Culture of feline pluripotent stem cells and development of hematopoietic differentiation methods to aim for clinically applying for blood transfusion

• Project/Area Number: 24780311
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Clinical veterinary science
• Research Institution: Osaka Prefecture University
• Principal Investigator: HATOYA Shingo 大阪府立大学, 生命環境科学研究科(系), 准教授 (40453138)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: iPS細胞 / ES細胞 / ネコ / 多能性幹細胞 / 再生医療 / 獣医学 / 血液分化 / 顕微授精

Outline of Final Research Achievements

The objective of this study was to establish feline ES (embryonic stem) cell lines and iPS (induced pluripotent stem) cell lines, and to induce hematopoietic cells from these stem cells. The following results were obtained.
① Blastocyst formation was observed using piezo-ICSI, and primary feline ES-like cell conolies were formed from blastocysts. ② We confirm the successful production for the first time of biologically active feline LIF within E.coli.③ We transduced four transcriptional factors (mouse Oct3/4, Sox2, Klf4, c-Myc) into fetal fibroblast cells of domestic cats with retrovirus vector, and could establish an iPSC-like cell line.④ We transduced four transcriptional factors into fetal fibroblast cells of domestic cats with sendai virus vector, and primary feline iPS-like cell colonies were formed.⑤ Feline iPS-like cells can be differentiated into macrophages and erythroid cells.

Research Products

• [Journal Article] Effect of ovarian hormones on maturation of dendritic cells from peripheral blood monocytes in dogs. (2015)
  • Author(s): Wijewardana V, Sugiura K, Wijesekera DP, Hatoya S, Nishimura T, Kanegi R, Ushigusa T, Inaba T.
  • Journal Title: J Vet Med Sci. Volume: -
  • NAID: 130005089954
  • Peer Reviewed
• [Journal Article] Effective enhancement of anti-tumor immune responses by transfection of IFNγ gene into tumor using a new type synthetic vector. (2014)
  • Author(s): Wijesekera, D.P.H., Sugiura, K., Yuba, E., Ueda, K., Wijewardana, V., Kanegi, R., Nishimura, T., Ushigusa, T., Hatoya, S., Kono, K., Inaba, T.
  • Journal Title: Vet. Immunol. Immunopathol. Volume: 162 Issue: 1-2 Pages: 59-64
  • DOI: 10.1016/j.vetimm.2014.08.016
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Profiling of serum metabolites in canine lymphoma using gas chromatography mass spectrometry. (2014)
  • Author(s): Tamai R, Furuya M, Hatoya S, Akiyoshi H, Yamamoto R, Komori Y, Yokoi S, Tani K, Hirano Y, Komori M, Takenaka S.
  • Journal Title: J Vet Med Sci. Volume: 76 Pages: 1513-1518
  • NAID: 130004677727
  • Peer Reviewed
• [Journal Article] Generation of functional platelets from canine induced pluripotent stem cells. (2013)
  • Author(s): Nishimura, T., Hatoya, S., Kanegi, R., Sugiura, K., Wijewardana, V., Kuwamura, M., Tanaka, M., Yamate, J., Izawa, T., Takahashi, M., Kawate, N., Tamada, H., Imai, H., Inaba, T.
  • Journal Title: Stem Cells Dev. Volume: 22 Issue: 14 Pages: 2026-2035
  • DOI: 10.1089/scd.2012.0701
  • Peer Reviewed
• [Presentation] 成犬体細胞からのiPS細胞樹立の試み (2014)
  • Author(s): 三埜晃佑、西村俊哉、鳩谷晋吾、杉浦喜久弥、今井 裕、高橋正弘、川手憲俊、玉田尋通、稲葉俊夫
  • Organizer: 第157回日本獣医学会学術集会
  • Place of Presentation: 北海道大学（北海道・札幌市）
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] センダイウイルスベクター（SeVdp）を用いたネコおよびイヌiPS細胞の作製 (2014)
  • Author(s): 淀江京平、鳩谷晋吾、金城凌二、杉浦喜久弥、大高真奈美、西村健、中西真人、高橋正弘、川手憲俊、玉田尋通、稲葉俊夫
  • Organizer: 第157回日本獣医学会学術集会
  • Place of Presentation: 北海道大学（北海道・札幌市）
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] イヌ・ネコiPS細胞の樹立と将来の展望 (2014)
  • Author(s): 鳩谷晋吾
  • Organizer: 平成25年度日本獣医師会獣医学術学会年次大会
  • Place of Presentation: 千葉
  • Invited
• [Presentation] イヌ・ネコiPS細胞の作製の試みと今後の展望 (2014)
  • Author(s): 鳩谷晋吾
  • Organizer: 第9回 日本獣医再生医療学会
  • Place of Presentation: 大阪
  • Invited
• [Presentation] Generation of induced pluripotent stem cells and production of leukemia inhibitory factor (LIF) in domestic cats (2013)
  • Author(s): Kanegi R, Hatoya S, Takenaka S, Nishimura T, Wijewardana V, Sugiura K, Takahashi M, Kawate N, Tamada H, Inaba T
  • Organizer: The International Society for Stem Cell Research (ISSCR) 11th Annual Meeting
  • Place of Presentation: Boston, USA.
• [Presentation] Generation of functional platelets from canine induced pluripotent stem cells (2013)
  • Author(s): Nishimura T, Hatoya S, Kanegi R, Sugiura K, Wijewardana V, Kuwamura M, Tanaka M, Takahashi M, Kawate N, Tamada H, Imai H, Inaba T
  • Organizer: The International Society for Stem Cell Research (ISSCR) 11th Annual Meeting
  • Place of Presentation: Boston, USA.
• [Presentation] ネコの体外受精法の改善に向けた酸素濃度および顕微授精についての検討 (2013)
  • Author(s): 藤木花奈、鳩谷晋吾、杉浦喜久弥、高橋正弘、川手憲俊、玉田尋通、稲葉俊夫
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 岐阜
• [Presentation] 獣医療における再生医療～iPS細胞の樹立 (2013)
  • Author(s): 鳩谷晋吾
  • Organizer: 内科学アカデミー
  • Place of Presentation: 横浜
  • Invited
• [Presentation] Generation of induced pluripotent stem cells and production of leukemia inhibitory factor (LIF) in domestic cats. (2013)
  • Author(s): Kanegi, R., Hatoya, S., Takenaka, S., Nishimura, T., Wijewardana, V., Sugiura, K., Takahashi, M., Kawate, N., Tamada, H., Inaba, T
  • Organizer: The International Society for Stem Cell Research (ISSCR) 11th Annual Meeting
  • Place of Presentation: ボストン
• [Presentation] Generation of functional platelets from canine induced pluripotent stem cells , International Society for Stem Cell Research (2013)
  • Author(s): Nishimura, T., Hatoya, S., Kanegi, R., Sugiura, K., Wijewardana, V., Kuwamura, M., Tanaka, M., Takahashi, M., Kawate, N., Tamada, H., Imai, H., Inaba, T
  • Organizer: The International Society for Stem Cell Research (ISSCR) 11th Annual Meeting
  • Place of Presentation: ボストン
• [Presentation] ネコiPS細胞株樹立と多能性維持タンパク質ネコLIFの作製 (2012)
  • Author(s): 金城綾二、鳩谷晋吾、杉浦喜久弥、竹中重雄、玉田尋通、川手憲俊、高橋正弘、稲葉俊夫
  • Organizer: 日本獣医学会
  • Place of Presentation: 岩手
• [Presentation] イヌiPS細胞の作製と血小板への分化誘導 (2012)
  • Author(s): 西村俊哉、鳩谷晋吾、杉浦 喜久弥、今井 裕、玉田尋通、川手憲俊、高橋正弘、稲葉 俊夫
  • Organizer: 日本獣医学会
  • Place of Presentation: 岩手
• [Remarks] http://www.vet.osakafu-u.ac.jp/cell/cell.htm
• [Remarks] 研究室ホームページ
  • URL: http://www.vet.osakafu-u.ac.jp/cell/cell.html
• [Remarks] 細胞病態学教室ホームページ
  • URL: http://www.vet.osakafu-u.ac.jp/cell/cell.htm