December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
The Proliferative Potential of Stem Cells From Fresh Versus Cadaveric Limbal Tissues
Author Affiliations & Notes
  • GK Vemuganti
    Ophthalmic Pathology Service L V Prasad Eye Institute Hyderabad India
  • S Kashyap
    Ophthalmic Pathology Service
    L V Prasad Eye Institute Hyderabad AP India
  • VS Sangwan
    Cornea and Anterior segment Service
    L V Prasad Eye Institute Hyderabad AP India
  • S Singh
    Electron Microscopy Centre for Cellular and Molecular Biology Hyderabad India
  • Footnotes
    Commercial Relationships   G.K. Vemuganti, None; S. Kashyap, None; V.S. Sangwan, None; S. Singh, None. Grant Identification: Dept of Biotechnology(BT/PR2305/Med/09/336/2001)
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 1623. doi:
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    • Get Citation

      GK Vemuganti, S Kashyap, VS Sangwan, S Singh; The Proliferative Potential of Stem Cells From Fresh Versus Cadaveric Limbal Tissues . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1623.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: To evaluate and compare the growth potential of limbal stem cells from the residual corneo-scleral rings, and fresh limbal tissues obtained from patients undergoing routine cataract surgery. Methods: After obtaining approval of Institutional Review Board and informed consent from the patients, 1-2mm of limbal tissues from 15 patients and 31 tissues from the cadaveric limbal ring preserved in MK medium (16 tissues) and Optisol (15 tissues) were used for the study. Donor data and duration of preservation was noted. The limbal tissue was explanted on the de-epithelized human amniotic membrane, using Human Corneal Epithelial cell medium. After adequate growth, the explant cultures were studied by stained whole mount preparations, routine histopathology examination, and immunohistochemical staining, using monoclonal antibodies to cytokeratin K3 and K19. Results: Forty six tissues were evaluated from individuals with a mean age of 55.3 years ± 21.23 years (range 18 years - 110 years). Explant cell cultures were observed in 100%(15/15) of limbal tissue obtained from patients, as against 56%(9/16) of MK medium preserved tissues and 46.7%(7/15) of Optisol preserved tissues (p=0.0131). The explant showed cells budding from the edge within 24- 72 hours, and a monolayer in 2-3 weeks, measuring 5 to 16mm. The cultured epithelium was 2-3 cell layered and immunopositive for K3. The time lapse interval between death to utilization and the donor age had no significant influence on the growth potential of the limbal stem cells. Conclusion: Fresh tissues from living subjects have a better growth potential of limbal stem cells as compared to cadaveric tissues, thereby suggesting a better outcome of allogenic transplantation using fresh tissues from living related donors.

Keywords: 370 cornea: basic science • 372 cornea: epithelium 
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