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

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...

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Bibliographic Details
Published in:Scientific reports Vol. 9; no. 1; p. 2520
Main Authors: Saborano, Raquel, Eraslan, Zuhal, Roberts, Jennie, Khanim, Farhat L., Lalor, Patricia F., Reed, Michelle A. C., Günther, Ulrich L.
Format: Journal Article
Language:English
Published: London Nature Publishing Group UK 21.02.2019
Nature Publishing Group
Subjects:
101/6
140/131
631/45/56
631/61/320
631/92/56
Animals
Carbon Isotopes - chemistry
Carbon Isotopes - pharmacology
Cell proliferation
Environmental changes
Hepatocytes
Humanities and Social Sciences
Humans
Inflammatory diseases
Isotope Labeling - methods
Liver cancer
Magnetic Resonance Imaging
Magnetic Resonance Spectroscopy
Mammalian cells
Mammals
Metabolic Networks and Pathways - genetics
Metabolism
Metabolism - genetics
Metabolites
multidisciplinary
NMR
Nuclear magnetic resonance
Science
Science (multidisciplinary)
Spectroscopy
Spectrum analysis
Tumor cell lines
Workflow
ISSN:2045-2322, 2045-2322
Online Access:Get full text
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Summary: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.
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ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-37525-3