Live monitoring of cellular metabolism and mitochondrial respiration in 3D cell culture system using NMR spectroscopy
Damian Hertig, Sally Maddah, Roman Memedovski, Sandra Kurth, Aitor Moreno, Matteo Pennestri, Andrea Felser, Jean‐Marc Nuoffer, Peter Vermathen
Abstract
BACKGROUND: F-NMR, with in depth description and validation by accompanying measures. METHODS: atmosphere for physiological pH in carbon based medium and is perfused by HPLC pump. RESULTS & DISCUSSION: Implemented perfused bioreactor allows detection of perfusion rate dependent metabolite content. We show interleaved dynamic profiling of 26 metabolites and mitochondrial respiration. During constant perfusion, sequential injection of rotenone/oligomycin and 2-deoxy-glucose indicated immediate activation and deactivation of glycolytic rate and full inhibition of oxygen consumption. We show sensitivity to detect substrate degradation rates of major mitochondrial fuel pathways and were able to simultaneously measure cellular oxygen consumption.