The potential for eye bank limbal rings to generate cultured corneal epithelial allografts

S.Elizabeth James, A. Rowe, L. Ilari, S. Daya, R. Martin

Research output: Contribution to journalArticle

Abstract

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).
Original languageEnglish
Pages (from-to)488-494
Number of pages7
JournalCornea
Volume20
Issue number5
DOIs
Publication statusPublished - 1 Jul 2001

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Eye Banks
Allografts
Biopsy
Epithelial Cells
Corneal Epithelium
Corneal Transplantation
Organ Culture Techniques
Cornea
Cultured Cells
Stem Cells
Tissue Donors

Keywords

  • Stems cells
  • Tissue engineering
  • Corneal epithelium
  • Cultured allografts

Cite this

James, S.Elizabeth ; Rowe, A. ; Ilari, L. ; Daya, S. ; Martin, R. / The potential for eye bank limbal rings to generate cultured corneal epithelial allografts. In: Cornea. 2001 ; Vol. 20, No. 5. pp. 488-494.
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The potential for eye bank limbal rings to generate cultured corneal epithelial allografts. / James, S.Elizabeth; Rowe, A.; Ilari, L.; Daya, S.; Martin, R.

In: Cornea, Vol. 20, No. 5, 01.07.2001, p. 488-494.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The potential for eye bank limbal rings to generate cultured corneal epithelial allografts

AU - James, S.Elizabeth

AU - Rowe, A.

AU - Ilari, L.

AU - Daya, S.

AU - Martin, R.

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Y1 - 2001/7/1

N2 - 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).

AB - 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).

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KW - Tissue engineering

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