The Study for Limbal Epithelial Cell Movement according to the Position of Limbal Transplant in Rabbits

Choi, Si Hwan; Choi, You Jin; Lee, Yang Won; Kim, Seung Young

Abstract

PURPOSE
This study aimed to evaluate the limbal epithelial cell movement according to the position of limbal transplant in rabbits. METHODS: We performed autologous conjunctival limbal transplantation in rabbits, in situ (directed corneal end toward cornea and conjunctival end toward conjunctiva) or in versus (directed corneal end toward conjunctiva and conjunctival end toward cornea) manner. We performed 360o whole autologous conjunctival limbal transplantation in quadrant fashion. Twelve months later, we observed the gross cornea under a slit lamp, and then observed corneal tissue with PAS staining using light microscopy and immunohistochemical and immunofluorescence staining using monoclonal antibody. As mentioned above, the conjunctival limbal tissue had been transplanted in situ and in versus manner for confocal microscopy. However, before transplantation, we performed organ culture of the limbal epithelial cells with CellTracker(TM) Green CMFDA dye (Molecular Probes, USA). Two weeks later, we observed the corneal and limbal epithelial cells using the confocal microscope. RESULTS: The slit lamp examinations of the cornea showed no new vessels and clear in both specimens. There were no goblet cells and the corneal epithelial arrangement was normal under light microscopy. There were positive immunohistochemical stains for cornea-specific AK2 and AE5, and negative immunofluorescence stains for goblet cell-specific AM3 patterns in both in situ and in versus specimens. CellTracker(TM) Green CMFDA dye up-taken epithelial cells were not presented in the conjunctival epithelial side, but were in the corneal epithelial side in both specimens. CONCLUSIONS: We confirmed that the epithelial cells with characteristics of corneal epithelium moved toward the corneal center at any positions of limbal transplants in rabbits.