Designing and Developing Innovative Use of Newly Discovered Colonic Epithelial Culture Method Applicable to Clinical Medicine.

• Project/Area Number: 22229005
• Research Category: Grant-in-Aid for Scientific Research (S)
• Allocation Type: Single-year Grants
• Research Field: Gastroenterology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: WATANABE Mamoru 東京医科歯科大学, 医歯(薬)学総合研究科, 教授 (10175127)
• Co-Investigator(Kenkyū-buntansha): NAKAMURA Tetsuya 東京医科歯科大学, 医歯学総合研究科, 教授 (70265809) ; 水島 昇 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (10353434)
• Project Period (FY): 2010-05-31 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥216,970,000 (Direct Cost: ¥166,900,000、Indirect Cost: ¥50,070,000); Fiscal Year 2014: ¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000); Fiscal Year 2013: ¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000); Fiscal Year 2012: ¥32,110,000 (Direct Cost: ¥24,700,000、Indirect Cost: ¥7,410,000); Fiscal Year 2011: ¥72,670,000 (Direct Cost: ¥55,900,000、Indirect Cost: ¥16,770,000); Fiscal Year 2010: ¥75,010,000 (Direct Cost: ¥57,700,000、Indirect Cost: ¥17,310,000)
• Keywords: 幹細胞移植 / 再生医療 / 大腸炎創傷治癒 / 大腸上皮 / 組織幹細胞 / 体外培養技術 / 細胞治療 / 大腸炎 / 創傷治癒 / 大腸上皮幹細胞 / 細胞増殖機構 / 細胞分化機構 / 炎症性腸疾患

Outline of Final Research Achievements

It has been expected that the elucidation of the mechanism for the regenerative regulation of intestinal epithelial cells would be applied to the gastrointestinal regenerative medicine.
It has however been technically impossible to assess the normal intestinal epithelial cells ex vivo. In this study, we therefore aimed to assess the function of normal intestinal epithelial cells and to establish the technical basis of clinical application for the primary culture of colonic epithelial cells by developing the long-term, serum-free, 3-dimential culture system of mice normal colon.

Assessment Rating

Verification Result (Rating)

A

Result (Rating)

A+: Progress in the research exceeds the initial goal. More than expected research results are expected.

Research Products

• [Journal Article] Comparison of magnetic resonance and balloon enteroscopic examination of the small intestine in patients with Crohn’s disease. (2014)
  • Author(s): Takenaka K, Ohtsuka K,Watanabe M et al
  • Journal Title: Gastroenterology. Volume: [Epub ahead of print] Pages: 334-342
  • DOI: 10.1053/j.gastro.2014.04.008
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Transplantation of expanded fetal intestinal progenitors contributes to colon regeneration after injury (2013)
  • Author(s): Fordham RP, Yui S, Nakamura T, Watanabe M et. al
  • Journal Title: Cell Stem Cell Volume: 13 Pages: 734-744
  • DOI: 10.1016/j.stem.2013.09.015
  • Peer Reviewed
• [Journal Article] Lineage-specific expression of bestrophin-2 and bestrophin-4 in human intestinal epithelial cells (2013)
  • Author(s): Ito G, Okamoto R, Nakamura T, Watanabe M et al
  • Journal Title: PLoS One. Volume: 8
  • DOI: 10.1371/journal.pone.0079693
  • Peer Reviewed
• [Journal Article] Th1/Th17-mediated interstitial pneumonia in chronic colitis mice independent of intestinal microbiota (2013)
  • Author(s): Nemoto Y, Watanabe M et al
  • Journal Title: J Immunol Volume: 190 Pages: 6616-25
  • DOI: 10.4049/jimmunol.1202930
  • Peer Reviewed
• [Journal Article] Bone marrow-mesenchymal stem cells are a major source of interleukin-7 and sustain colitis by forming the niche for colitogenic CD4 memory T cells (2013)
  • Author(s): Nemoto Y, Nakamura T, Watanabe M et al
  • Journal Title: Gut Volume: 62 Pages: 1142-1152
  • DOI: 10.1136/gutjnl-2012-302029
  • Peer Reviewed
• [Journal Article] The acquisition of malignant potential in colon cancer is regulated by the stabilization of Atonal homolog 1 protein (2013)
  • Author(s): Kano Y
  • Journal Title: Biochem Biophys Res Commun Volume: 432 Pages: 175-181
  • DOI: 10.1016/j.bbrc.2013.01.034
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5(+) stem cell. (2012)
  • Author(s): Yui S, Nakamura T, Sato T, Nemoto Y, Mizutani T, Zheng X, Ichinose S, Nagaishi T, Okamoto R, Tsuchiya K, Clevers H, and Watanabe M
  • Journal Title: Nat. Med. Volume: 18 Pages: 618-23
  • DOI: 10.1038/nm.2695
  • Peer Reviewed
• [Journal Article] The development of colitogenic CD4+ T cells is regulated by IL-7 in collaboration with NK cell function in a murine model of colitis. (2012)
  • Author(s): Yamaji O, Totsuka T
  • Journal Title: J. Immunol Volume: 188 Pages: 2524-2536
  • DOI: 10.4049/jimmunol.1100371
  • Peer Reviewed
• [Journal Article] Real-time analysis of P- gly coprotei n- mediated drug transport across primary intestinal epithelium three-dimensionally cultured in vitro (2012)
  • Author(s): Mizutani T, Nakamura T, et al
  • Journal Title: Biochem Biophys Res Commun. Volume: 419 Pages: 238-243
  • DOI: 10.1016/j.bbrc.2012.01.155
  • Peer Reviewed
• [Journal Article] Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5+ stem cell (2012)
  • Author(s): Yui S, Nakamura T, Watanabe M, et al.
  • Journal Title: Nature Medicine Volume: (in press)
  • Peer Reviewed
• [Journal Article] Luminal CD4+ T cells penetrate gut epithelial monolayers and egress from lamina propria to blood circulation. (2012)
  • Author(s): Nemoto Y, Watanabe M
  • Journal Title: Gastroenterology. Volume: 141 Pages: 2030-2039
  • DOI: 10.1053/j.gastro.2011.08.035
  • Peer Reviewed
• [Journal Article] Suppression ofhath1 gene expression directly regulated by hes1 via notch signaling is associated with goblet cell depletion in ulcerative colitis. (2012)
  • Author(s): Zheng X, Nakamura T, et al
  • Journal Title: Inflamm Bowel Di Volume: 17 Pages: 2251-2260
  • DOI: 10.1002/ibd.21611
  • Peer Reviewed
• [Journal Article] Upregulated IL-7Rα expression on colitogenic memory CD4+ T cells may participate in the development and persistence of chronic colitis (2011)
  • Author(s): Shinohara, T., et al
  • Journal Title: J Immunol Volume: 186 Pages: 2623-2632
  • DOI: 10.4049/jimmunol.1000057
  • Peer Reviewed
• [Journal Article] Suppression of hathl gene expression directly regulated by hesl via notch signaling is associated with goblet celldepletion in ulcerative colitis. (2011)
  • Author(s): Zheng X, Nakamura T, Watanabe M, et al.
  • Journal Title: Inflamm Bowel Dis. Volume: in press
  • Peer Reviewed
• [Journal Article] Longitudinal cell formation in the entire human smallintestine is correlated with the localization of Hathl and K1f4. (2011)
  • Author(s): Iwasaki M, Nakamura T, Watanabe M, etal.
  • Journal Title: J Gastroenterol Volume: 46 Pages: 191-202
  • Peer Reviewed
• [Journal Article] Upretulated IL-7 receptor (alpha) expression on colitogenicmemory CD4+ T cells may participate in the developmentand persistence of chronic colitis. (2011)
  • Author(s): Shinohara T, Nakamura T, Watanabe M, etal.
  • Journal Title: Journal of Immunology Volume: 186 Pages: 2623-2632
  • Peer Reviewed
• [Journal Article] Delta-like 1 expression promotes goblet cell differentiation in Notch-inactivated human colonic epithelial cells. (2010)
  • Author(s): Akiyama J, Nakamura T, Watanabe M, et al.
  • Journal Title: Biochem Biophys Res Commun. Volume: 393 Pages: 662-667
  • Peer Reviewed
• [Journal Article] IL-2 is positively involved in the development of colitogenic CD4+ IL-7R alpha high memory T cells inchronic colitis. (2010)
  • Author(s): Kameyama K, Nakamura T, Watanabe M, et al.
  • Journal Title: Eur J Immunol. Volume: 40 Pages: 2423-2436
  • Peer Reviewed
• [Journal Article] Cell culture and in-vivo analyses of plaque-derivedcytopathic hepatitis C virus mutants. (2010)
  • Author(s): Mishima K, Nakamura T, Watanabe M, etal.
  • Journal Title: Virology. Volume: 405 Pages: 361-369
  • Peer Reviewed
• [Presentation] Epithelial regeneration by cultured colonic cells expanded from a single adult Lgr5+ stem cell (2012)
  • Author(s): Nakamura T, Watanabe M
  • Organizer: The 6th Korea-Japan IBD Symposium
  • Place of Presentation: Tokyo
  • Year and Date: 2012-01-28
• [Presentation] Stem Cells (2012)
  • Author(s): Watanabe M
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] Real-time analysis of p-glycoprotein-mediated drug transport across primary intestinal epitherial cells three-dimensionally cultured in vitro (2012)
  • Author(s): Mizutani T, Watanabe M et.al.
  • Organizer: DDW2012
  • Place of Presentation: San Diego
• [Presentation] Regeneration of Damaged Colonic Tissue by Transplantation of Colonic Epithelial Stem Cells Maintained and Expanded In Vitro (2011)
  • Author(s): Yui S, Nakamura T, Watanabe M, et al.
  • Organizer: DDW2011
  • Place of Presentation: Chicago
  • Year and Date: 2011-05-07
• [Presentation] A long-term, mesenchyme-free in vitro culture system of colonic epithelial cells retaining stem cell compartment with proliferation/differentiation properties. (2010)
  • Author(s): Nakamura T, Watanabe M, et al.
  • Organizer: Digestive Disease Week 2010
  • Place of Presentation: New Orleans
• [Presentation] A Long-term, fully-defined culture system for colonic epithelial celis that allows efficient expansion of stem cell. (2010)
  • Author(s): Nakamura T, Watanabe M, et al.
  • Organizer: The 1st JSGE International Topic Conference
  • Place of Presentation: Kamakura
• [Presentation] JGF Marshal & Warren Lecture: Adult tissue stem cell therapy for gastrointestinal diseases.
  • Author(s): Watanabe M
  • Organizer: GASTRO 2013.
  • Place of Presentation: Shanghai, China
  • Invited
• [Presentation] Colonic Stem Cell Culture and Transplantation-Application in Mucosal Immunology.
  • Author(s): Watanabe M
  • Organizer: ICMI 2013
  • Place of Presentation: Vancouver, Canada
  • Invited
• [Presentation] Colonic Stem Cell Culture and Transplantation -Application in Mucosal Immunology-.
  • Author(s): Watanabe M
  • Organizer: RCAI-JSI International Symposium.
  • Place of Presentation: Yokohama, Japan
  • Invited
• [Presentation] Epidemiology of Asian IBD.
  • Author(s): Watanabe M
  • Organizer: 1st Annual Meeting of AOCC.
  • Place of Presentation: Tokyo, Japan
  • Invited
• [Presentation] State-of-the-Art lecture: Stem cell therapy in IBD?
  • Author(s): Watanabe M
  • Organizer: Falk Sympo 2013
  • Place of Presentation: Stuttgart, Germany
  • Invited
• [Presentation] Adult Stem Cell Therapy for Gastrointestinal Diseases
  • Author(s): Watanabe M
  • Organizer: International Ulcer Week 2012.
  • Place of Presentation: Tokyo
• [Presentation] Regeneration of damaged colon epithelium by transplanted colon Lgr5+ stem cells maintained and expanded in vitro.
  • Author(s): Yui S, Watanabe M et.al.
  • Organizer: 第10回 幹細胞シンポジウム.淡路島
  • Place of Presentation: Awajishima
• [Patent(Industrial Property Rights)] 大腸上皮幹細胞の単離・培養技術とこれを用いた大腸上皮移植技術 (2011)
  • Inventor(s): 渡辺守・中村哲也
  • Industrial Property Rights Holder: 国立大学法人東京医科歯科大学
  • Industrial Property Number: 2011-236469
  • Filing Date: 2011-10-27