Hi all, I am wondering whether the density of the cells cultured...

Macrophages expand a lot in morphology when we stimulate them. 5e5 works for us because the macrophages still maintain a good confluency and we get good metabolite signals. If you are using a smaller cell type, a higher density definitely makes sense. But I would recommend trying a few different plating densities to determine the optimal seeding for your analysis. Hope this helps!