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Measuring the Nutrient Metabolism of Adherent Cells in Culture

  • Martin P. Ogrodzinski
  • Shao Thing Teoh
  • Lei Yu
  • Deanna Broadwater
  • Elliot Ensink
  • Sophia Y. Lunt
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1862)

Abstract

Metabolite extraction from cells cultured in vitro enables the comprehensive measurement of intracellular metabolites. These extracts can be analyzed using techniques such as liquid chromatography-mass spectrometry (LC-MS). This chapter describes in detail a method for metabolite extraction from cultured adherent mammalian cells to collect both polar and nonpolar intracellular metabolites. This chapter also describes experimental design considerations for performing stable isotope labeling experiments, and the use of chemical derivatization to increase the number of compounds that can be detected using one chromatography method.

Key words

Adherent cell metabolism Amino acid chemical derivatization Isotope labeling LC-MS Metabolite extraction Metabolomics 

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Copyright information

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

Authors and Affiliations

  • Martin P. Ogrodzinski
    • 1
    • 2
  • Shao Thing Teoh
    • 1
  • Lei Yu
    • 1
  • Deanna Broadwater
    • 1
  • Elliot Ensink
    • 1
  • Sophia Y. Lunt
    • 1
    • 3
  1. 1.Department of Biochemistry and Molecular BiologyMichigan State UniversityEast LansingUSA
  2. 2.Department of PhysiologyMichigan State UniversityEast LansingUSA
  3. 3.Department of Chemical Engineering and Materials ScienceMichigan State UniversityEast LansingUSA

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