0.1 mg/ml DNaseI and 1xAntibiotic-Antimycotic (ThermoFisher) in aCSF at 37°C for 1h, with shaking of the tube every 15 min. The cells were gathered by centrifugation at 356 g for 5min followed by incubation in TrypLE for 30 min. The suspension was washed in aCSF, centrifuged at 356 g for 5 min and resuspended in aCSF, 1% BSA. After manual trituration using BSA-coated fire-polished Pasteur pipettes with decreasing opening size, the single cell suspension was filtered through 70μm filter (Miltenyi Biotec) and cleaned of debris by centrifugation through 1 mL FBS at 800 g for 10min. The cells were resuspended in oxygenated aCSF, 1%BSA and filtered through a 30μm filter (Miltenyi Biotec). Tom+ cells were FAC sorted on a BD FACSAria II and collected in ice-old aCSF.
