Penicillin streptomycin

Human neuroblastoma SH-SY5Y (ATCC CRL-2266) cells were maintained at 37℃ in a humidified incubator with 5% CO₂ and 95% relative humidity. SH-SY5Y cells were cultured in Dulbecco’s modified eagle medium (DMEM)/F12 supplemented with 10% of heat-inactivated fetal bovine serum (FBS) and 1% penicillin–streptomycin from Bovogen Biologicals (Victoria, Australia). SH-SY5Y cells were differentiated, as described previously [28 (link)]. Briefly, cells were seeded in MaxGel ECM (E0282, Sigma-Aldrich, Sydney)–coated 96-well plates with media containing 10 µM retinoic acid (RA, Sigma-Aldrich, Sydney) in the dark. Cells were observed for neurite outgrowth using the IncuCyte ZOOM live-cell imaging system (Essen BioScience, UK). Neurite integrity was quantified using the automated NeuroTrack module of the IncuCyte ZOOM system for live-cell analysis. Furthermore, differentiated cells were exposed to various concentrations of CBD (0.1, 0.5, 1, 2.5, 5, 10 µM) in 0.1% DMSO and incubated for 24, 48, and 72 h.