How does macrophages get activated?
Macrophages are activated by interferon-γ (IFN-γ) from various sources – including activated NK cells or T cells – or by non-immunologic stimuli such as endotoxin.
How do you differentiate THP-1 cells?
THP-1 cells were differentiated into macrophage-like cells (THP-1 macrophages) by incubation in the presence of PMA, which leads to a macrophage-like phenotype characterized by changes in morphology and increased cell surface expression of CD11 and CD14 [10,11,14,17,21].
What does THP-1 cells stand for?
acute monocytic leukemia
THP-1 designates a spontaneously immortalized monocyte-like cell line, derived from the peripheral blood of a childhood case of acute monocytic leukemia (M5 subtype) (3).
Do macrophages need to be activated?
These classically activated macrophages are vital components of host defence, but their activation must be tightly controlled because the cytokines and mediators that they produce can lead to host-tissue damage.
Why do macrophages need to be activated?
Activated macrophages undergo many changes which allow them to kill invading bacteria or infected cells. They release toxic chemicals and proteins which have toxic effects on other cells.
What type of cells are THP-1?
THP-1 is a human monocytic cell line derived from an acute monocytic leukemia patient. It is used to test leukemia cell lines in immunocytochemical analysis of protein-protein interactions, and immunohistochemistry.
How are THP-1 cells differentiated into macrophages?
THP-1 cells can be differentiated into macrophage-like cells that resemble properties of mature macrophages by activation of protein kinase C (PKC) with phorbol-12-myristate-13-acetate (PMA), ultimately resulting in cells with increased adherence and loss of proliferative activity ( Schwende et al., 1996 ).
How long to Prime THP-1 cells for differentiation?
However, priming THP-1 cells with 10 ng/ml PMA for 24 h and then resting for 24 h before exposing the primed cells to IL-4 for 48 h resulted in cells which transcribed MRC1 ( Morón-Calvente et al., 2018 ), which encodes the C-type lectin mannose receptor C 1 (CD206).
How are THP-1 cells polarized after PMA differentiation?
As Yaron mentioned, after PMA differentiation the THP-1 cells are polarized towards proinflammatory activation. For example, the cells stably express high levels of pro-interleukin-1beta without the need for additional stimulation with bacterial components, which was relevant to my own studies.
Which is the best treatment for THP-1 cells?
Treatment of THP-1 cells with PMA as a priming step, followed by rest and treatment with either proinflammatory or regulatory cytokines results in macrophage-like cells that express some of the marker genes typical of human monocyte-derived macrophages (hMDMs) ( Daigneault et al., 2010; Kohro et al., 2004; Shiratori et al., 2017 ).