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The Potential for Eye Bank Limbal Rings to Generate Cultured Corneal Epithelial Allografts

James, S. Elizabeth Ph.D.; Rowe, Andrea; Ilari, Luca M.D.; Daya, Sheraz M.D., F.A.C.P., F.A.C.S.; Martin, Robin Ph.D.

Basic Investigation

Purpose. 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.

Method. 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).

From Blond McIndoe Centre (S.E.J., R.M), the Corneo Plastic Unit (L.I., S.D.), and the Eye Bank (A.R., S.D.), Queen Victoria Hospital, East Grinstead, West Sussex, United Kingdom.

Submitted October 17, 2000.

Revision received February 13, 2001.

Accepted February 14, 2001.

Grants: The John Ellerman Foundation, The Band Trust, and Eye Bank Association of America.

Presented, in part, at the 38th Annual Scientific Session of the Eye Bank Association of America (Orlando 1999).

Address correspondence and reprint requests to Dr. S.E. James, Blond McIndoe Centre, Queen Victoria Hospital, East Grinstead, W. Sussex, RH19 3DZ, U.K. E-mail:

© 2001 Lippincott Williams & Wilkins, Inc.