Hi all,

I'm working with primary, human monocyte-derived macrophages and have been polarizing them in vitro with LPS+IFNy or IL-4 (or medium alone). I've done this many times before in human and mouse, and I know from the literature that with LPS+IFNy treatment, CD206 should be low and CD86 and MHCII should be high while with IL-4 (50ng/mL) the opposite should be true.

Sometimes though I find that CD86 and MHCII expression is higher after IL-4 treatment compared to LPS+IFNy!  I differentiate my macrophages in 50ng/mL M-CSF for 7 days in RPMI with 10% FBS.  On day 7 I wash the cells, add my treatments in the same medium as above, then after 24 hours detach the cells with accutase EDTA and stain for flow cytometry after blocking with 8mg/mL human IgG.

Any thoughts would be much appreciated.

Hefin

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