The potential for eye bank limbal rings to generate cultured corneal epithelial allografts.
OBJECTIVE
Patients with severe limbal deficiencies are unable to maintain a stable corneal surface. If sheets of cultured allogeneic corneal epithelium could be prepared from eye banked corneal limbal rings, which are normally discarded after keratoplasty, the sheets may be beneficial for grafting onto patients with limbal stem cell deficiencies.


METHODS
Biopsies of limbal tissue (2-3 mm2) removed from organ-cultured corneal limbal rings or from fresh whole globes were either trypsinized or set up as explants to assess their potential for corneal epithelial cell production.


RESULTS
Several biopsies were taken from each of 21 organ-cultured limbal rings and 10 fresh cadaveric globes. Cultures were generated from every cadaveric eye (10/10), although not all biopsies from the same eye gave rise to cultures. Confluent sheets of cultured cells were also produced successfully from limbal rings that had been in organ culture for up to 25 days, but the success rate from limbal ring material was variable (14/21). An analysis of parameters associated with each limbal ring was carried out in an attempt to identify the reasons for the different efficiencies of epithelial production. No obvious single parameter correlation was detected, although there was a trend to poorer efficiency with increased donor age.


CONCLUSIONS
Confluent sheets of cultured corneal epithelial cells, suitable for grafting, can be produced from limbal tissue taken from eye bank organ-cultured corneas, although it takes longer, on average, to reach confluence (17-21 days) than an equivalent sample from a fresh eye (9-12 days).
James
S E
SE
Blond McIndoe Centre, Queen Victoria Hospital, East Grinstead, West Sussex, United Kingdom. lizjamesmcindoe@hotmail.com
Rowe
A
A
Ilari
L
L
Daya
S
S
Martin
R
R
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018929:Cell Culture Techniques / Q000379:methods; D017690:Cell Transplantation; D002478:Cells, Cultured; D003316:Corneal Diseases / Q000601:surgery; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D005125:Eye Banks; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007633:Keratins / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology*; D008875:Middle Aged; D009924:Organ Culture Techniques; D010641:Phenotype; D033581:Stem Cell Transplantation; D014019:Tissue Donors; D014184:Transplantation, Homologous
D007633:Keratins