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Survival of mouse embryos frozen and thawed rapidly

Summary. Mouse morulae were frozen rapidly to –196°C in the presence of 1·0– 2·5 M-DMSO by a 3-step procedure; the samples were seeded at –4 to –8°C, held at –20°C in an ethanol bath for 10 min, suspended over liquid nitrogen at –100°C for 10 min and then plunged directly into liquid nitrogen at –196°C. The cooling rate between –20 and –75°C was 17°C/min. In all concentrations of DMSO significantly higher proportions of embryos developed to fully expanded blastocysts after 48 h in culture after rapid thawing (360°C/min) than after slow thawing (25°C/min). The highest survival rates were obtained for the embryos frozen rapidly in the presence of 1·5 and 2·0 M-DMSO (36 and 53% respectively). Various methods for removal of DMSO (2·0 M) were tested with the 3-step freezing and rapid thawing procedures. The best results for development to fully expanded blastocysts were obtained with PBS + 2·0 M-DMSO + 0·5 M-sucrose (2 min) followed by PBS + 0·5 M-sucrose (2 min) at room temperature (82%) and with stepwise dilution in PBS at 30°C (70%). When 26 embryos developed to blastocysts in culture after rapid freezing and thawing were transferred into 2 recipients, 11 newborn young (42%) were obtained.

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				M. Kasai, K. Ito, and K. Edashige Morphological appearance of the cryopreserved mouse blastocyst as a tool to identify the type of cryoinjury Hum. Reprod., July 1, 2002; 17(7): 1863 - 1874. [Abstract] [Full Text] [PDF]