45例不育男性精子核中二倍体和双染色体的发生率

对因男方精液因素(精子数目或/和活动力降低,精子形态异常)而致不育的夫妇,宫腔内配子移植和体外受精(IVF)可能使其受孕。然而,这些技术都要求精液质量至少达到最低标准。胞浆内单精子注射(ICSI)技术却不同,即使精子数目极少、无活力、未经顶体反应、无尾、形态异常或/和不成熟,仍可获得受精。为明确ICSI的应用是否可能增加产生     

常规体外受精失败患者卵浆内单精子注射治疗的结果分析

卵浆内单精子注射(ICSI)是治疗严重男性因素不育的有效方法。也有医师将它用于常规体外受精(IVF)失败的患者。在征得患者同意的情况下我们对以往常规IVF全部卵母细胞不受精的患者再次进入IVF周期时部分卵母细胞进行常规IVF，部分进行ICSI，取得了较好的效果。     

常规IVF中受精失败相关因素及后续处理结果分析

目的:分析常规IVF受精失败原因,以进一步提高IVF受精率。方法:回顾性分析常规IVF中受精率低下或受精失败患者的各种因素及后续处理结果。结果:438个常规IVF取卵周期中,受精率≤25%的周期30个(A组)(6.85%)。其中12个周期第1日行补救性ICSI或重复IVF,最终共20个周期进行了胚胎移植,但均未获得临床妊娠。与受精率正常的周期(B组)相比,A组精子密度、活率、a级精子比例、前向运动精子比例(PMS)及分离后的精子活率、PMS比例、精子密度等均显著降低(P均<0.05)。组间的获卵数无明显差别,但A组MI期及GV期卵的数量明显高于B组(P<0.01)。其他受精失败因素包括对精子因素导致受精失败的7例在随后的周期进行ICSI,5例获得临床妊娠。结论:受精失败的原因是多方面的,但精液各参数下降及卵子发育不成熟是其主要原因;第1日行补救性ICSI或重复IVF的妊娠结局差;对于精子因素造成的受精失败,在以后的周期中行ICSI能获得理想的结果。     

体外受精失败的原因及对策

在接受体外受精胚胎移植治疗的患者中,有时会出现全部获取的卵母细胞均未受精现象,即受精失败。其原因可能为精子与透明带结合失败和(或)精子穿透异常、卵母细胞激活失败等。目前尚缺乏准确的预测受精失败的方法,临床上一旦出现受精失败应采取积极的补救措施,并根据患者病史、临床特征、配子情况对患者以后的治疗措施提出建议。早补救ICSI的有效性和安全性尚需进一步的临床验证。     

透明带内精子冻存的初步实验研究

Cohen首次将精子冻存于人、小鼠、仓鼠空透明带内 ,经复苏后 ,回收率可达 4 4 %～ 94 % ,复苏后精子对体外成熟的卵母细胞行卵细胞浆内单精子注射 (intracytoplasmicspermin jection ,ICSI)后 ,可获正常双原核卵[1] 。本研究利用人受精前未成熟卵、ICSI后受精异常卵及发育停滞胚胎的透明带 ,以保存睾丸精子及取精困难的少弱精症患者的精子 ,同时比较了不同来源精子的冻融耐受性。1　材料与方法1.1　材料　精子取自接受ICSI治疗的男性不育患者 ,因女方原因行常规体外受精胚胎移植 (invitrofertilizationandembryotransfer,IVF ET)的…     

体外受精-胚胎移植周期透明带透亮致密患者的受精与临床结局

目的：探讨卵子透明带透亮致密患者行常规体外受精（IVF）或卵胞质内单精子显微注射（ICSI）后受精、胚胎发育情况及临床结局。方法：回顾性分析体外受精过程中卵子出现透明带透亮致密的43个周期,其中IvF周期27例,ICSI周期16例,比较受精率与正常受精比例、胚胎质量、临床妊娠结局。结果：27例IVF周期均未受精,行早期补救ICSI,其中23例完全不受精。补救后IVF总体受精率达64．83％,正常受精率59.32％;16例ICSI周期均受精,受精率和正常受精率分别为85．11％和79．01％,均显著高于IVF组（P〈0．05）。IVF组和ICSI组卵裂率分别为97．96％、95．65％,优质胚胎率分别为52．67％、43．75％,组间差异无统计学意义（P〉0．05）。IVF组妊娠率为37．04％,种植率为33．33％;ICSI组妊娠率为31．25％,种植率为25．00％,组间临床结局相近,无统计学差异（P〉0．05）。结论：透明带透亮致密患者的自然受精能力下降,需要采用ICSI方式授精。常规IVF受精失败后采用早期补救ICSI或直接行ICSI能明显改善透明带透亮致密患者的受精结局,但不能改善妊娠结局。     

在体外受精过程中引入胞浆内精子注射技术的模式

胞浆内精子注射(Intracytoplasmic sperm injec-tion,ICSI)是近4年来治疗严重男性不育症的重要进展,其成功超越了传统的体外受精(IVF)及其它显微外科受精技术。回顾性复习1994年～1995年在私人的社区IVF中心进行的75例ICSI的结果:     

不明原因不孕患者体外受精方式的探讨

目的探讨不明原因不孕患者行首次体外受精(IVF)治疗时受精方式的选择。方法对2004年6月至2005年12月在北京大学第三医院行IVF治疗的35例不明原因不孕患者,将其卵母细胞分为2组,同时行常规受精和卵胞浆内单精子注射受精(ICSI),比较2组的受精结局、妊娠率及着床率。结果常规受精组受精率(51.2%)低于ICSI组(61.3%),差异有统计学意义(P<0.05);而完全受精失败率(20.0%)明显高于ICSI组(2.8%),差异有统计学意义(P<0.05);2组的优质胚胎率、临床妊娠率、着床率差异无统计学意义。结论不明原因不孕患者行首次IVF治疗时,采用部分卵母细胞常规受精、部分卵母细胞ICSI的方法,可降低完全受精失败风险。     

精子与卵母细胞透明带结合对ICSI治疗结局的影响

目的:探讨使用透明带结合后的精子行ICSI,是否可改善ICSI治疗结局。方法:选择上一周期IVF完全不受精者或男方为少、弱、畸形精子症而接受ICSI治疗的不育患者,共收录106对不孕夫妇,将其随机分成常规ICSI对照组和利用未成熟卵透明带上结合的精子进行ICSI的试验组,每组53例,分析比较ICSI结局。结果:试验组的卵裂率、优质胚胎率、可使用胚胎率均显著高于对照组,差异有统计学意义(P<0.05)。虽然临床妊娠率试验组(36%,19/53)与对照组(49%,26/53)组间无统计学差异(P>0.05),但试验组种植率(29%)、多胎率(35.71%,10/26)明显高于对照组(17%、10.53%,2/19)(P<0.01)。结论:通过利用患者自体ZP结合的精子进行ICSI注射,能获取高质量胚胎和高种植率,这种改良的ZP结合ICSI方法或许能为今后ICSI治疗提供一个新的思路。     

非男性因素不孕体外受精失败行卵胞浆内单精子注射的结局分析

目的:探讨卵胞浆内单精子注射(ICSI)对非男性因素不孕IVF失败患者治疗结局的影响。方法:回顾性分析由于第一周期常规IVF治疗中卵子完全不受精或受精率≤25%,行补救性ICSI的10个周期(补救性ICSI组),以及因前次受精失败而在随后的治疗周期中采取ICSI方法受精的19个周期(后续性ICSI组)的ICSI治疗结局,并以因男方少弱精子症进行第1次ICSI治疗的133个周期为对照组。结果:后续性ICSI组受精率、植入率、妊娠率和分娩率均高于补救性ICSI组,但差异均无统计学意义(P>0.05)。后续性ICSI组优胚率显著高于补救性ICSI组(P<0.05);补救性ICSI组受精率(48.9%)、优质胚胎率(29.2%)、植入率(0%)、妊娠率(0%)、分娩率(0%)均显著低于对照组(分别为72.1%、46.6%、21.2%、45.1%、39.1%);后续性ICSI组受精率、植入率、妊娠率、分娩率分别为55.4%、8.8%、21.1%、15.8%,均低于对照组(P<0.05或P<0.01)。优质胚胎率后续性ICSI组(44.2%)低于对照组,但无统计学差异。结论:对于非男性因素不孕IVF失败患者,ICSI能避免受精失败,但是受精率以及妊娠结局受到卵母细胞隐匿性异常的影响。     

Clinical application of sperm-oocyte interaction tests in in vitro fertilization--embryo transfer and intracytoplasmic sperm injection programs

OBJECTIVE: To review the clinical value of sperm-oocyte interaction tests for the diagnosis and management of infertility by standard IVF or intracytoplasmic sperm injection (ICSI). DESIGN: Review of recent publications on relationships among sperm-oocyte interaction tests, sperm characteristics, and results of IVF and determination of frequency of defective sperm-oocyte interaction in infertile men. MAIN OUTCOME MEASURE(S): Fertilization rates with IVF, sperm characteristics, sperm-zona pellucida (ZP) binding, ZP-induced acrosome reaction (AR), and sperm-ZP penetration. RESULT(S): Sperm defects associated with low sperm-ZP binding or impaired ZP-induced AR and sperm-ZP penetration are the major causes of failure of fertilization when all or most oocytes from a couple do not fertilize in standard IVF. There is a high frequency of defective sperm-ZP interaction in men with oligozoospermia (<20 x 10(6)/mL) and severe teratozoospermia (strict normal sperm morphology < or =5%). Sperm morphology correlates with sperm-ZP binding, and sperm concentration correlates with ZP-induced AR in infertile men with sperm concentrations >20 x 10(6)/mL. Defective ZP-induced AR may cause infertility in up to 25% men with idiopathic infertility. These patients require ICSI despite the normal standard semen analyses. CONCLUSION(S): Sperm-oocyte interaction tests are useful for diagnosis of subtle sperm defects that cause infertility in men without severe abnormalities of semen analysis. Pre-IVF diagnosis of these sperm defects will assist in the clinical assignment of patients to treatment with either standard IVF or ICSI.     

Disordered zona pellucida-induced acrosome reaction and failure of in vitro fertilization in patients with unexplained infertility

OBJECTIVE: To determine the relationship between the zona pellucida (ZP)-induced acrosome reaction (AR) and fertilization rate and pregnancy rate in standard IVF and the frequency of disordered ZP-induced AR (DZPIAR) in patients with unexplained infertility. DESIGN: Prospective study. SETTING: Academic research and teaching tertiary hospital. PATIENTS: Patients with unexplained infertility with normal semen analysis. INTERVENTION: None. MAIN OUTCOME MEASURE: Semen analysis, the ZP-induced AR, and measurements of fertilization rate and pregnancy rate with in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). RESULT(S): A statistically significant correlation was found for the ZP-induced AR and fertilization rate with standard IVF (n = 65). Patients with DZPIAR (ZP-induced AR of 16%, who had an average fertilization rate of 61%. The sensitivity and specificity of DZPIAR for prediction of IVF rates <30% and >or=30% were 80% and 86%, respectively. Of 260 patients screened, the frequency of DZPIAR was 29%. Ten patients with DZPIAR had an average fertilization rate of 15% and no pregnancy with initial IVF cycles, and a fertilization rate of 61% (with three live-birth pregnancies) with subsequent ICSI cycles. Another 33 patients with DZPIAR were treated with ICSI alone, with an average fertilization rate of 71% and a live-birth pregnancy rate of 17% per embryo transfer. Sixteen of the patients had live-birth pregnancies (including one set of twins) after undergoing an average of 3.2 embryo transfers. CONCLUSION(S): Patients with DZPIAR have a low or zero fertilization rate with standard IVF but high fertilization and pregnancy rates with ICSI. Up to 29% of patients with unexplained infertility with normal semen analysis may have this condition, which should be diagnosed and treated with ICSI rather then standard IVF.     

Quinn's advantage fertilization medium enhances zona pellucida-induced acrosome reaction compared with human tubal fluid medium

The aim of this study was to compare sperm velocity, hyperactivation, zona pellucida (ZP) binding and ZP-induced acrosome reaction (AR) between Quinn's advantage fertilization (QAF), human tubal fluid (HTF) and Ham's F10 media. Semen samples were obtained from normozoospermic men and motile spermatozoa were prepared by gradient centrifugation (PureSperm). Unfertilized oocytes from clinical IVF were used for spermatozoa-oocyte interaction tests. Sperm velocity and hyperactivation were assessed using a Hamilton-Thorn motility analyser. When media were supplemented with human albumin, sperm motility and velocity and sperm binding were not significantly different between QAF and HTF. However, ZP-induced AR was significantly higher with QAF than HTF (42±22 versus 21±18, P<0.th001). Sperm velocity, hyperactivation and sperm binding were also significantly higher in QAF than Ham's F10 media. Supplementation of media with either human serum or human albumin showed no difference in effect on all sperm test results. In conclusion, QAF medium significantly enhances ZP-induced AR which is essential for sperm penetration. Thus QAF appears to be a better medium than HTF for sperm fertilizing ability in conventional IVF.     

Normal range and variation of the zona pellucida-induced acrosome reaction in fertile men

OBJECTIVE: To investigate the normal range of the zona pellucida (ZP)-induced acrosome reaction (AR) in fertile men. DESIGN: A study of normative data. SETTING: Academic research and teaching tertiary hospital.None.Fertile men (n = 111) who had children or whose female partner was currently pregnant. MAIN OUTCOME MEASURE (S): Semen analysis, sperm-ZP binding, and the ZP-induced AR. RESULT(S): There was wide range for the ZP-induced AR: 20%-98% with a mean of 48%. The ZP-induced AR was significantly correlated with sperm concentration and normal morphology. The ZP-induced AR was significantly lower in men with sperm concentration 60 x 10(6)/mL. Similarly, men with normal sperm morphology >or=15% had significantly higher ZP-induced AR than men with normal morphology <15%. CONCLUSION(S): There is a wide range of the ZP-induced AR in fertile men that is correlated with sperm concentration and normal morphology.     

Sperm nuclear chromatin normality: relationship with sperm morphology, sperm-zona pellucida binding, and fertilization rates in vitro.

OBJECTIVE: To study whether the results of tests of sperm chromatin and deoxyribonucleic acid (DNA) normality are related to fertilization rates in vitro. DESIGN: Normal morphology, nuclear maturity determined by acidic aniline blue stain, and DNA normality determined by acridine orange fluorescence of sperm in insemination medium and the number of sperm bound to the zona pellucida (ZP) of the oocytes that had failed to fertilize in vitro were determined. The relationship between sperm test results and fertilization rates were analyzed by logistic regression. SETTING: Samples were obtained from patients undergoing in vitro fertilization (IVF) treatment. RESULTS: The number of sperm bound to the ZP, the percentage of sperm with normal morphology, and the percentage of sperm with normal DNA were the most significant factors related to fertilization rates in vitro. In patients with normal morphology > or = 15% or with > 10 sperm bound per ZP, the percentage of sperm with normal DNA, the number of sperm bound to the ZP, and motility grade were significantly related to IVF rates. CONCLUSION: In patients with normal morphology > or = 15%, failure of fertilization may be because of defects of sperm-ZP binding or abnormal DNA. Assessment of DNA normality of motile sperm in the insemination medium may aid prediction of fertilization rates in addition to normal morphology and sperm-ZP binding.     

A prospective randomized trial of conventional in vitro fertilization versus intracytoplasmic sperm injection in unexplained infertility

PURPOSE: To compare outcomes in patients with unexplained infertility undergoing conventional in vitro fertilization (IVF) versus intracytoplasmic sperm injection (ICSI). METHODS: Sixty women with unexplained infertility in a Canadian tertiary-level clinic were randomized to IVF or ICSI. Subjects underwent downregulation with gonadotropin-releasing hormone agonist prior to initiation of recombinant human follicle-stimulating hormone. The primary outcome measure was fertilization rate. Secondary outcomes included implantation rate, embryo quality, clinical pregnancy rate, and live birth rate. RESULTS: There was no statistically significant difference in fertilization rate (77.2% IVF vs. 82.4% ICSI), implantation rate (38.2% IVF vs. 44.4% ICSI), clinical pregnancy rate (50% in each group), or live birth rate (46.7% IVF vs. 50% ICSI). There were two cases of failed fertilization in the IVF group. There was no significant difference in embryo quality between groups. CONCLUSIONS: There were no differences in clinical outcomes associated with IVF versus ICSI in the treatment of unexplained infertility.     

Birth after intracytoplasmic sperm injection with use of testicular sperm from men with Kartagener/immotile cilia syndrome

OBJECTIVE: To describe two cases of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with testicular sperm in men with immotile cilia syndromes. DESIGN: Case report. SETTING: A university-based male infertility clinic and assisted reproduction unit. PATIENT(S): Two couples with male factor infertility due to Kartagener/immotile cilia syndrome. INTERVENTION(S): IVF/ICSI with testicular sperm. MAIN OUTCOME MEASURE(S): Semen characteristics, sperm viability, fertilization rate, and pregnancy. RESULT(S): With testicular sperm, the two pronuclear fertilization rates were 63% and 60% in two cases. One case resulted in the birth of normal healthy girl. CONCLUSION(S): With testicular sperm, successful oocyte fertilization after ICSI in couples with male Kartagener/immotile cilia syndrome is possible despite the lack of sperm motility.     

Human sperm-oocyte recognition and infertility

The incidence of infertility related to both male and female factors continues to rise despite many advances in reproductive technologies. Some abnormalities in human gamete interaction have been shown to be due to defects in the sperm, and others have been attributed to defects in the zona pellucida (ZP). Our lack of understanding of molecular mechanisms involved in the interaction of human sperm with the ZP in fertile as compared with infertile females and males has been limited because of the unavailability of human oocytes and ethical restraints on experimental studies. It is becoming increasingly apparent that improved clinical assays are necessary for evaluating sperm-ZP interaction in order to assess the optimal procedures for successful fertilization and pregnancy. With advances in molecular biology, the genes encoding the three major human ZP proteins have been identified and complementary DNAs are available to begin to better evaluate the molecular basis of sperm-ZP interaction.     

Intracytoplasmic sperm injection increased fertilization and good-quality embryo formation in patients with non-male factor indications for in vitro fertilization: a prospective randomized study

OBJECTIVE: To compare the fertilization rate and formation of good-quality embryos with conventional IVF and ICSI in patients with non-male factor infertility. DESIGN: Prospective controlled study. SETTING: Infertility clinic. PATIENT(S): Thirty-five patients with non-male factor infertility. INTERVENTION(S): Retrieved sibling oocytes were randomly assigned to conventional IVF or ICSI. Of sibling oocytes assigned to ICSI, only metaphase II oocytes were injected with sperm. MAIN OUTCOME MEASURE(S): Fertilization rate and formation of good-quality embryos per retrieved oocyte. RESULT(S): Per retrieved oocyte, ICSI resulted in better fertilization rate compared with conventional IVF (71.3% [134 of 188] vs. 57.2% [107 of 187]). Per retrieved oocyte, ICSI also resulted in better formation of good-quality embryos at 48 hours after retrieval compared with conventional IVF (64.4% [121 of 188] vs. 47.1% [88 of 187]). CONCLUSION(S): In IVF patients with non-male factor infertility, subjecting some sibling oocytes to ICSI increased the fertilization rate and formation of good-quality embryos per retrieved oocyte. It also avoided the problem of total fertilization failure in almost all cases.     

Intracytoplasmic sperm injection and conventional in vitro fertilization for sibling oocytes in cases of unexplained infertility and borderline semen

Purpose: In a prospective study, conventional IVF and intracytoplasmic sperm injection (ICSI) were performed on sibling oocytes of 22 patients with unexplained infertility (Group A) and 24 patients with borderline semen (Group B).Results: In Group A, there was no significant difference (P=0.070) in the fertilization rate per oocyte between ICSI (63%) and conventional IVF (50.7%), however, there was total failure of fertilization in conventional IVF in 5 of the 22 patients with IVF and none in ICSI. In group B, there was a significant difference (P<0.001) between the fertilization rate per oocyte in ICSI (59%) and conventional IVF (27.1%). There was total failure of fertilization in 11 patients after conventional IVF and none after ICSI.Conclusions: The study showed that 22.7% of unexplained infertility and 45.8% of patients with borderline semen would have lost their chance of embryo transfer completely because of total failure of fertilization if ICSI was not performed on some oocytes in this cycle.Presented in part at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, Vienna, Austria, April 3–7, 1995.