Abstract
Purpose: :
The purpose of this study was to establish in vitro corneal neovascularization model using co-culture system of human corneal stromal cells (HCSCs) and human umbilical vein endothelial cells (HUVECs) and to examine the roles of HCSCs in the tube formation of HUVECs.
Methods: :
HCSCs and HUVECs (HCSCs + HUVECs) were co-cultured with or without VEGF-A165 (10ng/ml). In vitro tube formation has been established using co-culture system of human dermal fibroblasts (HDFs) and HUVECs (HDFs + HUVECs), and this co-culture assay was used as control. HUVECs were stained by anti-CD31 antibody, and the area of CD31-positive tube formation (vessel area/total field) was quantified by ImageJ software.
Results: :
The tube formation areas of co-cultured without VEGF were 6.81±1.07% in HCSCs + HUVECs and 10.37±2.52% in HDFs + HUVECs (P < 0.0001). The areas of co-cultured with VEGF-A165 were 12.28±3.41% in HCSCs + HUVECs, 33.87±9.10% in HDFs + HUVECs (P < 0.0001). The tube formation area in HCSCs + HUVECs was significantly less than that in HDFs + HUVECs with or without VEGF-A165.
Conclusions: :
We established a new in vitro corneal neovascularization assay. Our results suggest that HCSCs possess potent anti-angiogenic characteristic, which prevent angiogenesis.
Keywords: neovascularization • cornea: stroma and keratocytes