Microglia cell culture
Mixed primary glial cell cultures were prepared by enzymatic dissociation of postnatal P0-P5 mouse brains using Neural Tissue Dissociation Kit (P) (Miltenyi Biotec, 130-092-628). The cells were allowed to proliferate for 10 days prior to isolation of microglia by the “shake off” method (19). The isolated microglia were on average 95% pure, when characterized as CD11b+/CD45low on the flow cytometer (BDTM LSR II, BD biosciences).