›› 2008, Vol. 28 ›› Issue (6): 397-400.

所属专题: 实验动物资源开发与利用

• 论文 • 上一篇    下一篇

SD大鼠超排卯及胚胎冷冻方法比较

  

  1. 中国医科大学实验动物部,沈阳 110001
  • 收稿日期:2008-09-05 出版日期:2008-06-30 发布日期:2008-06-30
  • 基金资助:
    辽宁省科学技术厅攻关基金(2006408001-1);辽宁省重点实验室专项资金[辽科发(2005)36号]

Comparative Study on Superovulation and Embryo Freezing in SD Rats

  1. China Medical University, Shenyang 110001, China
  • Received:2008-09-05 Online:2008-06-30 Published:2008-06-30

摘要: 目的 优化超排卵效果及建立大鼠胚胎冷冻方法。方法 腹腔注射三种不同剂量PMSG-HCG(15IU/只、20IU/只、25IU/只)超排5周龄SD大鼠,当日与正常雄鼠1:1合笼交配,比较三种激素剂量的超排卵效果。应用输卵管吹卵法收集2-细胞胚胎,采用小鼠胚胎冷冻方法对大鼠胚胎进行冷冻,通过胚胎复苏率比较不同冷冻时间(方法Ⅰ:5min,方法Ⅱ:1min)的冷冻效果。结果 采用三种剂量进行超排分别获得13±12.7、31±14.2、41±25.4枚胚胎,其中激素剂量25IU/只超排效果最好;方法Ⅱ在操作简便性上明显优于方法Ⅰ,但冷冻效果无显著性差异(平均复苏率88.3%与85.0%)。采用方法Ⅱ冻存大鼠胚胎总计152枚,复苏后形态正常胚胎共计139枚(91.44%),培养20min后,对存活胚胎共计105枚(75.53%)进行胚胎移植,产仔39只(37.14%)。结论 成功优化大鼠超排及建立大鼠胚胎冷冻方法。

关键词: 大鼠, 超排卵, 胚胎冷冻

Abstract: Objective To improve superovulation and establish a method of embryo freezing for rats. Methods Different doses of PMSG-HCG (15 IU,20 IU,25 IU/rat) were injected (i.p.) into three groups(Group A,B,C) of 5 weeks SD rats, and each treated female was coupled with fertile male, the effect of each dose was compared to determine the best dose for superovulation. Two-cell was collected by clysising fimbriae tubae. The embryos were divided into 2 groups. Embryos in group 1 were frozen for 1 minute, and embryos in group 2 for 5 minutes. All the embryos of rats were frozen according to the standard method for mouse embryos frozen. The embryos were resuscitated, and the rate of resuscitation were compared to assess the two method. Results The average number of zygotes obtained from each rat in group A,B,C was 13 土 12.7,31 土 14.2,and 41 土 25.4. There were significant differences among the groups (P<0.05). The resuscitation rate of embryos in group 1 is 88.3%, and 84.9% in group 2. Of 152 rat embryos were frozen for 1 minute, 139 embryos were successfully resuscitated, and cultured for 20 minutes.The normal cultured 105 embryos were transplanted into pseudopregnant rats, and 39 offspring were obtained. Conclusion The method for rat superovulation has been successfully improved, and a method of embryo freezing for rats has been established.

Key words: Rat, Superovulation, Embryo freezing