Human umbilical vein-derived endothelial cells (HUVEC) were obtained by ATCC (PCS-100-010—ATCC USA). HUVECs were plated onto flask with EBM medium supplemented with 10% fetal calf serum and different endothelial growth factors as previously reported [29 (link)].
HUVECs were cultured for 24 h into an airtight humidified chamber flushed with a gas mixture containing 5% CO2, 94% N2, and 2% O2 at 20 atm, 37°C [30 (link)] or with the uremic toxins asymmetric dimethylarginine (ADMA 10 μg/ml from Sigma Aldrich, St. Louis, MO), p-cresyl sulfate (1 μg/ml from Alsachim, Illkirch Graffenstaden, France) and indoxyl sulfate (10 μg/ml from Sigma Aldrich, St. Louis, MO).
HUVECs were cultured for 24 h into an airtight humidified chamber flushed with a gas mixture containing 5% CO2, 94% N2, and 2% O2 at 20 atm, 37°C [30 (link)] or with the uremic toxins asymmetric dimethylarginine (ADMA 10 μg/ml from Sigma Aldrich, St. Louis, MO), p-cresyl sulfate (1 μg/ml from Alsachim, Illkirch Graffenstaden, France) and indoxyl sulfate (10 μg/ml from Sigma Aldrich, St. Louis, MO).