【作者】 王伟；

【导师】 邵小光；

【作者基本信息】 大连医科大学 ， 妇产科学， 2006， 硕士

【摘要】 自1992年Palermo等将胞浆内单精子显微注射(Intracytoplasmic sperm injection, ICSI)应用于人类并获成功分娩以来,该技术因其单精受精率高,多精受精率低以及不受精子浓度、形态影响等优点,而被广泛应用于临床。这是人类在人工辅助助孕技术(artificial reproduction technology, ART)方面的一个突破性进展。至今全世界采用ICSI技术已经超过35,000个治疗周期,超过10,319例妊娠,出生婴儿超过5,352名。该技术在治疗不孕症,尤其是男性不育中取得了令人瞩目的进展。ICSI为男性不育的治疗带来了革命性的突破,成为严重少精子症、阻塞性无精子症和一些非阻塞性无精子症的首选治疗方法。ICSI是借助显微操作仪将精子直接注入卵母细胞胞浆的一项辅助受精技术,近十年来广泛应用于人类辅助生育。ICSI是一种旁路受精方式,丰富了经典的受精理论,也可作为研究受精机制的实验模型。目的:探讨提高小鼠卵细胞浆内单精子注射受精率的方法及研究各种质量精子的鼠卵细胞激活能力。方法:选用鼠龄8~10周的健康昆明雌鼠作为卵子的供体,通过HMG/HCG方案行超促排卵,断髓处死后,从鼠输卵管中取卵。通过ICSI技术,了解不同采卵时间、不同前培养时间对ICSI的影响。精液标本来源于大连市妇产医院生殖中心不孕症门诊患者。用SWIM-UP法分别处理正常组、少精组、弱精组的精液,选择优质的精子进行ICSI ,计算卵母细胞激活率、卵细胞形态正常率,并进行各参数间对比。结果: 1、不同采卵时间的观察显示, hCG注射后18h获卵的成熟度高于其他各组,呈显著性差异(P<0.05)。2、卵细胞体外培养时间的选择:卵细胞体外培养2 h组卵细胞激活率、形态正常率均明显高于4 h组,差异具有统计学意义(P < 0. 05)。3、实验对照组、空白对照组卵细胞激活率远远低于实验组,差异有统计学意义(P < 0. 01)。4、空白对更多还原

【Abstract】 Objective: To study methods raising average percentage of ferti- lization by intracytoplasmic sperm injection (ICSI) in mouse eggs. To study oocyte activation capability of different parameters human semen samples.Methods: KunMing white mice (8~10 weeks of age) were used as donors of eggs. The influence of different phases of collecting eggs and different culture time on ICSI was investigated. Semen samples according to WHO (1994) recommendation were evaluated, and were treated by SWIM-UP.Results: 1. The mature rate of mouse oocyte, which the collection of oocytes is done 18 hours after the injection of HCG, is significant higher than the other groups (P<0.05).2. The oocyte activation rate and normal morphy rate in the group which culture time is 2 hours is higher than 4 hours. The difference is significant (P<0.05). 3. The activation rate of the experimental controlled group and the opening controlled group are significant lower than the experiment group (P<0.01).4. The survival rate of the opening controlled group is higher than the other groups (P<0.05).5. No significant difference on activation rate occurred between different groups of sperm (P>0.05).Conclusion:1. The proper time of the oocyte collecting and the oocyte culture is important for the result of ICSI.2. It might be suggested that the mouse oocyte activation capacity of human sperm do not directly relate to initial semen sample results.3. The fertilization rate of the mouse oocyte after ICSI is low. The volume and the ability of resistence of injury to mouse oocyte may result in the above results.4.The failure of oocyte更多还原