体外受精技术在大鼠胚胎冷冻保种中的应用

doi:10.3969/j.issn.1674-5817.2015.04.007

摘要/Abstract

摘要： 目的探讨大鼠体外受精(IVF)技术在大鼠胚胎冷冻应用的可行性。方法选用IVF-20溶液作为大鼠精子获能和受精培养液,改良大鼠1-细胞胚胎培养液(mR1ECM) 作为受精卵培养液,对SD 大鼠进行IVF,以输卵管灌流获得的2-细胞胚胎作为对照,并对发育良好的2-细胞胚胎实施胚胎冷冻,1周后复苏并对存活胚胎采用输卵管移植。结果采用灌流和IVF的方式分别获得胚胎463枚和588枚, 2-细胞胚胎301枚(65.01%)和 376枚(63.94%), 二者受精率相比无显著性差异(P>0.05); 采用两种方式获得的胚胎各冻存150枚, 复苏后形态正常胚胎分别为132枚(88.00%)和130枚(86.66%), 二者复苏率比较无显著性差异(P>0.05); 对存活的胚胎各移植5只受体, 分别产仔45只和22只, 二者的产仔率(32.14% vs 16.92%)有显著性差异(P<0.05)。结论 SD大鼠体外IVF技术在胚胎冷冻保种中应用是可行的, 将为以后基因工程SD大鼠的胚胎冷冻保种提供技术支持。

关键词: 大鼠, 超排卵, 体外受精(IVF), 胚胎冷冻

Abstract: Objective To investigate the feasibility of in vitro fertilization (IVF) technique in rat embryos cryopreservation. Methods IVF-20 was used as rat sperm capacitation and fertilization medium, mR1ECM as a culture solution for IVF fertilized egg of SD rats; 2-cell embryo were obtained by flushing as a control, and well-developed 2- embryos were cryopreserved, after a week the thawed embryos were transplanted to recipients of same strain rats. Results A total of 463 and 588 embryos were separately collected from IVF group and control group, the development rate of normal 2-cell embryos was 65.01% (301/463) and 63.94% (376/588) respectively, which showed no significant difference in the fertilization rate (P>0.05). In the 150 embryos in each group, morphologically normal embryos were 132 (88.00%) and 130 (86.66%) after the recovery, there was no significant difference (P>0.05); Surviving embryos were transplanted into five recipients of same strain rat, and newborn were 45 and 22. The litter rate (2.14% vs16.92%) showeda significant difference between the two groups (P<0.05). Conclusion IVF technology of rat is feasible in the embryo cryopreservation and breed preservation. It provides technical supports for cryopreservation of genetic manipulated rats in future.

Key words: Rat, Superovulation, In vitro fertilization(IVF), Embryo cryopreservation

中图分类号:

Q95-33

周生来, 王维, 于洋, 杨葳, 张梅英, 郑志红. 体外受精技术在大鼠胚胎冷冻保种中的应用[J]. 实验动物与比较医学, 2015, 35(4): 298-301.

ZHOU Sheng-lai, WANG Wei, YU Yang, YANG Wei, ZHANG Mei-ying, ZHENG Zhi-hong. Application of In Vitro Fertilization in Rat Embryos Cryopreservation[J]. Laboratory Animal and Comparative Medicine, 2015, 35(4): 298-301.

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