Three-dimensional construction of artificial seminiferoustubules for in vitro spermatogenesis

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K21656
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 56:Surgery related to the biological and sensory functions and related fields
• Research Institution: Okayama University
• Principal Investigator: Yamamoto Yuki 岡山大学, 環境生命科学学域, 准教授 (20645345)
• Co-Investigator(Kenkyū-buntansha): 木村 康二 岡山大学, 環境生命科学研究科, 教授 (50355070)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Keywords: 精細管 / 器官培養 / 3次元バイオプリンター

Outline of Final Research Achievements

In this study, we examined the isolation and culture conditions of testicular cells of immature bovine testis and the printing conditions with the aim of constructing seminiferous tubules using a 3D bioprinter. The culture medium containing 10% fetal bovine serum was used to maintain germ cells in the ducts, and the effects of hormones or retinoic acid were examined. Hormones had no effect on cell proliferation or germ cell localization, while the addition of retinoic acid increased the percentage of germ cells expressing Ki67.
The mixing ratio of commercially available polyethylene glycol diacrylate and gelatin methacrylate as bioink was also examined, but the conditions under which the cells settled and proliferated did not match those maintaining the morphology of the 3D structures, and further verification was required.

Free Research Field

動物生殖生理学

Academic Significance and Societal Importance of the Research Achievements

本研究成果から得られた情報をもとに、バイオプリンターを用いた精細管の構築条件や培養条件をさらに改良し、人工的に3次元構造を作りそれを用いて精子形成を体外で再現できれば、男性不妊治療の発展に大きく貢献することが期待される。完全な体外精子形成の手法は未だマウス以外で確立されておらず、マウスの手法でも精巣組織の培養が必要であり完全な in vitro システムではない。生体組織を要しない精子形成システムを開発することで、ヒトのみならずさまざまな動物の生殖学研究・繁殖・保全に寄与することが期待できる。