Determining Macrophage Polarization upon Metabolic Perturbation

  • Pu-Ste Liu
  • Ping-Chih HoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1862)


Metabolic reprograming controlling macrophage activation and function is emerging as new regulatory circuit on shaping immune responses. Generally, lipopolysaccharides (LPS)-induced pro-inflammatory activated macrophages, known as M1 macrophages, display higher glycolysis. In contrast, interleukin-4 (IL-4)-skewed anti-inflammatory activated macrophages, known as M2 macrophages, mainly rely on oxidative phosphorylation for their bioenergetic demands. Emerging evidence reveals that these metabolic preferences further fine-tune macrophage polarization process, including signaling cascades and epigenetic reprogramming. Thus, specific nutrient microenvironments may affect inflammatory responses of macrophages by intervening these metabolic machineries. How to measure the metabolic switch of macrophages both in vitro and in vivo is an important issue for understanding immunometabolic regulations in macrophages. Here, we describe a basic protocol for examining how glutamine metabolism affects macrophage polarization by using the Extracellular Flux (XF(e)96) Analyzer (Seahorse Bioscience), which takes real-time measurements of oxidative phosphorylation and glycolysis. We also present a detailed procedure for detecting the expression of inflammatory genes in polarized macrophages under glutamine-replete or -deprived conditions.

Key words

Macrophage polarization Glutamine metabolism Extracellular flux analyzer Oxygen consumption Extracellular acidification 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Fundamental Oncology, Faculty of Biology and MedicineUniversity of LausanneEpalingesSwitzerland
  2. 2.Ludwig Lausanne BranchEpalingesSwitzerland
  3. 3.Institute of Cellular and System MedicineNational Health Research InstitutesMiaoli CountyTaiwan

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