A framework for tracer-based metabolism in mammalian cells by NMR

Scientific Reports
Raquel SaboranoUlrich L Günther

Abstract

Metabolism changes extensively during the normal proliferation and differentiation of mammalian cells, and in cancer and inflammatory diseases. Since changes in the metabolic network reflect interactions between genetic, epigenetic and environmental changes, it is helpful to study the flow of label from isotopically labelled precursors into other metabolites rather than static metabolite levels. For this Nuclear Magnetic Resonance (NMR) spectroscopy is an attractive technique as it can quantify site-specific label incorporation. However, for applications using human cells and cell lines, the challenge is to optimize the process to maximize sensitivity and reproducibility. Here we present a new framework to analyze metabolism in mammalian cell lines and primary cells, covering the workflow from the preparation of cells to the acquisition and analysis of NMR spectra. We have applied this new approach in hematological and liver cancer cell lines and confirm the feasibility of tracer-based metabolism in primary liver cells.

References

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Citations

Apr 17, 2019·Chembiochem : a European Journal of Chemical Biology·Michelle A C ReedUlrich L Günther
Nov 16, 2019·Angewandte Chemie·Islam AlshamlehHarald Schwalbe
Aug 21, 2020·PLoS Pathogens·Philipp BrandtSlavena Vylkova
Oct 18, 2020·Disease Models & Mechanisms·Lozan SheriffPatricia F Lalor
Jan 26, 2021·JHEP Reports : Innovation in Hepatology·Emma L ShepherdPatricia F Lalor
Jun 2, 2020·Journal of Magnetic Resonance·Anne Birk FrahmMathilde Hauge Lerche
Jul 28, 2021·MBio·Laure Nicolas Annick RiesGustavo H Goldman

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Methods Mentioned

BETA
NMR
FCS
chemical shift

Software Mentioned

Metabolab
NMRPipe
NMRlab
MATLAB
hsqcetgpsp

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