Journal article
Analysis of mammalian cell proliferation and macromolecule synthesis using deuteratedwater and gas chromatography-mass spectrometry
VC Foletta, M Palmieri, J Kloehn, S Mason, SF Previs, MJ McConville, OM Sieber, CR Bruce, GM Kowalski
Metabolites | MDPI | Published : 2016
Abstract
Deuterated water (2H2O), a stable isotopic tracer, provides a convenient and reliable way to label multiple cellular biomass components (macromolecules), thus permitting the calculation of their synthesis rates. Here, we have combined 2H2O labelling, GC-MS analysis and a novel cell fractionation method to extract multiple biomass components (DNA, protein and lipids) from the one biological sample, thus permitting the simultaneous measurement of DNA (cell proliferation), protein and lipid synthesis rates. We have used this approach to characterize the turnover rates and metabolism of a panel of mammalian cells in vitro (muscle C2C12 and colon cancer cell lines). Our data show that in actively..
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Grants
Awarded by National Science Foundation
Funding Acknowledgements
This work was supported by the following grants: Diabetes Australia Research Trust (G.M.K.) and the Ludwig Institute for Cancer Research, and the Victorian Government's Operational Infrastructure Support Program (O.M.S.). M.J.M. is an NHMRC Principal Research Fellow (APP1059530). G.M.K. is supported by an Alfred Deakin Post-Doctoral Fellowship (Deakin University). O.M.S. is an NHMRC R.D. Wright Biomedical Fellow (APP1062226). G.M.K. is the guarantor of this work and, as such, has full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.