Comparison of Three Methods for Human Corneal Cryopreservation That Utilize Dimethyl Sulfoxide

• Published in: Cryobiology 1999-08, Vol.39 (1), p.47-57
• Main Authors: Bourne, William M., Nelson, Leif R., Hodge, David O.
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Biological and medical sciences; Cadaver; Cell Survival; Cornea; corneal endothelial cells; corneal storage; Corneal Transplantation; cryopreservation; Cryopreservation - methods; Cryoprotective Agents; Dimethyl Sulfoxide; Diseases of cornea, anterior segment and sclera; Endothelium, Corneal - cytology; Evaluation Studies as Topic; Humans; In Vitro Techniques; Medical sciences; Microscopy, Electron, Scanning; Middle Aged; Ophthalmology
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1006/cryo.1999.2182

We compared endothelial cell survival in human corneas after cryopreservation by three methods that utilize dimethyl sulfoxide. Twenty-eight human cadaver corneas were cryopreserved by one of three methods, stored briefly over liquid nitrogen, thawed, cultured at 37°C for 3 days, and fixed for scanning electron microscopy. Seventeen control corneas underwent identical cryoprotectant immersion and culture protocols but were not frozen. Endothelial photographs taken after 1 and 3 days of culture were analyzed. Endothelial cell losses in cryopreserved corneas by Methods 1, 2, and 3, respectively, were 36, 22, and 10% after 1 day of culture and 57, 36, and 27% after 3 days of culture. Cryopreservation by Method 3 had less cell loss than Methods 1 or 2 (P < 0.02) but greater cell loss than the control corneas for Method 3 (P < 0.001). No loss of cells occurred in the control corneas for Methods 1 and 3 but substantial cell loss (26%) occurred in the control corneas for Method 2. Polymegethism and pleomorphism of the endothelial cells were seen in the corneas that lost cells. The endothelial cell loss of 10% seen after 1 day of culture in human corneas cryopreserved by Method 3 is similar to the loss that occurs during organ culture storage as currently used clinically and therefore would be acceptable for clinical use. After 3 days of culture, however, the cell loss had increased significantly to 27%. This additional decrease in cell number that occurs in culture may represent latent cryodamage and must be understood and overcome in vivo before the technique can be used clinically.