Production of clinically applicable tissue-engineered cornea

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To produce clinically applicable tissue-engineered cornea, it is required to obtain tissue stem cells from each of three layers of the cornea. First, we showed that tissue stem cells can be obtained from each of three layers of the cornea by neuro-sphere assay. Then, we investigated the method of applying these tissue stem cells in animal models. We injected spheres from human corneal endothelium into the anterior chamber of rabbits and the spheres attached to the back surface of rabbit cornea, maintaining the transparency of the cornea. To evaluate the feasibility of a method of human corneal endothelial cells (HCEC) sheet transplantation using collagen membrane of 40 μm as a substitute carrier of Descemet's membrane. These results provide evidence for the feasibility of cultured HCEC sheet transplantation using collagen membrane for corneal endothelial decompensation. We seeded spheres from human corneal stroma onto the porous gelatin sheets of 40-50μm and transplanted the sheet into the corneal stromal pockets of rabbits. Then, the spheres from human corneal stroma produced abundant of extracellular matrix such as laminin and fibronectin, indicative of the clinical feasibility of spheres from human corneal stroma. We compared the distribution and self-renewal capacity of rabbit corneal endothelial cell precursors in the central and peripheral regions of the cornea. Our findings demonstrate that both peripheral and central rabbit corneal endothelium contain a significant number of precursors, but the peripheral endothelium contains more precursors with a stronger self-renewal capacity relative to the central region.