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Tetsuya Toyono, Tomohiko Usui, Seiichi Yokoo, Satoru Yamagami, Shiro Amano; In Vitro Corneal Neovascularization Model. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2401.
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© ARVO (1962-2015); The Authors (2016-present)
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.
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.
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.
We established a new in vitro corneal neovascularization assay. Our results suggest that HCSCs possess potent anti-angiogenic characteristic, which prevent angiogenesis.
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