Regeneration of the epithelium in organ‐cultured donor corneas with extended post‐mortem time

Abstract

ABSTRACT.Purpose: The maximum post-mortem time limit for obtaining donor corneas varies between eye banks. It is not known for how long a time the epithelial cells survive post-mortem, nor is it known if donor corneas with extended postmortem time are able to regenerate the epithelium. Therefore, we wanted to examine the epithelium in donor corneas for regenerative ability during storage in an eye bank organ culture system. Methods: Twenty-four paired donor corneas with post-mortem time from 28 to 163 hr were obtained. One cornea of a pair was fixed immediately to serve as a control, and the second was cultured in eye bank medium at 32°C for 3 days. Examination of the specimens was performed with light and scanning electron microscopy. Immunohistochemical staining methods with antibodies against K 3, K 19, vimentin and p63 were used to further characterize the cells. Results: The control corneas showed decreasing amounts of epithelial cells with increasing post-mortem time. All the cultured corneas demonstrated rapid regeneration of the epithelium. After 3 days in organ culture, 10 of 12 donor corneas were covered with epithelium. Conclusion: Even up to 7 days post-mortem, viable cells reside in the corneal epithelium. The study demonstrates the hardiness and enormous regenerative potential of peripheral corneal cells. Donor corneas processed in an eye bank organ culture storage system will obtain an intact epithelial layer within a few days.

Influence of donor age, post‐mortem time and cold storage on metabolic profile of human cornea

“…An absent or partially absent epithelium increases the potential for trauma from exposure, medications used in preparation of the donor or preservation medium. Disintegration of certain structural proteins or adhesion molecules contribute to the loosening of cells, especially if the basal cells are weakly attached to the central cornea (Van Meter et al 2005;Slettedal et al 2007). Such changes may lead to the apoptosis of anterior stromal keratocytes, and it has been shown that these effects might be prolonged (Wilson et al 2001;Erie 2003).…”

“…No general agreement exists on the maximum time limit for removing donor corneas, and this time limit varies greatly between eye banks. It is recommended that corneal preservation occurs as soon as possible after death (Slettedal et al 2007), but some researchers claim that there are no such upper time limits: as long as certain quality criteria are met, corneas can be used for grafting (Andersen & Ehlers 1988;Ehlers 2002). Another controversy exists regarding donor age as the factor that may determine the suitability of a cornea for transplantation (Gal et al 2008).…”

Levels of oxidative DNA damage are low in ex vivo engineered human limbal epithelial tissue

“…For organ culture, donor corneas were incubated as described previously (Slettedal et al 2007). For organ culture, donor corneas were incubated as described previously (Slettedal et al 2007).…”

Biomechanical Strengthening of the Human Cornea Induced by Nanoplatform-Based Transepithelial Riboflavin/UV-A Corneal Cross-Linking

“…The corneal storage protocol was effective to provide tissues with intact epithelium, 34,35 and no damage to the epithelial cells was found after transepithelial corneal crosslinking. In order to avoid any bias due to changes in the hydration state during the experiment, the corneal tissues were kept in 20% dextran solution overnight before AFM testing.…”