Identification of factors involved in the maintenance of haploidy and pluripotency of haploid ES cells

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K08578
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: LEE Jiyoung 東京医科歯科大学, 難治疾患研究所, 准教授 (20402860)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 1倍体ES細胞 / ES細胞 / 半数性

Outline of Final Research Achievements

In this study, we aimed to elucidate the signal cascade related to the maintenance of haploidy of haploid ES cells, and performed search for new targets to increase the haploid maintenance efficiency. As a result, it was revealed that the 14-3-3 protein inhibitor R18 exhibits high efficiency of cell proliferation and haploid maintenance ability, and that the ROCK inhibitor, Y27632 prevents the apoptosis of haploid ES cells. Thus, stable establishment of haploid ES cells and optimal culture conditions were established.
Also, the instability of epigenetic states such as DNA methylation in ES cells is one of the problems in using ES cells for regenerative medicine. In this study, we demonstrated that FBS+2i condition is particularly effective in preventing epigenetic instability during both ES establishment and maintenance. Therefore, this condition is useful as the culture condition of epigenetically stable ES cells.

Free Research Field

生物学

Academic Significance and Societal Importance of the Research Achievements

生物に見られる多様な表現型の解明には限界があるため、希望の表現型をもたらす遺伝子変異をスクリーニングするフォワードジェネティクスの方がより根本的な解決策となるため、１倍体ES細胞はこの実験系に大変有効である。本研究では半数性維持に効果的な阻害剤を見だすとともにエピジェネティックに安定なES樹立と維持条件を確立したため、これらの安定的な培養条件は幅広い有用性を持つと考えられる。