Aortic Ring Angiogenesis Assay
- Cover a 48-well plate with Matrigel (100μl/well) of and incubate for 30 min at 37°C, 5% CO2.
- Sacrifice 1-2 month old mice/rats and excise thoracic aorta.
- Transfer the aorta into a dish with sterilized 1x PBS buffer.
- After removing the fibroadipose tissue, section arteries into 1-1.5 mm long cross sections, rinse the sections five times with endothelial cell growth medium (ECGM), and place them on the Matrigel-coated wells.
- Cover the artery rings with an additional 100μl of Matrigel. If there are some bubbles, use pipette to remove them.
- Incubate the rings in 1.5 ml of ECGM medium for 24 hours.
- Replace the medium with 1.5 ml of endothelial cell growth medium (or containing with the interesting peptides/compounds/others).
- After 2 days, the medium was replaced with medium of the exact composition as described in step 4.
- At the day 4, take images of aortic rings. Delineate the outgrowth area and measure the outgrowth of micro-vessel.
Last Edited By: Protocol Monkey on 2008-09-28 [01:23:06]