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Abstract #4295

Application of Stable Isotope Labelling in Cell Culture Experiments: [2-13C]pyruvate as Novel and Superior Substrate for in Vitro Metabolic Studies in Primary Mouse Hepatocytes

Sven Gottschalk1, Dieter Leibfritz2, Claudia Zwingmann1,2, Marc Bilodeau1

1Dpartement de sciences biomdicales, Universit de Montral, Montral, Qubec, Canada; 2Department of Organic Chemistry, University of Bremen, Bremen, Germany

Application of stable isotope labeling in vitro is a potent technique to study metabolic pathways and fluxes. We applied [2-13C]pyruvate on primary mouse hepatocyte cultures to compare its usefulness for metabolic studies and flux analysis with other labeled substances. Our results show: [2-13C]pyruvate was metabolized by lactate dehydrogenase, alanine-/aspartate-aminotransferase, malic enzyme, PC, PDH and subsequent metabolic pathways through the TCA-cycle. Taken together with our previous findings, we conclude that [2-13C]pyruvate is a versatile physiological substrate to study hepatocellular pathways and the de novo synthesis of major metabolites in these cells under normal and pathological conditions.