The Characteristics of Limbal Epithelial Cells Auto-Cultivated In Vivo on Amniotic Membrane in Rabbits

Park, Jung Min; Woo, Jung Min; Cheon, Hyun Chul; Jeung, Woo Jin; Park, Woo Chan

Abstract

PURPOSE: To investigate the characteristics of the limbal epithelial cells auto-cultivated in vivo on amniotic membrane (LIVAMs) designed for the treatment of limbal stem cell deficiency.
METHODS
We removed the epithelium of AM with a No.15 blade after it was blotted with 20% ethanol and made a 360 degrees stromal flap along the epithelial defect. We then mounted over-sized AM (1 mm larger in diameter than the defect) over the defect with the border of AM inserted under the flap, and performed interrupted suture with 10-0 nylon. A therapeutic contact lens was fitted over the AM and a temporary tarsorrhaphy was performed. To examine whether the limbal epithelial cells grew well onto AM, we observed the cornea after fluorescein dye staining using a slit lamp. To explore the characteristics of LIVAMs, we performed hematoxylin-eosin (H&E) staining, immunochemical staining with AK-2, AE-5, AM-3 monoclonal antibodies, and transmission electron microscopy.
RESULTS
Three of four rabbits had successful epithelial growth on the amniotic membrane. The epithelial growth on the amniotic membrane was stained using immunohistochemical staining (AK-2, AE-5). Electron microscopy showed a structure similar to that of a normal corneal epithelium.
CONCLUSIONS
The technique of auto-cultivation of limbal epithelial cells in vivo on amniotic membrane can be an efficient and convenient method and preserves the characteristics of limbal epithelial cells for the treatment of limbal stem cell deficiency.