Extreme rapid warming yields high functional survivals of vitrified 8-cell mouse embryos even when suspended in a half-strength vitrification solution and cooled at moderate rates to 鈭?96 掳C

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• 作者：Shinsuke Seki ; Bo Jin ; Peter Mazur
• 关键词：Embryos ; Oocytes ; Mouse ; Vitrification ; Solute concentration in media ; Cooling rates ; Warming rates ; Survival
• 刊名：Cryobiology
• 出版年：February, 2014
• 年：2014
• 卷：68
• 期：1
• 页码：71-78
• 全文大小：702 K

文摘

To cryopreserve cells, it is essential to avoid intracellular ice formation during cooling and warming. One way to do so is to subject them to procedures that convert cell water into a non-crystalline glass. Current belief is that to achieve this vitrification, cells must be suspended in very high concentrations of glass-inducing solutes (i.e., 猢? molal) and cooled at very high rates (i.e., 鈮?000 掳C/min). We report here that both these beliefs are incorrect with respect to the vitrification of 8-cell mouse embryos. In this study, precompaction 8-cell embryos were vitrified in several dilutions of EAFS10/10 using various cooling rates and warming rates. Survival was based on morphology, osmotic functionality, and on the ability to develop to expanded blastocysts. With a warming rate of 117,500 掳C/min, the percentages of embryos vitrified in 1脳, 0.75脳, and 0.5脳 EAFS that developed to blastocysts were 93%, 92%, and 83%, respectively. And the percentages of morphological survivors that developed to expanded blastocysts were 100%, 92%, and 97%, 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.