SURVIVAL OF MOUSE EMBRYOS FROZEN AND THAWED RAPIDLY
SURVIVAL OF MOUSE EMBRYOS FROZEN AND THAWED RAPIDLY
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DOI：
10.1530/jrf.0.0590051
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发表时间：
1980-01-01
期刊：
影响因子：
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通讯作者：
IRITANI, A
中科院分区：
文献类型：
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作者：
KASAI, M;NIWA, K;IRITANI, A
将小鼠桑椹胚快速冷冻至-196 ℃。在1.0- 2. 5 M-DMSO [二甲基亚砜，一种细胞保护剂]存在下，通过3步程序将样品接种于-4至-8 ℃。温度保持在-20 ℃。在乙醇浴中于10 ℃下搅拌10分钟，在约100 ℃下悬浮于液体N上。100.degree. 10分钟，然后直接浸入-196 ℃的液体N中。C.冷却速率在-20 ℃和-75 ℃之间。C是. apprx。17.degree.在所有浓度的DMSO中，在快速解冻（360 ℃）后培养48小时后，显著更高比例的胚胎发育为完全扩张的胚泡。C/min）比缓慢解冻（25 ℃/min）后的温度低。C/min）。在1.5和2.0 M-DMSO存在下快速冷冻的胚胎获得最高的存活率（分别为36%和53%）。采用3步冷冻和快速解冻程序测试了各种去除DMSO（2.0 M）的方法。在室温（82%）下用PBS [磷酸盐缓冲盐水]+2.0M DMSO +0.5M-蔗糖（2分钟），然后用PBS +0.5M-蔗糖（2分钟），并在30 ℃下在PBS中逐步稀释，获得发育为完全扩张的胚泡的最佳结果。C（70%）。将26枚胚胎经快速冻融后培养至囊胚，移植到2只受体体内，获得新生幼仔11只（42%）。
Mouse morulae were frozen rapidly to -196.degree. C in the presence of 1.0-2.5 M-DMSO [dimethyl sulfoxide, a cytoprotective agent] by a 3-step procedure; the samples were seeded at -4 to -8.degree. C, held at -20.degree. C in an ethanol bath for 10 min, suspended over liquid N at .apprx. 100.degree. C for 10 min and then plunged directly into liquid N at -196.degree. C. The cooling rate between -20 and -75.degree. C was .apprx. 17.degree. 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.degree. C/min) than after slow thawing (25.degree. 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 [phosphate-buffered saline] + 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.degree. 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.