Autologous transplantation of conjunctival epithelial cells cultured on amniotic membrane in a rabbit model

PURPOSE To evaluate the feasibility of autologous transplantation in a rabbit model of conjunctival epithelial cells cultured on amniotic membrane for ocular surface reconstruction. METHODS Limbal stem cell deficiency was induced in the right eyes of 30 rabbits. This was done by performing a lamellar keratectomy of the entire cornea and a complete removal of the limbus and conjunctiva, extending 5 mm outside the limbus. Autologous conjunctival specimens were obtained from the left eyes of ten of those rabbits and cultured for four weeks on denuded amniotic membrane. Cultured epithelium was examined by transmission electron microscopy. Four weeks after lamellar keratectomy, conjunctivalized corneal surfaces were excised and autologous cultured conjunctival epithelial sheets transplanted (Conj-AM group, n=10). The controls were rabbits that underwent corneal surface removal but not transplantation (No Transplantation group, n=10) and those that underwent corneal surface removal but received only amniotic membrane (AM Alone group, n=10). A neovascularization and corneal opacity scoring system was used to evaluate each eye in the two months after surgery. RESULTS Cultured conjunctival epithelium formed three to four layers on denuded amniotic membrane. Averaged scores of corneal neovascularization and corneal opacity two months after transplantation were significantly low in the Conj-AM group as compared with those in the AM and no transplantation groups. CONCLUSIONS Transplantation of autologous conjunctival epithelial cells cultured on amniotic membrane should prove an effective strategy for treating total limbal stem cell deficiency.