The aims of this proposal are to establish short-term ocular surface epithelial cell cultures from rabbit and human tissue. The tissues to be studied are corneal epithelium from explants of rabbit and human cornea, and conjunctival epithelium which has resurfaced denuded corneal stromas. Once these cell cultures have been established, they will be characterized in terms of their viability, appearance, both light and electron microscopically and biochemically. Using cells from such cultures, epithelial cell-corneal stroma combinations will be created by incubating corneal stromas in suspensions of these cells. These epithelial cell-stromal cultures will also be characterized, with particular emphasis on cell: substrate interactions, such as spreading, adhesion, and viability. Manipulation of a number of factors, including stromal cell viability, growth factors, serum supplementation and surface active agents (e.g. fibronectins and mucins) will be undertaken to learn more about the nature of cell: substrate interdependencies. Finally, epithelial cell-stromal combination will be used to replace ocular surface components in rabbits, to determine the potential usefulness of this approach in the therapy of ocular surface disease and injury in humans.
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