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S. Jeon, M. Jang, H. Lee, K. Seo; Autologous Transplantation of Cornea Epithelial Cell Sheet Cultured from Human Limbus . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4867.
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Purpose: We sought to evaluate the effectiveness of cornea epithelial cell sheet on corneal wound repair. In this study, we have developed an approach involving autologous cornea epithelial stem cell sheets for treating damaged cornea due to alkali burn. Methods: Biopsies (1–2mm2) were taken from the limbus of 2∼75 years old donor. Human cornea limbal explants containing stem cells were cultured from cornea limbal tissue of 15 patients. Cornea epithelial cells were cultured on 3T3 feeder cells without fibrin. p63, which is believed to be a marker for keratinocyte stem cells, was identified in these cell populations by immunoblotting. Autologous cornea epithelial cell sheet was grafted onto damaged in a patients’ cornea along with amniotic membrane. Result: Cornea epithelial cell culture was successfully performed with limbal biopsies form 14 out of 15 patients, regardless of age. Cornea epithelial stem cells from the patients were cultured on 3T3 feeder cells for two weeks resulting in formation of a multilayered corneal epithelial sheet. We show by immunostaining that p63, a keratinocyte stem cell marker, is abundantly present in cultured cornea epithelial sheet whereas neither K3, a corneal differentiation marker, nor connexin 43, a corneal basal marker, is absent. When the corneal epithelial sheet was transplanted onto a patient, re–epithelialization occurred within the first week and the corneal surface was covered by a transparent, normal–looking epithelium. Conclusion: We were able to maintain corneal stem cells from patients up to more than 3 passages in culture independently of their age. We found that the use of the cultured cornea epithelial stem cell sheet accelerated re–epithelialization of corneal wound and that the resulting epithelium was smooth and transparent. Therefore, transplantation of cultured autologus cornea can reduce scarring and healing time compared to conventional epithelial transplantation.
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