Hcmec d3 cells

HUVECs (commercially purchased from Lonza, Walkersville, MD) and HCMEC/D3 cells (provided by Institut National de la Santé et del la Recherche Médicale [INSERM], Paris, France) were grown as previously described.16 (link),17 SKNSH human neuroblastoma cells (commercially purchased from American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle’s Medium supplemented with 10% heat-inactivated fetal bovine serum, penicillin (100 U/mL), streptomycin (100 µg/mL), and L-glutamine (2 mM) in a 5% CO₂ humidified incubator at 37°C. The cells were differentiated with 10 µM retinoic acid for 5 days.18 (link)

Human brain capillary endothelial hCMEC/D3 cells were obtained from Merck (Darmstadt, Germany). hCMEC/D3 cells were seeded in the collagen-I coated flasks and cultured in EGM-2 Endothelial Cell Growth Medium-2 Bullet Kit (Lonza, Verviers, Belgium) containing growth factors and 5% heat-inactivated fetal bovine serum (AusGene X, Brisbane, Australia) at 37 °C in 5% CO₂.