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The Conversion of Human Tissue‐Like Inflammatory Monocytes Into Macrophages
Author(s) -
Bsat Marwa,
Mehta Heena,
Rubio Manuel,
Sarfati Marika
Publication year - 2021
Publication title -
current protocols
Language(s) - English
Resource type - Journals
ISSN - 2691-1299
DOI - 10.1002/cpz1.74
Subject(s) - monocyte , cd14 , immunology , cd16 , macrophage , inflammation , biology , cell sorting , microbiology and biotechnology , in vitro , flow cytometry , immune system , cd8 , cd3 , biochemistry
Classical circulating LyC6 high murine monocytes differentiate progressively from inflammatory tissue monocytes to mature macrophages (Mϕ) after entry into gut mucosa. This protocol provides a two‐step in vitro culture method that replicates the human monocyte maturation cascade. First, purified circulating CD14 + CD16 − monocytes exposed to granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), interferon gamma (IFNγ), and interleukin 23 (IL‐23) differentiate into tissue‐like inflammatory monocytes. Next, addition of transforming growth factor beta (TGFβ) plus interleukin 10 (IL‐10) promotes their maturation into tissue‐like Mϕ. Methods to sort these cells after culture are also provided. The fine‐tuning of this system might open therapeutic avenues for chronic inflammatory disorders. © 2021 Wiley Periodicals LLC This article was corrected on 25 July 2022. See the end of the full text for details. Basic Protocol 1 : Isolation of human monocytes from peripheral blood Basic Protocol 2 : First step culture for generation of inflammatory monocyte‐like cells Basic Protocol 3 : Second step culture for differentiation of inflammatory monocyte‐like cells into macrophages Alternate Protocol : Sorting and culturing of inflammatory monocyte‐like cells