Analysis of proliferation and differentiation of primordial germ cell-like cells derived from ES cells

• Project/Area Number: 15H05636
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Integrative animal science
• Research Institution: Kyoto University
• Principal Investigator: Ohta Hiroshi 京都大学, 医学研究科, 助教 (50391892)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥22,750,000 (Direct Cost: ¥17,500,000、Indirect Cost: ¥5,250,000); Fiscal Year 2018: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2017: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2015: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: 始原生殖細胞 / ES細胞 / 配偶子形成 / 増殖 / 分化 / 細胞培養 / 培養 / 細胞増殖 / 細胞分化 / 試験管内

Outline of Final Research Achievements

Sperm and oocytes are originated from primordial germ cells (PGC), which arise at early embryonic development. In this research, we attempted to establish in vitro culture system of PGC by using PGC-like cells (PGCLC) derived from ES cells. We found that forskolin and PDE4 inhibitor (rolipram) synergistically enhance the proliferation of PGCLC, and addition of retinoic acid (vitamin A) and BMP2 is enough to induce female germ cell pathway in vitro.

Academic Significance and Societal Importance of the Research Achievements

始原生殖細胞を解析する上でこれまでの大きな問題点は、試験管内において適切な培養系が存在しないことであった。本研究計画の成果により、試験管内において始原生殖細胞を一定期間増殖させることが可能となり、また、雌の生殖細胞へ分化させるための因子の同定に成功した。今後、本研究により確立された培養系は、雄性生殖細胞の決定機構および減数分裂機構の解明など生殖細胞分化に関わる解析や他の種への応用など幅広い分野に貢献できると考えられる。

Research Products

• [Journal Article] Induction of fetal primary oocytes and the meiotic prophase from mouse pluripotent stem cells. (2018)
  • Author(s): Miyauchi H, Ohta H, Saitou M.
  • Journal Title: Methods Cell Biol. Volume: 144 Pages: 409-429
  • DOI: 10.1016/bs.mcb.2018.03.035
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] In vitro expansion of mouse primordial germ cell-like cells recapitulates an epigenetic blank slate. (2017)
  • Author(s): Ohta H, Kurimoto K, Okamoto I, Nakamura T, Yabuta Y, Miyauchi H, Yamamoto T, Okuno Y, Hagiwara M, Shirane K, Sasaki H, Saitou M
  • Journal Title: EMBO J Volume: 36 Issue: 13 Pages: 1888-1907
  • DOI: 10.15252/embj.201695862
  • Peer Reviewed / Open Access
• [Journal Article] Fertile offspring from sterile sex chromosome trisomic mice (2017)
  • Author(s): Hirota Takayuki、Ohta Hiroshi、Powell Benjamin E.、Mahadevaiah Shantha K.、Ojarikre Obah A.、Saitou Mitinori、Turner James M. A.
  • Journal Title: Science Volume: 357 Issue: 6354 Pages: 932-935
  • DOI: 10.1126/science.aam9046
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bone morphogenetic protein and retinoic acid synergistically specify female germ‐cell fate in mice (2017)
  • Author(s): Miyauchi Hidetaka、Ohta Hiroshi、Nagaoka So、Nakaki Fumio、Sasaki Kotaro、Hayashi Katsuhiko、Yabuta Yukihiro、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: The EMBO Journal Volume: 36 Issue: 21 Pages: 3100-3119
  • DOI: 10.15252/embj.201796875
  • Peer Reviewed / Open Access
• [Journal Article] Expansion of mouse primordial germ cell-like cells in culture reconstitutes an epigenetic blank slate. (2017)
  • Author(s): Hiroshi Ohta, Kazuki Kurimoto, Ikuhiro Okamoto, Tomonori Nakamura, Yukihiro Yabuta, Hidetaka Miyauchi, Takuya Yamamoto, Yukiko Okuno, Masatoshi Hagiwara, Kenjiro Shirane, Hiroyuki Sasaki, Mitinori Saitou.
  • Journal Title: The EMBO Journal Volume: 印刷中
  • Peer Reviewed / Acknowledgement Compliant