Establishment of chimeric mice having a primatized intervertebral discs and comprehensive research of the primatized intervertebral disc.

• Project/Area Number: 25670653
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Orthopaedic surgery
• Research Institution: Kumamoto University
• Principal Investigator: SEMBA KEI 熊本大学, 学内共同利用施設等, 研究員 (00398190)
• Co-Investigator(Renkei-kenkyūsha): ARAKI KIMI 熊本大学生命資源研究, 支援センター・疾患モデル分野, 教授 (90211705)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2015: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 椎間板 / 霊長類化椎間板キメラ / チンパンジーiPS / Sd変異マウス / Ptf1a / CRISPR-Cas9 / 椎間板髄核マーカー分子 / 椎間板線維輪マーカー分子 / マウス / 霊長類 / 細胞移植 / 細胞間ネットワーク / 霊長類iPS / 霊長類ES細胞 / 椎間板髄核 / 椎間板線維輪 / 椎間板髄核欠損 / 臓器欠損 / キメラマウス

Outline of Final Research Achievements

This research goal is the establishment of the experimental animal having a humanized intervertebral disc. However, nowadays, the production of a humanized chimeric animal is strongly restricted. In this study, a chimpanzee was used to be established such as an experimental animal having primatized intervertebral discs. At first, we microinjected chimpanzee induced pluripotent stem cells (induced pluripotent stem cells: iPS cells) into mutant mouse fertilized eggs. And, we examined whether isolated single chimpanzee iPS cell contributed to development of organ in chimeric mice. However, unfortunately we found that isolated chimpanzee iPS cell did not contribute to mouse development, using the Ptf1a-null homogenous fertilized egg. These results suggested that the transplant of isolated cells into the different species is difficult to engraft in a primordium during development because of mismatch neighboring environment of intercellular communication between a primate and a rodent.

Research Products

• [Journal Article] GNAS and Kras cooperate to promote murine pancreatic tumorigenesis recapitulating human intraductal papillary mucinous neoplasm. (2016)
  • Author(s): Taki, K., Ohmuraya, M., Tanji, E., Komatsu, H., Hashimoto, D., Semba, K., Araki, K., Kawaguchi, Y., Baba, H. and Furukawa, T.
  • Journal Title: Oncogene Volume: in press Issue: 18 Pages: 1-6
  • DOI: 10.1038/onc.2015.294
  • Peer Reviewed / Open Access
• [Journal Article] Etiology of caudal regression syndrome. (2013)
  • Author(s): Semba K. & Yamamura, K.
  • Journal Title: Human Genet. Embryol. Volume: 3 Issue: 02 Pages: 1-7
  • DOI: 10.4172/2161-0436.1000107
  • Peer Reviewed
• [Journal Article] Ectopic expression of Ptf1a induces spinal defects, urogenital defects and anorectal malformations in Danforth's short tail mice. (2012)
  • Author(s): Semba, K., Araki, K., Matsumoto, K.-I., Suda, H., Ando, T. Sei, A., Mizuta, H., Takagi, K., Nakahara, M., Muta, M., Yamada, G., Nakagata, N., Iida, A., Ikegawa, Y., Araki, M., Abe, K. and Yamamura, K.
  • Journal Title: PLoS Genet. Volume: Vol.9 Issue: 2 Pages: 1-17
  • DOI: 10.1371/journal.pgen.1003204
  • Peer Reviewed
• [Presentation] 発生期の脊索・中腎・総排泄腔でのPtf1a異所性発現が尾部退行症候群を発症させる (2015)
  • Author(s): 仙波 圭
  • Organizer: 第31回日本糖尿病・妊娠学会
  • Place of Presentation: リーガロイヤルホテル東京
  • Year and Date: 2015-11-20
• [Presentation] 椎間板線維輪細胞・髄核細胞のマーカー分子になり得るPax1・Sktの変性椎間板・発生異常椎間板における発現動態の解析 (2015)
  • Author(s): 仙波 圭
  • Organizer: 第30回日本整形外科学会基礎学術集会
  • Place of Presentation: 富山国際会議場
  • Year and Date: 2015-10-22
• [Presentation] Ptf1a遺伝子の脊索細胞での異所性発現が脊椎欠損・椎間板変性を発症させる (2015)
  • Author(s): 仙波 圭
  • Organizer: 第88回日本整形外科学会総会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2015-05-21
• [Presentation] 椎間板髄核細胞・線維輪細胞のマーカ分子になり得る Skt・Pax1 の加齢における 発現動態の解析 (2015)
  • Author(s): 仙波 圭
  • Organizer: 第28回日本軟骨代謝学会
  • Place of Presentation: 東京医科歯科大学鈴木章夫記念講堂(東京都)
  • Year and Date: 2015-03-06 – 2015-03-07
• [Presentation] 軟骨細胞内のオートファジー発生とその病態解析 (2014)
  • Author(s): 仙波 圭
  • Organizer: 第29回日本整形外科学会基礎学術集会
  • Place of Presentation: 鹿児島市城山観光ホテル(鹿児島市)
  • Year and Date: 2014-10-09 – 2014-10-10
  • Invited
• [Presentation] 椎間板変性モデルマウス(Danforth’s short tail)の発症原因の解明 (2014)
  • Author(s): 仙波 圭
  • Organizer: 第29回日本整形外科学会基礎学術集会
  • Place of Presentation: 鹿児島市城山観光ホテル(鹿児島市)
  • Year and Date: 2014-10-09 – 2014-10-10
• [Presentation] 脊索欠損から椎間板変性を引き起こすPtf1a異所性発現の機能解析 (2014)
  • Author(s): 仙波 圭
  • Organizer: 第27回日本軟骨代謝学会
  • Place of Presentation: 京都府医師会館, 京都市
• [Presentation] Caudal regression in Danforth’s short tail is caused by the ectopic expression of Ptf1a induced by the insertion of a transposon. (2013)
  • Author(s): Kei Semba
  • Organizer: The 27th International Mammalian Genome Conference
  • Place of Presentation: Colegio Fonseca, Salamanca, Spain
• [Presentation] Ectopic expression of Ptf1a induces spinal defects, urogenital defects, and anorectal malformations in Danforth’s short tail mice. (2013)
  • Author(s): Kei Semba
  • Organizer: The 26th annual Mouse Molecular Genetics meeting
  • Place of Presentation: Wellcome Trust Genome Campus, Cambridge, UK
• [Presentation] 尾部退行症候群の疾患モデル動物であるDanforth’s short tail変異マウスの解析 (2013)
  • Author(s): 仙波 圭
  • Organizer: 第60回日本実験動物学会
  • Place of Presentation: つくば国際会議場, つくば市
• [Remarks] 熊本大学生命資源・研究支援センター・疾患モデル分野
  • URL: http://irda-genetics.kuma-u.jp/publication/p02.html
• [Remarks] http://irda-genetics.kuma-u.jp/index.html
• [Remarks] 仙波圭 原著論文、学会発表
  • URL: http://irda-genetics.kuma-u.jp/publication/p07.html
• [Remarks] 仙波圭 研究内容
  • URL: http://irda-genetics.kuma-u.jp/research/r06.html