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。     

未成熟卵母细胞体外培养在体外受精中对卵巢低反应性者的补救性应用

目的:探讨未成熟卵母细胞体外培养(IVM)在体外受精(IVF)中对卵巢低反应性者的补救性治疗效果。方法:30例行IVF时表现为卵巢低反应者(实验组)在B超引导下经阴道行卵泡穿刺术,取卵后对不同期未成熟卵母细胞行IVM,再行胞浆内单精子注射(ICSI)受精;对成熟卵母细胞则进行常规受精,胚胎形成后行移植术。取同期进行ICSI周期治疗的63例作为对照组。结果:应用IVM行IVF-ET补救治疗的30个周期中有29个周期获胚胎移植,临床妊娠9例(31%),胚胎种植率18.47%,但该组未成熟卵培养成熟后的受精率、卵裂率均低于对照组;总妊娠率及胚胎种植率比常规ICSI组低。结论:IVM技术联合IVF为卵巢低反应性者提供了一种有效的补救措施,明显减少了IVF周期的取消率,提高了妊娠机会。     

体外受精失败后补行卵胞浆内单精子注射的临床价值探讨

目的:探讨卵胞浆内单精子注射(ICSI)在体外受精完全失败或受精率低于25%的常规IVF周期中的临床价值。方法:回顾分析2001.01-2004.12在我院生殖医学中心接受常规IVF治疗的35例非男性因素不育患者,取卵后体外受精培养16-18h,发现卵母细胞完全未受精或受精率低于25%,立即行ICSI再授精。结果:在24个常规IVF低于25%的周期中,共有197个未受精卵,其中159个MⅡ期卵,显微注射159个,受精123个,最终形成胚胎96个,受精率为77.4%,卵裂率为78.1%,在22个新鲜移植周期(每周期的移植胚胎由来源于常规体外受精卵和补救ICSI后受精卵的胚胎组成),共有4例临床妊娠;在8个冷冻移植周期中(每周期的移植胚胎完全来源于补救ICSI后的受精卵),有1例临床妊娠。在11个常规IVF完全失败周期中,共有89个未受精卵,其中78个MⅡ期卵,显微注射78个,受精63个,卵裂51个,受精率为80.7%,卵裂率为80.9%,在10个新鲜移植周期(每周期的移植胚胎完全来源于补救ICSI后的受精卵)中共有2例临床妊娠;在2个冷冻移植周期(每周期的移植胚胎完全来源于补救ICSI后的受精卵)中有1例单胎妊娠,妊娠早期流产。胚胎来源于常规体外受精卵和补救ICSI后受精卵的移植周期临床妊娠率为18%;胚胎完全来源于补行ICSI后受精卵的移植周期临床妊娠率为20%。结论:ICSI可作为常规IVF失败后的有效补救措施。     

同周期同胞卵研究较早期实施补救ICSI价值的探讨

目的:比较同周期同胞卵不同时间补救卵细胞浆内单精子显微注射(ICSI)的结果。方法:选取常规体外受精(IVF)治疗失败的26个周期312枚卵子(每周期未受精卵≥8枚,受精率<25%),分为两组:观察组受精后8h实施补救ICSI,对照组20h实施补救ICSI,对比两组受精率、优质胚胎形成率、囊胚形成率。结果:观察组和对照组受精率分别为81.4%、74.36%,两者差异无统计学意义(P>0.05),观察组优质胚胎形成率和囊胚形成率分别为48.82%、38.58%,明显高于对照组的33.62%,22.41%(P<0.01)。结论:对常规IVF受精失败及受精率较低的周期受精后8h实施补救ICSI得到的结果优于受精后20h的结果。     

原发不孕患者行短时受精和half-ICSI结果的比较

目的:探讨女性原发不孕患者行短时受精和half-ICSI 2种受精方式对受精率和胚胎发育潜能的影响。方法:回顾性分析175个周期女性原发不孕患者体外助孕资料,分为短时受精和/或联合早期补救ICSI周期组(A组,114个周期,其中A1组103个短时IVF受精周期,A2组11个早期补救ICSI周期)及half-ICSI周期组(B组,61个周期),分析比较各组IVF/ICSI-ET结局。结果:B组较A组不孕年限长,获卵数多,正常受精率低,差异有统计学意义(P<0.05)。但异常受精率、优质胚胎率和临床妊娠率差异无统计学意义(P>0.05)。A1组正常受精率明显低于A2组(P<0.05),异常受精率、优质胚胎率和临床妊娠率组间差异无统计学意义(P>0.05)。不同病因对A、B组患者的妊娠结局无显著影响。结论:短时受精和/或联合早期补救ICSI对原发不孕患者是一种较为合理而安全的受精方式。     

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。     

早期补救卵胞浆内单精子注射妊娠结局及新生儿出生结局评估

目的:对常规体外受精(IVF)失败的周期行早期补救性卵胞浆内单精子注射(ICSI)后的妊娠结局及新生儿出生情况进行探讨和分析。方法:常规IVF后6小时未发现明确双极体者判断为完全受精失败,予以早期补救ICSI,纳入早期补救ICSI组(204个周期),随机选择与早期补救ICSI组患者同一日或前一日因男性因素而直接进行ICSI的204例患者纳入ICSI组(204个周期)。回顾性分析两组患者的累计临床妊娠率、流产率、异位妊娠率、累计活产率、分娩孕周及新生儿的单双胎比例、男女性别比例、出生体质量、出生缺陷等指标的差异性。结果:早期补救ICSI组与ICSI组的累计临床妊娠率、流产率、异位妊娠率、累计活产率、新生儿性别比例、新生儿出生体质量等指标的比较差异均无统计学意义(P0. 05)。早期补救ICSI组产妇的分娩孕周较ICSI组略长,双胎比率较ICSI组少,差异有统计学意义(P0. 05)。将分娩孕周分为单胎和双胎分别进行比较,则发现两组间差异无统计学意义(P0. 05)。早期补救ICSI组的活婴中出现1例先天性心脏病出生缺陷,ICSI组的活婴中未发现出生缺陷。结论:早期补救ICSI没有增加胎儿流产及新生儿出生缺陷的风险,双胎率明显下降,怀孕周期略长,可作为常规IVF失败时的一种安全有效的补救方法。     

短时体外受精、及时补救ICSI的临床应用

目的:探讨可行的体外受精方式以期避免IVF完全受精失败及提高补救性ICSI妊娠率。方法:回顾性分析了556个IVF-ET周期,精卵共培养2h后捞出取母细胞,单独孵育4h观察,对周期获得的所有MII期卵均未见双极体者,及时实施ICSI补救。结果:502个IVF周期部分或全部MII卵可见双极体,新鲜周期临床妊娠率44.02%;54个周期所获的MII卵均未见双极体,即时行ICSI补救后,新鲜周期临床妊娠率31.48%。结论:短时受精、观察第2极体(Pb2)、补救ICSI三者相结合的方法应是更可行的选择。     

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。     

对体外受精失败后不同授精时间进行补救ICSI的结局比较

目的:探讨初次体外受精(in vitro fertilization,IVF)失败后不同时间进行补救单精子卵胞浆内注射(intracytoplasmic sperm injection,ICSI)授精对胚胎发育潜力及妊娠结局的影响。方法:将104例完全不受精或低受精率的常规IVF周期分为6 h后行补救ICSI短时授精组(A组,再将A组按补救ICSI时间分为6～8 h的A1组和8.1～12 h的A2组)及晚时授精(22 h后行补救ICSI)组(B组),比较各组间的治疗结局。结果:A组总的卵裂率、可移植胚胎率、优质胚胎率、妊娠率与B组相比差异显著(P<0.05)。A1组与A2组相比,受精率、卵裂率、可移植胚胎率、优质胚胎率无显著差异,妊娠率差异显著(P<0.05)。结论:缩短卵龄有利于补救ICSI结局的改善。     

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.     

A strategy for treatment of couples with unexplained infertility who failed to conceive after intrauterine insemination

Couples with unexplained infertility treated unsuccessfully with intrauterine insemination often receive further treatment with IVF or intracytoplasmic sperm injection (ICSI). The aim of this study was to evaluate the frequency of fertilization and fertilization failure with respect to the method of fertilization used, when half of the sibling oocytes were fertilized by conventional IVF and insemination and the remainder by ICSI. Included was the first IVF/ICSI treatment of 248 unexplained infertile couples who had failed to conceive after three IUI cycles. An overall pregnancy rate per embryo transfer of 57% was observed. A significantly better fertilization rate was obtained after ICSI as compared with IVF (68 versus 46%) (P < 0.005), and total fertilization failure following ICSI and IVF treatment was seen in 4.4 and 25% of the cycles respectively. The group who experienced total fertilization failure after IVF had normal semen parameters, although significantly lower sperm concentration and motility as compared with the entire study group. Transfer of their ICSI-fertilized oocytes subsequently resulted in a pregnancy rate of 49.2% per transfer. The policy of splitting the sibling oocytes can effectively minimize complete fertilization failure while maintaining high chances of achieving a pregnancy. At the same time, the optimal fertilization method for subsequent treatment cycles can be determined.     

Cryptic sperm defects may be the cause for total fertilization failure in oocyte donor cycles

In conventional IVF cycles with total fertilization failure, rescue intracytoplasmic sperm injection (ICSI) performed 24h after insemination has yielded poor results. However, when ICSI is used, total fertilization failure is a rare event. The aim of the present study is to investigate the degree of sperm contribution to fertilization failures using the egg-sharing model in oocyte donor cycles. The study included only the oocyte donor cycles of sibling oocytes with total fertilization failure in at least one of the matched recipients. Oocytes from 49 oocyte donor cycles were equally shared among 98 recipients undergoing conventional IVF. Due to total fertilization failure in half of the recipients, rescue ICSI was carried out. Compared with the conventional IVF only group, the rescue ICSI group had a lower pregnancy rate (30.61% versus 71.43%), clinical pregnancy rate (28.57% versus 67.35%) and ongoing pregnancy rate (28.57% versus 63.27%) (all P<0.01). Cryptic sperm defects in apparently normal spermatozoa may be the cause of total fertilization failure, indicating the need for simple routine tests to detect them.     

Intracytoplasmic sperm injection is not associated with poor outcome in couples with normal semen parameters and previous idiopathic fertilization failure in conventional in vitro fertilization

In this study, we compared the results of intracytoplasmic sperm injection (ICSI) in patients with normal semen parameters and a history of failed fertilization with conventional IVF (study group) and in patients with male factor infertility (control group). Patient and cycle characteristics were similar in both groups. The mean number of retrieved and metaphase II oocytes, fertilized oocytes, embryos developed, embryos transferred and the number of cycles with fertilization failure also did not differ between groups. Although differences were not statistically significant, pregnancy rate (56.3% vs. 31.5%), implantation rate (14.2% vs. 12%) and ongoing pregnancy rate (37.5% vs. 17.7%) per embryo transfer were higher in the study group than the ones in the control group. We concluded that previous idiopathic fertilization failure with conventional IVF is not associated with poor outcome in subsequent ICSI treatment.     

In vitro fertilization-intracytoplasmic sperm injection split: an insemination method to prevent fertilization failure

OBJECTIVE: To determine whether the performance of intracytoplasmic sperm injection (ICSI) in cases of unexplained infertility and of borderline semen parameters can prevent fertilization failure. DESIGN: Prospective cohort study. SETTING: Hospital-based assisted reproductive technology program. PATIENT(S): Sixty couples with unexplained infertility (group 1) and 50 couples with borderline semen (group 2). INTERVENTION(S): Conventional IVF and ICSI were performed on sibling oocyte cumulus complexes. MAIN OUTCOME MEASURE(S): Rate of fertilization according to patient diagnosis. RESULT(S): The fertilization rate per oocyte cumulus complex in couples with unexplained infertility (group 1) was significantly higher with ICSI (65.3%) compared with IVF insemination (48.1%). Fertilization rates per OCC in couples with borderline semen (group 2) were not significantly different between ICSI (58.1%) and IVF (56.7%). In group 1, failure of fertilization after IVF insemination occurred in 10 of the 60 couples (16.7%) and in none with ICSI. In group 2, 2 of 50 couples (4.0%) failed fertilization by IVF, and 1 (2.0%) failed with ICSI. No couple in group 2 failed both IVF insemination and ICSI. CONCLUSION(S): In couples with unexplained infertility, a higher fertilization rate was achieved through ICSI compared with conventional IVF. No such benefit could be demonstrated for couples with borderline semen parameters. The use of ICSI rescued 12 of 110 cycles (10.9%) where IVF failed. Adoption of the ICSI-IVF insemination split in cases of unexplained infertility may help eliminate fertilization failures.     

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 Are Complementary Techniques in Management of Unexplained Infertility

Purpose : To evaluate the role of ICSI in unexplained infertility. Methods : In 125 cycles with six or more oocytes retrieved per cycle, sibling oocytes were randomly allocated to IVF or ICSI (group A). In 74 cycles with less than six oocytes retrieved per cycle, cycles were allocated to IVF or ICSI (group B). Results : In group A, ICSI fertilization rate of 61% per allocated oocyte was higher than IVF fertilization rate of 51.6% (P < 0.001). Complete fertilization failure occurred in 19.2 and 0.8% of cycles in IVF and ICSI, respectively (P < 0.001). In group B, fertilization rate in IVF cycles was 53.3% as compared to 60.7% per allocated oocyte in the ICSI cycles (P = 0.29). Complete fertilization failure was higher (P = 0.02) in conventional IVF (34.3%) than ICSI cycles (10.3%). Conclusions : Allocation of sibling oocytes to IVF and ICSI in the first cycle minimizes risk of fertilization failure. For patients with limited number of oocytes, ICSI technique is recommended.     

精子质量和数量对体外受精率,卵裂率和卵裂胚胎的影响

目的观察精子质量和数量对体外受精－胚胎移植（ＩＶＦ－ＥＴ）的卵细胞受精、卵裂和卵裂胚胎的影响。方法观察常规ＩＶＦ－ＥＴ９２个周期的精液和授精精子的密度和活动力与孕卵细胞分裂的关系。结果对４８１个成熟卵及２７３个未成熟卵的观察表明,快速直线运动（ａ级）精子和快速非直线或慢速直线运动（ｂ级）精子的密度下降,使成熟卵和未成熟卵的细胞受精率和受精卵的卵裂率明显下降;ａ级和ｂ级精子的密度增高,可明显降低形态不良卵裂胚胎的比率。结论精子的密度和活动力对体外受精过程非常重要。     

Intracytoplasmic sperm injection as a treatment for unexplained total fertilization failure or low fertilization after conventional in vitro fertilization

OBJECTIVE: To determine whether IVF or intracytoplasmic sperm injection (ICSI) should be the choice of treatment in case of a previous IVF attempt with unexplained total fertilization failure or low fertilization (<25%). DESIGN: Prospective study. SETTING: Leiden University Medical Center. PATIENT(S): Thirty-eight couples undergoing IVF and ICSI on sibling oocytes after a first IVF attempt with total fertilization failure or with low fertilization (<25%). INTERVENTION(S): Performing IVF and ICSI on sibling oocytes. MAIN OUTCOME MEASURE(S): Fertilization and (ongoing) pregnancy rate. RESULT(S): A total of 271 oocytes were collected in 24 oocyte retrievals in the total fertilization failure group. Hundred nine oocytes were randomly allocated to IVF and 12 were fertilized (11%); 162 sibling oocytes were allocated to ICSI and 78 were fertilized (48%). In 8 of the 24 patients fertilization occurred after IVF. The pregnancy rate after transfer of 1 IVF and 1 ICSI embryo (n = 3) was 67% and after the transfer of 2 ICSI embryos (n = 21) this was 52%. In the low fertilization group 169 oocytes were collected in 14 oocyte retrievals. Seventy-two oocytes were randomly allocated to IVF and 16 were fertilized (22%). Ninety-seven sibling oocytes were allocated to ICSI and 58 were fertilized (60%). In 7 of 14 patients fertilization occurred after IVF. The pregnancy rate after the transfer of 1 IVF and 1 ICSI embryo (n = 5) was 80% and after the transfer of 2 ICSI embryos (n = 9) this was 33%. CONCLUSION(S): Performing ICSI on some oocytes of a cohort may avoid total fertilization failures both in patients with a history of total fertilization failure and in patients with a history of low fertilization, as the percentage of fertilization is higher after ICSI compared to IVF and the recurrence of total fertilization failure and low fertilization is high after IVF treatment.     

Comparison of piezo-assisted micromanipulation with conventional micromanipulation for intracytoplasmic sperm injection into human oocytes

OBJECTIVE: To compare the efficacy of piezo-assisted micromanipulation with conventional micromanipulation for intracytoplasmic sperm injection (ICSI) into oocytes in patients with impaired semen parameters and no success with in vitro fertilization (IVF). STUDY DESIGN: A retrospective randomized study was conducted on 204 cycles for 104 couples with piezo-assisted ICSI and 122 cycles for 96 couples with conventional ICSI. Piezo-assisted ICSI consists of two steps, namely penetration of the zona pellucida alone with a piezo-pulse and then puncturing of the oolemma with a light negative pressure without piezo, as with conventional ICSI. The tips of injection pipettes were prepared after pulling by breakage with a scalpel under a microscope, so that the inner diameter at and near the tip was 5 microm, as for conventional ICSI. RESULTS: Piezo-assisted ICSI demonstrated significantly more favorable results, with a fertilization rate of 90.3% (conventional ICSI: 83.1%, p < 0.01) and a cleavage rate of 88.1% (conventional ICSI: 84.6%, p < 0.01). CONCLUSION: Piezo-assisted ICSI is easy to incorporate a spermatozoa exactly into the ooplasm with little deformation of the oocyte during insertion. Piezo-assisted ICSI can be used effectively for human oocytes to improve the fertilization, cleavage rates.