激光透明带穿孔和激光透明带削薄对冻融小鼠卵子受精率的影响(1)
摘要：目的 探讨激光透明带穿孔和激光透明带削薄对玻璃化冷冻复苏后小鼠卵子受精率的影响。方法 将玻璃化冷冻复苏后的KM小鼠卵子分为4组，前3组为经透明质酸消化处理后脱去颗粒细胞的卵子，其中第1组对透明带进行激光穿孔处理，第2组对透明带进行激光削薄处理，第3组未做激光处理，而第4组为未经透明质酸消化带有颗粒细胞的卵子，同样未做激光处理。此后，在同一时间点使4组卵子与雄鼠精子进行体外受精，次日观察受精结果，比较各组受精率。结果 4组冻融卵子的受精率分别为52.1%(25/48)，25.0%(12/48)，21.3%(10/47)和17.0%(8/47)。激光透明带穿孔组的卵子受精率要显著高于其余3组，P<0.05。而激光透明带削薄组的受精率与未作激光处理的两组相比没有显著差异，P>0.05。结论 激光透明带削薄虽然对卵子造成的损伤较小，但不能有效提高受精率，而激光透明带穿孔能克服冻融造成的透明带硬化及糖蛋白结构改变等问题，显著提高冻融卵子复苏后的受精率。
关键词：激光；透明带穿孔；透明带削薄；冻融小鼠卵子；受精率
, 百拇医药
The Effects of Laser Zona Drilling and Laser Zona Pellucida Thinning on the Fertilization Rates of Mouse Vitrified-thawed Oocytes
RAO Jin-peng，QIU Feng，QIAN Xiao-hong，CAI Yi-ting
(Reproductive Medicine Center， The Second Affiliated Hospital of Zhejiang University School of Medicine， Hangzhou 310052，Zhejiang，China)
Abstract：Objective To evaluate the effects of laser zona drilling and laser zona pellucida thinning on the fertilization rates of mouse vitrified-thawed oocytes. Methods The Kunming mouse vitrified-thawed oocytes were divided into four groups： the first three groups were occytes without cumulus (DO)， but the forth group were oocytes with cumulus (COC). Among the three DO groups， the first one were treated by laser zona drilling and the second one were treated by laser zona pellucida thinning， however， the third DO group and the COC group were not treated by laser shooting. All the oocytes with or without cumulus were fertilized in vitro at the same time， and the fertilization rate were compared on the next day. Results The fertilization rates of four groups were 52.1%(25/48)，25.0%(12/48)，21.3%(10/47) and 17.0%(8/47) respectively. The laser zona drilling group's rates were significantly higher than the other three groups' (P<0.05). However， there is no significant different among the laser zona pellucida thinning group and no laser treating groups(P>0.05).Conclusion The methods of laser zona drilling could improve the fertilization rates of mouse vitrified-thawed oocytes. However， the laser zona pellucida thinning method seems could reduce the damage caused by laser， but could not overcome the problems of zona pellucida hardening and glycoprotein hurting during vitrification， and could not improve fertilization rates.
, http://www.100md.com
Key words：Laser； Zona drilling； Zona pellucida thinning； Mouse vitrified-thawed oocytes；Fertilization rates
激光破膜輔助孵化(Laser assisted hatching)因具有精确，快速，便捷，安全的特点，近年来已经在辅助生殖技术(Assisted reproductive technology，ART)中广泛应用，对因胚胎冷冻造成的透明带变硬、反复着床失败患者的作用已经得到了普遍认可[1-3]。而人类卵子玻璃化冷冻技术因在生育力保存上具有重大的意义而成为近年来研究的热点。但有研究表明卵母细胞在培养过程中，不理想的体外条件会导致透明带变硬，而玻璃化冻融过程中温度的急剧变化则使得透明带硬化的现象进一步加剧，且冻融造成透明带糖蛋白损伤，使顶体反应进行困难，最终致使精子无法穿透透明带而受精失败[4]。本研究将激光破膜法运用到冻融小鼠卵子上，探讨激光透明带穿孔和激光透明带削薄两种方法对冻融卵子复苏后受精率的影响，为辅助生殖技术提供新的思路。, 百拇医药(饶金鹏 邱枫 钱小红 蔡益婷)
关键词：激光；透明带穿孔；透明带削薄；冻融小鼠卵子；受精率
, 百拇医药
The Effects of Laser Zona Drilling and Laser Zona Pellucida Thinning on the Fertilization Rates of Mouse Vitrified-thawed Oocytes
RAO Jin-peng，QIU Feng，QIAN Xiao-hong，CAI Yi-ting
(Reproductive Medicine Center， The Second Affiliated Hospital of Zhejiang University School of Medicine， Hangzhou 310052，Zhejiang，China)
Abstract：Objective To evaluate the effects of laser zona drilling and laser zona pellucida thinning on the fertilization rates of mouse vitrified-thawed oocytes. Methods The Kunming mouse vitrified-thawed oocytes were divided into four groups： the first three groups were occytes without cumulus (DO)， but the forth group were oocytes with cumulus (COC). Among the three DO groups， the first one were treated by laser zona drilling and the second one were treated by laser zona pellucida thinning， however， the third DO group and the COC group were not treated by laser shooting. All the oocytes with or without cumulus were fertilized in vitro at the same time， and the fertilization rate were compared on the next day. Results The fertilization rates of four groups were 52.1%(25/48)，25.0%(12/48)，21.3%(10/47) and 17.0%(8/47) respectively. The laser zona drilling group's rates were significantly higher than the other three groups' (P<0.05). However， there is no significant different among the laser zona pellucida thinning group and no laser treating groups(P>0.05).Conclusion The methods of laser zona drilling could improve the fertilization rates of mouse vitrified-thawed oocytes. However， the laser zona pellucida thinning method seems could reduce the damage caused by laser， but could not overcome the problems of zona pellucida hardening and glycoprotein hurting during vitrification， and could not improve fertilization rates.
, http://www.100md.com
Key words：Laser； Zona drilling； Zona pellucida thinning； Mouse vitrified-thawed oocytes；Fertilization rates
激光破膜輔助孵化(Laser assisted hatching)因具有精确，快速，便捷，安全的特点，近年来已经在辅助生殖技术(Assisted reproductive technology，ART)中广泛应用，对因胚胎冷冻造成的透明带变硬、反复着床失败患者的作用已经得到了普遍认可[1-3]。而人类卵子玻璃化冷冻技术因在生育力保存上具有重大的意义而成为近年来研究的热点。但有研究表明卵母细胞在培养过程中，不理想的体外条件会导致透明带变硬，而玻璃化冻融过程中温度的急剧变化则使得透明带硬化的现象进一步加剧，且冻融造成透明带糖蛋白损伤，使顶体反应进行困难，最终致使精子无法穿透透明带而受精失败[4]。本研究将激光破膜法运用到冻融小鼠卵子上，探讨激光透明带穿孔和激光透明带削薄两种方法对冻融卵子复苏后受精率的影响，为辅助生殖技术提供新的思路。, 百拇医药(饶金鹏 邱枫 钱小红 蔡益婷)
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