The aim of my study is to cultivate different types of macrophages, and to put them in co-culture with fibroblasts. I already arrived to differentiate monocytes in M1 and M2a macrophages. But my problem is that, when I put macrophages on my fibroblasts, I don't see any effect. Maybe it's because my macrophages are not activated ? What should I use to activate and to make them synthesize cytokines ?
Two questions for you:
E. coli LPS (I use O111:B4) at a concentration of 1-10 ng/mL will induce several cytokines to release from macrophages within a few hours. A great example is TNF. However, other cytokines will be expressed but not released, such as IL-1b. It will remain intracellular upon LPS stimulation until induced to release by an inflammasome activator.