FIG. 4.

Decrease in endothelia cell damage in corneas stored in Optisol-GS after organ cultured with 15-epi-LXA4. (A) A pair of corneas from a 47-year-old donor was stored in 20 mL Optisol-GS with or without 100 nM 15-epi-LXA4 for 12 days at 4°C as explained in Methods section. At the end of storage, the corneas were stained with a Live/Dead Viability/Cytotoxicity Kit. A fluorescence microscope was used to evaluate the cell damage. There were areas of endothelial cell loss with multiple red fluorescent cells representing dead cells; 8–10 pictures were taken from each cornea and the average±SD of the damaged area was calculated. (B) A pair of corneas from a 58-year-old donor was organ cultured with or without 100 nM 15-epi-LXA4 for 24 h at 37°C. Afterward, the corneas were transferred to Optisol-GS with or without 15-epi-LXA4 and stored for 12 days at 4°C. The endothelial cell damage was detected with a combination of 2 stains: trypan blue and alizarin red S. The percentage of cell damage was calculated as explained in Methods section. (C) One pair of corneas from a 59-year-old donor incubated as in (B) was fixed with 4% paraformaldehyde, embedded with optimal cutting temperature (OCT), cut into 6 μm serial sections, and immunostained with Ki-67 antibody. Ki-67 positive cells are shown in the endothelium. *Significant difference with respect to control.