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

  • Daniel R. Crooks
  • Teresa W.-M. FanEmail author
  • W. Marston Linehan
Part of the Methods in Molecular Biology book series (MIMB, volume 1928)


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.

Key words

Tissue culture Stable isotopes Metabolomics Metabolite extraction Glutamine Glucose 



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Daniel R. Crooks
    • 1
  • Teresa W.-M. Fan
    • 2
    Email author
  • W. Marston Linehan
    • 1
  1. 1.Urologic Oncology BranchNational Cancer Institute, National Institutes of HealthBethesdaUSA
  2. 2.Department of Toxicology and Cancer BiologyCenter for Environmental and Systems Biochemistry, Markey Cancer Center, and University of KentuckyLexingtonUSA

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