104 cells were plated on a 96-well plate with endothelial cell medium containing 5% FBS (Cell Biologics #M1168). After 24 h incubation under normoxia or hypoxia, media was collected and briefly spun down at 200 × g for 5 min to remove cell debris. 50 μl of Corning® Matrigel® Growth Factor-Reduced basement membrane matrix (Corning 354230) was added to a 96-well plate on ice. After 1 h incubation to polymerize at 37 °C, 0.5 × 104 GFP-labeled human umbilical vein endothelial cells (HUVEC) (Angio-Proteomie #cAP-0001GFP) in 100 μl of collected cell media were plated onto matrigel. 4 h after tube formation was imaged using a fluorescent microscope (Leica DMi8) with 10-power magnification.
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