I've been treating microglia with Il-4, and Il-13 (separately), to induce M2, but I'm not seeing any obvious response. I've tried 0.01 up to 100 ng/ml, and both 24 and 72 h exposures.
Both are recombinant rat, used on rat cells. Medium contains 10% FBS.
We've looked at qPCR (Il1B, TNFa, Il6, TGFB, Fizz, Ym1, Cxcl10, Il4r) without seeing any clear changes.
So, just wondered whether you may spot anything aberrant in my expt design..?
Hello Sama,
As you can see in the litterature a lot of people use 20 ng/ml of rIL4 for up to 72 h to polarize microglia toward a M2a phenotype. I don't think there is anything wrong in your experiment design but maybe you can look at other targets or shorter time points...There are a lot of studies in the litterature describing the expression profile of mRNA in polarized microglia that may help you choosing new targets. 2 papers that I really like are one from Chhor Vibol published in 2013 in Brain, Behavior and Immunity untitled " Characterization of phenotype markers and neuronotoxic potential of polarised primary microglia in vitro" and an other from Freilich Robert published in 2013 in Plus one called "Integrated Expression Profiles of mRNA and miRNA in Polarized Primary Murine Microglia".
In my own experiments I mainly use Arginase-1 as a marker for M2-activated microglia/macrophages as you can see in my paper published in 2015 in Stroke "Delayed pituitary adenylate cyclase-activating polypeptide delivery after brain stroke improves functional recovery by inducing m2 microglia/macrophage polarization".
Good luck with your experiments!
Article Delayed Pituitary Adenylate Cyclase-Activating Polypeptide D...
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