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Analysis of mammalian cell proliferation and macromolecule synthesis using deuterated water and gas chromatography-mass spectrometry

Foletta, Victoria C, Palmieri, Michelle, Kloehn, Joachim, Mason, Shaun, Previs, Stephen F, McConville, Malcolm J, Sieber, Oliver M, Bruce, Clinton R and Kowalski, Greg M 2016, Analysis of mammalian cell proliferation and macromolecule synthesis using deuterated water and gas chromatography-mass spectrometry, Metabolites, vol. 6, no. 4, pp. 1-19, doi: 10.3390/metabo6040034.

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Title Analysis of mammalian cell proliferation and macromolecule synthesis using deuterated water and gas chromatography-mass spectrometry
Author(s) Foletta, Victoria CORCID iD for Foletta, Victoria C orcid.org/0000-0002-5209-6134
Palmieri, Michelle
Kloehn, Joachim
Mason, ShaunORCID iD for Mason, Shaun orcid.org/0000-0002-6138-2239
Previs, Stephen F
McConville, Malcolm J
Sieber, Oliver M
Bruce, Clinton R
Kowalski, Greg MORCID iD for Kowalski, Greg M orcid.org/0000-0002-1599-017X
Journal name Metabolites
Volume number 6
Issue number 4
Start page 1
End page 19
Total pages 19
Publisher MDPI
Place of publication Basel, Switzerland
Publication date 2016-10-13
Keyword(s) C2C12
DNA synthesis
GC-MS
biomass
colon cancer
deuterated water
lipogenesis
protein synthesis
stable isotopes
Summary 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-proliferating cells, biomass synthesis rates are strongly linked to the rate of cell division. Furthermore, in both proliferating and non-proliferating cells, it is the lipid pool that undergoes the most rapid turnover when compared to DNA and protein. Finally, our data in human colon cancer cell lines reveal a marked heterogeneity in the reliance on the de novo lipogenic pathway, with the cells being dependent on both 'self-made' and exogenously-derived fatty acid.
Language eng
DOI 10.3390/metabo6040034
Field of Research 060101 Analytical Biochemistry
060104 Cell Metabolism
110199 Medical Biochemistry and Metabolomics not elsewhere classified
Socio Economic Objective 970106 Expanding Knowledge in the Biological Sciences
HERDC Research category C1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2016, The Authors
Free to Read? Yes
Use Rights Creative Commons Attribution licence
Persistent URL http://hdl.handle.net/10536/DRO/DU:30088277

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Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.