Tube formation assay
Ninety-six-well plates were precoated with 50
μL
liquid
Matrigel
per well. After incubation at
37°C
for 1 hour,
HUVESs
(1.5 × 104 cells/well), suspended in DMEM medium with
PBS,
10 nmol/L thrombin, 25 nmol/L r-hirudin, 50 nmol/L DTIP, 10 nmol/L
thrombin + 25 nmol/L r-hirudin and 10 nmol/L thrombin + 50 nmol/L DTIP
and were seeded and cultured for 8 hours.
Five
randomly chosen fields were photographed and counted.