Differentiation and Polarization of Monocyte-Derived Cells into Macrophage-Like Cells
Published: July 31, 2023
Abstract
Source: Mily, A., et al. Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection. J. Vis. Exp. (2020).
In this video, we demonstrate a procedure to induce the differentiation and polarization of peripheral blood-derived monocytes into M1 or M2 macrophages. Incubation with granulocyte-macrophage colony-stimulating factor, lipopolysaccharide, and interferon‐gamma yields pro-inflammatory M1 macrophages, while incubation with macrophage colony-stimulating factor and interleukin-4 induces anti-inflammatory M2 macrophage production.
Protocol
All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Differentiation and polarization of monocyte-derived cells Use plastic adherence for isolation of monocytes. Briefly, seed freshly isolated peripheral blood mononuclear cells (PBMCs) in a 6-well culture plate at an appropriate concentration, e…
Divulgazioni
The authors have nothing to disclose.
Materials
| Falcon 6-well Flat Bottom plates | Corning Life Sciences | 353046 | |
| GM-CSF | Peprotech | 300-03 | |
| IFN-γ | Peprotech | 300-02 | |
| IL-4 | Peprotech | 200-04 | |
| L-Glutamine | GE Healthcare Life Sciences | SH30034.01 | |
| M-CSF | Peprotech | 300-25 | |
| RPMI 1640 | Life Technologies Corporation | SH30096.01 | |
| Fetal bovine serum (FBS) | Sigma-Aldrich | F7524 | |
| HEPES | GE Healthcare Life Sciences | SH30237.01 | |
| LPS (Escherichia coli O55:B5) | Sigma-Aldrich | L6529 | |
| Na-pyruvate | GE Healthcare Life Sciences | SH300239.01 |
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Citazione di questo articolo
Differentiation and Polarization of Monocyte-Derived Cells into Macrophage-Like Cells. J. Vis. Exp. (Pending Publication), e21520, doi: (2023).