13C NMR spectroscopy and mass spectrometry analysis of intermediary metabolism in cultured neural cells

Ursula Sonnewald*, Arne Schousboe, Helle S. Waagepetersen

*Corresponding author for this work

    Abstract

    The use of 13C and 15N labeled precursors in combination with adequate analytical tools makes it possible to study metabolic pathways in cultured neural cells. The most commonly used precursors are 13C labeled glucose, lactate, glutamate and acetate. For a dynamic evaluation of intermediary metabolism of cell cultures, incubation with 13C containing substrates followed by nuclear magnetic resonance spectroscopy (NMRS) and mass spectrometry (MS) is excellent. NMRS can be used on cell extracts or living cells if a sufficient quantity of labeled atoms is present. MS is the more sensitive of the two methods but often it requires derivatization and separation of the components before analysis. The review provides descriptions of the basic and practical aspects of culturing neural cells, incubation and superfusion experiments and NMRS and MS analyses. It focuses on the analytical tools and the use of primary cultures of neurons and astrocytes for the elucidation of metabolic interactions between neurons and astrocytes.

    Original languageEnglish
    Title of host publicationCell Culture Techniques
    EditorsMichael Aschner, Cristina Sunol, Anna Bal-Price
    Number of pages13
    Publication date18 Mar 2011
    Pages403-415
    ISBN (Print)9781617790768
    DOIs
    Publication statusPublished - 18 Mar 2011
    SeriesNeuromethods
    Volume56
    ISSN0893-2336

    Keywords

    • [1-C]glucose
    • Cell culture
    • Mass spectrometry
    • Neurons
    • Nuclear magnetic resonance spectroscopy

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