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T. Nakamura, K. Endo, T. Inatomi, C. Sotozono, T. Amemiya, N. Kanamura, S. Kinoshita; Development of Human Cultivated Oral Mucosal Epithelium on Amniotic Membrane for Ocular Surface Reconstruction . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3153.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To determine the feasibility of using human amniotic membrane (AM) as a substrate for culturing human oral epithelial cells. To investigate the possibility of adapting human cultivated oral epithelial cells for clinical use in severe ocular surface disease. Methods: Normal human oral mucosa obtained from 5 patients as superfluous tissue debrided during oral surgery was cultivated for 2 - 3 weeks on denuded AM carrier with 3T3 fibroblast co-culture and airlifting. The cultivated epithelium was examined by light microscopy and immunohistochemically labeled for several keratins. The superficial keratectomized corneas of 5 rabbits were surgically reconstructed by xeno-transplanting human oral epithelial cells cultivated on AM. Results: A confluent primary culture of human oral epithelial cells was established on acellular AM after 7 days. After 2-3 weeks in culture, the human cultivated oral epithelial sheet had 5-7 layers of stratified, well-differentiated cells. LM revealed that the epithelial cells were very similar in appearance to normal corneal epithelium, and corneal epithelium cultivated using our established technique. Immunohistochemistry confirmed the presence of keratins 4, 13 and 3 in the cultivated oral epithelial cells. Rabbit corneas grafted with the human oral epithelial cells cultivated on an AM carrier were clear, and were all epithelialized at 2 days after surgery. Conclusions: We have generated confluent cultures of human oral epithelial cells on AM expanded ex vivo from biopsy-derived oral mucosal tissues. We have successfully xeno-transplanted these cultivated oral epithelial cells onto the ocular surfaces of keratectomized rabbit eyes. We believe that cultivated human oral epithelium transplantation is a feasible method of ocular surface reconstruction.
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