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T Nakamura, K Endo, C Leanne, NJ Fullwood, N Tanifuji, M Tsuzuki, N Koizumi, T Inatomi, Y Sano, S Kinoshita; Successful Culture of Rabbit Oral Epithelial Cells on Amniotic Membrane - A New Strategy for Ocular Surface Reconstruction . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4183.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Many attempts, such as cultivated corneal stem cell transplantation, have been made to establish a surgical treatment for severe ocular surface diseases. However, despite the relative success of these surgical procedures, several problems remain, including the high risk of rejection. The purpose of this study is to examine the viability of using human amniotic membrane (AM) as a substrate for culturing oral epithelial cells and to investigate the possibility using oral epithelial cells as a substitute for corneal epithelial cells. Methods:Oral mucosal biopsies were taken from 8 adult white rabbits and cultivated for 3 weeks on a denuded AM carrier with 3T3 fibroblast co-culture and air-lifting. The cultivated epithelium was examined by electron microscopy (EM) and immunohistochemically labelled for several keratins. Results:The cultivated oral epithelial sheet had 4-5 layers of stratified, well differentiated cells. EM examination revealed that the epithelial cells were very similar in appearance to those of normal corneal epithelium and corneal epithelium, cultivated using our established technique. EM revealed that the cultivated oral epithelial cells were differentiated into columnar, wing and squamous cells, had numerous desmosmal junctions and were attached to a basement membrane with hemi-desmosomes. Immunohistochemistry confirmed the presence of keratins 4/13 and cornea-specific keratin 3 in the cultivated oral epithelial cells. Conclusion:We have successfully generated on AM confluent cultures of oral epithelial cells expanded ex vivo from biopsy-derived oral mucosal tissues for ocular surface reconstruction. Our ultrastructural and immunohistochemical findings indicate that cultivated oral epithelial sheets could be used as a substitute for corneal epithelial cells.
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