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Kyoung-pil Lee, Man-Il Huh, Jeongho Kim, Myung-Jun Kim, Hong Kyun Kim; Hypoxia induce endothelial invasion by using 3-deimensional choroidal neovascularization model. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4991.
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© ARVO (1962-2015); The Authors (2016-present)
Aged-related macular degeneration, the biggest cause of blindness in the world, were connected angiogenesis between retinal pigment epithelial (RPE) cells and choroid. However, there was no an appropriate 3D model, 2D in vitro model has a limitation reflecting the disease. We developed 3D retina-vascular mimetic system reflecting angiogenesis in AMD by stacking with each cell layer.
Human RPE cells and peripheral micro-vascular endothelial (PMVE) cells were co-cultured in normoxia and hypoxia condition for 24 hours. The hypoxia conditions were induced chemically using CoCl2. We examined the invasion assay of endothelial cells under 3D co-culture system and the expression of VEGF and HIF-1α in retinal pigment epithelial cells.
HIF-1a and VEGF mRNA level were increased in RPE cells under the hypoxia condition. ELISA assay data showed that higher level of the secretion of VEGF than a normoxia a CoCl2 dose-dependent manner.In the co-culture system combined RPE cells and PMVE cells, we observed PMVE cell invasion (35 ± 3.08 cells/unit area) into RPE cells in hypoxia condition. In the 50 ng/ml VEGF treated cells, also invaded toward RPE cells (13.3 ± 0.81 cells/unit area). Invaded cells of endothelial cells were detected by a CoCl2 concentration-dependent manner.
In this study, data showed hypoxia induced VEGF expression and then PMVE cell invasion in 3D co-culture model. We established new 3D model, an angiogenesis mimetic model related with AMD, would be able to detect invasion of endothelial cells under a disease condition. This model would be to employ for a candidate selection before a pre-clinical trial.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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