Establishment of culture system for inducing oocytes from primordial germ cell-like cells

• Project/Area Number: 19H03618
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Kyoto University
• Principal Investigator: Ohta Hiroshi 京都大学, 医学研究科, 准教授 (50391892)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,030,000 (Direct Cost: ¥13,100,000、Indirect Cost: ¥3,930,000); Fiscal Year 2022: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2020: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2019: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
• Keywords: 卵原細胞 / 卵母細胞 / 卵子形成 / 再構成卵巣 / 始原生殖細胞 / 試験管内誘導 / ヒト / 多能性幹細胞 / 始原生殖細胞様細胞 / 試験管内 / 胎生期生殖巣 / iPS細胞 / in vitro / 細胞分化

Outline of Research at the Start

精子や卵子の元となる細胞は胎生期に発生する始原生殖細胞に由来するが、ヒトを含めた霊長類では実験的に研究が困難なことから不明な点が数多く残されている。本研究ではヒト多能性幹細胞から誘導した始原生殖細胞を卵母細胞へ分化させることが可能な実験系を確立する。本研究計画が達成される事により、胎児医学、ヒト発生学、生殖医療など、幅広い分野において重要な知識基盤を形成する事が可能となる。

Outline of Final Research Achievements

For inducing in vitro oogenesis from human iPS cells via primordial germ cell-like cells (PGCLC), in this study, we attempted to establish the culture system that possible to induce human oogenesis in vitro. We tested and optimized the culture conditions and found that the primordial follicles can be induced from ooginia by culturing reconstituted ovaries for 15 weeks. In vitro induced primordial follicles exhibited the gene expression profiles similar with in vivo primordial follicles. Further improvement will enable us to induce in vitro oogenesis from human PGCLCs.

Academic Significance and Societal Importance of the Research Achievements

生殖細胞は次世代へ遺伝情報を伝えることができる唯一の細胞系譜であり、種の継続性や進化を担う重要な細胞種である。マウスにおいては多能性幹細胞から誘導した始原生殖細胞様細胞を用いて卵子形成を試験管内誘導することが可能となっており、生殖細胞の研究が飛躍的に進んでいる。しかしながら、ヒトにおいては卵子形成を試験管内で誘導する実験系が確立されておらず大きな研究の障壁となっていた。本研究では、あらゆる培養条件を検討することによりヒト卵原細胞から原始卵胞を誘導することに成功した。今後本培養系を用いることにより始原生殖細胞様細胞からの卵子形成誘導が期待できる。

Research Products

• [Journal Article] Generation of functional oocytes from male mice in vitro (2023)
  • Author(s): Murakami Kenta、Hamazaki Nobuhiko、Hamada Norio、Nagamatsu Go、Okamoto Ikuhiro、Ohta Hiroshi、Nosaka Yoshiaki、Ishikura Yukiko、Kitajima Tomoya S.、Semba Yuichiro、Kunisaki Yuya、Arai Fumio、Akashi Koichi、Saitou Mitinori、Kato Kiyoko、Hayashi Katsuhiko
  • Journal Title: Nature Volume: 615 Issue: 7954 Pages: 900-906
  • DOI: 10.1038/s41586-023-05834-x
  • Peer Reviewed
• [Journal Article] Nucleome programming is required for the foundation of totipotency in mammalian germline development (2022)
  • Author(s): Nagano Masahiro, Hu Bo, Yokobayashi Shihori et al.
  • Journal Title: The EMBO Journal Volume: 41 Issue: 13
  • DOI: 10.15252/embj.2022110600
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] <i>Ex vivo</i> reconstitution of fetal oocyte development in humans and cynomolgus monkeys (2022)
  • Author(s): Mizuta Ken、Katou Yoshitaka、Nakakita Baku、Kishine Aoi、Nosaka Yoshiaki、Saito Saki、Iwatani Chizuru、Tsuchiya Hideaki、Kawamoto Ikuo、Nakaya Masataka、Tsukiyama Tomoyuki、Nagano Masahiro、Kojima Yoji、Nakamura Tomonori、Yabuta Yukihiro、Horie Akihito、Mandai Masaki、Ohta Hiroshi、Saitou Mitinori
  • Journal Title: The EMBO Journal Volume: 41 Issue: 18
  • DOI: 10.15252/embj.2022110815
  • Peer Reviewed / Open Access
• [Journal Article] The embryonic ontogeny of the gonadal somatic cells in mice and monkeys (2021)
  • Author(s): Sasaki Kotaro、Oguchi Akiko、Cheng Keren、Murakawa Yasuhiro、Okamoto Ikuhiro、Ohta Hiroshi、Yabuta Yukihiro、Iwatani Chizuru、Tsuchiya Hideaki、Yamamoto Takuya、Seita Yasunari、Saitou Mitinori
  • Journal Title: Cell Reports Volume: 35 Issue: 5 Pages: 109075-109075
  • DOI: 10.1016/j.celrep.2021.109075
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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] Long‐term expansion with germline potential of human primordial germ cell‐like cells in?vitro (2020)
  • Author(s): Murase Yusuke、Yabuta Yukihiro、Ohta Hiroshi、Yamashiro Chika、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: The EMBO Journal Volume: 39 Issue: 21 Pages: 104929-104929
  • DOI: 10.15252/embj.2020104929
  • Peer Reviewed
• [Journal Article] Cyclosporin A and FGF signaling support the proliferation/survival of mouse primordial germ cell-like cells in vitro† (2020)
  • Author(s): Ohta Hiroshi、Yabuta Yukihiro、Kurimoto Kazuki、Nakamura Tomonori、Murase Yusuke、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Biology of Reproduction Volume: 104 Issue: 2 Pages: 344-360
  • DOI: 10.1093/biolre/ioaa195
  • Peer Reviewed / Open Access
• [Journal Article] Induction of the germ cell fate from pluripotent stem cells in cynomolgus monkeys. (2020)
  • Author(s): Sakai Y, Nakamura T, Okamoto I, Gyobu-Motani S, Ohta H, Yabuta Y, Tsukiyama T, Iwatani C, Tsuchiya H, Ema M, Morizane A, Takahashi J, Yamamoto T, Saitou M.
  • Journal Title: Biol Reprod Volume: 102(3) Issue: 3 Pages: 620-638
  • DOI: 10.1093/biolre/ioz205
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ZGLP1 is a determinant for the oogenic fate in mice (2020)
  • Author(s): Nagaoka So I.、Nakaki Fumio、Miyauchi Hidetaka、Nosaka Yoshiaki、Ohta Hiroshi、Yabuta Yukihiro、Kurimoto Kazuki、Hayashi Katsuhiko、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Science Volume: 367 Issue: 6482 Pages: 4115-4115
  • DOI: 10.1126/science.aaw4115
  • NAID: 120006850276
  • Peer Reviewed / Int'l Joint Research