BeWo cells (ATCC CCL-98) were grown at 37°C under a humidified 5% CO2/95% air atmosphere in F-12K Medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. Experiments were performed under serum-starved conditions using cells at 80% confluence between passages 30 and 40. BeWo cells were grown under low glucose (7 mM) or HG (25 mM) conditions. Supernatants were harvested, submitted to centrifugation, aliquoted, and frozen at −80°C until use. The coculture experiments were performed in 6-well cell culture plates with 12 mm coverslips (Corning® BioCoatTM Coverslips). When indicated, neutrophils were directly added to the BeWo cells at a density of 0.5 × 106 neutrophil/ml for 3 h. In neutralization studies, 5 µg/ml of chimeric anti-TNF-α IgG1 antibody infliximab (Remicade, MSD) was given simultaneously to neutrophil addition.