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要旨:
Glycolysis is a metabolic pathway comprising a sequence of biochemical key reactions to generate energy and precursors for numerous associated metabolic pathways in mammalian cells. Obviously, there are several levels of control required for a cellular system of such importance. However, despite considerable efforts to investigate regulation of energy metabolism, the dynamics of glycolysis in mammalian cells remains rather unclear. Our group investigates the energy metabolism of adherently growing Madin Darby canine kidney (MDCK) cells, which serve as host cells for influenza vaccine production. After inoculation, cells grow exponentially until they reach confluency and eventually enter a stationary phase before they are infected. To understand the dynamics of energy metabolism, pulse experiments in stationary growth phase are performed that alter the metabolite levels in glycolysis and citric acid cycle within seconds. Most likely this highly dynamic behaviour is based on enzyme-metabolite interactions, which allows to obtain insight in the inherent regulation of the biochemical reaction sequences. Based on a method recently developed for the quantification of most of the intracellular metabolites involved in these pathways, detailed experimental data is available for establishment and validation of mathematical models [1]. Therefore, data obtained in high temporal resolution experiments using MDCK cells are integrated into a mathematical model to investigate the inherent dynamics of glycolysis. The model comprises a set of differential equations and aims at identifying crucial regulatory mechanisms as well as elucidating the role of associated metabolic pathways. The approach focuses on the design of new experiments for a better understanding of energy and carbon metabolism of mammalian cells, and subsequently the identification of process parameters relevant for optimisation of vaccine production processes.
