应用Time-lapse对IMSI技术中精子头部空泡的研究

发布时间：2018-02-08 17:10

  本文关键词： 精子头部空泡 IMSI 受精率 time-lapse　出处：《大连医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:在辅助生殖技术(assisted reproductive technology,ART)中,对怎样提高临床妊娠率、减少移植胚胎数目的研究主要针对女性因素,但精子在胚胎发育中的作用同样重要。怎样挑选更优质的精子以及怎样挑选更具有发育潜能的胚胎用以移植成为了关注的焦点。随着胞质内形态学选择性精子注射(intracytoplasmic morphologically selected sperm injection,IMSI)技术的应用,研究者在精子核区经常能观察到至少1个空泡,并发现精子头部的这些空泡与IMSI的预后不良有关。本研究的目的是应用time-lapse探究精子头部空泡的数量和大小在IMSI中对胚胎发育的影响。资料和方法:选自2015年1月至12月期间于大连市妇女儿童医疗中心生殖门诊就诊的患者,按纳入标准收集选取了 21对不孕夫妇的共243枚MII卵母细胞进行IMSI注射。同时根据精子头部空泡情况将胚胎分为四组,A组(n=48)注射精子不含空泡,B组(n=79)注射精子含有1-2个小空泡(d1umm),C组(n=75)注射精子含有2个以上小空泡(d1um),D组(n=41)注射精子含有1个以上大空泡(d≥1um)。分析入组患者的一般情况,如男女方年龄、不孕年限、不孕因素、Gn总量及天数、精液常规参数;对四组分别进行实验室一般情况的分析比较,如平均获卵数、受精率、卵裂率、D3优胚率;应用time-lapse对四组胚胎发育动力学参数进行统计分析。应用SPSS 17.0系统软件进行数据分析,计量资料数据应用方差检验,计数资料应用卡方检验,以P0.05为差异有统计学意义。结果:对ABCD四组进行统计学分析:①入组患者一般情况:女方平均年龄(33.8±2.1)周岁,男方平均年龄(35.6±4.2)周岁;患者平均不孕年限(2.0± 1.4)年;其中原发不孕患者14例,继发不孕7例;仅男性因素8例,男性因素合并卵管因素 11 例;平均Gn总量(2645.4±216.3)IU;平均Gn天数(11.5士 1.2)天。21例精液样本的总体参数:精液体积(3.4±0.6)ml,精子密度(6.1±1.2)10/ml,精子活力(26.9±2.6)%;②四组实验室一般情况,如2PN受精率、卵裂率无统计学差异(P0.05),但D组的D3优胚率显著低于A组(66.66%vs 23.53%,P0.001);③A、B、C、D四组的雌雄原核出现时间、消失时间、分裂成2细胞、3细胞时间均无统计学差异;④A组到达4细胞的时间(t4,h)显著早于D组(40.14±4.33 vs 51.16±11.16,P=0.012);A组到达5细胞阶段的时间(t5,h)显著早于D组(46.61±3.43 vs 59.74±10.34,P=0.009);A组到达8细胞阶段的时间(t8,h)显著早于D组(55.37±3.56 vs 70.12±13.41,P=0.037);⑤A组2细胞阶段(CC2,h)显著短于D组(9,91±2.23 vs 16.33士6.78,P=0.043);A组从3细胞到4细胞的同步性(S2,h)显著优于D组(0.33 ±0.06 vs 1.99±0.06,P=0.024);四组胚胎从3细胞到5细胞(CC3)的时间没有统计学差异;从5细胞到8细胞(S3)的时间没有统计学差异。结论:精子头部空泡对胚胎早期发育的影响不能被排除,注射没有空泡精子到达4细胞的时间显著短于注射大空泡精子,含有大空泡的精子染色体或DNA的损伤可能已经超出了胚胎能够修复的能力;在IMSI注射前应用高倍放大系统对精子头部空泡的大小和数量进行筛选对早期胚胎的动力学发育有重要影响。
[Abstract]:Objective: in assisted reproductive technology (assisted reproductive technology, ART), on how to improve the clinical pregnancy rate, reduce the number of embryos for research purposes the main factors of female, but the role of sperm during embryonic development are equally important. How to choose a better quality of sperm and how to choose more with the developmental potential of embryos for transplantation into the focus of attention. With intracytoplasmic sperm injection (intracytoplasmic morphologically selective morphology of selected sperm injection, IMSI) technology, the researchers often observed at least 1 void in sperm nuclei, and found that these vacuoles and IMSI sperm head of bad prognosis. The purpose of this study is to influence the number and size of the application of time-lapse on sperm head cavity on embryonic development in IMSI. Materials and methods: from January 2015 to December in Dalian city. Women and children's Medical Center reproductive clinic patients, according to the inclusion criteria collect from 21 infertile couples were a total of 243 MII oocytes were injected with IMSI. At the same time according to the situation of the embryo sperm head were divided into four groups, group A (n=48) injection of sperm containing vacuoles, group B (n=79) containing sperm injection 1-2 small vacuoles (d1umm), C group (n=75) injection of sperm containing more than 2 small vacuoles (d1um), D group (n=41) injection of sperm containing more than 1 large vacuoles (d = 1um). Analysis of general groups, such as women age, duration of infertility, infertility factors. The total Gn and the number of days, semen parameters; comparative analysis of four groups of laboratory conditions, such as the average number of oocytes, fertilization rate, cleavage rate, good embryo rate D3; time-lapse application development on the kinetic parameters of the four groups of embryos were analyzed. The application of SPSS 17 software was used for data analysis, measurement data 鏂欐暟鎹�搴旂敤鏂瑰樊妫�楠，

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