Establishment of a Cultivated Human Conjunctival Epithelium as an Alternative Tissue Source for Autologous Corneal Epithelia Transplantation

Abstract

Abstract: : Purpose: Corneal epithelium is essential for transparency of cornea. Several efforts have been made to establish an improved technique in the corneal epithelial transplantation for patients with a total limbal failure. In order to assess the availability of the conjunctival epithelium as an alternative corneal epithelium, the histological properties and the applicability for transplantation on the ocular surface of the cultivated human conjunctival epithelium (HCjE) were investigated. Methods: The HCjE cells were cultivated on a human amniotic membrane (HAM) according to the previously established culture protocol. The cultivated HCjE were transplanted on the rabbit cornea and were observed 2 or 5 days after the transplantation. The cultivated HCjE was also analyzed by immunostaining against keratin4/13, ABCG2/p63 or integrin alpha6/beta4, which represents a conjunctival abundant keratin pair, generally accepted stem cell markers or hemidesmosome component proteins, respectively. Results: The cultivated HCjE demonstrated a well–stratified epithelial structure. Its basal cells expressed ABCG2 and p63, suggesting that the HCjE still maintained its stem cells even after the culture process. The keratin 4/13 pair and the integrin alpha6/beta4 pair were expressed as observed in in vivo conjunctival epithelium, suggesting that the cultivated HCjE has quite equivalent tissue/cellular properties as in vivo conjunctival epithelium. The transplanted HCjE cells were well maintained at least five days after the surgery and demonstrated a gradual cell–migration on the recipient’s corneal surface, suggesting that the cultivated HCjE has enough robustness and viability for the standard transplantation. Conclusions: The cultivated HCjE on the HAM seems to have potential to be used as an alternative corneal epithelium.

Keywords: conjunctiva • cornea: epithelium