Regeneration of the epithelium in organ-cultured donor corneas with extended post-mortem time.
OBJECTIVE
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 post-mortem 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 degrees 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.


CONCLUSIONS
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.
Slettedal
Jon Klokk
JK
Center for Eye Research, Department of Ophthalmology, Ullevaal University Hospital, Oslo, Norway. jon.klokk.slettedal@gmail.com
Lyberg
Torstein
T
Ramstad
Hanne
H
Beraki
Kahsai
K
Nicolaissen
Bjørn
B
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival / Q000502:physiology; D003315:Cornea; D019573:Epithelium, Corneal / Q000502:physiology* / Q000648:ultrastructure; D005260:Female; D006801:Humans; D008297:Male; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D009924:Organ Culture Techniques; D009926:Organ Preservation; D012038:Regeneration / Q000502:physiology*; D013997:Time Factors; D014019:Tissue Donors
D015415:Biomarkers