Culturing Human Umbilical Vein Endothelial Cells

The HUVECs originated from human umbilical cords that were collected from normal-term placenta and obtained from the Department of Obstetrics and Gynecology, Clinical Centre, University of Debrecen, Debrecen, Hungary. HUVECs were maintained according to the method previously described [24 (link)]. Cells were cultured in M199 medium (Biosera, Nuaille, France) supplemented with 10% (v/v) fetal bovine serum (Biosera, Nuaille, France), 10% (v/v) endothelial cell growth (EGM)-2 complex medium (Lonza, Basel, Switzerland), 1.2% (v/v) 2 mM glutamine (1:500; Biosera, Nuaille, France) 1.2% (v/v) 1X penicillin/streptomycin (Biosera, Nuaille, France), 1.2% (v/v) 1X penicillin/streptomycin (Biosera, Nuaille, France), and 1% amphotericin B. Cells were subcultured at 80–100% confluence and incubated at 37 °C with 5% CO₂ level.