31P Magnetic Resonance Spectroscopy of Endothelial Cells Grown in Three-Dimensional Matrigel Construct as an Enabling Platform Technology: I. The Effect of Glial Cells and Valproic Acid on Phosphometabolite Levels

• Published in: Endothelium (New York, N.Y.) Vol. 15; no. 5-6; pp. 288 - 298
• Main Authors: Sterin, M., Ringel, I., Lecht, S., Lelkes, P. I., Lazarovici, P.
• Format: Journal Article
• Language: English
• Published: England Informa UK Ltd 01.09.2008; Taylor & Francis
• Subjects: Animals; Anticonvulsants - pharmacology; Antitumor agents; Biochemistry - methods; Biological Assay - instrumentation; Biological Assay - methods; Bioreactors; Cell Communication - physiology; Cell Line; Cell Line, Tumor; Coculture Techniques; Collagen - pharmacology; Degradation products; Drug Combinations; Endothelial cells; Endothelial Cells - drug effects; Endothelial Cells - metabolism; Endothelium; Energy metabolism; Energy Metabolism - drug effects; Energy Metabolism - physiology; Glial cells; Laminin - pharmacology; Lipid Metabolism - drug effects; Magnetic resonance spectroscopy; Magnetic Resonance Spectroscopy - methods; Matrigel; Neuroglia - metabolism; Neuroglia - secretion; Organ Culture Techniques - methods; P MRS; Phosphocreatine; Phosphocreatine - metabolism; Phospholipids; Phospholipids - metabolism; Phosphometabolites; phosphomonoesters; Phosphorus Isotopes - chemistry; Proteoglycans - pharmacology; Rats; Tissue culture; Tissue engineering; Valproic acid; Valproic Acid - pharmacology
• ISSN: 1062-3329; 1029-2373
• DOI: 10.1080/10623320802487841

Very few studies describe endothelial cell (EC) properties under three-dimensional (3D) conditions using 31P magnetic resonance spectroscopy (MRS). The authors developed a model in which living ECs growing in Matrigel threads (3D conditions) for 5 days are monitored by 31P MRS, providing the fingerprint of the major EC phosphometabolites. Organic extracts of membranal phospholipids were also analyzed by 31P MRS. For comparison and as a model for two-dimensional (2D) tissue culture conditions, 31P MRS spectra of aqueous extracts of EC phosphometabolites grown under 2D conditions were also evaluated. The phosphometabolites fingerprint of the cells cultured under 3D was significantly different from that of ECs maintained under 2D. Moreover, the pattern of phosphometabolites was affected by coculture with C6-glioma cells and upon treatment with valproic acid, which is under clinical investigation as an antioangiogenic anticancer drug. The major effects were modulation of (i) energy metabolism intermediates such as phosphocreatine, (ii) precursors of phospholipids such as phosphomonoesters, and (iii) degradation products of phospholipids such as glycerophosphocholine. This endothelial model will be usefull as an enabling platform technology for tissue engineering.