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K. Haug, E. W. Gulliksen, A. Shahdadfar, O. Ringen, L. K. Drolsum, M. C. Moe, B. Nicolaissen; Human Cornea in Long Term Optisol-gs Storage, Proliferation, Phenotype and Ultra Structure of the Limbal Epithelium. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1943. doi: https://doi.org/.
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Cornas used for transplantation are commonly stored in Optisol GS at +4 °C and recommended upper limit for such storage is approximately two weeks. The medium is primarily composed to preserve endothelial viability and prolonged storage may increase the likelihood of persistent post operative epithelial defects, such defects could hypothetically be due to basement membrane damage inflicted by the medium with a decrease in ability to support migration and cellular attachment. The purpose of the study was to test this hypothesis.
Corneas stored in Optisol GS for 10-14 days (n=40) where the central part had been removed for transplant purposes, were fixed and processed for light microscopy (H&E) or transferred to MEM with 8% FBS at 32°C with 5% CO2 for one week. Samples were fixed in buffered formalin and processed for light micorcopy (H&E) and Immunohistochemistry (Ki67, PCNA, p63, ABCG2 ). Cell counting and assessment of p63 and ABCG2 was preformed by three independent observers, triplicate samples were examined for each parameter. For TEM, the samples were fixed and processed according to routine procedures.
Light Microscopy (LM) of tissue fixed after storage showed extensive loss of peripheral epithelial cells. LM and TEM of cultured tissue showed the peripheral cornea to be covered with a bilayered epithelium in all triplicate samples from all donors. Both markers for proliferation (Ki67, PCNA) and for stemness (ABCG2, p63) were positive in the limbal region. In some sections, epithelial cells seemed to invade the stroma (LM), TEM of such areas however showed an evident basement membrane to separate these cells from underlying tissue.
Revitalization of epithelium after long term storage resulted in complete covering of the corneal surface, and a hypothesis involving Optisol GS as a cause for persistent epithelial defects could not be confirmed in our study.
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