Abstract
Purpose: :
To examine the human conjunctiva during eye bank storage.
Methods: :
Twelve donor corneas with a conjuntival brim were obtained by excision with a 16 mm trephine. The samples were stored in an eye bank medium containing 8% serum at 32°C for 3, 6 or 18 days. Examination was performed with light microscopy, immunostaining for the cytokeratins K3, K4, K7, K13 and K19, MUC5AC, connexin 43, E-cadherin, proliferation markers PCNA and Ki-67 and with transmission electron microscopy. Apoptosis were evaluated with the TUNEL method.
Results: :
The cells were positive for connexin 43, E-cadherin, K19, K13 and K4, some superficial cells were positive for K7 and MUC5AC and some cells, predominantly in the basal cell layer, were positive for Ki-67. The cells were negative for K3. The epithelium covering the cornea showed a corneal phenotype.
Conclusions: :
All the samples showed a multilayered epithelium peripheral to the limbus with a characteristic conjunctival phenotype after storage. Our study demonstrates that the conjunctiva survives and the expression of cytokeratins and goblet cells is maintained in the eye bank medium. This is relevant in relation to development of new techniques for storage of cultured conjunctival epithelium for transplantation purposes and as well as for the developmement of new conjunctival storage media.
Keywords: conjunctiva • microscopy: light/fluorescence/immunohistochemistry • transplantation