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Y. Hayashida, K. Nishida, M. Yamato, K. Watanabe, N. Maeda, H. Watanabe, A. Kikuchi, T. Okano, Y. Tano; Autologous Transplantation of a Bioengineered Cultivated Oral Epithelial Cell Sheet using a Temperature-responsive Culture Surface for Ocular Surface Reconstruction . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4695.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: The limbal epithelium is used as an autologous cell source for treating unilateral ocular surface diseases, but not bilateral diseases. In this study, we have developed a novel tissue engineering approach for treating bilateral diseases using autologous oral epithelial cells expanded on temperature-responsive cell culture surfaces. Methods: Oral mucosal epithelial cells from rabbits were cultured on a temperature-responsive culture dish with 3T3 feeder cells for 2 weeks. Cultivated cells were then harvested by reducing the temperature of the culture insert. Tissues were observed by light and electron microscopy. Autologous transplantation of cultivated oral epithelial cells was also performed in rabbits with total limbal stem cell deficiencies. Results: The rabbit oral epithelial sheet was harvested from the dish as a transplantable cell sheet simply by reducing temperature without enzymatic treatment. The bioengineered sheets were three to five cell layers thick with good evidence of tight junction and desmosome formation. By autologous transplantation, the rabbit corneal surface was successfully reconstructed and its transparency was restored. Cornea-specific keratin K3 was expressed in the bioengineered oral epithelial cell sheet when grafted. Conclusions:The corneal surface can be successfully reconstructed by autologous transplantation of our bioengineered oral epithelial cell sheet. With promising implications for bilateral ocular surface diseases.
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