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1991 Fiscal Year Final Research Report Summary

Nuclear Magnetic Resouance Study of Intact Cells Cultured in Hollow-Fiber

Research Project

Project/Area Number 01870045
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field Pediatrics
Research InstitutionKyoto Prefectural University of Medicine

Principal Investigator

SAWADA Tadashi  Kyoto Prefectural Univ. of Med. Prof., 医学部, 教授 (10079874)

Project Period (FY) 1989 – 1991
Keywordshollow - fiber / high cell density in Hure / ^<31>P-MRS
Research Abstract

The human neuroblastoma cell line, KP-N-RT, 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 24 hours after inoculation. A peristaltic pump was used to maintain the flow rate of 25 ml/min in ICS and 8 ml/hr in ECS. Phosphorus-31 nuclear magnetic resonance ( ^<31>P-NMR) spectra were obtained on CSIII (GE) operating at 81 MHz for phosphorus. The spectral conditions were a 60゚ pulse, a 2.0 recycle time. Starling 14 days after inoculation of the culture unit, prominent spectra could be obtained by 2000 scans on intact cells aerated by continuous medium perfusion 37゚C. Peaks were phosphomonoesters (PME), inorganic phosphate (Pi), phosphopdiesters (PDE), phosphocreatine (PCr), gamma -adennosine triphosphate (ATP), alpha -ATP and beta -ATP.
The ^<31P>-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 energy metabolism and phospholipid metabolism in culture cells.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] T.Sugimoto: "Alpha-smooth-muscle actin and desmin expressions in human neuroblastoma cell lines" Int.J.Cancer. 48(1). 277-283 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Kuroda: "Different drug sensitivity in two neuroblastoma cell lines established from the same patient before and after chemotherapy" Int.J.Cancer. 47(2). 732-737 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Sugimoto: "Neuronal differentiation of human neuroblastoma cells by retinoid" J.Nutritional Science Vitaminology. (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 杉本 徹ら: "細胞表面膜抗原,細胞骨格蛋白とNーmyc癌遺伝子による小児固形悪性腫痛の診断と問題点" Human Cell. 5(1). 11-11 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 杉本 徹ら: "平滑筋細胞マ-カ-を持つヒト神経芽腫細胞株" Human Cell. 4(3). 248-256 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Sugimoto, H. Ueyama, H. Hosoi, J. Jnazawa, T. Kato, J. T. Kemshead, C. P. Reynols, A. M. Gown, H. Mine and T. Sawada: "Alpha - smooth - muscle actin and desmin expressions in human neuroblastoma cell lines." Int. J. Cancer. 48(1). 277-283 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H. Kuroda, T. Sugimoto, K. Ueda, S. Tsuchida, Y. Horii, J. Inazawa, K. Sato and T. Sawada: "Different drug sensitivity in two neuroblastoma cell lines established from the same patient before and after chemotherapy." Int. J. Cancer. 47(2). 732-737 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Sugimoto, Y. Horii, T. Sawada, S. Fushuki, Y. Suzuki, O. Tagaya: "Neuronal differentiation of human neuroblastoma cells by retinoid." J. Nutritional Science Vitaminology. (1992)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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