一种非Piezo依赖的高卵子存活率小鼠显微授精技术

目的对已建立的非Piezo依赖的小鼠显微授精技术作进一步改良,以获得更高的卵子存活率。方法将卵胞浆内单精子显微注射(ICSI)技术中对卵子深吸持后直接深穿刺(一步法非Piezo-ICSI组)的步骤分成两步进行(两步法非Piezo-ICSI组):先浅吸持后浅穿刺,再进一步深吸持后深穿刺;另以孤雌激活发育的卵子作为卵子质量对照组。比较两步法非Piezo-ICSI组与一步法非Piezo-ICSI组卵子的存活率、原核出现率和卵裂率。结果两步法非Piezo-ICSI组的卵子存活率(92.2%)显著高于一步法非Piezo-ICSI组(83.7%)(P<0.01);而原核出现率和卵裂率与一步法非Piezo-ICSI组比较,差异无统计学意义(P>0.05)。结论小鼠卵子胞膜在被新型持卵管深吸持时的变形造成了部分卵子胞膜变脆,两步法吸持穿刺在深吸持前进行了穿刺,避免了脆性敏感时的穿刺,有利于卵子存活。

A new technique of mouse intracytoplasmic sperm injection without assistance of Piezo for high survival rate of oocytes

Objective To achieve higher survival rate of mouse oocytes through modification of established technique of mouse intracytoplasmic sperm injection(ICSI) without assistance of Piezo.Methods The step of deeply sucking and deeply insertion to mouse oocytes in the established method(one-step method without Piezo-ICSI group) was modified to two steps(two-step method without Piezo-ICSI group),in which slightly sucking and slightly insertion was first performed before deeply sucking and deeply insertion.Besides,oocytes of parthenogenetic activation were served as control group.The survival rate,fertilization rate and cleavage rate of oocytes were compared between one-step method without Piezo-ICSI group and two-step method without Piezo-ICSI group.Results The survival rate of oocytes in two-step method without Piezo-ICSI group was significantly higher than that in one-step method without Piezo-ICSI group(92.2% vs 83.7%,P<0.01),while there was no significant difference in both fertilization rate and cleavage rate of oocytes between two group(P>0.05).Conclusion Some oolemma becomes fragile when the shape of the oocyte membrane is changed by drawing deeply with new type of holding pipette.The new technique of mouse ICSI avoids this sensitive fragile by two-step method for oocyte holding and insertion,which is beneficial to achieve higher survival rate of mouse oocytes.