Human umbilical vein endothelial cells huvecs

Human umbilical vein endothelial cells (HUVECs) were purchased from Cell Applications (San Diego, CA). The cells were cultured in 100x20 or 60x15 mm cell culture dishes (Corning, 430167 and 430166) in 5% CO₂ at 37°C and passaged when achieved 80% confluency or more. Medium was changed every other day or when the confluency reached 60% or more. The cells were used up to 16 doublings, the minimum number of doublings guaranteed by Cell Applications. The growth medium used was Endothelial Cell Growth Medium (Cell Applications, 211–500).

The SKOV-3 ovarian cancer cell line was obtained from the American Type Culture Collection. A2780 and OVCAR429 ovarian cancer cells were kindly provided by Dr. Danny Dhanasekharan (Stephenson Cancer Center, OUHSC). The A2780 ovarian cancer cell line was initially obtained from Sigma-Aldrich (St. Louis, MO). OVCAR429 are ovarian cancer cells that have been previously published [18 (link), 19 (link)]. A2780 and OVCAR429 cells were maintained in RPMI medium (Invitrogen). SKOV-3 cells were maintained in McCoy’s 5A medium (Invitrogen). Media were supplemented with 10% fetal bovine serum (Invitrogen), 100 IU/mL of penicillin, and 100 μg/mL of streptomycin (Invitrogen) at 37°C in a humidified incubator containing 5% CO₂. Human umbilical vein endothelial cells (HUVECs) and endothelial cell media were purchased from Cell Applications (San Diego, CA). Sorafenib was obtained from LC laboratories (Woburn, MA). SB225002 (N-(2-hydroxy-4-nitrophenyl)-N′-(2-bromophenyl)urea) is a potent and selective non-peptide inhibitor of CXCR2 (IL-8R) chemokine receptor. SB225002 was procured from Tocris Bioscience (Bristol, UK). Drug preparations for Sorafenib and SB225002 were performed according to methods described elsewhere [20 (link), 21 (link)].