DAP 玻璃化冷冻液对小鼠2-细胞胚胎的冷冻保存

摘要: 目的探索和优化小鼠2-细胞胚胎冷冻效果，建立小鼠胚胎冷冻保存技术以及超低温保存模型动物胚胎实验室所需的相关技术。方法选择3 个品系( C57BL/6、ApoE － / －、NOS3 － / － ) SPF 级4 周龄雌性小鼠，运用超数排卵、体外受精、玻璃化冷冻法，以及二细胞期胚胎复苏、体外培养等技术方法，观察上述实验效果。结果C57BL/6、ApoE － / －、NOS3 － / － 3 个品系小鼠超数排卵平均值分别为42. 98 个/只、27. 93 个/只、23. 11 个/只，体外受精2-细胞期胚胎率分别为68. 79%、32. 70%、23. 08 %，胚胎冷冻复苏后胚胎回收率72. 77%、80. 87%、83. 33%，存活率分别为56. 92%、54. 84%、70%，存活胚胎体外培养发育到囊胚比率分别为72. 37%、60. 78%、25%。结论选择4 周龄小鼠，按预定相同的技术方法做超数排卵、体外受精、胚胎冻存、胚胎复苏，呈现出较好效果，C57BL/6 小鼠优于基因工程小鼠，基本建立起小鼠2-细胞胚胎冷冻保存技术。

Cryopreservation of Mice 2-cell Embryos by Vitrification Based on DAP

Abstract: Objective To explore the frozen effect of mice 2-cells embryo，to establish embryo cryopreservation technique of mice． Method Three strains mice of SPF female in 4 weeks old ( including C57BL /6，ApoE － / － ，NOS3 － / － ) were selected to carry out experiments of superovulation，in vitro fertilization，vitrification，2-cell embryo recovery and in vitro culture technique． Then，observed the experiment effects． Result The average number of superovulation of the three strains including C57BL /6，ApoE － / － ，NOS3 － / － mice was 42. 98，27. 93，23. 11 per mouse，respectively． The mean 2-cell embryo rate of in vitro fertilization was 68. 79%，32. 70%，23. 08%，respectively． The survival rate after freezing-thawing was 56. 92%，54. 84%，70%，respectively． The embryo recovery rate after freezing-thawing was 72. 77%、80. 87%、83. 33%，and the rate of embryos developed to blastocyst in vitro was 72. 37%，60. 78%，25%，respectively． Conclusion Superovulation，in vitro fertilization，embryo cryopreservation，embryo recovery，of 4 week old mice showing a better effect by the same technology method，and C57BL /6 was better than genetically engineered mice． Thus the basic establishment of mouse 2-cell embryos cryopreservation technology．