Metabolite Profiling and Stable Isotope Tracing in Sorted Subpopulations of Mammalian Cells.

Détails

ID Serval
serval:BIB_E934B2F0A47D
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Metabolite Profiling and Stable Isotope Tracing in Sorted Subpopulations of Mammalian Cells.
Périodique
Analytical chemistry
Auteur(s)
Roci I., Gallart-Ayala H., Schmidt A., Watrous J., Jain M., Wheelock C.E., Nilsson R.
ISSN
1520-6882 (Electronic)
ISSN-L
0003-2700
Statut éditorial
Publié
Date de publication
01/03/2016
Volume
88
Numéro
5
Pages
2707-2713
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Résumé
Biological samples such as tissues, blood, or tumors are often complex and harbor heterogeneous populations of cells. Separating out specific cell types or subpopulations from such complex mixtures to study their metabolic phenotypes is challenging because experimental procedures for separation may disturb the metabolic state of cells. To address this issue, we developed a method for analysis of cell subpopulations using stable isotope tracing and fluorescence-activated cell sorting followed by liquid chromatography-high-resolution mass spectrometry. To ensure a faithful representation of cellular metabolism after cell sorting, we benchmarked sorted extraction against direct extraction. While peak areas differed markedly with lower signal for amino acids but higher signal for nucleotides, mass isotopomer distributions from sorted cells were generally in good agreement with those obtained from direct extractions, indicating that they reflect the true metabolic state of cells prior to sorting. In proof-of-principle studies, our method revealed metabolic phenotypes specific to T cell subtypes, and also metabolic features of cells in the committed phase of the cell division cycle. Our approach enables studies of a wide range of adherent and suspension cell subpopulations, which we anticipate will be of broad importance in cell biology and biomedicine.

Pubmed
Création de la notice
19/04/2017 18:08
Dernière modification de la notice
21/08/2019 6:34
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