Abstract
Purpose: :
We previously reported that cultivated human conjunctival epithelium (HCjE) may potentially be used as an alternative corneal epithelium. To further validate these findings, transplanted HCjE and cultivated human corneal epithelium (HCE) were compared.
Methods: :
The HCjE cells (n=8) and HCE cells (n=7) were cultivated on a human amniotic membrane (HAM) according to our previously established culture protocol. The organotypically–cultured HCjE or HCE cells were transplanted onto rabbit corneas. Their ocular surfaces were examined by slit–lamp microscopy at 4 or 14 days after the transplantation. After enucleation at 14 days, expression of keratins (K3, 4, 12, and 13) and mucins (Muc4 and Muc5AC) were analyzed by immunostaining. Also, ultra–microstructure of these tissues was analyzed by transmission electron microscopy (TEM).
Results: :
The transplanted HCjE and HCE were well maintained on the recipient’s cornea at 4 days and 14 days. Both of the engrafted HCjE and HCE demonstrated 5 – 6 layers of stratified squamous epithelium. These cells exhibited microvilli, desmosomes and hemidesmosomes as seen in in vivo corneal epithelium. CK4, CK13, CK3, and CK12 were found in both of the engrafted HCjE and HCE epithelium. Goblet cells were not observed in both engrafted epithelia. The engrafted HCjE expressed MUC4 on its surface, while the engrafted HCE did not.
Conclusions: :
Engrafted HCjE demonstrated similar cellular features as the engrafted HCE. These findings support the use of organotypically–cultured HCjE as an alternative tissue for corneal epithelium.
Keywords: conjunctiva • cornea: epithelium • immunohistochemistry