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Julie Albon, Andrew B. Tullo, Saeed Aktar, Michael E. Boulton; Apoptosis in the Endothelium of Human Corneas for Transplantation. Invest. Ophthalmol. Vis. Sci. 2000;41(10):2887-2893.
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purpose. To determine whether endothelial cell loss of human corneas stored in
organ culture before transplantation is due to apoptosis.
methods. The corneal endothelium of human corneas, stored in organ culture at
34°C for varying periods of time, were analyzed for the presence of
apoptotic cells using the TdT-mediated dUTP nick-end labeling (TUNEL)
technique. Corneal endothelial cell apoptosis was confirmed by Hoechst
staining and immunolabeling with anti–caspase 3 active antibody.
results. Apoptotic cells were identified in the corneal endothelium of human
organ cultured corneas: their number and distribution demonstrated a
close correlation with corneal folding and overall quality of the
corneal endothelium. TUNEL-positive labeling of cells was confirmed as
apoptotic by the presence of morphologic nuclear alterations identified
by Hoechst staining and the presence of immunostaining for caspase-3
activity. Corneal endothelial cell apoptosis was independent of cause
of donor death, death to enucleation time, and death to culture times.
conclusions. Corneal endothelial cell apoptosis appears to determine the suitability
of a cornea for transplantation.
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