Development of new procedure to make chimeric animals without using naive pluripotent stem cells

• Project/Area Number: 15K20948
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry; Laboratory animal science
• Research Institution: The University of Tokyo
• Principal Investigator: Masaki Hideki 東京大学, 医科学研究所, 助教 (20571988)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: ES細胞 / iPS細胞 / 再生医療 / 臓器再生 / キメラ / 遺伝子改変動物 / ウサギ / 多能性幹細胞 / 臓器

Outline of Final Research Achievements

Our group had shown that forced expression of anti-apoptotic factor enables developmental stage mismatched cells, such as primed state pluripotent stem cells or endodermal progenitor cells, to form chimera with pre-implantation mouse embryos. In this research, we tried to clarify this approach is also applicable for non-rodent cells, and/or other lineage progenitor cells except for endodermal progenitor cells. As a result, we observed that rabbit ESCs formed chimera with mouse and rabbit pre-implantation embryo by forced expression of anti-apoptotic gene, whereas control rabbit ESCs did not form any chimeras. We also observed heart region specific chimera formation with Nkx2.5 expressing cardiomyocyte progenitor cells by preventing apoptosis. Therefore, we conclude that promotion of chimera formation via inhibition of apoptosis can be applicable not only for rodent, and not only for endodermal progenitors.

Research Products

• [Journal Article] Interspecific in vitro assay for the chimera-forming ability of human pluripotent stem cells. (2015)
  • Author(s): Masaki H, Kato-Itoh M, Umino A, Sato H, Hamanaka S, Kobayashi T, Yamaguchi T, Nishimura K, Ohtaka M, Nakanishi M, Nakauchi H.
  • Journal Title: Development. Volume: 142(18) Pages: 3222-30
  • DOI: 10.1242/dev.124016
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] A UNIQUE SYSTEM TO ASSESS THE IN VIVO LINEAGE POTENTIALS OF DEVELOPMENTALLY-ADVANCED CELLS BY BLASTOCYST INJECTION (2016)
  • Author(s): Hideki Masaki
  • Organizer: Annual Meeting of International Society for Stem Cell Research
  • Place of Presentation: Moscone Center, San Francisco CA, USA
  • Year and Date: 2016-06-22
  • Int'l Joint Research
• [Presentation] Predict the unpredictable: using the in vivo niche to assess lineage potential and functionality of PSC-derived cells (2016)
  • Author(s): Hideki Masaki
  • Organizer: Annual Meeting of International Society for Stem Cell Research
  • Place of Presentation: Moscone Center, San Francisco CA, USA
  • Year and Date: 2016-06-22
  • Int'l Joint Research
• [Presentation] A unique system to assess the in vivo lineage potentials of developmentally-advanced progenitors (2016)
  • Author(s): 正木英樹
  • Organizer: 日本再生医療学会年会
  • Place of Presentation: 大阪国際展示場（大阪府大阪市）
  • Year and Date: 2016-03-17
• [Presentation] 幹細胞研究における生体環境利用の有用性 (2016)
  • Author(s): 正木英樹
  • Organizer: 日本再生医療学会
  • Place of Presentation: 大阪国際展示場（大阪府大阪市）
  • Year and Date: 2016-03-17
• [Presentation] Novel chimera forming technology by preventing cell death (2015)
  • Author(s): 正木英樹
  • Organizer: 日本分子生物学会年会
  • Place of Presentation: 神戸国際展示場（兵庫県神戸市）
  • Year and Date: 2015-12-01