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Metabolic Labeling of Cultured Mammalian Cells for Stable Isotope-Resolved Metabolomics: Practical Aspects of Tissue Culture and Sample Extraction

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Cancer Metabolism

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1928))

Abstract

Stable isotope-resolved metabolomics (SIRM) methods are used increasingly by cancer researchers to probe metabolic pathways and identify vulnerabilities in cancer cells. Analytical and computational advances are being made constantly, but tissue culture and sample extraction procedures are often variable and not elaborated in the literature. This chapter discusses basic aspects of tissue culture practices as they relate to the use of stable isotope tracers and provides a detailed metabolic labeling and metabolite extraction procedure designed to maximize the amount of information that can be obtained from a single tracer experiment.

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Acknowledgments

This work was supported by the Intramural Research Program of the National Cancer Institute, NIH, NIH 1U24DK097215-01A1, 1P01CA163223-01A1, and 3R01ES022191-04S1.

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Authors and Affiliations

  1. Urologic Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Daniel R. Crooks & W. Marston Linehan

  2. Department of Toxicology and Cancer Biology, Center for Environmental and Systems Biochemistry, Markey Cancer Center, and University of Kentucky, Lexington, KY, USA

    Teresa W.-M. Fan

Authors
  1. Daniel R. Crooks
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  2. Teresa W.-M. Fan
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  3. W. Marston Linehan
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Corresponding author

Correspondence to Teresa W.-M. Fan .

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Editors and Affiliations

  1. National Cancer Institute, North Bethesda, MD, USA

    Majda Haznadar

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Crooks, D.R., Fan, T.WM., Linehan, W.M. (2019). Metabolic Labeling of Cultured Mammalian Cells for Stable Isotope-Resolved Metabolomics: Practical Aspects of Tissue Culture and Sample Extraction. In: Haznadar, M. (eds) Cancer Metabolism. Methods in Molecular Biology, vol 1928. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9027-6_1

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  • DOI: https://doi.org/10.1007/978-1-4939-9027-6_1

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