两种玻璃化胚胎冷冻方法对不同品系小鼠胚胎冷冻.docVIP

研究报告 两种玻璃化胚胎冷冻方法对不同品系小鼠胚胎冷冻效果比较 王俊风1，李 淼2，张艺宝3，郁丽丽1，魏 勤2，刘丽均1，施美莲1,2徐 平1,2 （1. 上海斯莱克实验动物有限责任公司，上海 201615；2.中国科学院上海生命科学研究院（实验动物中心），上海 201615；3. 南京农业大学动物医学院，南京 210006） 【摘要】 目的 探讨EFS和DAP两种玻璃化冷冻方法对不同品系小鼠胚胎冷冻的效果。方法6个品系小鼠（KM、ICR、BALB/c、C57BL/6J、OB/OB、LAP/rtTA0F59）的2-cell胚胎分别用EFS和DAP两种玻璃化冷冻方法进行冷冻和复苏，比较两种冷冻方法的胚胎复苏率和着床率。结果 6个品系小鼠冷冻胚胎EFS方法的平均复苏率为69.97%（47.9%～83.6%），DAP方法的平均复苏率47.23% （26.3%～76.7%）, EFS方法明显优于DAP方法。其中KM、ICR和BALB/c小鼠EFS方法的冷冻复苏率显著高于DAP方法（P0.01）；冻融胚胎移植后EFS方法的平均着床率27.23%（1.75%～45.0%），DAP方法的平均着床率31.43%（7.0%～46.3%）。除KM、ICR小鼠外，其他4个品系小鼠的着床率DAP方法高于EFS方法。结论 KM和ICR远交群小鼠胚胎适合用EFS方法冷冻保存；C57BL/6J、OB/OB、LAP/rtTA0F59三个品系小鼠DAP方法优于EFS方法，但差异不大； BALB/c小鼠两种玻璃化冷冻方法的冻融胚胎着床率均较低，需进一步研究。 【关键字】 胚胎；玻璃化冷冻；复苏率；着床率【中图分类号】R33【文献标识码】A【文章编号】1671-7856（201）-0000-00 Doi:10.3969/j.issn.1671-7856.2013.006.000 Comparison of two vitrification-cryopreservation methods of embryos of various mouse strains WANG Jun -feng1, LI Miao2, ZHANG Yi-bao3, YU Li-li1, WEI Qin2, LIU Li-jun1, SHI Mei-lian1，2, XU Ping1，2 Shanghai SLAC Laboratory Animal Co. Ltd., Shanghai 201615, China; 2. Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201615; 3. College of Veterinarian Medicine, Nanjing Agriculture University, Nanjing 210006) 【Abstract】 Objective To compare the effect of two vitrification-cryopreservation methods of embryos of several mouse strains. Methods The two-cell embryos from six mouse strains (KM, ICR, BALB/c, C57BL/6J, OB/OB, LAP/rtTA0F59) were cryopreserved and thawed, using EFS and DAP cryoprotectants, respectively. The recovery rate in vitro and implantation rate in vivo were determined to evaluate the efficacy of the two methods. Results The average post-thaw survival rates of the embryos of different strains preserved in EFS were 69.97% (47.9% to 83.6% ), and preserved in DAP 47.23% (26.3% to 76.7%). After 2-cell embryos were transferred to the oviducts of pseudopregnant recipient in all the strains, the implantation rates of EFS-vitrified embryos were ranged f