Unraveling the metabolism of HEK-293 cells using lactate isotopomer analysis

• Published in: Bioprocess and biosystems engineering Vol. 34; no. 3; pp. 263 - 273
• Main Authors: Henry, Olivier, Jolicoeur, Mario, Kamen, Amine
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01-03-2011; Springer-Verlag; Springer; Springer Nature B.V
• Subjects: Acetyl Coenzyme A - metabolism; Amino Acids - metabolism; Biological and medical sciences; Biosynthesis; Biotechnology; C-metabolic flux analysis; Carbon; Carbon - metabolism; Carbon Isotopes - metabolism; Cell culture; Cell Line; Cell Survival - physiology; Cellular biology; Chemistry; Chemistry and Materials Science; Chromatography, Liquid - methods; Citric Acid Cycle - physiology; Environmental Engineering/Biotechnology; Food Science; Fundamental and applied biological sciences. Psychology; Glucose - metabolism; Glutamine - metabolism; HEK-293 cells; HEK293 Cells - metabolism; Humans; Industrial and Production Engineering; Industrial Chemistry/Chemical Engineering; Isotopic tracer; Kidneys; Lactic Acid - metabolism; Lipids; Lipids - biosynthesis; Magnetic Resonance Spectroscopy - methods; Malates - metabolism; Metabolism; Metabolites; Original Paper; Oxaloacetic Acid - metabolism; Pentose Phosphate Pathway - physiology; C-metabolic flux analysis; Isotopic tracer; Metabolism; HEK-293 cells; Lactates; Metabolic flux analysis
• ISSN: 1615-7591; 1615-7605
• DOI: 10.1007/s00449-010-0468-9

HEK-293 is the most extensively used human cell line for the production of viral vectors and is gaining increasing attention for the production of recombinant proteins by transient transfection. To further improve the metabolic characterization of this cell line, we have performed cultures using ¹³C-labeled substrates and measured the resulting mass isotopomer distributions in lactate by LC/MS. Simultaneous metabolite and isotopomer balancing allowed improvement and validation of the metabolic model and quantification of key intracellular pathways. We have determined the amounts of glucose carbon channeled through the PPP, incorporated into the TCA cycle for energy production and lipids biosynthesis, as well as the cytosolic and mitochondrial malic enzyme fluxes. Our analysis also revealed that glutamine did not significantly contribute to lactate formation. An improved and quantitative understanding of the central carbon metabolism is greatly needed to pursue the rational development of engineering approaches at both the cellular and process levels.