M199 medium

HUVEC/TERT 2 was obtained from ATCC (ATCC, Manassas, VA, USA). The cell line was isolated from the vascular endothelium of a white female patient. To culture the cells, M199 medium (Biosera, Nuaille, France) was used, which was supplemented with 10% heat-inactivated FBS, 1% penicillin/streptomycin, 1% amphotericin B, 2 mM glutamine (Biosera, Nuaille, France) and endothelial cell growth medium-2 (Lonza, Basel, Switzerland). The cells were maintained in an Esco CelCulture^® CO₂ Incubator (Esco Lifesciences Group, Singapore) at 37 °C, under 5% CO₂. Before seeding, to support the adhesion of the cells, 0.1% gelatin solution was used. To create the inflammatory model, LPS (eBioscienc, San Diego, CA, USA) was added to the M199 medium to a final concentration of 100 ng/mL. The cells were divided into four groups, 24 h of incubation with basic medium (control), 24 h incubation with 100 ng/mL of LPS (LPS), 24 h incubation with 100 μg/mL of HAE (100 μg/mL HAE) and 24 h incubation with 100 ng/mL of LPS plus 100 μg/mL of HAE (LPS + 100 μg/mL HAE).

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.