Our lab usually uses the shaking method to isolate primary mouse microglia from mixed glial cultures. Till now, I have been collecting microglia, plating them in isolation, and then activating them for further experiments. I'm curious to test whether I will be able to shake 'activated' microglia off if I activate the mixed glial culture instead? Has anyone tried this?
Hi Tanya,
how are you going to stimulate your cells?
We stimulated mixed cultures with different concentrations of LPS, INFy, Ab, as well as with oxygen-glucose-deprivation and isolated microglia by shaking. However, the problem was that astrocytes and oligos were also stimulated with the aforementioned stimulations and were present in the isolated cultures. I think that a sorting method following stimulation would be more adequate (e.g. MACS).
Cheers,
Pardes