Ocular surface reconstruction using cultivated mucosal epithelial stem cells

PURPOSE: To investigate the possibility of using cultivated oral epithelial cells in ocular surface reconstruction. METHODS: An ocular surface injury was created in adult albino rabbits by a lamellar keratectomy. Oral mucosal biopsy specimens taken from both adult albino rabbits and human volunteers were cultivated for 2-3 weeks on a denuded amniotic membrane (AM) carrier. The cultivated epithelium was examined by histologic and immunohistochemical analysis. At 3-4 weeks after the ocular surface injury, the rabbit conjunctivalized corneal surfaces were surgically reconstructed by transplanting both the rabbit and human cultivated oral epithelial cells on the AM carrier. RESULTS: Both the rabbit and human cultivated oral epithelial sheets had 5 or 6 layers of stratified, well-differentiated cells. Histologic examination revealed that the cultivated epithelial cells were similar in appearance to those of in vivo normal corneal epithelium. Immunohistochemistry confirmed the presence of the keratin pair 4 and 13 and keratin 3 in the cultivated oral epithelial cells. Corneas that were grafted with rabbit and human cultivated oral epithelial cells on an AM carrier were clear and were epithelialized 10 and 2 days after surgery, respectively. CONCLUSIONS: We have generated confluent cultures of both rabbit and human oral epithelial cells on AM expanded ex vivo from biopsy-derived oral mucosal tissues. We have successfully carried out xeno- and autologous transplantation of these cultivated oral epithelial cells onto the ocular surfaces of keratectomized rabbit eyes. We believe that xeno- and autologous transplantation of cultivated oral epithelium is a feasible method for ocular surface reconstruction.