Development of new methods for producing chimeric common marmosets

• Project/Area Number: 15H02360
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Central Institute for Experimental Animals
• Principal Investigator: Sasaki Erika 公益財団法人実験動物中央研究所, マーモセット医学生物学研究部, 部長 (70390739)
• Co-Investigator(Kenkyū-buntansha): 高島 康弘 京都大学, iPS細胞研究所, 特定拠点講師 (70469930)
• Project Period (FY): 2015-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥42,380,000 (Direct Cost: ¥32,600,000、Indirect Cost: ¥9,780,000); Fiscal Year 2019: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000); Fiscal Year 2018: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2017: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2016: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000); Fiscal Year 2015: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
• Keywords: マーモセット / 幹細胞 / 初期胚 / 初期発生 / 胚発生 / エピブラスト / ナイーブ / キメラ / 霊長類 / 発生工学

Outline of Final Research Achievements

To produce allogenic ICM injected chimeric marmoset embryos, efficient processing method of ICM has been examined and contributions of the donor ICM cells into the host ICM have been observed by live cell imaging technic.
OCT4 distal enhancer is known as a marker for human naive pluripotent stem cells. Therefore marmoset ES cells with reporter gene at homologous region to human OCT4 distal enhancer in marmoset were established. Using this ES cell line, marmoset primed ES cells were converted morphologically similar to human naive pluripotent stem cells. To analyze kinetics of chimeric marmoset embryos, a pseudo post-implantation embryo culture method that allows in vitro observation of post-implantation embryo development has been developed. Finally, using allogenic ICM transplanted chimeric embryo production method, marmoset naive ES cells were injected into marmoset host embryos and kinetics of the naive ES cells in the embryo was analyzed by pseudo post-implantation embryo culture.

Academic Significance and Societal Importance of the Research Achievements

本研究を通じて、マーモセットnaive細胞の樹立、疑似着床後胚培養法、同種キメラ胚作製技術など多くの技術が開発された。これにより、非ヒト霊長類の初期発生の理解がより進むことが期待される。更にこれら研究技術を応用し、研究が発展することにより、習慣性流産や不育症などの原因究明に資すると期待される。

Research Products

• [Int'l Joint Research] University of Cambridge(英国)
• [Presentation] Genome editing in non-human primates (2016)
  • Author(s): Erika Sasaki
  • Organizer: The Wellcome Trust Sanger Institute AZ CRISPR Conference
  • Place of Presentation: The Wellcome Trust Sanger Institute, Cambridge,英国
  • Year and Date: 2016-01-17
  • Int'l Joint Research / Invited