%0 Journal Article %J Ophthalmic Res %D 2003 %T Improved preparation and preservation of human keratoplasty lenticules %A Salvalaio,G. %A Fasolo,A. %A Bruni,A. %A Frigo,A. C. %A Favaro,E. %A Ponzin,D. %K *Cornea %K Cell Survival / physiology %K Chondroitin Sulfates %K Complex Mixtures %K Corneal Stroma / *cytology %K Cryopreservation / *methods %K Cryoprotective Agents / therapeutic use %K Culture Media, Serum-Free %K Debridement %K Dextrans %K Fibroblasts / *cytology %K Gentamicins %K Humans %K Keratoplasty, Penetrating %K Organ Preservation / *methods %K Staining and Labeling / methods %X PURPOSE: To improve the preparation of lenticules from human cornea and to obtain their preservation without loss of viable keratocytes. METHODS: The epithelium was manually removed after bathing the surface of the cornea with a solution of trypsin and EDTA. Lenticules were prepared by microkeratome resection and viable keratocytes were visualized by staining with thiazolyl blue (MTT). RESULTS: The pretreatment with trypsin-EDTA allowed the removal of the epithelium without damage to the keratocytes and the stroma. When these lenticules were incubated in Optisol-GS for 7 days at 4 degrees C, they showed a limited thickness increase and a preservation of keratocyte viability. CONCLUSION: This procedure allows the preparation of lenticules with viable keratocytes that can be preserved in the cold for at least 1 week. %B Ophthalmic Res %7 37978 %V 35 %P 313 - 8 %8 Nov-Dec %G eng %N 6 %M 14688420 %R 10.1159/000074069 ORE2003035006313 [pii]