Latent endothelial cell damage after experimental corneal cryopreservation.
Ninety porcine corneas were evaluated by vital staining with alizarin red S and trypan blue in a three-step experiment. Central cell densities were counted (a) on freshly dissected corneas (n = 30), (b) on cryopreserved corneas directly after thawing (n = 30), and (c) after a postthawing organ culture interval of 24 h (n = 30). Two freezing methods were used: (a) minimum essential medium--containing 20% fetal calf serum and (b) the same but containing additionally 2% chondroitin sulfate. Directly after thawing neither method showed significant cell loss (3.9% and 3%) compared to fresh tissue. After postthawing organ culture, however, tissue that had been frozen without chondroitin sulfate displayed a cell loss of 73.5% compared to corneas of the same freezing protocol directly after thawing. Corneas in chondroitin sulfate containing medium showed a cell loss of only 33.2%. We conclude that reliable morphologic evaluation should not be obtained from cryopreserved corneas examined directly after thawing.
Hagenah
M
M
Universitäts-Augenklinik, Hamburg, Germany.
Böhnke
M
M
D000818:Animals; D000880:Anthraquinones; D002452:Cell Count; D002470:Cell Survival; D002809:Chondroitin Sulfates; D004396:Coloring Agents; D003315:Cornea / Q000473:pathology*; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media; D004728:Endothelium, Corneal / Q000473:pathology*; D009924:Organ Culture Techniques; D013552:Swine; D014343:Trypan Blue
D000880:Anthraquinones; D004396:Coloring Agents; D003470:Culture Media; C004468:Alizarin Red S; D002809:Chondroitin Sulfates; D014343:Trypan Blue