| Project/Area Number |
03454268
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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 |
Pediatrics
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
SAWADA Tadashi Kyoto Prefectural Univ.of Med.Prof, 医学部, 教授 (10079874)
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| Co-Investigator(Kenkyū-buntansha) |
SAWADA Tadashi Kyoto Prefectural Univ.of Med.Prof (10079874)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥3,000,000 (Direct Cost: ¥3,000,000)
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| Keywords | ^<31>P-MRS / Hollow fiber / Perfusion culture / PME / PCr / metabolite / metabolism / growth / 灌流培養 / ^<31>P‐MRS測定 / スペクトル / 低酸素負荷 / 高エネルギーリン酸化合物 / ATP / ^<31>PーMRS / 神経芽腫 / 培養細胞 / ホロ-ファイバ- / 高密度細胞培養 |
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| Research Abstract |
The human neuroblastoma cell line was used. The medium was supplemented with 10 % fetal bovine serum. Minivitafiber unit ( Grace Japan, Amicon ) was chosen in this study. Cells were introduced into the hollow-fiber unit and allowed to deposit and reattach for a few hours, and then the intracapillary space ( ICS ) was begun to be perfused. The perfusion of the extracapillary space ( ECS ) was started 48 hours after inoculation. A peristaltic pump was used to maintain the flow rate of 22 ml/min in ICS and 0.3 ml/min in ECS.Phosphorus-31 nuclear magnetic resonance ( ^<31>P-NMR ) spectra were obtained on CSI II operating at 81 MHz for phosphorus. The spectral conditions were a 60ﾟ pulse, a 2.0 recycle time. Starling 10 days after inocuation of the culture unit, prominent spectra could be obtained by 2000 scans on intact cells aerated by continuous medium perfusion at 37ﾟC.Peaks were phosphomonoesters ( PME ), inorganic phosphate ( Pi ), phosphodiesters ( PDE ), phosphocreatine ( PCr ), gamma -adennosine triphosphate ( ATP ), alpha -ATP and beta -ATP.
The ^<31>P-NMR spectra of human neuroblastoma cells were obtained on intact cells cultured in hollow fiber. The spectra were dominated by phosphomonoester peaks. The perfusion system was developed to achieve the high cell density and maintain viable intact cells for NMR study.
This system would be useful for study of growth and metabolism in neuroblastoma.
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