| Project/Area Number |
62480105
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurophysiology and muscle physiology
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| Research Institution | University of Tokyo |
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Principal Investigator |
FUKUDA Jun Assoocicte Professor, Medical School, University of Tokyo, 医学部, 助教授 (50010062)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | Hybridoma / Neuson / Epithelial cells / Ekctro-cell-fusion Jonicchannel Gene / ハイブリッド細胞 / 高圧パルス法による細胞融合 / ハイブリッド細胞イオンチャネル形質 / 電気的細胞融合 / SZOY / ハイブリッド |
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| Research Abstract |
This project is aimed to develop new hybridoma cells being obtained by electric fusion of neuronal cells and epithelial cells, in which typical neurogenic gene properties such as ionic channels and neurofilament fibers are expressed in their cell membranes and in their processes. The first step of the project was started to obtain 8-azaguanine-resistant epithelial cells so as to be able to remove during culture in the HAT selection medium. We obtained 2 species of epithelial cells and 1 neuroblastoma cells having sufficiently high resistance against 8-AZ. Electrofusion was made between the 8-AZ resistant epithelial cells and serum-free cultured dorsal root ganglion cells from either the embryonic chick or the adult mouse grown on poly-L-lysine coated plastic dishes.
Gene expression on the hybridoma cells was studied, 1) by immunostaining with anti-neurofilament antibody, and 2) by intracellular recording with glass microelectrode. Neurofilaments staining indicated that the gene of the neurofilaments were well transformed into the hybridoma cells, although some cells lost such gene activities. 2) Ionic channel properties were also transformed to these hybridoma cells; they were able to generate Na, Ca skikes and the presence of K channels which were originated from the neuronal cells were recognized in the plasma membranes of the hybridoma cells.
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