Trial of non-invasive picking of donor cells and production of cloned embryos for conservation of small wild cats

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K22311
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 39:Agricultural and environmental biology and related fields
• Research Institution: Kyushu University
• Principal Investigator: Soh Tomoki 九州大学, 農学研究院, 助教 (90221340)
• Co-Investigator(Kenkyū-buntansha): 山内 伸彦 九州大学, 農学研究院, 准教授 (00363325)
• Project Period (FY): 2019-06-28 – 2023-03-31
• Keywords: 核移植胚 / 非侵襲 / 糞 / フィルター法 / 除核

Outline of Final Research Achievements

The purpose of this study is to collect basic information for developing somatic cell cloning technology as a means of conserving small wild cats. We investigated the use of feces or urine for non-invasive picking of donor cells. It was shown that somatic cells can be collected from feces and urine of cats or from urine of cows and can be obtained efficiently by the double filter method. Recipient oocytes were obtained from neutered discarded ovaries of cats at a veterinary hospital. It was confirmed that normal and good oocytes were matured and easily enucleated by drug treatment. Due to the corona crisis, I was able to improve my manipulator manipulation skills by substituting bovine ovaries. As a result, nuclear transfer was performed using cumulus cells as donor cells, and cleavage was observed.

Free Research Field

動物繁殖生理

Academic Significance and Societal Importance of the Research Achievements

繁殖力が衰えた絶滅危惧種の個体数を増やすという点では体細胞クローン動物作製技術が有効である。本研究では絶滅危惧小型野生ネコの繁殖のためドナー細胞を非侵襲的に得られるイエネコの糞から、レシピエントとなる近縁種のイエネコの卵巣を動物病院の廃棄卵巣から採取して体細胞核移植卵作製の可能性を試みた。
その結果、糞由来の体細胞を効率よく単離する方法は2重フィルター法が有効であることと、卵巣から得られる卵丘卵母細胞複合体の選別法及び成熟卵母細胞から効率よく除核する方法を見出した。このことより小型野生ネコから非侵襲的にクローンを作製する糸口が示され、絶滅危惧種の、特に老齢化した個体の増殖に貢献できる。