4.2. In-Vitro Cell Culture

Immortalized mouse brain endothelial cells (bEnd3) from ATCC (Manassas, VA, USA) were used as a representative model for a mouse BBB endothelium. bEnd3, passage 25–35, were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), penicillin G (100 units/mL) and streptomycin (100 μg/mL); all were purchased from ATCC (Manassas, VA, USA). Cultures were maintained in a humidified atmosphere (5% CO₂) at 37 °C incubator (VWR International, Radnor, PA, USA).