In vitro reconstitution of male germ cell development from pluripotent stem cells

• Project/Area Number: 17H06802
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: General medical chemistry
• Research Institution: Kyoto University
• Principal Investigator: Ishikura Yukiko 京都大学, 医学研究科, 特定研究員 (00807943)
• Project Period (FY): 2017-08-25 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 精原幹細胞 / 多能性幹細胞 / 始原生殖細胞 / 生殖細胞 / リプログラミング / 試験管内再構成

Outline of Final Research Achievements

The mammalian germline lineage begins at the fertilized egg and differentiates into gametes via PGCs capable of differentiating male and female.In the process, important controls such as epigenome reprogramming, sexual differentiation and meiosis are performed.
In recent years, an extracorporeal culture system has been reported in which oocyte-like cells and spermatogonia stem cell-like cells are induced from mouse pluripotent stem cells via PGC-like cells (PGC-like cells; PGCLCs).On the other hand, cells induced by these in vitro culture systems exhibited various abnormalities caused by epigenome abnormalities and the like.
In this study, we will develop a methodology to induce GSCLCs with high spermatogenic efficiency from pluripotent stem cells by improving the reconstructed testis method to one more compliant with the conditions of the living body.

Academic Significance and Societal Importance of the Research Achievements

本実験系が確立すれば、雄性生殖細胞分化機構、雄性エピゲノム獲得・制御機構、精原幹細胞の分化機構などの、雄性生殖細胞発生過程に附随する、生物学的に重要な様々な事象が、多能性幹細胞を用いたゲノム編集技術と組み合わせ、全て試験管内で解析可能となると期待される。また、本実験系は、医学生物学的に重要な、サルやヒト、もしくは他の希少哺乳類の多能性幹細胞を起点とした、雄性生殖細胞系譜の誘導研究とその応用の基盤となると期待される。

Research Products

• [Journal Article] In vitro reconstitution of the whole male germ-cell development from mouse pluripotent stem cells (2021)
  • Author(s): Ishikura Yukiko、Ohta Hiroshi、Sato Takuya、Murase Yusuke、Yabuta Yukihiro、Kojima Yoji、Yamashiro Chika、Nakamura Tomonori、Yamamoto Takuya、Ogawa Takehiko、Saitou Mitinori
  • Journal Title: Cell Stem Cell Volume: 28 Issue: 121 Pages: 2167-2179.e9
  • DOI: 10.1016/j.stem.2021.08.005
  • Peer Reviewed / Open Access
• [Journal Article] Generation of human oogonia from induced pluripotent stem cells in vitro. (2018)
  • Author(s): 30.Yamashiro C, Sasaki K, Yabuta Y, Kojima Y, Nakamura T, Okamoto I, Yokobayashi S, Murase Y, Ishikura Y, Shirane K, Sasaki H, Yamamoto T, Saitou M.
  • Journal Title: Science Volume: 362(6412) Issue: 6412 Pages: 356-360
  • DOI: 10.1126/science.aat1674
  • Peer Reviewed
• [Journal Article] In Vitro Spermatogenesis (2018)
  • Author(s): Yukiko Ishikura, Mitinori Saitou
  • Journal Title: Reference Module in Biomedical Sciences Volume: 3 Pages: 1-10
  • DOI: 10.1016/b978-0-12-801238-3.64441-0
  • ISBN: 9780128151457
  • Peer Reviewed / Int'l Joint Research
• [Book] 雄性生殖細胞全発生過程の試験管内再構成 (2022)
  • Author(s): 石藏 友紀子
  • Pages: 1440
  • Total Pages: 131
  • Publisher: 羊土社
  • DOI: 10.18958/7011-00003-0000154-00
• [Patent(Industrial Property Rights)] 多能性幹細胞から精子幹細胞を誘導する方法論 (2021)
  • Inventor(s): 斎藤通紀、石藏友紀子
  • Industrial Property Rights Holder: 京都大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2021
• [Patent(Industrial Property Rights)] 多能性幹細胞から生殖系列幹細胞様細胞への分化誘導方法 (2019)
  • Inventor(s): 斎藤通紀、石藏友紀子
  • Industrial Property Rights Holder: 京都大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-523955
  • Filing Date: 2019
  • Overseas