小鼠卵母细胞和胚胎不同冷冻方法的比较研究

Comparative Study on Different Freezing Methods of Mouse Oocytes and Embryos

探讨程序化冷冻与玻璃化冷冻对小鼠GV期卵母细胞及二细胞期胚胎的复苏率及其发育潜能的影响。通过小鼠的卵母细胞与早期胚胎的不同冷冻方法的比较,为后续阿旺绵羊的胚胎冷冻保存提供参考。采用程序化冷冻与玻璃化冷冻技术,分别冷冻小鼠GV期卵母细胞及二细胞期胚胎,复苏后培养,比较不同冷冻处理后的复苏率、成熟率与囊胚率。小鼠GV期卵母细胞程序化冷冻复苏率(48.00%±5.29%)显著低于玻璃化冷冻复苏率(65.00%±5.00%),有统计学差异(P=0.0147<0.05);而程序化冷冻后复苏卵母细胞的发育成熟率略高于玻璃化冷冻组,但无统计学意义。小鼠二细胞期胚胎程序化冷冻组复苏率(76.00%±2.00%)显著高于玻璃化冷冻组复苏率(70.00%±2.00%),有统计学差异(P=0.0213<0.05);冷冻后复苏胚胎发育的囊胚率程序化冷冻略低于玻璃化冷冻及对照组,但无统计学意义。

This paper explores the effects of programmed freezing and vitrification on the recovery rate and developmental potential of mouse GV stage oocytes and two-cell stage embryos.We compared different cryopreservation methods of mouse oocytes and early embryos to provide a reference for subsequent Avang sheep embryo cryopreservation.In this study,programmed freezing and vitrification techniques were used to freeze mouse GV stage oocytes and two-cell stage embryos respectively,and culture them after recovery.The recovery rate,maturity rate and blastocyst rate after different freezing treatments were compared.The programmed freezing recovery rate(48.00%±5.29%)of mouse GV stage oocytes was significantly lower than the vitrification recovery rate(65.00%±5.00%),with statistical difference(P=0.0147<0.05).The development and maturation rate of revived oocytes after programmed freezing was slightly higher than that of vitrification group,but there was no statistical significance.The recovery rate(76.00%±2.00%)of the mouse two-cell stage embryo programmed freezing group was significantly higher than that of the vitrification group(70.00%±2.00%),with statistical difference(P=0.0213<0.05).The rate of blastocyst blastocysts in thawed embryos developed by programmed freezing was slightly lower than that of vitrification and control group,but there was no statistical significance.