13C-Metabolic flux analysis (13C-MFA) to quantify cellular metabolism is classified with stationary and instationary 13C-MFA in terms of labeling dynamics at intracellular metabolites by introduction of isotopic tracers. As mammalian cell metabolism shows isotopic non-steady state due to complex metabolism, instationary 13C-MFA should be applied to study cellular metabolism. However, instationary 13C-MFA requires time course data and high computational time. Thus, we introduced a parallel labeling strategy using two isotopic tracers for stationary 13C-MFA: [1,2-13C]glucose tracer for glycolysis pathway and [U-13C]glutamine for TCA cycle to determine metabolic fluxes in CHO cells. We computed the two types of measurements for flux analysis using one metabolic model. In the end, flux results revealed significant metabolic rewiring from growth phase to non-growth during culture in oxidative pentose phosphate pathway, anaplerosis, amino acid metabolism, and fatty acid biosynthesis. In addition, the parallel labeling strategy needs less computational time and experimental loads.

Estimation of metabolic rewiring in CHO cell culture using parallel labeling experiment

245th National Meeting (2013)

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