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Development of a cure for intractable diseases using canine induced pluripotent stem cell-derived mesenchymal stem cells

Research Project

Project/Area Number 18H02349
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 42020:Veterinary medical science-related
Research InstitutionOsaka Prefecture University

Principal Investigator

Hatoya Shingo  大阪府立大学, 生命環境科学研究科, 准教授 (40453138)

Co-Investigator(Kenkyū-buntansha) 西田 英高  大阪府立大学, 生命環境科学研究科, 准教授 (00622804)
杉浦 喜久弥  大阪府立大学, 生命環境科学研究科, 教授 (30171143)
井上 徳光  和歌山県立医科大学, 医学部, 教授 (80252708)
Project Period (FY) 2018-04-01 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2021: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2020: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2018: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
KeywordsiPS細胞 / イヌ / 間葉系幹細胞 / MSC / 血液 / 多能性幹細胞 / 再生医療 / フィーダーフリー / PBMC / センダイウイルスベクター / 血液細胞 / iXEN細胞
Outline of Final Research Achievements

We attempted to differentiate canine iPS cells into mesenchymal stem cells for use in the treatment of intractable diseases.
1. Canine iPS cell lines were generated from canine fetal fibroblasts by introducing four specific genes (OCT3/4, SOX2, C-MYC, KLF4) with Sendai virus vector. 2. We determined the reprogramming efficiency of canine PBMCs under several conditions involving several types of media supplemented with small-molecule compounds, and successfully generated canine iPS cell lines. 3. Enzyme passage and feeder-free culture of canine iPS cells were examined, enabling the efficient establishment and mass culture of canine iPS cells. 4. The differentiation of canine iPS cells into MSC-like cells was successfully induced.

Academic Significance and Societal Importance of the Research Achievements

現在、研究に使用できる質の良いイヌiPS細胞株は世界的にもほとんどなく、獣医学領域におけるiPS細胞を用いた再生医療研究はほとんど進んでいない。本研究成果から、線維芽細胞や血液細胞を使用して安定的にイヌiPS細胞株の作製に成功し、これを大量に培養することも可能となった。以上の結果は、イヌiPS細胞を用いた研究・臨床応用を推進する研究成果である。さらに、イヌiPS細胞からMSC様細胞への分化に成功しており、将来的には、難治性疾患の治療薬として応用できる可能性を示した。

Report

(5 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (43 results)

All 2022 2021 2020 2019 2018 Other

All Journal Article (17 results) (of which Int'l Joint Research: 8 results,  Peer Reviewed: 16 results,  Open Access: 10 results) Presentation (20 results) (of which Int'l Joint Research: 9 results,  Invited: 3 results) Remarks (6 results)

  • [Journal Article] Effects of the preservation medium and storage duration of domestic cat ovaries on the maturational and developmental competence of oocytes <i>in vitro</i>2022

    • Author(s)
      Takumi Yoshida, Md Emtiaj Alam, Keisuke Hanafusa, Yasunori Tsujimoto, Masaya Tsukamoto, Ryoji Kanegi, Toshio Inaba, Kikuya Sugiura, Shingo Hatoya
    • Journal Title

      Journal of Reproduction and Development

      Volume: 68 Issue: 2 Pages: 160-164

    • DOI

      10.1262/jrd.2021-084

    • NAID

      130008139942

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Potent adjuvant effect elicited for tumor immunotherapy by a liposome conjugated pH-sensitive polymer and dendritic cell-targeting Toll-like-receptor ligand2022

    • Author(s)
      Watanabe Shunichi、Yuba Eiji、Akazawa Takashi、Wijewardana Viskam、Kakihara Yuka、Azuma Ayaka、Hagimori Kenji、Kanegi Ryoji、Hatoya Shingo、Inoue Norimitsu、Inaba Toshio、Sugiura Kikuya
    • Journal Title

      Vaccine

      Volume: 40 Issue: 10 Pages: 1448-1457

    • DOI

      10.1016/j.vaccine.2022.01.048

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Quality of life-improving effect of autologous ex vivo expanded cytotoxic and opioid-producing lymphocytes for dogs with cancers2021

    • Author(s)
      Mitani Kosuke、Ito Yuki、Takene Yukio、Hatoya Shingo、Sugiura Kikuya、Inaba Toshio
    • Journal Title

      Veterinary Immunology and Immunopathology

      Volume: 238 Pages: 110292-110292

    • DOI

      10.1016/j.vetimm.2021.110292

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Canine induced pluripotent stem cell maintenance under feeder‐free and chemically‐defined conditions2021

    • Author(s)
      Kimura Kazuto、Tsukamoto Masaya、Yoshida Takumi、Tanaka Miyuu、Kuwamura Mitsuru、Ohtaka Manami、Nishimura Ken、Nakanishi Mahito、Sugiura Kikuya、Hatoya Shingo
    • Journal Title

      Molecular Reproduction and Development

      Volume: 88 Issue: 6 Pages: 395-404

    • DOI

      10.1002/mrd.23478

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Retrospective evaluation of nimustine use in the treatment of feline lymphoma2021

    • Author(s)
      Sakai K, Hatoya S, Furuya M, Nabetani T, Kanegi R, Shimamura S, Tani H, Shimada T
    • Journal Title

      Veterinary Medicine and Science

      Volume: 8 Issue: 1 Pages: 3-8

    • DOI

      10.1002/vms3.652

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 獣医再生医療における iPS 細胞、ES 細胞の作製と将来展望2021

    • Author(s)
      鳩谷晋吾
    • Journal Title

      JSAVBR News Letter(動物用ワクチン-バイオ医薬品研究会)

      Volume: 23 Pages: 15-21

    • Related Report
      2021 Annual Research Report
  • [Journal Article] Long-Term Trypsin Treatment Promotes Stem Cell Potency of Canine Adipose-Derived Mesenchymal Stem Cells2021

    • Author(s)
      Mitani Kosuke、Ito Yuki、Takene Yukio、Hatoya Shingo、Sugiura Kikuya、Inaba Toshio
    • Journal Title

      Stem Cells and Development

      Volume: 30 Issue: 6 Pages: 337-349

    • DOI

      10.1089/scd.2020.0175

    • NAID

      120007001724

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Efficient Reprogramming of Canine Peripheral Blood Mononuclear Cells into Induced Pluripotent Stem Cells2021

    • Author(s)
      Kimura Kazuto、Tsukamoto Masaya、Tanaka Miyuu、Kuwamura Mitsuru、Ohtaka Manami、Nishimura Ken、Nakanishi Mahito、Sugiura Kikuya、Hatoya Shingo
    • Journal Title

      Stem Cells and Development

      Volume: 30 Issue: 2 Pages: 79-90

    • DOI

      10.1089/scd.2020.0084

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Generation of Footprint‐Free Canine Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells Using Sendai Virus Vector2020

    • Author(s)
      Tsukamoto Masaya、Kimura Kazuto、Tanaka Miyuu、Kuwamura Mitsuru、Ohtaka Manami、Nakanishi Mahito、Sugiura Kikuya、Hatoya Shingo
    • Journal Title

      Molecular Reproduction and Development

      Volume: 87 Issue: 6 Pages: 663-665

    • DOI

      10.1002/mrd.23349

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Canine mesenchymal stromal cell-conditioned medium promotes survival and neurite outgrowth of neural stem cells2020

    • Author(s)
      Nakamura Michi, Nishida Hidetaka, Yoshizaki Karin, Akiyoshi Hideo, Hatoya Shingo, Sugiura Kikuya, Inaba Toshio
    • Journal Title

      Journal of Veterinary Medical Science

      Volume: 82 Issue: 5 Pages: 668-672

    • DOI

      10.1292/jvms.19-0141

    • NAID

      130007844440

    • ISSN
      0916-7250, 1347-7439
    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Development of feline embryos produced using freeze-dried sperm2020

    • Author(s)
      Tsujimoto Yasunori, Fujiki Kana, Alam Md Emtiaj, Tsukamoto Mssaya, Azuma Rika, Kanegi Ryoji, Anzai Masayuki, Inaba Toshio, Sugiura Kikuya, Hatoya Shingo
    • Journal Title

      Theriogenology

      Volume: 147 Pages: 71-76

    • DOI

      10.1016/j.theriogenology.2020.02.021

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Decreased serum zinc concentration in dogs with lymphocytic-plasmacytic enteritis, and its associations with disease severity and prognosis2020

    • Author(s)
      Sakai K, Hatoya S, Furuya M, Shimamura S, Nabetani T, Tani H, Shimada T.
    • Journal Title

      Journal of Veterinary Medical Science

      Volume: 82 Issue: 6 Pages: 759-763

    • DOI

      10.1292/jvms.20-0109

    • NAID

      130007863927

    • ISSN
      0916-7250, 1347-7439
    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Manipulation of the tumor microenvironment by cytokine gene transfection enhances dendritic cell‐based immunotherapy2019

    • Author(s)
      Wijesekera Daluthgamage Patsy Himali、Yuba Eiji、De Silva Nadeeka Harshini、Watanabe Shun‐ichi、Tsukamoto Masaya、Ichida Chihiro、Izawa Takeshi、Itoh Kazuyuki、Kanegi Ryoji、Hatoya Shingo、Yamate Jyoji、Inaba Toshio、Sugiura Kikuya
    • Journal Title

      FASEB BioAdvances

      Volume: 2 Issue: 1 Pages: 5-17

    • DOI

      10.1096/fba.2019-00052

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Feline embryo development in commercially available human media supplemented with fetal bovine serum2019

    • Author(s)
      ALAM Md. Emtiaj、IWATA Jun、FUJIKI Kana、TSUJIMOTO Yasunori、KANEGI Ryoji、KAWATE Noritoshi、TAMADA Hiromichi、INABA Toshio、SUGIURA Kikuya、HATOYA Shingo
    • Journal Title

      Journal of Veterinary Medical Science

      Volume: 81 Issue: 4 Pages: 629-635

    • DOI

      10.1292/jvms.18-0335

    • NAID

      130007638716

    • ISSN
      0916-7250, 1347-7439
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Development of feline embryos produced by Piezo-actuated intracytoplasmic sperm injection of elongated spermatids2019

    • Author(s)
      TSUJIMOTO Yasunori、FUJIKI Kana、ALAM MD Emtiaj、TSUKAMOTO Masaya、AZUMA Rika、KANEGI Ryoji、ANZAI Masayuki、INABA Toshio、SUGIURA Kikuya、HATOYA Shingo
    • Journal Title

      Journal of Reproduction and Development

      Volume: 65 Issue: 3 Pages: 245-250

    • DOI

      10.1262/jrd.2018-119

    • NAID

      130007662981

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Correlation between toll-like receptor 4 and nucleotide-binding oligomerization domain 2 (NOD2) and pathological severity in dogs with chronic gastrointestinal diseases2019

    • Author(s)
      Aono Kimiya、Azuma Yasu-Taka、Nabetani Tomoyo、Hatoya Shingo、Furuya Masaru、Miki Mariko、Hirota Kana、Fujimoto Yasuyuki、Nishiyama Kazuhiro、Ogata Yoshiyuki、Mochizuki Tomofumi、Tani Hiroyuki
    • Journal Title

      Veterinary Immunology and Immunopathology

      Volume: 210 Pages: 15-22

    • DOI

      10.1016/j.vetimm.2019.03.003

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Generation of Footprint-Free Canine Induced Pluripotent Stem Cells Using Auto-Erasable Sendai Virus Vector2018

    • Author(s)
      Tsukamoto Masaya、Nishimura Toshiya、Yodoe Kyohei、Kanegi Ryoji、Tsujimoto Yasunori、Alam Md Emtiaj、Kuramochi Mizuki、Kuwamura Mitsuru、Ohtaka Manami、Nishimura Ken、Nakanishi Mahito、Inaba Toshio、Sugiura Kikuya、Hatoya Shingo
    • Journal Title

      Stem Cells and Development

      Volume: 27 Issue: 22 Pages: 1577-1586

    • DOI

      10.1089/scd.2018.0084

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] シンポジウム「腎臓再生へのニューテクノロジー」iPS細胞研究と異種移植研究の融合は、猫の慢性腎不全に対する移植医療を実現できるのか? 猫iPS細胞開発の現状と未来2021

    • Author(s)
      鳩谷晋吾
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] イヌiPS細胞から間葉系幹細胞の誘導の試み2021

    • Author(s)
      塚本雅也、川端千晶、木村和人、杉浦喜久弥、鳩谷晋吾
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 末梢血単核球を用いた効率的なイヌiPS細胞の樹立および大量培養2021

    • Author(s)
      木村和人、塚本雅也、吉田拓海、田中美有、桑村充、大高真奈美、西村健、中西真人、杉浦喜久弥、鳩谷晋吾
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 4.臨床免疫検討会 間葉系幹細胞による治療は難治性疾患の画期的な治療なのか?2021

    • Author(s)
      Thierry Olivry、小林哲郎、田村勝利、鳩谷晋吾、福田威、水野拓也
    • Organizer
      獣医アトピー・アレルギー・免疫学会 第13回ベーシックセミナー
    • Related Report
      2021 Annual Research Report
  • [Presentation] サイトカイン遺伝子の腫瘍細胞への導入による微小環境内免疫状態の改変がもたらす治療効果の検討2021

    • Author(s)
      渡邊 駿一、高木 彩夏、弓場 英司、桑村 充、井澤 武史、鳩谷 晋吾、金城 綾二、稲葉 俊夫、杉浦 喜久弥
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report
  • [Presentation] ネコ体外胚産生法の低コスト化および簡易化の検討2021

    • Author(s)
      吉田 拓海、辻本 恭典、木村 和人、杉浦 喜久弥、鳩谷 晋吾
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 低酸素環境を標的とした犬の消化管型リンパ腫に対する新たな治療法の開発;マウスモデルによる前臨床試験2021

    • Author(s)
      山崎 裕毅、西田 英高、鳩谷 晋吾、秋吉 秀保
    • Organizer
      第164回日本獣医学会学術集会
    • Related Report
      2021 Annual Research Report
  • [Presentation] CHEMICALLY-DEFINED AND FEEDER-FREE MAINTENANCE OF CANINE INDUCED PLURIPOTENT STEM CELLS2021

    • Author(s)
      K Kimura, M Tsukamoto, T Yoshida, Miyuu Tanaka, Mitsuru Kuwamura, M Ohtaka, K Nishimura, M Nakanishi, K Sugiura, S Hatoya
    • Organizer
      The International Society for Stem Cell Research (ISSCR) 2021 Virtual
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] DIFFERENTIATION OF CANINE INDUCED PLURIPOTENT STEM CELLS INTO DEFINITIVE ENDODERM2021

    • Author(s)
      M Tsukamoto, K Kimura, K Sugiura, S Hatoya
    • Organizer
      The International Society for Stem Cell Research (ISSCR) 2021 Virtual
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] CHEMICALLY-DEFINED AND FEEDER-FREE MAINTENANCE OF CANINE INDUCED PLURIPOTENT STEM CELLS2020

    • Author(s)
      S Hatoya, S Inoue, K Kimura, M Tsukamoto, M Ohtaka, K Nishimura, M Nakanishi, K Sugiura.
    • Organizer
      The International Society for Stem Cell Research (ISSCR) 2020 Virtual
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] REPROGRAMMING OF CANINE EMBRYONIC FIBROBLASTS TO PRIMED AND NAIVE LIKE PLURIPOTENT STATES2020

    • Author(s)
      M Tsukamoto, K Kimura, R Kanegi, M Ohtaka, M Nakanishi, T Inaba, K Sugiura, S Hatoya
    • Organizer
      he International Society for Stem Cell Research (ISSCR) 2020 Virtual
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] EFFICIENT GENERATION OF CANINE INDUCED PLURIPOTENT STEM CELLS FROM PERIPHERAL BLOOD MONONUCLEAR CELLS USING A SMALL MOLECULE COCKTAIL2020

    • Author(s)
      K Kimura, M Tsukamoto, M Ohtaka, K Nishimura, M Nakanishi, K Sugiura, S Hatoya.
    • Organizer
      he International Society for Stem Cell Research (ISSCR) 2020 Virtual
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 末梢血単核球からのイヌiPS細胞作製に向けた培養条件の検討2020

    • Author(s)
      木村和人,塚本雅也,大高真奈美,西村健,中西真人,杉浦喜久弥,鳩谷晋吾
    • Organizer
      第19回日本再生医療学会総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 末梢血単核球を利用したイヌiPS細胞作製の試み2020

    • Author(s)
      塚本雅也,木村和人,大高真奈美,西村健,中西真人,杉浦喜久弥,鳩谷晋吾.
    • Organizer
      第19回日本再生医療学会総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Challenges to Stem Cell Therapy Development and Regulation in Dogs and Cats: 「Generation of canine and feline pluripotent stem cells for application in regenerative medicine」2018

    • Author(s)
      Shingo Hatoya
    • Organizer
      The 5th TERMIS World Congress(国際学会)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Induction of transgene free canine induced pluripotent stem cells by using Sendai virus vectors2018

    • Author(s)
      TSUKAMOTO M, KANEGI Ryouji, NISHIMURA Toshiya, OHTAKA Mana, NISHIMURA Ken, NAKANISHI Mahito, INABA Toshio, SUGIURA Kikuya, HATOYA Singo
    • Organizer
      The 5th TERMIS World Congress(国際学会)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Isolation and qualification of canine and feline mesenchymal stem cells by monitoring of glutathione levels2018

    • Author(s)
      MITANI Kosuke, ITO Yuki, TAKENE Yukio, JEONG Eui Man, KANG Heun-Soo, KIM In-Gyu, INABA Toshio, HATOYA Singo, SUGIURA Kikuya
    • Organizer
      Tissue Engineering & Regenerative Medecine Exposition 2018 (TERMEX 2018)(国際学会)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 治療や病態解明へ応用可能なイヌiPS細胞の作製2018

    • Author(s)
      塚本雅也、西村俊哉、金城綾二、倉持瑞樹、桑村充、大高真奈美、西村健、中西真人、稲葉俊夫、杉浦喜久弥、鳩谷晋吾
    • Organizer
      第161回日本獣医学会学術集会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Developmental competence of domestic cat oocytes from ovaries stored at various durations at 4 &ordm;C in different solutions2018

    • Author(s)
      ALAM Md. Emtiaj、花房佳祐、鳩谷普吾、辻本恭典、稲葉俊夫、杉浦喜久弥
    • Organizer
      第161回日本獣医学会学術集会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Clinical application andfuture directionsof mesenchymal stem/stromal celltherapy forspinalcord injuryin dogs2018

    • Author(s)
      Hidetaka Nishida
    • Organizer
      The 5th TERMIS World Congress(国際学会)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Remarks] 細胞病態学教室ホームページ

    • URL

      https://www.omu.ac.jp/vet/cellpatho/

    • Related Report
      2021 Annual Research Report
  • [Remarks] 細胞病態学教室ホームページ

    • URL

      http://www.vet.osakafu-u.ac.jp/cell/

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
  • [Remarks] 犬の血液細胞からイヌiPS細胞の作製に成功

    • URL

      https://www.osakafu-u.ac.jp/news/nws20210202/

    • Related Report
      2020 Annual Research Report
  • [Remarks] EurekAlert! 掲載

    • URL

      https://www.eurekalert.org/pub_releases/2021-02/opu-nsc020221.php

    • Related Report
      2020 Annual Research Report
  • [Remarks] Asia Research News 掲載(トップページ)

    • Related Report
      2020 Annual Research Report
  • [Remarks] Laboratory of Cell Pathology

    • URL

      http://www.vet.osakafu-u.ac.jp/english/cell/

    • Related Report
      2018 Annual Research Report

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Published: 2018-04-23   Modified: 2023-12-25  

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