Targeted LC-MS/MS-based metabolomics and lipidomics on limited hematopoietic stem cell numbers

Metabolism is important for the regulation of hematopoietic stem cells (HSCs) and drives cellular fate. Due to the scarcity of HSCs, it has been technically challenging to perform metabolome analyses gaining insight into HSC metabolic regulatory networks. Here, we present two targeted liquid chromat...

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Bibliographic Details
Published in:STAR protocols 2022-06, Vol.3 (2), p.101408-101408, Article 101408
Main Authors: Schönberger, Katharina, Mitterer, Michael, Buescher, Joerg M., Cabezas-Wallscheid, Nina
Format: Article
Language:English
Subjects:
Chromatography, Liquid
Hematopoietic Stem Cells
Lipidomics
Mass Spectrometry
Metabolomics
Metabolomics - methods
Protocol
Stem Cells
Tandem Mass Spectrometry - methods
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Summary:Metabolism is important for the regulation of hematopoietic stem cells (HSCs) and drives cellular fate. Due to the scarcity of HSCs, it has been technically challenging to perform metabolome analyses gaining insight into HSC metabolic regulatory networks. Here, we present two targeted liquid chromatography–mass spectrometry approaches that enable the detection of metabolites after fluorescence-activated cell sorting when sample amounts are limited. One protocol covers signaling lipids and retinoids, while the second detects tricarboxylic acid cycle metabolites and amino acids. For complete details on the use and execution of this protocol, please refer to Schönberger et al. (2022). [Display omitted] •Isolation of rare hematopoietic cell types from the murine bone marrow•Metabolite extraction of limited sample amounts after FACS purification•Detection of signaling lipids, retinoids, and polar metabolites Publisher's note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Metabolism is important for the regulation of hematopoietic stem cells (HSCs) and drives cellular fate. Due to the scarcity of HSCs, it has been technically challenging to perform metabolome analyses gaining insight into HSC metabolic regulatory networks. Here, we present two targeted liquid chromatography–mass spectrometry approaches that enable the detection of metabolites after fluorescence-activated cell sorting when sample amounts are limited. One protocol covers signaling lipids and retinoids, while the second detects tricarboxylic acid cycle metabolites and amino acids.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2022.101408