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Cell death during corneal storage at 4 degrees C
A Komuro1, D O Hodge, G J Gores
1Department of Ophthalmology, Mayo Clinic, and Mayo Foundation, Rochester, Minnesota 55905, USA.
Investigative Ophthalmology & Visual Science
|November 5, 1999
Summary
Cell death, including apoptosis and necrosis, occurs in human corneas during cold storage. The TUNEL assay can identify apoptosis but not necrosis, potentially aiding in extending corneal storage viability.
Area of Science:
- Ophthalmology
- Cell Biology
- Tissue Engineering
Background:
- Human donor corneas are stored at 4 degrees C for transplantation.
- Understanding cell death mechanisms during storage is crucial for optimizing graft survival.
Purpose of the Study:
- To evaluate cell death in human donor corneas stored at 4 degrees C.
- To determine if the terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick-end labeling (TUNEL) assay can differentiate between apoptosis and necrosis in stored corneas.
Main Methods:
- Human corneas were stored at 4 degrees C for up to 21 days.
- Corneal sections were analyzed using transmission electron microscopy (TEM) and the TUNEL assay.
- Keratocytes, epithelial, and endothelial cells were graded as normal, apoptotic, or necrotic.
Main Results:
- TEM revealed apoptosis in 23% and necrosis in 12% of keratocytes.
- The TUNEL assay identified apoptosis in 11% of keratocytes, correlating with TEM findings for apoptosis.
- Apoptosis was detected in 13% of epithelial cells and 8% of endothelial cells by TUNEL assay.
Conclusions:
- Both apoptosis and necrosis occur in corneal cells during cold storage, with apoptosis being more prevalent.
- The TUNEL assay is effective in identifying apoptosis but not necrosis in corneal tissue.
- Inhibiting apoptosis may extend the viable storage time of donor corneas.