Cell culture and primary neurons

Human neuroblastoma cell lines SHSY5Y were grown at 37 ^oC with 5 % CO₂ in a 1:1 mixture of minimum essential medium MEM (Gibco 41090036) and Ham's F-12 medium (Gibco 21127002), supplemented with 10% fetal bovine serum (Gibco 10270106). The cells were plated in 96 well black plates with clear bottoms (Corning Incorporated 3603) at a density of 5 000 cells per well. Primary Hippocampal/Striatal neurons were isolated from C57Bl/6 mice at embryonic day 14. The embryonic brain tissue was dissected in Hank's buffered salt solution (HBSS, Invitrogen 88284) containing 50 U/ml penicillin and 50 mg/ml streptomycin (Thermofisher 15140122) and 8 mM Hepes buffer (Invitrogen 15630-080). Following centrifugation for 3 min at 150 x g, the pellet was resuspended in HBSS and any remaining blood vessels were let to sediment for 15 min at RT. The supernatant was centrifuged for 5 min at 150 x g and the cell pellet was resuspended in neurobasal medium (Invitrogen 21103049) supplemented with 1x B-27 (Invitrogen 17504001), 50 U/ml penicillin, 50 mg/ml streptomycin (Thermofisher 15140122) and 20 mM L-glutamine (Invitrogen 25030081). The cells were plated in 96 well black plates with clear bottoms (Corning Incorporated 3603) at a density of 10 000 cells per well. The media was changed completely the day after seeding and half of the media was changed every 2-4 days after that. All the procedures with primary neurons were conducted according to the Swedish ethical policies regarding animal experiments and were approved by the Uppsala County Animal Ethics Board (#5.8.18-08472/18).