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목적 : In this study, four decellularization protocols for acellular lenticules were evaluated to determine suitability as a scaffold for bio-engineered cornea. We tested the decellularized lenticules in vitro and to be useful for multilayered epithelium sheet for selective allograft.
방법 : With 0.25% trypsin-EDTA, 0.1 % SDS, or 0.1 % Triton X-100 in hypotonic Tris buffer (10mM, pH 7.6), human corneal lenticules were decellularized. We evaluated and compared the efficiency and safety of the methods. To test attachment of epithelial cells on the lenticule, the lenticules were coated with various extracellular matrices (ECM) composing basement membrane, such as type I collagen, type IV collagen, or laminin. The epithelial sheets were analyzed by histological methods.
결과 : Histological analysis of the decellularized corneal lenticules showed that trypsin-EDTA treated group is completely acellular and microstructure of the cornea was intact. Multilayered corneal epithelial cell sheets were able to be obtained. Immunohistochemistry showed that multilayered epithelium on the lenticule is composed of cells expressing markers such as CK15, and CK3/12.
결론 : The decellularized corneal lenticule from human corneal stroma was successfully obtained by our developed method. With this decellularized lenticules, multilayered epithelial cell sheets were successfully achieved.
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