Profiling of mouse macrophage lipidome using direct infusion shotgun mass spectrometry

Wei-Yuan Hsieh; Kevin J Williams; Baolong Su; Steven J Bensinger

doi:10.1016/j.xpro.2020.100235

Profiling of mouse macrophage lipidome using direct infusion shotgun mass spectrometry

STAR Protoc. 2020 Dec 19;2(1):100235. doi: 10.1016/j.xpro.2020.100235. eCollection 2021 Mar 19.

Authors

Wei-Yuan Hsieh¹, Kevin J Williams^{2

3}, Baolong Su^{2

3}, Steven J Bensinger^{1

3}

Affiliations

¹ Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ UCLA Lipidomics Lab, University of California, Los Angeles, Los Angeles, CA 90095, USA.

Abstract

Immune cells, such as macrophages, reprogram their lipid metabolism in response to the activation of pattern recognition receptors (e.g., TLRs, NLRs) and cytokine receptors (e.g., interferons, interleukins). Profiling these changes can be achieved with shotgun mass spectrometry. This protocol provides step-by-step instructions on the generation and stimulation of bone marrow-derived macrophages (BMDMs), sample collection, and lipid extraction for profiling the macrophage lipidome. For complete details on the use and execution of this protocol, please refer to Hsieh et al. (2020).

Keywords: Cell culture; High Throughput Screening; Immunology; Mass Spectrometry; Metabolism; Model Organisms.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Lipid Metabolism*
Lipidomics*
Macrophages / metabolism*
Mass Spectrometry*
Mice

Grants and funding

P01 HL146358/HL/NHLBI NIH HHS/United States