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Korean Journal of Obstetrics & Gynecology 2003;46(5):907-912.
Published online May 1, 2003.
The Effect of Equilibration Temperature and Time Prior to Cryopreservation on the Outcome of Cryopreservation of 2-Cell Mouse Embryos.
Hee Eun Ko, Kyung Nam Chung, Yong Bong Kim
1Department of Obstetrics and Gynecology, College of Medicine, Inje University, Seoul Paik Hospital, Seoul, Korea.
2Infertility Clinic, College of Medicine, Inje University, Seoul Paik Hospital, Seoul, Korea.
One-cell human zygotes have been successfully frozen and thawed using 1,2-propanediol (PROH) during freezing and thawing. This study was performed to assess the effect of equilibration temperature and time on cryopreservation of 2-cell mouse embryos by investigating the equilibration temperature and time during initial PROH exposure prior to cryopreservation. MATERIALS AND METHODS: The late 2-cell mouse embryos were obtained from 5-6 week old ICR mice and were exposed to 1.5 M PROH in phosphate-buffered saline (PROH-PBS) at 37degrees C, room temperature (22- 24degrees C), and 4degrees C for 30 min. The PROH was washed off the 2-cell mouse embyos by incubating them for 5 min each in 1, 0.5, and then 0 M PROH-PBS in the order named. The 2-cell mouse embryos were subsequently cultured in Ham's F-10 medium and embryo development was assessed at 24, 48, and 96 hours. RESULTS: Incubation of 2-cell mouse embryos at 37degrees C for 30 min significantly impaired embryo development to blastocysts. Embryo development after exposure to PROH at 37degrees C for 10 min was 8.3% (P=0.047) and embryo development for 30 min was 5.4% (P=0.038). Incubation of 2-cell mouse embryos at room temperature or 4degrees C for up to 30 min did not significantly reduce embryo development. Cryosurvival of 2-cell mouse embryos exposed to PROH at room temperature or 4degrees C was similar. CONCLUSION: These findings suggested that pronanediol is toxic to 2-cell mouse embryos in a temperature- and time-dependent fashion. Cryopreservation of 2-cell mouse embryos after exposure at 4degrees C appears to be no better than after exposure at room temperature.
Key Words: Mouse embryo, Propanediol, Equilibration temperature, Equilibration time

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