Hello,
I want to differentiate Monocytes isolated from PBMCs into M0 state Macrophages. We can either use CSF-1 (M-CSF) or GM-CSF for this purpose. But in many papers I found that monocytes treated with M-CSF exhibits M2 like phenotype and those treated with GM-CSF shows M1 type phenotype. Few groups are using cocktail of both of these cytokines. I need to get Macrophages at M0 state which can then treated with INF- γ+LPS to get M1 or with IL-4 or IL-13 to differentiate into M2.
Dear Abishek,
According to this Paper (https://www.atsjournals.org/doi/full/10.1165/rcmb.2015-0012OC), both GM-CSF and M-CSF should work. It will be important to thoroughly characterise your cells throughout the Process to follow their development. It is also important to note that the Isolation protocol might influence cell fate, which could explain different findings by different Groups.How Do you Plan to isolate the monocytes and how will their purity be determined?
Best
Andreas
I would recommend using M-CSF. The resultant macrophages are said to be more M2 like compared to macrophages that are differentiated with GM-CSF. Regardless of which cytokine you use to differentiate your monocytes, you would still have to polarize the macrophages to be M1 or M2 with the cytokine mixes you mentioned. This is a widely accepted protocol and the reason why I recommend using M-CSF is to get a purer/more homogeneous macrophage population in the end. GM-CSF would cause your monocytes to differentiate into a mix of macrophages and monocyte derived dendritic cells.
MCSF (10 ng/ml) will work. If you do not have MCSF, alternatively you can use GM-CSF but at very low concentrations (1-5 ng/ml). Compared to GM-CSF, MCSF induces the most effective differentiation into macrophages.
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