
    • Organizer
      第162回日本獣医学会学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Trial of canine corneal endothelial cell cultivation2018

    • Author(s)
      Koji Hayashimoto, Naoki Fujita, keiko Tuzuki Ryohei Nishimura
    • Organizer
      8th Annual Congress of Asian Society of Veterinary Surgery
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Remarks] 東京大学獣医外科学研究室HP

    • URL

      http://www.vm.a.u-tokyo.ac.jp/geka/

    • Related Report
      2019 Annual Research Report
  • [Remarks] 獣医外科学研究室 - 東京大学獣医学専攻HP

    • URL

      http://www.vm.a.u-tokyo.ac.jp/geka/

    • Related Report
      2018 Research-status Report
  • [Remarks] 東京大学大学院農学生命科学研究科獣医外科学研究室HP

    • URL

      http://www.vm.a.u-tokyo.ac.jp/geka/

    • Related Report
      2017 Research-status Report

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Published: 2017-04-28   Modified: 2021-02-19  

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