Development of cell type-specific in vitro gene function analysis system in mouse ovary

2019 Fiscal Year Final Research Report

• Project/Area Number: 18K14571
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 42010:Animal production science-related
• Research Institution: Tokyo University of Agriculture
• Principal Investigator: SASAKI Keisuke 東京農業大学, 生命科学部, 研究員 (10737159)
• Project Period (FY): 2018-04-01 – 2020-03-31
• Keywords: レンチウイルスベクター / 遺伝子導入 / 卵巣 / 卵母細胞 / 顆粒層細胞

Outline of Final Research Achievements

To develop a research tool to analyze the cell type-specific gene function in mouse ovary, an attempt to establish an oocyte-specific gene delivery system using modified lentiviral vector was subjected. Although this tool has not been completely established so far, the causes of the failure were narrowed down in several points. Moreover, reconstituted ovarian culture system has been established. Dispersed ovarian cells from newborn mice were transduced to modified lentivirus, and were aggregated to form reconstituted ovary in vitro. This method was able to apply for ovarian reconstitution by transduced non-growing oocytes and non-transduced ovarian somatic cells.

Free Research Field

生殖生物学

Academic Significance and Societal Importance of the Research Achievements

本研究の成果から、レンチウイルスを改変することで少なくとも卵巣細胞に対する感染性が変化することがわかった。これを利用して今後細胞種特異的な遺伝子導入法を確立できる可能性がある。また、新生仔マウス由来の卵母細胞と体細胞をそれぞれ別個体から分取し、それらから体外で卵巣を再構成する卵巣再構築培養法を確立できた。今後この培養法を改変して卵母細胞のみ、あるいは、卵巣体細胞のみに遺伝子導入を行い、正常な卵形成を支持できない疾患モデルマウス等のレスキュー法として応用できると考えられる。