Importance of Rapid Warming for Successful Vitrification of Mouse Embryos

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  • マウス初期胚のガラス化保存における急速融解の重要性
  • マウス ショキハイ ノ ガラスカ ホゾン ニ オケル キュウソク ユウカイ ノ ジュウヨウセイ

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Abstract

To cryopreserve cells, it is important to prevent intracellular ice formation (IIF) during cooling and warming. One way to prevent IIF is to subject cells to procedures that convert cell water into a non-crystalline glass. Current belief is that to achieve this vitrification, cells must be cooled at high rates to prevent IIF during cooling and cells must be suspended in very high concentrations of cryoprotectants. We report here that these beliefs are incorrect with respect to the vitrification of 8-cell mouse embryos. We emphasize that it is important to prevent IIF (recrystallization) during warming by rapid warming. In this study, 8-cell embryos are vitrified in several dilutions of EAFS10/10 using various cooling rates and warming rates. Survival was assessed by their developmental ability to blastocysts. With a warming rate of 117,500oC/min, the percentages of embryos vitrified in 1Å~, 0.75Å~, and 0.5Å~ EAFS that developed to blastocysts were 93%, 92%, and 83%, respectively. Even when the solute concentration of the EAFS was reduced to 33% of normal, we obtained 40% functional survival of these 8-cell embryos.

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