A kinetic-metabolic model based on cell energetic state: study of CHO cell behavior under Na-butyrate stimulation
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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.
KeywordsMetabolic modeling CHO cells Kinetic model Metabolic regulation Energy regulation Sodium butyrate
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.
- 16.Hendrick V, Winnepenninckx P, Abdelkafi CV, Andeputte O, Cherlet M, Marique T, Renemann G, Loa A, Kretzmer G, Werenne J (2001) Increased productivity of recombinant tissular plasminogen activator (t-PA) by butyrate and shift of temperature: a cell cycle phases analysis. Cytotechnology 36:71–83CrossRefGoogle Scholar
- 24.Zamoranoa F, Vande Wouwera A, Bastin G (2010) A detailed metabolic flux analysis of an underdetermined network of CHO cells. J Biotechnol 150:495–508Google Scholar
- 33.Dash RK, DiBella JA, Cabrera ME (2007) A computational model of skeletal muscle metabolism linking cellular adaptations induced by altered loading states to metabolic responses during exercise. Biomol Eng 6:14Google Scholar
- 42.Maier K, Hofmann U, Reuss M, Mauch K (2007) Identification of metabolic fluxes in hepatic cells from transient 13C-labeling experiments: part II. Flux estimation. Biotechnol Bioeng 100:344–354Google Scholar
- 49.Rizhaupt A, Ellis A, Hosie KB, P Shirazi-Beechey S (1998) The characterization of butyrate transport across pig and human colonic luminal membrane. J Physiol 507:819–830Google Scholar
- 50.Beauvieux MC, Tissier P, Gin H, Canioni P, Gallis JL (1986) Butyrate impairs energy metabolism in isolated perfused liver of fed rats. J Nutr 131:1986–1992Google Scholar
- 51.Wlaschin KF, Hu WS (2006) Fedbatch culture and dynamic nutrient feeding. Adv Biochem Eng Biotechnol 101:43–74Google Scholar
- 56.Van der Valk P, Gille JJP, van der Plas LHW, Jongkind JF, Verkerk A, Konings AWT, Joenje H (1988) Characterization of oxygen-tolerant Chinese hamster ovary cells: II. Energy metabolism and antioxidant status 4:345–356Google Scholar