• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development of in vitro culture system for expansion of Leydig stem cells

Research Project

Project/Area Number 15K15013
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field General anatomy (including histology/embryology)
Research InstitutionKyoto University

Principal Investigator

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

Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywords幹細胞 / 生殖 / 精子 / 精巣 / ライディッヒ細胞 / Leydig細胞
Outline of Final Research Achievements

Recent studies suggest a possibility that not only germ cell, but Leydig cells of testes is supported by stem cell system. This study aimed at the establishment of in vitro culture system to maintain and expand Leydig stem cell for a long-term period. To enrich Leydig stem cell population, we isolated side population (SP) fraction from testes cells, based on the ability of stem cells to effectively exclude the dye Hoechst 33342. After SP fraction was cultured with serum-free medium for 2 weeks, a part of the cultured cells expressed PDGFRA, a marker of Leydig cells. Cultured cells derived from Green mice, which ubiquitously express EGFP, were dissociated by enzyme digestion and injected into the interstitial space of wild-type mouse testes. At 3 months after injection, host testes had EGFP expressing cells, which was detected by UV light excitation, however, immunofluorescence study showed that Leydig cell marker PDGFRA was not detected in EGFP expressing cells.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (25 results)

All 2018 2017 2016 2015

All Journal Article (11 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 11 results,  Open Access: 7 results,  Acknowledgement Compliant: 8 results) Presentation (12 results) (of which Int'l Joint Research: 2 results,  Invited: 2 results) Book (2 results)

  • [Journal Article] In vivo genetic manipulateon of spermatogonial stem cells and their microenvironment by adeno-associated viruses.2018

    • Author(s)
      Watanabe S., Kanatsu-Shinohara, M., Ogonuki N., Matoba S., Ogura A., Shinohara T.
    • Journal Title

      Stem Cell Reports

      Volume: S2213-6711 (18) Issue: 5 Pages: 30132-2

    • DOI

      10.1016/j.stemcr.2018.03.005

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Nonrandom contribution of left and right testes to germline transmission from mouse spermatogonial stem cells.2017

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

      Biol. Reprod.

      Volume: 92(6) Pages: 902-910

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Transfer of a mouse artificial chromosome into spermatogonial stem cells genetates transchromosomic mice,2017

    • Author(s)
      Shinohara T, Kazuki K, Ogonuki N, Morimoto H, Matoba S, Hiramatsu K, Honma K, Suzuki T, Hara T, Ogura A, Oshimura M, Kanatsu-Shinohara M, Kazuki Y.
    • Journal Title

      Stem Cell Reports.

      Volume: 9(4) Issue: 4 Pages: 1180-1191

    • DOI

      10.1016/j.stemcr.2017.08.012

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Myc/Mycn-mediated glycolysis enhances mouse spermatogonial stem cell self-renewal.2016

    • Author(s)
      Kanatsu-Shinohara M., Tanaka T., Ogonuki N., Ogura A., Morimoto H., Cheng PF., Eisenman RN., Trumpp A., Shinohara T.
    • Journal Title

      Genes Dev.

      Volume: 30(23) Issue: 23 Pages: 2637-2648

    • DOI

      10.1101/gad.287045.116

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] The luteinizing hormone-testosterone pathway regulates mouse spermatogonial stem cell self-renewal by suppressing WNT5A expression in Sertoli cells.2016

    • Author(s)
      Tanaka T., Kanatsu-Shinohara M., Lei Z., Rao CV., Shinohara T.
    • Journal Title

      Stem Cell Reports

      Volume: 7(2) Issue: 2 Pages: 279-291

    • DOI

      10.1016/j.stemcr.2016.07.005

    • NAID

      120005825924

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Nonrandom germline transmission of mouse spermatogonial stem cells.2016

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

      Dev Cell

      Volume: 38 (3) Issue: 3 Pages: 248-261

    • DOI

      10.1016/j.devcel.2016.07.011

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Fertility of male germline stem cells following spermatogonial transplantation in infertile mouse models.2016

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

      Biol Reprod.

      Volume: 94(5) Issue: 5 Pages: 112-112

    • DOI

      10.1095/biolreprod.115.137869

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Enrichment of Mouse Spermatogonial Stem Cells by the Stem Cell Dye CDy1.2015

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

      Biology of reproduction

      Volume: 94 (1) 13 Issue: 1 Pages: 1-10

    • DOI

      10.1095/biolreprod.115.135707

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Pluripotent cell derivation from male germline cells by suppression of <i>Dmrt1</i> and <i>Trp53</i>2015

    • Author(s)
      Tanaka, T., Kanatsu-Shinohara, M., Hirose, M., Ogura, A., Shinohara, T.
    • Journal Title

      Journal of Reproduction and Development

      Volume: 61 Issue: 5 Pages: 473-484

    • DOI

      10.1262/jrd.2015-059

    • NAID

      130005104118

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] The CDKN1B-RB1-E2F1 pathway protects mouse spermatogonial stem cells from genomic damage2015

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

      Journal of Reproduction and Development

      Volume: 61 Issue: 4 Pages: 305-316

    • DOI

      10.1262/jrd.2015-027

    • NAID

      130005094647

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] ROS-Generating Oxidase Nox3 Regulates the Self-Renewal of Mouse Spermatogonial Stem Cells2015

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

      Biology of reproduction

      Volume: 92 (6) 147 Issue: 6 Pages: 1-10

    • DOI

      10.1095/biolreprod.114.127647

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] 精子幹細胞の寿命と精子形成への寄与の動態解明2017

    • Author(s)
      篠原美都
    • Organizer
      JST 戦略的創造研究推進事業(さきがけ)第10回「細胞機能の構成的な理解と制御」領域会議
    • Place of Presentation
      淡路夢舞台国際会議場
    • Year and Date
      2017-02-21
    • Related Report
      2016 Research-status Report
  • [Presentation] 精子幹細胞の寿命と精子形成への寄与の動態解明2017

    • Author(s)
      篠原美都
    • Organizer
      生命動態システム科学四拠点 合同シンポジウム
    • Related Report
      2017 Annual Research Report 2016 Research-status Report
    • Invited
  • [Presentation] 精子幹細胞の寿命制御とアンチエイジング機構2017

    • Author(s)
      篠原美都
    • Organizer
      第7回細胞再生医療研究会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 精子幹細胞の寿命と精子形成への寄与の動態解明2016

    • Author(s)
      篠原美都
    • Organizer
      JST 戦略的創造研究推進事業(さきがけ)H25年度採択(第三期)研究成果報告会 「細胞機能の構成的な理解と制御」
    • Place of Presentation
      東京大学・理学部化学講堂
    • Year and Date
      2016-11-01
    • Related Report
      2016 Research-status Report
  • [Presentation] 長期培養系を用いた精子幹細胞の老化メカニズムの解明2016

    • Author(s)
      篠原美都
    • Organizer
      新学術領域「幹細胞老化と疾患」第三回領域会議
    • Place of Presentation
      東京大学 伊藤国際学術研究センター
    • Year and Date
      2016-06-28
    • Related Report
      2016 Research-status Report
  • [Presentation] 精子幹細胞の寿命と精子形成への寄与の動態解明2016

    • Author(s)
      篠原美都
    • Organizer
      JST 戦略的創造研究推進事業(さきがけ)第9回「細胞機能の構成的な理解と制御」領域会議
    • Place of Presentation
      福岡県福岡市 ホテル ザ・ルイガンズ
    • Year and Date
      2016-06-21
    • Related Report
      2016 Research-status Report
  • [Presentation] Nonrandom germline transmission of mouse spermatogonial stem cells2016

    • Author(s)
      Takashi Shinohara, Honda Naoki, and Mito Kanatsu-Shinohara
    • Organizer
      文部科学省 科学研究費補助金 新学術領域研究 「生殖細胞のエピゲノムダイナミクスとその制御」による国際シンポジウムEpigenome Dynamics and Regulation in Germ cells
    • Place of Presentation
      Clock Tower Centennial Hall, Kyoto University
    • Year and Date
      2016-02-17
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] Enrichment of mouse spermatogonial stem cells by a stem cell dye CDy12016

    • Author(s)
      Mito Kanatsu-Shinohara, Hiroko Morimoto, and Takashi Shinohara
    • Organizer
      文部科学省 科学研究費補助金 新学術領域研究 「生殖細胞のエピゲノムダイナミクスとその制御」による国際シンポジウム Epigenome Dynamics and Regulation in Germ cells
    • Place of Presentation
      Clock Tower Centennial Hall, Kyoto University
    • Year and Date
      2016-02-17
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] 精子幹細胞の寿命と精子形成への寄与の動態解明2015

    • Author(s)
      篠原美都
    • Organizer
      JST 戦略的創造研究推進事業(さきがけ)第8回「細胞機能の構成的な理解と制御」領域会議
    • Place of Presentation
      東京白金台・八芳園
    • Year and Date
      2015-11-26
    • Related Report
      2015 Research-status Report
  • [Presentation] 精子幹細胞の寿命と精子形成への寄与の動態解明2015

    • Author(s)
      篠原美都
    • Organizer
      JST 戦略的創造研究推進事業(さきがけ)H24年度採択(第二期)研究成果報告会「細胞機能の構成的な理解と制御」
    • Place of Presentation
      東京大学・伊藤謝恩ホール
    • Year and Date
      2015-11-25
    • Related Report
      2015 Research-status Report
  • [Presentation] 精子幹細胞の寿命と精子形成への寄与の動態解明2015

    • Author(s)
      篠原美都
    • Organizer
      JST 戦略的創造研究推進事業(さきがけ)第7回「細胞機能の構成的な理解と制御」領域会議
    • Place of Presentation
      IBM天城ホームステッド
    • Year and Date
      2015-07-14
    • Related Report
      2015 Research-status Report
  • [Presentation] 長期培養系を用いた精子幹細胞の老化メカニズムの解明2015

    • Author(s)
      篠原美都
    • Organizer
      新学術領域「幹細胞老化と疾患」第二回領域会議
    • Place of Presentation
      新潟グランドホテル
    • Year and Date
      2015-06-19
    • Related Report
      2015 Research-status Report
  • [Book] 実験医学 特集 「再発見!MYCの多機能性-グローバル転写因子として見直される古典的がん遺伝子」2018

    • Author(s)
      田中敬・篠原美都・篠原隆司
    • Total Pages
      6
    • Publisher
      羊土社
    • ISBN
      9784758125055
    • Related Report
      2017 Annual Research Report
  • [Book] 実験医学 特集 「臓器老化の本質に迫るステムセルエイジング:幹細胞の変遷と加齢関連疾患のメカニズム」2017

    • Author(s)
      篠原美都・本田直樹・篠原隆司
    • Total Pages
      6
    • Publisher
      羊土社
    • ISBN
      9784758101639
    • Related Report
      2017 Annual Research Report

URL: 

Published: 2015-04-16   Modified: 2019-03-29  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi