RRC ID 34775
Author Okahashi N, Kohno S, Kitajima S, Matsuda F, Takahashi C, Shimizu H.
Title Metabolic characterization of cultured mammalian cells by mass balance analysis, tracer labeling experiments and computer-aided simulations.
Journal J Biosci Bioeng
Abstract Studying metabolic directions and flow rates in cultured mammalian cells can provide key information for understanding metabolic function in the fields of cancer research, drug discovery, stem cell biology, and antibody production. In this work, metabolic engineering methodologies including medium component analysis, (13)C-labeling experiments, and computer-aided simulation analysis were applied to characterize the metabolic phenotype of soft tissue sarcoma cells derived from p53-null mice. Cells were cultured in medium containing [1-(13)C] glutamine to assess the level of reductive glutamine metabolism via the reverse reaction of isocitrate dehydrogenase (IDH). The specific uptake and production rates of glucose, organic acids, and the 20 amino acids were determined by time-course analysis of cultured media. Gas chromatography-mass spectrometry analysis of the (13)C-labeling of citrate, succinate, fumarate, malate, and aspartate confirmed an isotopically steady state of the cultured cells. After removing the effect of naturally occurring isotopes, the direction of the IDH reaction was determined by computer-aided analysis. The results validated that metabolic engineering methodologies are applicable to soft tissue sarcoma cells derived from p53-null mice, and also demonstrated that reductive glutamine metabolism is active in p53-null soft tissue sarcoma cells under normoxia.
Volume 120(6)
Pages 725-31
Published 2015-12-1
DOI 10.1016/j.jbiosc.2015.04.003
PII S1389-1723(15)00153-X
PMID 25936961
MeSH Amino Acids / metabolism Animals Cell Line, Tumor Computer Simulation* Gas Chromatography-Mass Spectrometry Glucose / metabolism Glutamine / metabolism* Isocitrate Dehydrogenase / metabolism Male Metabolic Engineering Mice Oxygen / metabolism Sarcoma / enzymology Sarcoma / metabolism* Sarcoma / pathology* Time Factors Tumor Suppressor Protein p53 / deficiency
IF 2.366
Times Cited 6
Mice RBRC01361