Real Time Movie of In Vitro Tube Formation
In vitro tube formation was assessed using a Matrigel endothelial cell tube formation assay (Chemicon International, Chemicon International, Temecula, CA) (Yahata, Shirakata et al. 2003). Matrigel, a solid gel of basement membrane proteins, prepared from Engelbreth-Holm-Swarm mouse tumor was added to 6 well plate and allowed to gel for >1 h at 37C. PAEC were seeded on the surface plates at 18 x 104 cells/well. Angiogenesis was observed by phase contrast microscopy and documented by collecting digital images over time (0 to 8 h). The time course of angiogenesis was evaluated by collecting and quantitating time-lapse movies of cells, using a live-cell imaging system and Metamorph software (Universal Imaging, Downingtown, PA).
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