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Long-term culture of human spermatogonial stem cells

Research Project

Project/Area Number 18H02935
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 56030:Urology-related
Research InstitutionKyoto University

Principal Investigator

Shinohara Mito  京都大学, 医学研究科, 助教 (10372591)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000)
Fiscal Year 2020: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2019: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2018: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Keywords精子 / 幹細胞 / 生殖 / 不妊症 / 不妊
Outline of Final Research Achievements

Spermatogonial stem cells (SSCs) continuously proliferate by self-renewal division and produce sperm after injection into the seminifelous tubules of testes. This study aimed at the development of new infertility treatment, and tried to develop a method to manipulate human SSCs. CXCR4 was identified as a marker of human SSCs, through screening of cell surface antigens. Factors which promote proliferation of spermatogonia were examined by screening of small molecule chemical libraries and ARTN, NRTN, PSPN, the family molecules of GDNF, the self-renewal factor of SSCs, as well as FGF family molecules, and BMP family molecules, which are considered to be involved in survival of spermatogonia. Using culture conditions modified from the conventional culture condition of mouse SSCs by adding ARTN, a ligand for GFRA3, human testes cells produced colonies in vitro, which were maintained for > 2 months period.

Academic Significance and Societal Importance of the Research Achievements

小児悪性腫瘍患者の生存率は近年向上したが、放射線治療や薬物療法の副作用として不妊になるケースが多い。精子凍結ができない小児患者の妊孕性を担保する方法として、あらかじめ精子幹細胞を凍結保存し治療後に精細管内へ移植する試みが海外にて進められている。しかし小児精巣から採取できる精子幹細胞は極めて少なく、試験管内にて増幅する技術が必要とされている。また培養を行うことで悪性腫瘍細胞の混入を防ぐことができるという利点もある。本研究の成果はヒト精子幹細胞の試験管内増幅技術の確立のために不可欠であるヒト精子幹細胞のマーカー分子や増殖を促す因子を同定した点で、学術的・社会的意義がある。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (16 results)

All 2021 2020 2019 2018 Other

All Journal Article (9 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 9 results,  Open Access: 4 results) Presentation (6 results) (of which Invited: 1 results) Remarks (1 results)

  • [Journal Article] OGG1 protects mouse spermatogonial stem cells from reactive oxygen species in culture2021

    • Author(s)
      Mori Y, Ogonuki N, Hasegawa A, Kanatsu-Shinohara M, Ogura A, Wang Y, McCarrey JR, Shinohara T.
    • Journal Title

      Biol Reprod

      Volume: 104 Issue: 3 Pages: 706-716

    • DOI

      10.1093/biolre/ioaa216

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] An interplay of NOX1-derived ROS and oxygen determines the spermatogonial stem cell self-renewal efficiency under hypoxia.2021

    • Author(s)
      Hiroko Morimoto, Takuya Yamamoto, Takehiro Miyazaki, Harumi Ogonuki, Atsuo Ogura, Takashi Tanaka, Mito Kanatsu-Shinohara , Chihiro Yabe-Nishimura, Hongliang Zhang, Yves Pommier, Andreas Trumpp, Takashi Shinohara
    • Journal Title

      Genes Dev.

      Volume: 35(3-4) Issue: 3-4 Pages: 250-260

    • DOI

      10.1101/gad.339903.120

    • NAID

      120006951673

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Autologous transplantation of spermatogonial stem cells restores fertility in congenitally infertile mice2020

    • Author(s)
      Kanatsu-Shinohara Mito、Ogonuki Narumi、Matoba Shogo、Ogura Atsuo、Shinohara Takashi
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 117 Issue: 14 Pages: 7837-7844

    • DOI

      10.1073/pnas.1914963117

    • NAID

      120006826644

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] CD2 is a surface marker for mouse and rat spermatogonial stem cells.2020

    • Author(s)
      Kanatsu-Shinohara M, Chen G, Morimoto H, Shinohara T.
    • Journal Title

      J Reprod Dev.

      Volume: Mar 26. Issue: 4 Pages: 1-32

    • DOI

      10.1262/jrd.2020-019

    • NAID

      130007888738

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Transgenesis and genome editing of mouse spermatogonial stem cells by lentivirus pseudotyped with Sendai virus F protein.2020

    • Author(s)
      Shinohara, T. and Kanatsu-Shinohara, M.
    • Journal Title

      Stem Cell Reports

      Volume: 14 Issue: 3 Pages: 447-461

    • DOI

      10.1016/j.stemcr.2020.02.001

    • NAID

      120006936408

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Expression and functional analyses of EPHA2 in mouse spermatogonial stem cells.2020

    • Author(s)
      Morimoto, H., Kanatsu-Shinohara, M., Orwig, K. E. and Shinohara, T.
    • Journal Title

      Biol. Reprod.

      Volume: 102 Issue: 1 Pages: 220-232

    • DOI

      10.1093/biolre/ioz156

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Aging of spermatogonial stem cells by Jnk-mediated glycolysis activation.2019

    • Author(s)
      Kanatsu-Shinohara, M., Yamamoto, T., Toh, H., Kazuki, Y., Kazuki, K., Imoto, J., Ikeo, K., Oshima, M., Shirahige, K., Iwama, A., Nabeshima, Y., Sasaki, H. and Shinohara, T.
    • Journal Title

      PNAS

      Volume: 116 Issue: 33 Pages: 16404-16409

    • DOI

      10.1073/pnas.1904980116

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Sendai virus-mediated transduction of mammalian spermatogonial stem cells.2019

    • Author(s)
      Watanabe S., Kanatsu-Shinohara, M., Shinohara T.
    • Journal Title

      Biol. Reprod.

      Volume: 100 Issue: 2 Pages: 523-534

    • DOI

      10.1093/biolre/ioy192

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Reversible inhibition of the blood-testis barrier protein improves stem cell homing in mouse testes2018

    • Author(s)
      Kanatsu-Shinohara M, Morimoto H, Watanabe S, Shinohara T.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 64 Issue: 6 Pages: 511-522

    • DOI

      10.1262/jrd.2018-093

    • NAID

      130007530501

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Presentation] 睡眠構築の変化が精子形成に及ぼす影響の解析2020

    • Author(s)
      篠原美都・林悠
    • Organizer
      AMED-CREST機能低下 領域会議
    • Related Report
      2019 Annual Research Report
  • [Presentation] 精子幹細胞の老化機構の解明2020

    • Author(s)
      篠原美都・篠原隆司
    • Organizer
      京都大学・医薬系研究交流サロン メディカルイノベーション卓越大学院プログラム キックオフ企画
    • Related Report
      2019 Annual Research Report
  • [Presentation] 精子幹細胞の自己複製と維持のメカニズムの解明2018

    • Author(s)
      篠原美都
    • Organizer
      第23回日本女性科学者の会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 精子幹細胞の老化において精巣支持環境が果たす役割の解明2018

    • Author(s)
      篠原美都
    • Organizer
      新学術領域「幹細胞老化と疾患」第5回領域会議
    • Related Report
      2018 Annual Research Report
  • [Presentation] 長期培養における精子幹細胞の老化2018

    • Author(s)
      篠原美都
    • Organizer
      第1回AMED-CREST機能低下・睡眠グループ分科会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 精子を作る幹細胞とその操作技術2018

    • Author(s)
      篠原美都
    • Organizer
      日本女性科学者の会 60周年記念シンポジウム
    • Related Report
      2018 Annual Research Report
  • [Remarks] 精子幹細胞の自家移植により先天性男性不妊症が回復することを発見-先天性不妊症にも柔軟性

    • URL

      http://www.kyoto-u.ac.jp/ja/research/research_results/2019/200331_1.html

    • Related Report
      2020 Annual Research Report

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Published: 2018-04-23   Modified: 2022-01-27  

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