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Developing a scalable-coculturing system for gut anaerobes and intestinal epithelium

Research Project

Project/Area Number 19K22544
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 49:Pathology, infection/immunology, and related fields
Research InstitutionKeio University

Principal Investigator

SASAKI Nobuo  慶應義塾大学, 医学部(信濃町), 特任助教 (30777769)

Project Period (FY) 2019-06-28 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywordsオルガノイド / 腸内細菌 / 嫌気性細菌 / 共培養システム / プロバイオティクス / 嫌気性腸内細菌 / 共培養システム開発 / 嫌気性培養
Outline of Research at the Start

本課題研究は,正常腸管上皮細胞培養法のオルガノイドを腸内細菌研究分野へ応用させることを目的としているが,腸管上皮を含む生体の上皮細胞の生存には酸素は不可欠であり,反対に腸内細菌のほとんどは酸素存在下では生育できない.したがって,このような相反する酸素要求性を示す腸管上皮細胞と腸内細菌の共培養は原理的に不可能である.そこで我々は,生体内の腸管の生理条件をシャーレ上で再現することで,消化管感染を中心とした様々な疾患の発症メカニズムの理解につながる感染学や腸内細菌学の発展に貢献できる,新規の低酸素環境におけるオルガノイド培養法の確立を目指す.

Outline of Final Research Achievements

The aim of this study is to develop a novel culturing system of healthy human intestinal epithelium together with anaerobic gut bacteria. We modified the classical culture method for 3D adult-tissue stem cell, organoid firstly. Using the Boyden chamber, we succeeded in culturing human normal colon epithelial cells as the monolayer condition for long periods. Next, we also figure out the suitable conditioned medium to enable to generate anaerobic environments at apical surface of their epithelium. Actually, the obligate anaerobic bacteria can grow to form their colonies on the human colonic epithelial cells in our culture condition. Here, we demonstrated a novel coculturing system in modeling competitive and mutually beneficial relationship between human epithelium and gut microbes in vitro.

Academic Significance and Societal Importance of the Research Achievements

最近では企業における機能細菌(プロバイオティクス)研究開発現場において,動物愛護の観点から動物試験が禁止されるようになってきたため,モデル動物に変わる細菌の機能を試験する新規モデルの開発が急務であった.我々が開発した新規ヒト腸管上皮-腸内細菌共培養システムは,動物試験を不要とするだけではなく,細菌が与えるヒト細胞への影響を直接検証できるといった特徴も有している.そのため本研究成果は,製薬や食品メーカーが直面している商品開発の“死の谷”を克服するための一翼を担うことが期待されているので,今後積極的に本技術を産業界へ導出したいと考えている.

Report

(3 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • Research Products

    (15 results)

All 2021 2020 2019

All Journal Article (7 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 6 results,  Open Access: 3 results) Presentation (6 results) (of which Int'l Joint Research: 2 results,  Invited: 6 results) Book (1 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] 腸内細菌-宿主細胞の相互作用メカニズムを紐解く新規共培養系の開発2021

    • Author(s)
      佐々木 伸雄
    • Journal Title

      臨床免疫・アレルギー科

      Volume: 75 Pages: 77-81

    • NAID

      40022452487

    • Related Report
      2020 Annual Research Report
  • [Journal Article] Development of a scalable co-culture system for gut anaerobes and human colon epithelium2020

    • Author(s)
      Sasaki Nobuo、Miyamoto Kentaro、Maslowski Kendle M.、Ohno Hiroshi、Kanai Takanori、Sato Toshiro
    • Journal Title

      Gastroenterology

      Volume: in Press Issue: 1 Pages: 388-390

    • DOI

      10.1053/j.gastro.2020.03.021

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Somatic inflammatory gene mutations in human ulcerative colitis epithelium.2020

    • Author(s)
      Nanki K, Fujii M, Shimokawa M, Matano M, Nishikori S, Date S, Takano A, Toshimitsu K, Ohta Y, Takahashi S, Sugimoto S, Ishimaru K, Kawasaki K, Nagai Y, Ishii R, Yoshida K, Sasaki N, Hibi T, Ishihara S, Kanai T, Sato T.
    • Journal Title

      Nature

      Volume: 7789 Issue: 7789 Pages: 254-259

    • DOI

      10.1038/s41586-019-1844-5

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Gut pathobionts underlie intestinal barrier dysfunction and liver T helper 17 cell immune response in primary sclerosing cholangitis.2019

    • Author(s)
      Nakamoto N, Sasaki N, Aoki R, Miyamoto K, Suda W, Teratani T, Suzuki T, Koda Y, Chu PS, Taniki N, Yamaguchi A, Kanamori M, Kamada N, Hattori M, Ashida H, Sakamoto M, Atarashi K, Narushima S, Yoshimura A, Honda K, Sato T, Kanai T.
    • Journal Title

      Nat Microbiol.

      Volume: 4 Issue: 3 Pages: 492-503

    • DOI

      10.1038/s41564-018-0333-1

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Ongoing chromosomal instability and karyotype evolution in human colorectal cancer organoids2019

    • Author(s)
      Bolhaqueiro A.C.F., Ponsioen B., Bakker B., Klaasen S., Kucukkose E., van Jaarsveld R.H., Vivie J., Verlaan-Klink I., Hami N., Spierings D.C.J., Sasaki N., Dutta D., Boj S.F., Vries R.G.J., Lansdorp P.M., van de Wetering M. van Oudenaarden A., Clevers H., Kranenburg O., Foijer F., Snippert H.J.G., Kops G.J.P.
    • Journal Title

      Nature Genetics

      Volume: 51 Issue: 5 Pages: 824-834

    • DOI

      10.1038/s41588-019-0399-6

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Defining the Identity and Dynamics of Adult Gastric Isthmus Stem Cells2019

    • Author(s)
      Han S, Fink J, Jorg DJ, Lee E, Yum MK, Chatzeli L, Merker SR, Josserand M, Trendafilova T, Andersson-Rolf A, Dabrowska C, Kim H, Naumann R, Lee JH, Sasaki N, Mort RL, Basak O, Clevers H, Stange DE, Philpott A, Kim JK, Simons BD, Koo BK
    • Journal Title

      Cell Stem Cell

      Volume: 25 Issue: 3 Pages: 342-356.e7

    • DOI

      10.1016/j.stem.2019.07.008

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Enteroendocrine and tuft cells support Lgr5 stem cells on Paneth cell depletion2019

    • Author(s)
      van Es Johan H.、Wiebrands Kay、L?pez-Iglesias Carmen、van de Wetering Marc、Zeinstra Laura、van den Born Maaike、Korving Jeroen、Sasaki Nobuo、Peters Peter J.、van Oudenaarden Alexander、Clevers Hans
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 116 Issue: 52 Pages: 26599-26605

    • DOI

      10.1073/pnas.1801888117

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Presentation] The study of epithelial crosstalk at the host-microbes interface using organoid2020

    • Author(s)
      Nobuo Sasaki
    • Organizer
      8th Global Network Forum on Infection and Immunity: Microbiome
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 嫌気性腸内細菌とヒト正常大腸上皮細胞の共培養を可能にする新規培養系の開発2020

    • Author(s)
      佐々木 伸雄
    • Organizer
      第43回日本分子生物学会年会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Organoid unravels the interaction mechanism between mucosal barrier and bacteria2019

    • Author(s)
      Nobuo Sasaki
    • Organizer
      Keio International Symposium, The Crossroads of Intestinal Microbiology, Mucosal Immunology and Epithelial Biology
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 大腸がんにおいて1細胞レベルで明らかにされた単一腫瘍内における不均一性の網羅的解析2019

    • Author(s)
      佐々木 伸雄
    • Organizer
      第46回日本毒性学会学術年会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] オルガノイドが切り拓く腸内細菌研究の新時代2019

    • Author(s)
      佐々木 伸雄
    • Organizer
      日本乳酸菌学会2019年度秋季セミナー
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] オルガノイドが切り拓く次世代の腸内細菌研究2019

    • Author(s)
      佐々木 伸雄,佐藤 俊朗
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Research-status Report
    • Invited
  • [Book] 決定版 オルガノイド実験スタンダード (佐藤,武部,永楽/編)2019

    • Author(s)
      佐々木 伸雄,金井 隆典,佐藤 俊朗
    • Total Pages
      372
    • Publisher
      羊土社
    • ISBN
      9784758122399
    • Related Report
      2019 Research-status Report
  • [Patent(Industrial Property Rights)] 上皮細胞培養用培養容器及びその使用2020

    • Inventor(s)
      佐藤俊朗,佐々木伸雄
    • Industrial Property Rights Holder
      佐藤俊朗,佐々木伸雄
    • Industrial Property Rights Type
      特許
    • Filing Date
      2020
    • Related Report
      2020 Annual Research Report

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Published: 2019-07-04   Modified: 2022-01-27  

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