The stimulation of capillary endothelial cell migration by aqueous humor

We have developed a quantitative in vitro assay of aqueous humor based on capillary endothelial cell migration. Bovine adrenal capillary endothelial cells are plated onto glass coverslips that have previously been coated with colloidal gold and incubated for 18 hr. During this time, cells that migrate sweep up the gold and leave a bare track that serves as a record of their movement. When the coverslips are projected by phase microscopy onto a television screen, the tracks can be traced and their areas are measured by a Zeiss MOP-3 digital image analyzer. V-2 carcinoma was implanted onto the iris of each eye of 15 NZ white rabbits and 200 μl of aqueous humor was aspirated from the anterior chamber. The aqueous humor was added to nonconditioned culture medium (DME with 10% calf serum) in concentrations of 6.25, 25, 50, and 100 μl. The tracks of 100 cells were traced for each concentration. The aqueous humor from these tumor-bearing animals stimulated capillary endothelial cell migration in a statistically significant, dose-dependent fashion. Aqueous humor from the eyes of 10 control rabbits (5 sham operated and 5 with heat killed, washed tumor implants) did not stimulate capillary endothelial cell migration at any concentration. This in vitro quantitative capillary endothelial cell migration assay of aqueous humor might improve the clinical diagnosis of intraocular tumors, guide subsequent treatment regimens, and identify those patients at a high risk of developing malignant eye tumors.