We studied freezing rates, cryoprotectants, and storage times on keratocyte viability, using rabbit corneal buttons incubated in either KM-26, CPTES, K-Sol, or TC 199 for 30 minutes at 4 degrees C. Using a controlled freezing rate (2 degrees/min to -40 degrees C), viabilities were 69 +/- 7% for KM-26, 113 +/- 21% for CPTES, 0.1 +/- 0.08% for K-Sol, and 0 +/- 0% for TC 199. The KM-26 and CPTES were further studied in corneas stored one to 30 days at -80 or -196 degrees C; CPTES had a better cryoprotective efficacy over one, three, and seven days of -80 degrees storage, and at liquid nitrogen storage temperature (-196 degrees) over one, 14, and 30 days storage. The findings demonstrate the superiority of CPTES. It provides better viability than KM-26 under similar conditions, and may enable long-term frozen storage of lenticules for later use in cryorefractive surgeries, with minimal loss of keratocyte viability.