Amniotic membrane as a substrate for cultivating limbal corneal epithelialcells for autologous transplantation in rabbits

Purpose. To examine the viability of using human amniotic membrane as subst rate for culturing corneal epithelial cells and transplanting them onto sev erely injured rabbit eyes. Methods. An ocular-surface injury was created in the right eye of eight rabbits by a lamellar keratectomy extending 5 mm ou tside the limbus. Next, from the limbal region of the uninjured left eyes o f five of these animals, a small biopsy of corneal epithelial cells was tak en and cultured on acellular human amniotic membrane. One month later, the invading conjunctiva that covered the corneal surface of all eight injured eyes was surgically removed. Five of the eyes then received grafts of amnio tic membrane containing autologous cultured epithelial cells, whereas the o ther three received grafts of acellular amniotic membrane alone. Results, A confluent primary culture of limbal corneal epithelial cells was establish ed on acellular human amniotic membrane after 14 days. Cells were partially stratified and fairly well attached to the underlying amniotic membrane, a lthough a fully formed basement membrane was not evident. The three rabbits that received amniotic membrane transplantation alone all had total epithe lial defects on the graft in the early postoperative period. Eyes that were grafted with amniotic membrane that contained cultivated epithelial cells, however, were all successfully epithelialized up to 5 days after surgery. Conclusion. Autologous transplantation of cultivated corneal epithelium is feasible by using acellular amniotic membrane as a carrier.