透明带激光操作对玻璃化冻融囊胚复苏及孵出动态模式的影响

Effects of laser-assisted zona pellucid micromanipulation on development and hatching mode of vitrified-warmed blastocysts

目的 采用Time-lapse系统探讨激光人工皱缩及辅助孵化对玻璃化冻融囊胚复苏效率以及孵出动态模式的影响。 方法 收集2017年4月至2018年6月期间93例体外受精周期患者移植及冷冻后的剩余囊胚共212个,随机分组处理:采用激光打孔使囊胚腔完全皱缩后冷冻(AS组,n=106)和囊胚未人工皱缩直接冷冻(NAS组,n=106);2组囊胚解冻后随机再分为辅助孵化组(AH组,激光法溶解约1/4透明带)和未辅助孵化组(NAH组,直接囊胚培养)2个亚组;4组囊胚在Primovision时差监测系统培养48h,记录囊胚复苏率、囊胚复张时间、孵出开始时间、完全孵出时间、孵出模式、囊腔回缩次数等动态参数,分析比较各组间差异。 结果 AS+AH组及AS+NAH组囊胚解冻后复苏率显著高于NAS+AH组及NAS+NAH组(P<0.05),而AS+AH组与AS+NAH组、NAS+AH组与NAS+NAH组间囊胚解冻后复苏率无显著差异(P>0.05);与AS+AH组相比,AS+NAH组及NAS+NAH组的囊胚孵出率、完全孵出率及U型孵出率均显著下降(P<0.05),AS+NAH组8字型孵出率显著升高(P<0.05);囊胚解冻后复张时间上4组间均无显著性差异(P>0.05);囊胚孵出开始时间比较,NAS+NAH组显著晚于其他3组(P<0.05),而AS+NAH组及NAS+NAH组囊胚完全孵出时间显著晚于AS+AH组与NAS+AH组,而囊腔回缩次数显著增多(P均<0.05)。 结论 激光人工皱缩联合激光辅助孵化能够改善囊胚玻璃化冻融复苏效率,并且有利于囊胚正常孵出。

Objective : To investigate the effects of laser-assisted artificial shrinking and assisted hatching on the survival efficiency of vitrified-warmed blastocysts and hatching dynamic mode by using Time-lapse monitoring system. Methods : A total of 212 blastocysts were collected from 93 patients undergoing IVF cycles after the embryos transferred and cryopreservation from April 2017 to June 2018.These blastocysts were randomly divided into laser-assisted artificial shrinking group (AS group, n =106) and non-artificial shrinking group (NAS group, n =106) according to whether laser-assisted artificial shrinking was used before vitrification.And then the blastocysts of the two groups were randomly divided into assisted hatching group (AH group) and non-assisted hatching group (NAH group) according to whether laser-assisted hatching was used after warmed.In AH group,about 1/4 of zona pellucida was dissolved by laser.In NAH group,blastocysts were cultured directly.All blastocysts were cultured in Primo vision Time-lapse monitoring system for 48 hours,and dynamic parameters such as embryo survival rate,blastocyst re-expansion time,hatching start time,complete hatching time,hatching mode,contraction frequency of blastocyst,etc.were recorded,and the differences among groups were analyzed. Results : The survival rate of blastocysts in AS+AH group and AS+NAH group were significantly higher than that in NAS+AH group and NAS+NAH group ( P <0.05),but there was no significant difference between AS+AH group and AS+NAH group,NAS+AH group and NAS+NAH group ( P >0.05).Compared with AS+AH group,the hatching rate,complete hatching rate and U-type hatching rate of blastocysts in AS+NAH group and NAS+NAH were significantly decreased ( P <0.05),while the 8-shaped hatching rate of blastocyst in AS+NAH was significantly increased ( P <0.05).There was no significant difference in the re-expansion time of blastocysts among the four groups ( P >0.05).The blastocyst hatching start time in NAS+NAH group was significantly later than that in other three groups ( P <0.05).The blastocyst complete hatching time of AS+NAH group was significantly later than that of the other three groups.The complete hatching time of AS+NAH group and NAS+NAH group was significantly later than AS+AH group and NAS+AH group,while the contraction frequency of blastocyst was significantly increased ( P <0.05). Conclusions : Laser-assisted artificial shrinkage combined with laser-assisted hatching can improve the survival efficiency of vitrified-warmed blastocysts,and which is conducive to the normal hatching of blastocysts.