The development of three-dimensional cell culture system imitating in vivo niche and architecture during the formation of seminiferous tubules to generate functional testicular organoid

• Project/Area Number: 17H05098
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: Urology
• Research Institution: Yokohama City University
• Principal Investigator: Komeya Mitsuru 横浜市立大学, 医学研究科, 客員研究員 (60721082)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥25,610,000 (Direct Cost: ¥19,700,000、Indirect Cost: ¥5,910,000); Fiscal Year 2019: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2018: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2017: ¥13,910,000 (Direct Cost: ¥10,700,000、Indirect Cost: ¥3,210,000)
• Keywords: 男性不妊 / 精子形成 / オルガノイド / マイクロ流体システム / トランスクリプトーム / 細胞培養 / 細胞・組織培養 / 再生医学 / マイクロ流体デバイス

Outline of Final Research Achievements

This study used two approaches to generate functional testicular organoid. First, the architecture of seminiferous tubule was imitated by the sheet device in which germ cells were incubated on Sertoli cell sheet. By the device, Sertoli cell sheet was maintained over 3months and germ cell were divided into the early meiotic stages. Second, testicular cells were incubated by the self-organization technique. Germ cells were arranged in a chain at the outer area of organoids and were divided into the early meiotic stages.

Academic Significance and Societal Importance of the Research Achievements

不妊症の50～60%に関与する男性不妊の病態解明や治療法の開発をすすめるうえで、ヒトin vitro実験系の確立が重要である。特に細胞培養系によりiPS細胞を用いたin vitro精子形成が実現できれば研究推進が強く期待できる。本研究は、精細管の基本である層構造および精細管の管腔形成過程を再現することで精子形成を減数分裂初期まで進展させることができた。細胞培養下でのヒトin vitro精子形成の実現に向けた貴重な知見を得ることが出来た。

Research Products

• [Journal Article] Antioxidant Vitamins and Lysophospholipids Are Critical for Inducing Mouse Spermatogenesis Under Organ Culture Conditions (2020)
  • Author(s): Sanjo H, Yao T, Katagiri K, Sato T, Matsumura T, Komeya M, Yamanaka H, Yao M, Matsuhisa A, Asayama Y, Ikeda K, Kano K, Aoki J, Arita M, Ogawa T.
  • Journal Title: The FASEB Journal Volume: online ahead of print Issue: 7 Pages: 9480-9497
  • DOI: 10.1096/fj.202000245r
  • Peer Reviewed / Open Access
• [Journal Article] In vitro spermatogenesis in two‐dimensionally spread mouse testis tissues (2019)
  • Author(s): Komeya Mitsuru、Yamanaka Hiroyuki、Sanjo Hiroyuki、Yao Masahiro、Nakamura Hiroko、Kimura Hiroshi、Fujii Teruo、Sato Takuya、Ogawa Takehiko
  • Journal Title: Reproductive Medicine and Biology Volume: 18 Issue: 4 Pages: 362-369
  • DOI: 10.1002/rmb2.12291
  • Peer Reviewed / Open Access
• [Journal Article] Neonatal testis growth recreated in vitro by two-dimensional organ-spreading. (2018)
  • Author(s): Kojima K, Nakamura H, Komeya M, Yamanaka H, Makino Y, Okada Y, Akiyama H, Torikai N, Sato T, Fujii T, Kimura H, *Ogawa T.
  • Journal Title: Biotechnol Bioeng. Volume: 115 Issue: 12 Pages: 3030-3041
  • DOI: 10.1002/bit.26822
  • Peer Reviewed / Open Access
• [Journal Article] In vitro spermatogenesis: A century-long research journey, still half way around. (2018)
  • Author(s): Komeya M, Sato T, *Ogawa T.
  • Journal Title: Reprod Med Biol. Volume: 17 Issue: 4 Pages: 407-420
  • DOI: 10.1002/rmb2.12225
  • Peer Reviewed / Open Access
• [Journal Article] A monolayer microfluidic device supporting mouse spermatogenesis with improved visibility. (2018)
  • Author(s): Yamanaka H, Komeya M, Nakamura H, Sanjo H, Sato T, Yao M, Kimura H, Fujii T, Ogawa T.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 印刷中 Issue: 4 Pages: 885-891
  • DOI: 10.1016/j.bbrc.2018.04.180
  • Peer Reviewed
• [Journal Article] In vitro mouse spermatogenesis with an organ culture method in chemically defined medium. (2018)
  • Author(s): Sanjo H, Komeya M, Sato T, Abe T, Katagiri K, Yamanaka H, Ino Y, Arakawa N, Hirano H, Yao T, Asayama Y, Matsuhisa A, Yao M, Ogawa T.
  • Journal Title: PLoS One Volume: 12 Issue: 2 Pages: 1-13
  • DOI: 10.1371/journal.pone.0192884
  • Peer Reviewed / Open Access
• [Journal Article] Pumpless microfluidic system driven by hydrostatic pressure induces and maintains mouse spermatogenesis in vitro. (2017)
  • Author(s): Komeya M, Hayashi K, Nakamura H, Yamanaka H, Sanjo H, Kojima K, Sato T, Yao M, Kimura H, Fujii T, Ogawa T.
  • Journal Title: Sci Rep. Volume: 13 Issue: 1 Pages: 15459-15459
  • DOI: 10.1038/s41598-017-15799-3
  • Peer Reviewed / Open Access
• [Presentation] 静水圧を用いて血流を再現した精巣組織培養システムの開発 (2019)
  • Author(s): 古目谷 暢、小川 毅彦
  • Organizer: 日本アンドロロジー学会第38回学術大会
  • Invited
• [Presentation] in vitro精子形成システムの開発 (2018)
  • Author(s): 古目谷 暢
  • Organizer: 日本泌尿器科学会東部総会
  • Invited
• [Presentation] シリコン樹脂カバーを併用した精巣組織培養法による精子形成効率の向上 (2018)
  • Author(s): 古目谷 暢
  • Organizer: 第106回 日本泌尿器科学会総会
• [Presentation] シリコン樹脂カバーによる精巣組織培養法の改良 (2017)
  • Author(s): 加藤 美樹、古目谷 暢、小島 一晃、加藤 佐樹子、林 功晃、山中 弘行、佐藤 卓也、三條 博之、矢尾 正祐、中村 寛子、木村 啓志、藤井 輝夫、小川 毅彦
  • Organizer: 第36回日本アンドロロジー学会
• [Remarks] 公立大学法人 横浜市立大学生命医科学研究科創薬再生科学小川グループ（生殖再生医学）
  • URL: http://www.tsurumi.yokohama-cu.ac.jp/proteome/ogawa/index.html