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Establishment of novel culture method of canine intestinal organoid and development of organoid library

Research Project

Project/Area Number 17K08092
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Veterinary medical science
Research InstitutionHokkaido University

Principal Investigator

Ohta Hiroshi  北海道大学, 獣医学研究院, 講師 (50431333)

Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords犬 / 消化管 / オルガノイド / 3次元培養 / ミニチュア・ダックスフンド / 炎症性結直腸ポリープ / 一塩基多型 / MMP / 蛋白喪失性腸症 / 三次元培養 / 疾患ライブラリー / 消化管上皮細胞 / 培養
Outline of Final Research Achievements

in this study, we tried to establish a novel culture method of canine intestinal organoid. Firstly, we tried to isolate colonic crypt that contain colonic epithelia stem cells. We successfully isolated colonic crypt from about five colonic mucosal biopsy samples obtained by endoscopy. Next, we tried to establish the 3D culture method of canine colonic organoid using these colonic crypt, Matrigel, and several growth factors. We successfully cultured canine colonic organoid for 5 days, and these colonic organoid formed luminal structure. Using same culture methods, we successfully cultured the colonic organoid from three Miniature Dachshund with inflammatory colorectal polyps (ICRPs) and two dogs with colorectal adenocarcinoma. We cryopreserved organoids obtained from normal dogs, ICRPs and dogs with colorectal adenocarcinoma.
In addition, we examined the single nucleotide polymorphisms of ICRPs and matrix metalloproteinase activity within the colorectal mucosa of ICRPs.

Academic Significance and Societal Importance of the Research Achievements

医学領域では、オルガノイドを用いた消化管疾患の研究が急速に進展している。一方で、獣医領域においてはこれまで報告がなされて来なかった。本研究では、他の研究施設に先駆けて犬のオルガノイドの培養を試み、犬の結腸由来のオルガノイドの培養手法を確立した。また同手法を用いて、炎症性結直腸ポリープおよび結直腸腺癌由来のオルガノイドの培養にも成功した。本研究で確立した培養手法には改善の余地が多く残されてはいるものの、獣医療におけるオルガノイドを用いた研究の先駆けとなった。

Report

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

    (6 results)

All 2020 2019 Other

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

  • [Int'l Joint Research] Utrecht University/Clinical Sciences of Companion Animals/Regenerative Medicine Centre Utrecht(オランダ)

    • Related Report
      2019 Annual Research Report
  • [Journal Article] Activities of matrix metalloproteinase-2, matrix metalloproteinase-9, and serine proteases in samples of the colorectal mucosa of Miniature Dachshunds with inflammatory colorectal polyps.2020

    • Author(s)
      Nagata N, Ohta H, Yamada A, Teoh YB, Ichii O, Morishita K, Sasaki N, Takiguchi M.
    • Journal Title

      Am J Vet Res

      Volume: -

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Clinical characteristics of dogs with food-responsive protein-losing enteropathy.2020

    • Author(s)
      Nagata N, Ohta H, Yokoyama N, Teoh YB, Nisa K, Sasaki N, Osuga T, Morishita K, Takiguchi M.
    • Journal Title

      J Vet Intern Med

      Volume: 34 Issue: 2 Pages: 659-668

    • DOI

      10.1111/jvim.15720

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genome-wide DNA methylation analysis in canine gastrointestinal lymphoma2020

    • Author(s)
      OHTA Hiroshi、YAMAZAKI Jumpei、JELINEK Jaroslav、ISHIZAKI Teita、KAGAWA Yumiko、YOKOYAMA Nozomu、NAGATA Noriyuki、SASAKI Noboru、TAKIGUCHI Mitsuyoshi
    • Journal Title

      Journal of Veterinary Medical Science

      Volume: 82 Issue: 5 Pages: 632-638

    • DOI

      10.1292/jvms.19-0547

    • NAID

      130007844434

    • ISSN
      0916-7250, 1347-7439
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] 蛋白喪失性腸症の犬40例からの腸リンパ管拡張症の診断と治療に関する考察2019

    • Author(s)
      永田矩之、大田寛、横山望、佐々木東、大菅辰幸、森下啓太郎、滝口満喜
    • Organizer
      日本獣医師会 獣医学術学会 年次大会(神奈川)
    • Related Report
      2019 Annual Research Report
  • [Remarks] 第2回One Health Relay Report

    • URL

      https://onehealth.vetmed.hokudai.ac.jp/activity/one-health-relay-report/3/

    • Related Report
      2019 Annual Research Report

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

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