Choroidal endothelial cells transmigrate across the retinal pigment epithelium but do not proliferate in response to soluble vascular endothelial growth factor

摘要: The purpose of this study was to investigate the effects soluble VEGF on human choroidal endothelial cell (CEC) transmigration across an RPE monolayer as it relates neovascularization in AMD. In coculture assays, ARPE-19 (ARPE) plated undersides Transwell inserts having 0.4 microm pores. Primary CECs were then into insert. counted after 72 hr growth. CEC proliferation also measured culturing ARPE-CEC coculture-conditioned media or with exogenous VEGF121 and/or VEGF165 added. Transmigration assays performed Transwells 8.0 pores: green-labelled without red-labelled ARPE some wells provide a chemotactic gradient for transmigration. After plated, green cells either within well that transmigrated epithelial monolayer, underside insert but not beyond monolayer. A neutralizing antibody added at time and counted. protein conditioned assays. Compared control, significantly increased when cultured (p=0.001) (p<0.001). However, there no effect VEGF121, VEGF165, both solo CECs. Antibody did reduce proliferative CEC. (p<0.001) compared well. greater than (p<0.004). Exogenous reduced transmigration, dose-dependent. provides transmigrate ARPE. Neutralization partially reduces Whereas does increase CECs, enhances proliferation, suggesting growth factors other cause proliferation. These findings may have relevance transformation occult CNV neurosensory retina