The effective method for generation of platelets from canine iPS cells by perfusion cocultivation

• Project/Area Number: 15K14888
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Integrative animal science
• Research Institution: Osaka Prefecture University
• Principal Investigator: INABA TOSHIO 大阪府立大学, 生命環境科学研究科, 教授 (00137241)
• Co-Investigator(Kenkyū-buntansha): 杉浦 喜久弥 大阪府立大学, 生命環境科学研究科, 准教授 (30171143) ; 鳩谷 晋吾 大阪府立大学, 生命環境科学研究科, 准教授 (40453138)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 再生医療 / 統合動物科学 / ｉＰＳ細胞 / 血小板 / バイオテクノロジー / トランスレーショナルリサーチ / iPS細胞

Outline of Final Research Achievements

The objective of this study was in the development of the effective method for generation of platelets from canine iPS cells, and the following results were obtained. We generated canine hematopoietic cytokine-secreting feeder cells by transfection of the genes to OP9 cells. While we established a stable and long-term culture of canine iPS cells, we could not differentiate them into platelets. We generated new canine stem cell lines that closely resemble to extraembryonic endoderm cells. These cell lines will apply to platelet production.

Research Products

• [Journal Article] Feeder-independent canine induced pluripotent stem cells maintained under serum-free conditions (2017)
  • Author(s): Nishimura T, Hatoya S, Kanegi R, Wijesekera DP, Sanno K, Tanaka E, Sugiura K, Hiromitsu Tamada NK, Imai H, Inaba T.
  • Journal Title: Molecular Reproduction and Development Volume: 印刷中 Issue: 4 Pages: 329-339
  • DOI: 10.1002/mrd.22789
  • Peer Reviewed / Open Access
• [Journal Article] Generation of canine induced extraembryonic endoderm-like cell line that forms both extraembryonic and embryonic endoderm derivatives (2017)
  • Author(s): Nishimura T, Unezaki N, Kanegi R, Wijesekera DPH, Hatoya S, Sugiura K, Kawate N, Tamada H, Imai H, Inaba T.
  • Journal Title: Stem Cells and Development Volume: 印刷中 Issue: 15 Pages: 1111-1120
  • DOI: 10.1089/scd.2017.0026
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Production of feline leukemia inhibitory factor with biological activity in Escherichia coli. (2016)
  • Author(s): Kanegi R, Hatoya S, Tsujimoto Y, Takenaka S, Nishimura T, Wijewardana V, Sugiura K, Takahashi M, Kawate N, Tamada H, Inaba T.
  • Journal Title: Theriogenology Volume: 印刷中 Issue: 2 Pages: 604-611
  • DOI: 10.1016/j.theriogenology.2016.02.013
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Methylprednisolone sodium succinate reduces spinal cord swelling but does not affect recovery of dogs with surgically treated thoracolumbar intervertebral disk herniation (2016)
  • Author(s): Nishida H, Tanaka H, Kitamura M, Inaba T, Nakayama M.
  • Journal Title: Japanese Journal of Veterinary Research Volume: 64 Issue: 3 Pages: 191-196
  • DOI: 10.14943/jjvr.64.3.191
  • NAID: 120005828519
  • ISSN: 0047-1917
  • Peer Reviewed / Open Access
• [Journal Article] Exosomes derived from tumor cells genetically modified to express Mycobacterium tuberculosis antigen: a novel vaccine for cancer therapy. (2016)
  • Author(s): Koyama Y, Ito T, Hasegawa A, Eriguchi M, Inaba T, Ushigusa T, Sugiura K.
  • Journal Title: Biotechnol Lett Volume: 38 Issue: 11 Pages: 1857-1866
  • DOI: 10.1007/s10529-016-2185-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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 DPH, Hatoya S, Nishimura T, Kanegi R, Ushigusa T, Inaba T.
  • Journal Title: Journal of Veterinary Medical Science Volume: 77 Issue: 7 Pages: 771-775
  • DOI: 10.1292/jvms.14-0558
  • NAID: 130005089954
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Highly effective non-viral antitumor gene therapy system comprised of biocompatible small plasmid complex particles consisting of pDNA, anionic polysaccharide, and fully deprotected linear polyethylenimine (2015)
  • Author(s): Yoshiyuki Koyama, Kikuya Sugiura, Chieko Yoshihara, Toshio Inaba, Tomoko Ito
  • Journal Title: Pharmaceutics Volume: 7 Issue: 3 Pages: 152-164
  • DOI: 10.3390/pharmaceutics7030152
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ヒト再生医療の実現に向けたイヌiPS細胞の創製 (2015)
  • Author(s): 稲葉俊夫
  • Journal Title: ケミカルエンジニアリング Volume: 61 Pages: 119-125
• [Presentation] "Artificial neoepitope"-bearing exosomes as novel vaccine for cancer immunotherapy (2016)
  • Author(s): Koyama Y, Ito T, Inaba T, Sugiura K
  • Organizer: 第45回日本免疫学会学術集会
  • Place of Presentation: 沖縄コンベンションセンター（沖縄県宜野湾市）
  • Year and Date: 2016-12-05
• [Presentation] 結核菌抗原遺伝子を用いた腫瘍治療メカニズムの解明 (2016)
  • Author(s): 長谷川綾、牛草貴博、小山義之、伊藤智子、鳩谷晋吾、杉浦喜久弥、稲葉俊夫
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 日本大学生物資源科学部（神奈川県藤沢市）
  • Year and Date: 2016-09-06
• [Presentation] ネコiPS細胞株のセンダイウイルスベクターを用いた作製 (2016)
  • Author(s): 沖田良太、鳩谷晋吾、杉浦喜久弥、大高真奈美、西村健、中西真人、稲葉俊夫
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 日本大学生物資源科学部（神奈川県藤沢市）
  • Year and Date: 2016-09-06
• [Presentation] ウシ卵子の体外成熟培養時におけるMAPKK阻害剤 (U0126) 添加の胚発生に及ぼす影響 (2016)
  • Author(s): 篠原健志、太田祥弘、川手憲俊、高橋正弘、坂上信忠、稲葉俊夫、玉田尋通
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 日本大学生物資源科学部（神奈川県藤沢市）
  • Year and Date: 2016-09-06
• [Presentation] イヌ体細胞を用いた人工誘導胚体外内胚葉細胞の作製と肝細胞への分化誘導 (2016)
  • Author(s): 畦﨑直哉、西村俊哉、金城綾二、Daluthgamage Patsy Himali Wijesekera、鳩谷晋吾、杉浦喜久弥、川手憲俊、玉田尋通、今井裕、稲葉俊夫
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 日本大学生物資源科学部（神奈川県藤沢市）
  • Year and Date: 2016-09-06
• [Presentation] 「人工ネオエピトープ」を担持した腫瘍由来エクソソームによるガン免疫治療 (2016)
  • Author(s): 小山義之、伊藤智子、長谷川綾、杉浦喜久弥、稲葉俊夫、江里口正純
  • Organizer: 第3回日本細胞外小胞学会
  • Place of Presentation: グランドプリンスホテル広島（広島県広島市）
  • Year and Date: 2016-08-31
• [Presentation] Manipulation of tumor microenvironment by cytokine gene transfection enhances dendritic cell-based immunotherapy (2016)
  • Author(s): Wijesekera D.P.H., Sugiura K, Ichida C, Yuba E, Kono K, Yamate J, Kanegi R, Nishimura T, Nadeeka H.D., Hatoya S, Inaba T.
  • Organizer: ICI 2016-International Congress of Immunology 2016
  • Place of Presentation: Melbourne Convention and Exhibition Centre （Melbourne, Australia）
  • Year and Date: 2016-08-21
  • Int'l Joint Research
• [Presentation] Manipulation of tumor microenvironment by cytokine gene transfection enhances dendritic cell-based immunotherapy (2016)
  • Author(s): Wijesekera DPH, Sugiura K, Ichida C, Yuba E, Kono K, Yamate J, Kanegi R, Nishimura T, De Silva NH, Hatoya S, Inaba T.
  • Organizer: 16th International Congress of Immunology
  • Place of Presentation: Melbourne Convention and Exhibition Centre, Melbourne, Australia
  • Year and Date: 2016-08-21
  • Int'l Joint Research
• [Presentation] Generation of multipotent canine extraembryonic endoderm-like cells via the formation of induced pluripotent stem cells. (2016)
  • Author(s): Unezaki N, Nishimura T, Kanegi R, Wijesekera DPH, Hatoya S, Sugiura K, Kawate N, Tamada H, Imai H, Inaba T.
  • Organizer: The International Society for Stem Cell Research (ISSCR) 14th Annual Meeting
  • Place of Presentation: Moscone West（San Francisco, USA）
  • Year and Date: 2016-06-22
  • Int'l Joint Research
• [Presentation] Endoderm-like cells via the formation of induced pluripotent stem cells (2016)
  • Author(s): Unezaki N, Nishimura T, Kanegi R, Wijesekera, DPH, Hatoya S, Sugiura K, Kawate N, Tamada H, Imai H, Inaba T.
  • Organizer: The International Society for Stem Cell Research 14th Annual Meeting
  • Place of Presentation: Moscone Center West, San Francisco, USA
  • Year and Date: 2016-06-22
  • Int'l Joint Research
• [Presentation] "Artificial neoepitope"- bearing exosomes derived from tumor cells being genetically modified to express Mycobacterium tuberculosis antigen: a novel vaccine for cancer therapy (2016)
  • Author(s): Koyama Y, Ito T, Eriguchi M, Hasegawa A, Inaba T, Sugiura K.
  • Organizer: The 5th Annual Meeting of the International Society for Extracellular Vesicles
  • Place of Presentation: De Doelen Concert and Congress Center （Rotterdam, Netherlands）
  • Year and Date: 2016-05-04
  • Int'l Joint Research
• [Presentation] "Artificial neoepitope"- bearing exosomes derived from tumor cells being genetically modified to express Mycobacterium tuberculosis antigen: a novel vaccine for cancer therapy (2016)
  • Author(s): Koyama Y, Ito T, Eriguchi M, Hasegawa A, Inaba T, Sugiura K.
  • Organizer: The 5th Annual Meeting of the International Society for Extracellular Vesicles
  • Place of Presentation: De Doelen International Congress Centre, Rotterdam, Netherlands
  • Year and Date: 2016-05-04
  • Int'l Joint Research
• [Presentation] イヌiPS細胞のマトリゲル上での培養と間葉系幹細胞への分化誘導 (2016)
  • Author(s): 西村俊哉、田中恵里菜、鳩谷晋吾、金城綾二、Wijesekera, D.P.H.、杉浦喜久弥、玉田尋通、川手憲俊、今井 裕、稲葉俊夫
  • Organizer: 第15回日本再生医療学会総会
  • Place of Presentation: 大阪国際会議場(大阪府大阪市)
  • Year and Date: 2016-03-17
• [Presentation] Feeder-independent and serum-free canine induced pluripotent stem cells reprogrammed by a drug-inducible system (2015)
  • Author(s): Nishimura T, Kanegi R, Wijesekera, DPH, Sanno K, Tanaka E, Sugiura K, Hatoya S, Kawate N, Tamada H, Imai H, Inaba T.
  • Organizer: The International Society for Stem Cell Research 13th Annual Meeting
  • Place of Presentation: Stockholmsmassan Exhibition and Convention Center, Stockholm, Sweden
  • Year and Date: 2015-06-24
  • Int'l Joint Research
• [Remarks] Laboratory of Cell Pathobiology
  • URL: http://www.vet.osakafu-u.ac.jp/english/cell/
• [Patent(Industrial Property Rights)] イヌ人工誘導胚体外内胚葉細胞様株の作製方法 (2016)
  • Inventor(s): 稲葉俊夫、西村俊哉、畦﨑直哉、鳩谷晋吾、杉浦喜久弥
  • Industrial Property Rights Holder: 大阪府立大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-116612
  • Filing Date: 2016-06-10
• [Patent(Industrial Property Rights)] イヌｉＰＳ細胞の製造方法 (2015)
  • Inventor(s): 稲葉俊夫、西村俊哉、田中恵里菜、鳩谷晋吾、杉浦喜久弥
  • Industrial Property Rights Holder: 公立大学法人大阪府立大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-123775
  • Filing Date: 2015-06-19