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Br J Ophthalmol 2004;88:422-427 doi:10.1136/bjo.2003.026054
  • Laboratory science - Extended reports

Correlation of long term phenotypic and clinical outcomes following limbal epithelial transplantation cultivated on amniotic membrane in rabbits

  1. S-E Ti1,3,
  2. M Grueterich1,2,
  3. E M Espana1,
  4. A Touhami1,
  5. D F Anderson2,
  6. S C G Tseng1,2
  1. 1TissueTech, Inc, and Ocular Surface Center, Miami, FL, USA
  2. 2Bascom Palmer Eye Institute, University of Miami School of Medicine, Miami, FL, USA
  3. 3Singapore National Eye Centre, Singapore
  1. Correspondence to: Scheffer C G Tseng MD, PhD, Ocular Surface Center, 7000 SW 97th Avenue, Suite 213, Miami, FL 33173, USA; stsengocularsurface.com
  • Accepted 1 August 2003

Abstract

Aim: To determine the epithelial phenotype in rabbits with total limbal stem cell deficiency (LSCD) after reconstruction with autologous limbal epithelial stem cells ex vivo expanded on rabbit amniotic membrane (AM).

Methods: Left eyes of 52 rabbits were rendered total LSCD, verified by impression cytology. The fibrovascular pannus of each cornea was removed. Group I (n = 10) received rabbit AM transplantation alone, while groups II–IV (n = 42) underwent transplantation of LSC cultured on rabbit AM (LSC-AM) from a small limbal biopsy taken from the right eye. Clinical outcome was graded as “success,” “partial success,” or “failure” depending on the corneal smoothness and avascularity. Epithelial phenotype was determined by immunostaining and graded as “corneal (K),” “conjunctival (J),” or “mixed (M)” depending on expression of K3 and Muc5AC.

Results: After 1 year follow up, group I showed 100% failure and groups II–IV showed 26% success (p<0.001). Clinical failure correlated with J phenotype p = 0.001), while clinical success correlated with K phenotype p = 0.01). When the phenotypic outcome was used for comparison, J phenotype was significantly high in group I (p = 0.003), while K phenotype was significantly high in groups II–IV (p<0.05).

Conclusion: There is a strong correlation between clinical success and resultant corneal epithelial phenotype. Ex vivo expanded LSC can successfully reconstruct corneal surfaces with unilateral total LSCD.

Footnotes

  • Proprietary interest: SCGT and his family are more than 5% shareholders of TissueTech, Inc, which owns US patents Nos 6,152,142 and 6,326,019 on the method of preparation and clinical uses of human amniotic membrane distributed by Bio-Tissue, Inc.

  • Presented in part at the annual meeting of ARVO in Ft Lauderdale, FL, USA, in May 2003

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