| Project/Area Number |
07558125
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Osaka University |
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Principal Investigator |
SUGA Ken-ichi Faculty of Engineering, Osaka University, Professor, 工学部, 教授 (20029250)
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| Co-Investigator(Kenkyū-buntansha) |
OMASA Takeshi Faculty of Engineering, Osaka University, Assistant Professor, 工学部, 助手 (00252586)
KATAKURA Yoshio Faculty of Engineering, Osaka University, Assistant Professor, 工学部, 助手 (50263207)
KISHIMOTO Michimasa Faculty of Engineering, Osaka University, Associate Professor, 工学部, 助教授 (00144436)
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| Project Period (FY) |
1995 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 1995: ¥6,700,000 (Direct Cost: ¥6,700,000)
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| Keywords | hybrid artificial liver support system / recombinant DNA / ammonia removal / drug metabolism / gene amplification / glutamine synthetase / 人工肝臓 / 組伝子組換え / 人工肝 / MSX / GS / アンモニア |
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| Research Abstract |
In order to establish the cell line which was suitable for hybrid artificial liver support system, an ammonia metabolizing activity was given to Chinese Hamster Ovary (CHO) and HepG2 cell line using genetic engineering techniques. CHO-K1 and HepG2 cells were transformed with pSV2-GS vector which contains glutamine synthetase gene connected with SV40 or CMV promoter. The recombinant cell line was selected under the various concentrations of glutamine synthetase inhibitor, methionine sulfoximine (MSX). The obtained MSX tolerable cell line had the high glutamine synthetase activity and could remove the ammonium from medium. The kinetic parameters of this cell line were determined under 0,1, or 2mM ammonia concentration. The specific ammonium consumption rates of established CHO and HepG2 cell line during growth phase were about one fourth and one seventh of that of primary hepatocyte respectively.
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