Compartment-Specific Metabolic Investigation of 3D Cell Culture by Real-time NMR for Investigating Metabolic Diseases
Christian Urzi1,2,3, Damian Hertig1,2, Christoph Meyer1,2,3, Jean-Marc Nuoffer2,4, and Peter Vermathen1
1Magnetic Resonance Methodology, Institute of Diagnostic and Interventional Neuroradiology, University of Bern, Bern, Switzerland, 2Institute of Clinical Chemistry, University Hospital Bern, Bern, Switzerland, 3Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland, 4Department of Pediatric Endocrinology, Diabetology and Metabolism, University Children’s Hospital of Bern, Bern, Switzerland
In this study was shown the possibility to perform compartment-specific metabolic investigations of living fibroblasts in real-time using a bioreactor system within an NMR spectrometer. Cell volume and the ratio cells to extracellular medium within the sensitive region of NMR coil were evaluated for quantification purposes. The needed sensitivity to detect metabolite changes in the extracellular footprint at different flow rates was shown. The capability of diffusion technique to distinguish between compartment-specific contributions was revealed, and to detect changes in compartment-specific metabolite pools under standard or selective culture conditions, and upon inhibitor challenges.
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