Culturing Human Cerebral Endothelial and HEK293T Cells

Human cerebral microvascular endothelial cells (HBEC-5i) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured on 0.1% gelatin solution-coated (ATCC) flasks in Dulbecco's modified Eagle's medium (DMEM): F12 medium (ATCC) supplemented with 10% fetal bovine serum (FBS, Gibco/Thermo Fisher, Waltham, MA, USA), 40 μg/mL endothelial cell growth supplement (ECGS, ATCC), and 100 U/mL penicillin as well as 0.1 mg/mL streptomycin (Beyotime, Shanghai), maintained at 37°C in a humidified atmosphere with 5% CO₂ as described previously [20 (link)].
Human embryonic kidney (HEK) 293T cells (ATCC, Manassas, VA, USA) were grown in DMEM containing 10% FBS, 100 U/mL penicillin, and 0.1 mg/mL streptomycin (Beyotime, Shanghai), maintained at 37°C in a humidified atmosphere with 5% CO₂.