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Protocols and Applications of Cellular Metabolomics in Safety Studies Using Precision-Cut Tissue Slices and Carbon 13 NMR

  • Gabriel BaverelEmail author
  • Maha El Hage
  • Guy Martin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1641)

Abstract

Numerous xenobiotics are toxic to human and animal cells by interacting with their metabolism, but the precise metabolic step affected and the biochemical mechanism behind such a toxicity remain often unknown. In an attempt to reduce the ignorance in this field, we have developed a new approach called cellular metabolomics. This approach, developed in vitro, provides a panoramic view not only of the pathways involved in the metabolism of physiological substrates of any normal or pathological human or animal cell but also of the beneficial and adverse effects of xenobiotics on these metabolic pathways. Unlike many cell lines, precision-cut tissue slices, for which there is a renewed interest, remain metabolically differentiated for at least 24–48 h and allow to study the effect of xenobiotics during short-term and long-term incubations. Cellular metabolomics (or metabolic flux analysis), which combines enzymatic and carbon 13 NMR measurements with mathematical modeling of metabolic pathways, is illustrated in this brief chapter for studying the effect of insulin on glucose metabolism in rat liver precision-cut slices and of valproate on glutamine metabolism in human renal cortical precision-cut slices. The use of very small amounts of test compounds allows to predict their toxic effect and eventually their beneficial effects very early in the research and development processes. Cellular metabolomics is complementary to other omics approaches, but, unlike them, provides functional, mechanistic, and dynamic pieces of information by measuring enzymatic fluxes.

Key words

Toxicology Liver Kidney Cellular metabolomics Carbon 13 NMR 

Notes

Acknowledgements

The authors would like to thank Claudie Pinteur, Rémi Nazaret, and Lara Koneckny for technical assistance as well as Claire Morel for secretarial assistance. This work was supported by grants from the European Commission [project numbers: BIO4-CT97-2145 (Euroslice) and STREP 032731 (CellNanoTox)], and from INSERM.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Metabolys Inc.LyonFrance

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