Differentiation and Polarization of Monocyte-Derived Cells into Macrophage-Like Cells

Specific growth factors and cytokines regulate the differentiation of monocytes — innate immune system cells — into pro-inflammatory M1 or anti-inflammatory M2 macrophages.

To study monocyte differentiation, seed peripheral blood mononuclear cells in a plastic multi-well plate containing a suitable medium. Incubate to allow the monocytes to attach to the plate surface while the non-adherent mononuclear cells remain suspended. Remove the non-adherent cells.

Add medium containing growth factor granulocyte-macrophage colony-stimulating factor, GM-CSF, to one set of wells and medium containing macrophage colony-stimulating factor, M-CSF, to the other.

GM-CSF binds to its heterodimeric receptor on the monocyte cell surface, leading to monocyte expansion and differentiation into M1-like macrophages, coupled with pro-inflammatory cytokine release.

M-CSF binds to its homodimeric receptor on the monocyte cell surface, triggering monocyte proliferation, differentiation into M2-like macrophages, and anti-inflammatory cytokine release.

Incubate the M1-like macrophages with lipopolysaccharide, LPS, and the cytokine Interferon‐gamma, IFN‐γ.

LPS and IFN‐γ treatment yields fully polarized and matu