| Project/Area Number |
17K19285
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Forestry and Forest Products Science, Applied aquatic science, and related fields
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| Research Institution | Tokyo University of Marine Science and Technology |
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Principal Investigator |
SANO MOTOHIKO 東京海洋大学, 学術研究院, 教授 (00372053)
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| Project Period (FY) |
2017-06-30 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2017: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
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| Keywords | 細胞融合 / 魚類細胞 / ウイルス培養 / 組織・細胞 / 魚病 / 魚類培養細胞 / 魚介類ウイルス / ポリエチレングリコール / ウイルス / 水産学 / ウイルス感受性 / 魚類 / 細胞・組織 / ウィルス |
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| Outline of Final Research Achievements |
Cell fusion protocol using polyethylene glycol (PEG) was studied for fish cells. For selective culture of the fusion cells, eel ovary cell line EO-2 and Chinook salmon embryo cell line CHSE-214 were tolerized to nucleotide synthesis inhibitor 5-bromodeoxyuridine and 8-azaguanin respectively to induce nucleic acid synthesis enzyme deficiency. Since there two tolerized cell lines were not killed by an inhibitory medium, selective culture of the fused cells cannot be conducted. The mechanism of the tolerance to the drugs in fish cells may different from those in mammalian cell lines. Fusion protocol using EO-2 cells might be of PEG treatment at 25℃ for 1 min. The results suggest that new cell line can be developed using PEG cell fusion method as long as selective culture works well on fused cells.
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| Academic Significance and Societal Importance of the Research Achievements |
PEGによる魚類の細胞融合自体は可能であることがはじめて分かったが、融合細胞の選択培養はできなかった。選択培養のため、核酸合成阻害剤である5-bromodeoxyuridine (BUdR)あるいは8-azaguanin(8-AZ)への魚類培養細胞の薬剤耐性化を進めたが、アミノプテリンを含む選択培地で細胞が死滅することはなく、ほ乳類の培養細胞とは異なり、魚類培養細胞株独自の耐性メカニズムが存在する可能性が示された。薬剤選別がうまく機能するようになれば、細胞融合により魚類の新たな細胞株は作出可能と考えられ、魚類ウイルス培養に新たな道を拓くものと期待される。
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