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INTEGRITY OF HUMAN CORNEAL EPITHELIUM MAINTAINED IN ORGAN-CULTURE USINGCORNEAMAX®F. Majo¹, M. Deprez², M. Nicolas¹¹Jules-Gonin Eye Hospital, Switzerland; ²University of Liege, BelgiumPurpose: Development of medium for organ-culture during eye banking is based on endotheliumintegrity. Nothing is described in the literature about conservation of corneal epithelium withCorneaMax® during banking. Therefore, we wanted to examine the integrity of human cornealepithelium maintained in CorneaMax®.Methods: All procedures conformed to the tenets of the Declaration of Helsinki for biomedicalresearch involving human subjects. Human corneas, considered unsuitable for transplantation, wereobtained from the Eye Bank in Lausanne and maintained in organ-culture in Corneamax® at 32°C.Average post-mortem time was 14 hours. Different time points were analysed from 0 to 35 days (N=5for each time points). Epithelial integrity was evaluated by H-E staining and by immunostaining withantibodies against E-cadherin and ZO-1. Proliferation and apoptosis were assessed by immunostainingwith antibodies against Ki67 and Caspase3 respectively.Results: During the first three days of culture the epithelium lost its adherence to the basal laminaof the cornea creating a large epithelial sheet. Some remaining limbal basal cells could be detected,allowing regeneration of the epithelium between day 2 to day 10. From day 10, the depth of theepithelium is reduced, consisting of only two to three cell layers. No change is observed in thedistribution of E-cadherin and ZO-1. Ki 67 staining demonstrated that the whole cornea proliferatedduring the 35 days of organ-culture. Apoptosis was rarely detected in the corneal epithelium.Conclusions: Corneas maintained in CorneaMax® showed a complete disappearance of the cornealepithelium during the two first days and a conservation of limbal basal cells in the limbal region. Theseremaining cells allowed a full regeneration of the tissue, leading to an atrophic epithelium, composedof only two to three cell layers. Following regeneration, adherens and tight-junctions proteins aredetected, suggesting that the epithelium integrity is maintained. This study is a first step to developmedium in organ-culture in order to conserve corneal epithelial cells.XXV ANNUAL MEETING OF THE EUROPEAN EYE BANK ASSOCIATION Zagreb, Croatia 18/19 January 2013 49