We are currently trying to investigate some phenomenon of macrophage observed in human tissue sample, using cultured M1/M2 polarized macrophages in vitro. Therefore, I am designing my experiments as follows:
1. Polarization of differentiated macrophages towards M1/M2 phenotypes
Monocytes are initially cultured for 6 days with human AB serum to obtain differentiated macrophages. And then, in order to promote differentiation into M1 or M2 macrophages, cells are treated with IFNγ plus LPS, or IL-4, respectively,for 12-24 hours.
2. Polarization of monocytes towards M1/M2 phenotypes
Monocytes are induced to differentiate for 6 days in the presence of either IFNγ plus LPS to obtain M1 phenotype, or IL-4 to obtain M2 phenotype, both supplemented with human AB serum.
Which method should I choose?
I use the method 1 for polarization macrophages always, I think that method is the most correct. I doubt that the method 2 work for you goal.
Addition of IL-4 and GM-CSF will differentiate Monocytes to Dendritic cells so please be cautious in this case. We generally differentiate monocytes in 11 days to Macrophages with human serum with out adding any external M-CSF.
I followed this publication's methods successfully in the past:
Proc Natl Acad Sci U S A. 2004 Mar 30;101(13):4560-5. Epub 2004 Mar 19.
Hope this helps.
Thank you very much for all suggestions, I will look into these!
@Denis Polancec
Can you tell me what are the differences between immature and mature macrophages?
Sorry, I have a question instead of an answer. My question is that can you induce M1 macrophages into M2 macrophages? Thanks!
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