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

Katharina Schönberger; Michael Mitterer; Joerg M Buescher; Nina Cabezas-Wallscheid

doi:10.1016/j.xpro.2022.101408

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

STAR Protoc. 2022 May 19;3(2):101408. doi: 10.1016/j.xpro.2022.101408. eCollection 2022 Jun 17.

Authors

Katharina Schönberger^{1

2

3}, Michael Mitterer⁴, Joerg M Buescher⁴, Nina Cabezas-Wallscheid¹

Affiliations

¹ Max Planck Institute of Immunobiology and Epigenetics, Department of Cellular and Molecular Immunology, Stübeweg 51, 79108 Freiburg, Germany.
² Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany.
³ International Max Planck Research School for Molecular and Cellular Biology (IMPRS-MCB), Freiburg, Germany.
⁴ Max Planck Institute of Immunobiology and Epigenetics, Metabolomics Core Facility, Stübeweg 51, 79108 Freiburg, Germany.

Abstract

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).

Keywords: Mass Spectrometry; Metabolomics; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography, Liquid
Hematopoietic Stem Cells
Lipidomics*
Metabolomics / methods
Tandem Mass Spectrometry* / methods