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Establish a culture system for somatic ovarian cells that can support in vitro oogenesis

Research Project

Project/Area Number 20K21500
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 48:Biomedical structure and function and related fields
Research InstitutionKyoto University

Principal Investigator

Ohta Hiroshi  京都大学, 医学研究科, 准教授 (50391892)

Project Period (FY) 2020-07-30 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2021: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2020: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords卵子形成 / 再構成卵巣 / 雌性生殖巣体細胞 / 前顆粒膜細胞 / 細胞培養 / PGCLC / マウス / 生殖巣 / 体細胞 / 培養 / 支持細胞 / 始原生殖細胞
Outline of Research at the Start

本研究の目的は、マウスにおいて、卵子形成の支持が可能な胎生期卵巣生殖支持細胞の増殖培養系を確立し、その細胞を用いてES細胞由来始原生殖細胞様細胞からの卵子形成誘導を可能にする事である。本研究が達成される事により、ES細胞由来始原生殖細胞様細胞からの卵子形成誘導が全て特定の培養細胞を用いた培養系で可能となり、生殖細胞の基礎研究のみならず、発生工学、不妊症の機構解明など幅広い分野への応用が期待できる。

Outline of Final Research Achievements

This study aims to establish a culture system for mouse ovarian somatic cells that can support in vitro oogenesis from primordial germ cell-like cells (PGCLCs). Among the ovarian somatic cells, we attempted to establish the culture condition for pre-granulosa cells,as we found that pre-granulosa cells could support in vitro oogenesis from PGCLCs. In addition to this, we analyzed the transcriptomes of pre-granulosa cells in mouse, monkey, and human to find the common cytokine receptors and cell signalling. We expect to improve the current culture condition based on the results obtained from transcriptome analysis.

Academic Significance and Societal Importance of the Research Achievements

哺乳類の卵子形成を理解する上で重要な実験系は試験管内再構成系である。マウスにおいては再構成卵巣を体外培養することにより、卵子形成の全過程を試験管内で再構成することが可能となり、飛躍的に研究が進んでいる。本研究の目的は再構成卵巣に必要な細胞種を同定し、さらにその細胞種を培養することである。本研究によりマウスの再構成卵巣には前顆粒膜細胞が必要であることが明らかとなり、さらに、マウス、カニクイザル、ヒトの前顆粒膜細胞に共通して発現する栄養因子受容体および細胞内シグナルを同定した。今後これらの情報を基に、試験管内で卵子形成を支持可能な細胞種の培養系の確立が期待できる。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • Research Products

    (8 results)

All 2023 2022 2021 2020

All Journal Article (8 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 8 results,  Open Access: 6 results)

  • [Journal Article] Generation of functional oocytes from male mice in vitro2023

    • 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

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] <i>Ex vivo</i> reconstitution of fetal oocyte development in humans and cynomolgus monkeys2022

    • 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

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Optimized protocol to derive germline stem-cell-like cells from mouse pluripotent stem cells2022

    • Author(s)
      Ishikura Yukiko、Ohta Hiroshi、Nagano Masahiro、Saitou Mitinori
    • Journal Title

      STAR Protocols

      Volume: 3 Issue: 3 Pages: 101544-101544

    • DOI

      10.1016/j.xpro.2022.101544

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Nucleome programming is required for the foundation of totipotency in mammalian germline development2022

    • Author(s)
      Nagano Masahiro, Hu Bo, Yokobayashi Shihori et al.
    • Journal Title

      The EMBO Journal

      Volume: 41 Issue: 13

    • DOI

      10.15252/embj.2022110600

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] The embryonic ontogeny of the gonadal somatic cells in mice and monkeys2021

    • 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

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] In vitro reconstitution of the whole male germ-cell development from mouse pluripotent stem cells2021

    • 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

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Long‐term expansion with germline potential of human primordial germ cell‐like cells in?vitro2020

    • 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

    • Related Report
      2020 Research-status Report
    • 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

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access

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Published: 2020-08-03   Modified: 2024-01-30  

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