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A kinetic-metabolic model based on cell energetic state: study of CHO cell behavior under Na-butyrate stimulation

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Abstract

A kinetic-metabolic model approach describing and simulating Chinese hamster ovary (CHO) cell behavior is presented. The model includes glycolysis, pentose phosphate pathway, TCA cycle, respiratory chain, redox state and energetic metabolism. Growth kinetic is defined as a function of the major precursors for the synthesis of cell building blocks. Michaelis–Menten type kinetic is used for metabolic intermediates as well as for regulatory functions from energy shuttles (ATP/ADP) and cofactors (NAD/H and NADP/H). Model structure and parameters were first calibrated using results from bioreactor cultures of CHO cells expressing recombinant t-PA. It is shown that the model can simulate experimental data for all available experimental data, such as extracellular glucose, glutamine, lactate and ammonium concentration time profiles, as well as cell energetic state. A sensitivity analysis allowed identifying the most sensitive parameters. The model was then shown to be readily adaptable for studying the effect of sodium butyrate on CHO cells metabolism, where it was applied to the cases with sodium butyrate addition either at mid-exponential growth phase (48 h) or at the early plateau phase (74 h). In both cases, a global optimization routine was used for the simultaneous estimation of the most sensitive parameters, while the insensitive parameters were considered as constants. Finally, confidence intervals for the estimated parameters were calculated. Results presented here further substantiate our previous findings that butyrate treatment at mid-exponential phase may cause a shift in cellular metabolism toward a sustained and increased efficiency of glucose utilization channeled through the TCA cycle.

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Acknowledgments

This project was funded by the MabNet Research Network of the Natural Sciences and Engineering Research Council of Canada (NSERC) (MJ) and by an NSERC Discovery grant to MJ and OH.

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Authors and Affiliations

  1. Canada Research Chair in Applied Metabolic Engineering, Department of Chemical Engineering, École Polytechnique de Montréal, Centre-ville Station, P.O. box 6079, Montréal, QC, H3C 3A7, Canada

    Atefeh Ghorbaniaghdam, Olivier Henry & Mario Jolicoeur

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  1. Atefeh Ghorbaniaghdam
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Correspondence to Atefeh Ghorbaniaghdam.

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449_2012_804_MOESM2_ESM.tif

Supplementary Fig. 1 Simulated data of intracellular metabolites for Control (solid line), and cultures with sodium butyrate addition at 48 h (NaBu-48h) (dashed line) and at 74 h (NaBu-74h) (dotted line). Axis units are mmol (106 cells)−1. Simulated values of intracellular concentrations showed to be within the range found in literature for TCA cycle intermediates (10−7–10−6 mmol (106 cells)−1) [39, 40], intracellular sugar phosphates (10−7–10−6 mmol (106 cells)−1) (CHO cells: [41]; Hepatic cells: [42]; MDCK cells: [43]), intracellular glutamine (10−7–10−6 mmol (106 cells)−1) and glutamate (10−6–10−5 mmol (106 cells)−1) (CHO cells: [44, 45]; HEK-293 cells: [45]). (TIFF 560 kb)

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Ghorbaniaghdam, A., Henry, O. & Jolicoeur, M. A kinetic-metabolic model based on cell energetic state: study of CHO cell behavior under Na-butyrate stimulation. Bioprocess Biosyst Eng 36, 469–487 (2013). https://doi.org/10.1007/s00449-012-0804-3

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