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

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K08092
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Hokkaido 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
• 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

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.

Free Research Field

獣医学・臨床獣医学

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。