In vitro tube formation assay

EM Eui Jung Moon
SM Stephano S. Mello
CL Caiyun G. Li
JC Jen-Tsan Chi
KT Kaushik Thakkar
JK Jacob G. Kirkland
EL Edward L. Lagory
IL Ik Jae Lee
AD Anh N. Diep
YM Yu Miao
MR Marjan Rafat
MV Marta Vilalta
LC Laura Castellini
AK Adam J. Krieg
EG Edward E. Graves
LA Laura D. Attardi
AG Amato J. Giaccia
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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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