Figure 1.

Primary microglia culture and main usages to assess the phagocytic process. Schematic figure depicting newborn mice (P0–P4) from pregnant female used to obtain primary microglia culture. After dissection and enzymatic digestion of cortices and hippocampi, cells are resuspended in growth medium (usually composed by either DMEM or EMEM with 10 to 20% of FBS), to sustain microglial growth. Cells are plated in T75 flasks and cultured for at least 10 days at 37°C. Microglia are subsequently collected either by vigorously tapping the flasks, by agitation at 230–245 rpm for 45 min or through mild trypsinization. In some assays, microglia are cultured alone or co-cultured with neurons and the cells are analyzed by fluorescent microscopy (e.g. to quantify neuronal spine number) or by electrophysiology (top panel); in other assays, microglia phagocytic properties are assessed by feeding the cells with specific substrates: fluorescent beads, synaptosomes, liposomes, or apoptotic neurons (bottom panel).