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C.J. Lee, S.F. Bent, M.S. Blumenkranz, H.A. Fishman; RPE Cell Adhesion on Spatially Patterned Lens Capsule . Invest. Ophthalmol. Vis. Sci. 2003;44(13):501.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Retinal pigment epithelial (RPE) cells are of uniform morphology and size in the retina; however, when cultured on lens capsule, they adhere and spread readily, while exhibiting a variety of morphologies. As a potential replacement for damaged RPE and Bruch’s membrane in patients with age-related macular degeneration, lens capsule was micropatterned to control the attachment and growth of RPE cells. The cell attachment probability on micropatterned lens capsule was examined to determine the parameters necessary to obtain the organized, high density layer necessary to mimic the native epithelium. Methods: Photolithography was utilized to create silicon molds. Elastomer stamps, cured on these molds, were inked with polyvinyl alcohol (PVA) and then pressed against the lens capsule to transfer the polymer to create a micropattern on the surface of the lens capsule. Human RPE cells (ARPE-19) cultured on the modified lens capsules were examined for viability and morphology by light, fluorescence, and electron microscopy. Results: We successfully micropatterned inhibitory molecules such as PVA on lens capsule and examined RPE cells cultured on the modified surface. Micropatterning PVA confined RPE cells to cuboidal structures that closely mimic the natural RPE layer. We found that pattern size strongly affects the probability of cell adhesion and subsequent cell spreading. The cells have been successfully patterned on lens capsule to hexagonal patches of diameters varying from 15 µm to 50 µm, with the smaller hexagons occupied by a single cell and the larger hexagons by multiple cells. Thus, all pattern sizes lead to a high density layer of RPE cells. The hexagonal patches have separations of only 2, 5, and 10 µm, which allow for the eventual spreading of the cells and for the possible formation of tight junctions between neighboring cells. Conclusions: Overall, micropatterning PVA appears to be a promising and reproducible method for confining cells to high density and to a single morphology. The ability to organize the RPE cells into a single sheet on a biocompatible substrate may aid in the creation of an implant that will restore function to patients suffering from age-related macular degeneration.
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