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X. Nie, C.L. Berglin, J. Wen, N. L'Hernault, H. Yang, J. Kapp, H.E. Grossniklaus; Murine model of choroidal neovascularization with subretinal beads and heterologous retinal pigment epithelial (RPE) cells . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1864.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To induce a CNV model in mice that recapitulates the temporal cellular and cytokine expression of human CNV. Methods: C57BL/6 mice and Balb/C mice were injected with a 2 µl mixture of heterologous RPE and microbeads in the subretinal space. Injections of beads or RPE separately were used as controls. A time curve of CNV growth and cytokine expression was recorded up to 28 days. The actual thickness of the CNV complex was estimated indirectly by measuring the difference between the thickness from the outer border of the pigmented choroidal layer to the top of the CNV complex (T) and the thickness of the intact, pigmented choroid adjacent to the lesion (C). Five to 10 serial sections from each membrane were measured and the highest value (representing the top of a given CNV complex) for each membrane was stored. The stored data from each mouse were pooled, and a mean T and C was calculated. The relative thickness of the CNV membrane complex in each mouse was then calculated using the formula R = T–C/C. Light microscopy, immunohistochemistry, confocal scanning laser microscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to examine the CNV. Results: The CNV grew in the subretinal space (type 2 pattern) with an outer reflected layer of RPE. A time curve revealed a typical growth pattern with increased growth from day three (R = 4.9) and day five (R = 7) to maximal size at day seven (R = 8.5) and day 10 (R = 7.9) with a gradual decline thereafter to day 28 similar to other CNV models. Findings included ingrowth of endothelial cells, fibroblasts macrophages and lymphocytes demonstrated by immunohistochemical staining and TEM. Confocal microscopy with dual labelling revealed expression of MCP and TGF–ß by CK–18 positive cells. The cytokine expression exhibited a temporal course associated with the CNV growth. Conclusions: CNV in this murine model recapitulates human CNV with initiation, active inflammatory and regression stages. The CNV grows in a type 2 pattern in this model.
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