Open AccessEffect of stimulation time on the expression of human macrophage polarization markers
Author(s) -
Duygu Unuvar Purcu,
Asli Korkmaz,
Sinem Gunalp,
Derya Goksu Helvaci,
Yonca Erdal,
Yavuz Doğan,
Aslı Süner,
Gerhard Wingender,
Duygu Sağ
Publication year - 2022
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0265196
Subject(s) - macrophage polarization , ccl22 , cd163 , microbiology and biotechnology , macrophage , cd64 , biology , cxcl10 , in vitro , stimulation , immunology , m2 macrophage , chemokine , inflammation , flow cytometry , neuroscience , genetics
Macrophages are highly plastic cells that can polarize into functionally distinct subsets in vivo and in vitro in response to environmental signals. The development of protocols to model macrophage polarization in vitro greatly contributes to our understanding of macrophage biology. Macrophages are divided into two main groups: Pro-inflammatory M1 macrophages (classically activated) and anti-inflammatory M2 macrophages (alternatively activated), based on several key surface markers and the production of inflammatory mediators. However, the expression of these common macrophage polarization markers is greatly affected by the stimulation time used. Unfortunately, there is no consensus yet regarding the optimal stimulation times for particular macrophage polarization markers in in vitro experiments. This situation is problematic, (i) as analysing a particular marker at a suboptimal time point can lead to false-negative results, and (ii) as it clearly impedes the comparison of different studies. Using human monocyte-derived macrophages (MDMs) in vitro , we analysed how the expression of the main polarization markers for M1 (CD64, CD86, CXCL9, CXCL10, HLA-DR, IDO1, IL1β, IL12, TNF), M2a (CD200R, CD206, CCL17, CCL22, IL-10, TGM2), and M2c (CD163, IL-10, TGFβ) macrophages changes over time at mRNA and protein levels. Our data establish the most appropriate stimulation time for the analysis of the expression of human macrophage polarization markers in vitro . Providing such a reference guide will likely facilitate the investigation of macrophage polarization and its reproducibility.