Increase of GPC Levels in Cultured Mammalian Cells during Acidosis. A 31P MR Spectroscopy Study Using a Continuous Bioreactor System

The purpose of this study was to study the metabolic events during a slow acidosis in three different cell lines by combining 31P magnetic resonance spectroscopy and hollow fiber bioreactor technology. The rate of change in intracellular pH, glycerophosphorylcholine (GPC), phophorylcholine (PCho), and nucleoside‐triphosphate (NTP) levels were measured during 8 h of acidosis and 16 h of recovery in EPO, EAT, and RN1a cells, three cultured mammalians cell lines. Our results show a significant increase in GPC levels to 330 ± 21, 540 ± 25, and 220 ± 21% of their initial value correlated to a decrease of PCho levels to 57 ± 14, 58 ± 17 and 45 ± 15% of their initial value in EAT, RN1a, and EPO cells, respectively. These changes are discussed in terms of perturbation of energetic metabolism in cells undergoing a slow acidosis.

[1]  R. Gillies,et al.  31P-MRS measurements of extracellular pH of tumors using 3-aminopropylphosphonate. , 1994, The American journal of physiology.

[2]  R. Gillies,et al.  Dimethyl methylphosphonate (DMMP): a 31P nuclear magnetic resonance spectroscopic probe of intracellular volume in mammalian cell cultures. , 1993, Biochemistry.

[3]  J. Galons,et al.  Design and application of NMR‐compatible bioreactor circuits for extended perfusion of high‐density mammalian cell cultures , 1993, NMR in biomedicine.

[4]  R. Watson,et al.  In vitro methods in toxicology , 1993 .

[5]  H. Degani,et al.  Lipid metabolism in T47D human breast cancer cells: 31P and 13C-NMR studies of choline and ethanolamine uptake. , 1991, Biochimica et biophysica acta.

[6]  R. Gillies,et al.  Iteration of hybridoma growth and productivity in hollow fiber bioreactors using 31P NMR , 1991, Magnetic resonance in medicine.

[7]  R. Gillies,et al.  Nuclear magnetic resonance spectroscopy of dense cell populations for metabolic studies and bioreactor engineering: a synergistic partnership. , 1991, Biotechnology.

[8]  S. Muallem,et al.  Intracellular pH-regulatory mechanisms in pancreatic acinar cells. I. Characterization of H+ and HCO3- transporters. , 1990, The Journal of biological chemistry.

[9]  G. Zugmaier,et al.  Regulation of the cytidine phospholipid pathways in human cancer cells and effects of 1-beta-D-arabinofuranosylcytosine: a noninvasive 31P nuclear magnetic resonance study. , 1990, Cancer research.

[10]  G. Mandel,et al.  Effect of pH on growth of mouse renal cortical tubule cells in primary culture. , 1989, The American journal of physiology.

[11]  J. Vion-Dury,et al.  Metabolic changes in undifferentiated and differentiated human colon adenocarcinoma cells studied by multinuclear magnetic resonance spectroscopy. , 1989, Biochimie.

[12]  R. Gillies,et al.  Oxygen transfer properties of a bioreactor for use within a nuclear magnetic resonance spectrometer , 1988, Biotechnology and bioengineering.

[13]  B. Dale,et al.  Optimum fiber spacing in a hollow fiber bioreactor , 1988, Biotechnology and bioengineering.

[14]  R. Serrano,et al.  Increased pH and tumorigenicity of fibroblasts expressing a yeast proton pump , 1988, Nature.

[15]  R. Tsien,et al.  A new generation of Ca2+ indicators with greatly improved fluorescence properties. , 1985, The Journal of biological chemistry.

[16]  J. Haveman,et al.  The relevance of tumour pH to the treatment of malignant disease. , 1984, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[17]  D. Vance,et al.  Regulation of phosphatidylcholine biosynthesis. , 1984, Biochimica et biophysica acta.

[18]  R G Shulman,et al.  31P nuclear magnetic resonance evidence for the regulation of intracellular pH by Ehrlich ascites tumor cells , 1982, The Journal of cell biology.

[19]  Levin Bl.  Intracellular pH: Its Measurement, Regulation, and Utilization in Cellular Functions. , 1982 .

[20]  M. Nunnally,et al.  Filamin concentration in cleavage furrow and midbody region: frequency of occurrence compared with that of alpha-actinin and myosin , 1980, The Journal of cell biology.

[21]  A. Warner,et al.  Intracellular pH in early Xenopus embryos: its effect on current flow between blastomeres. , 1980, The Journal of physiology.