Cellular Fatty Acid Analysis in Macrophage Using Stable Isotope Labeling

Kevin J Williams; Steven J Bensinger

doi:10.1007/978-1-0716-0802-9_4

Cellular Fatty Acid Analysis in Macrophage Using Stable Isotope Labeling

Methods Mol Biol. 2020:2184:47-60. doi: 10.1007/978-1-0716-0802-9_4.

Authors

Kevin J Williams^{1

2}, Steven J Bensinger^{3

4}

Affiliations

¹ Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA. kevinwilliams@mednet.ucla.edu.
² UCLA Lipidomics Laboratory, Los Angeles, CA, USA. kevinwilliams@mednet.ucla.edu.
³ UCLA Lipidomics Laboratory, Los Angeles, CA, USA. sbensinger@mednet.ucla.edu.
⁴ Department of Microbiology, Immunology and Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA. sbensinger@mednet.ucla.edu.

PMID: 32808217
DOI: 10.1007/978-1-0716-0802-9_4

Abstract

Fatty acids (FAs) are essential for building complex lipids, posttranslational modifications, and energetics. FAs can be imported from extracellular sources or synthesized by cells. The analysis of fatty acid methyl esters (FAMEs) by gas chromatography-mass spectrometry (GC-MS) allows for the quantitative analysis of long-chain and very-long-chain fatty acid content of cells. When coupled with isotopic labeling, this approach can elucidate the synthetic pathways being engaged by the cells, and the relative contribution of synthesis and import to maintain lipid content. Here, we describe a method for total cellular fatty acid analysis in macrophages.

Keywords: FAME analysis; Fatty acids; GC–MS; Macrophages; Stable isotope labeling.

MeSH terms

Biosynthetic Pathways / physiology
Fatty Acids / metabolism*
Gas Chromatography-Mass Spectrometry / methods
Isotope Labeling / methods
Lipids
Macrophages / metabolism*

Substances

Fatty Acids
Lipids