concentration 50,000/cm2 for further differentiation into microglia-like cells in Advanced DMEM/F12:Neurobasal medium (both Gibco) supplemented with 1x B27 supplement plus Vitamin A (Thermo Fisher Scientific), 2 mM GlutaMAX, 0.055 μM β-mercaptoethanol, 20 ng/ml M-CSF and 100 ng/ml IL-34 (PeproTech) for 14 days with medium change twice a week. The cells were stimulated on day 13 of culture with either 500 U/ml TNF-α or 40 ng/ml IL-4 (Peprotech).
