Hello,
i would like to ask you how it is possible to differentiate freshly isolated monocyte to M2d subtype (TAMs). I studied about M2a,b,c differentiation but it is not clear to me how is it possible to obtain TAMs. Maybe using IL-6 or LIF? or there is a cytokines cocktail?
Thank you!
Hi Meroopi
We performed either co-culture with cancer cells or used conditioned media from cancer cells (A549; Müller-Quernheim. Tumor-cell co-culture induced alternative activation of macrophages is modulated by interferons in vitro. J Interferon Cytokine Res 2012: 32(4): 169-177.). Both work nicely. Fibroblasts might also induce such a phenotype (Prasse, A vicious circle of alveolar macrophages and fibroblasts perpetuates pulmonary fibrosis via CCL18. Am J Respir Crit Care Med 2006: 173(7): 781-792.). We tried to identify the responsible factor(s), However, this is ongoing work and we have no conclusive results yet. The important point is time! We allow monocytes at least three days to differentiate as you may know from your previous work.
good luck
Gernot
Hello Gernot,
thank you very much for your answer. i will try to choose the most appropriate method for my experiment!
have a nice day and good luck also to you!
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