Figure 4.
The effect of SLPI on in vitro growth and migration of HUVECs. (A) 1 × 104 cells/well were seeded in triplicate in 12-well plates and cultivated in growth medium containing recombinant SLPI at the indicated concentration. Cell numbers were determined by direct counting, using a haemocytometer, after detachment of adherent cells. (B) Effect of SLPI on endothelial cell migration. A total of 1 × 105 HUVECs were seeded in the upper chamber of a culture insert coated with Matrigel containing rSLPI (0, 0.1 and 1 μg/ml). After 6 h incubation, the migrating cells were counted. (C) Effect of SLPI on an in vivo Matrigel plug assay. Various doses of rSLPI were mixed with Matrigel, with or without bFGF (100 ng/ml). A total of 0.5 ml Matrigel was implanted into the mouse. After 7 days, the Matrigel plug was removed and neovessel formation was quantified by calculating the vascular density. The dose-dependent effects of rSLPI in (B) and (C) were validated by one-way ANOVA, followed by Fisher’s PLSD procedure. Each value in (A–C) represents mean ± SD of triplicate measurements