Incidence and Perinatal Outcome of Multiple Pregnancies After Intracytoplasmic Sperm Injection Compared to Standard In Vitro Fertilization

Purpose:Our purpose was to assess the incidence of multiple pregnancies and their obstetric outcome after intracytoplasmic sperm injection.Methods:The study group comprised women who delivered twins or triplets after intracytoplasmic sperm injection and standard in vitro fertilization. The incidence and main perinatal outcome of 140 multiple pregnancies resulting from intracytoplasmic sperm injection or standard in vitro fertilization treatment were analyzed.Results:A total of 60 multiple pregnancies was obtained after intracytoplasmic sperm injection (3.4 ± 1.1 embryos/cycle) and 80 after standard in vitro fertilization (3.3 ± 2.0 embryos/cycle). The incidence of multiple pregnancy, i.e., 22.6 compared to 20.7%, respectively, was calculated. The obstetric outcome of 47 multiple pregnancies after intracytoplasmic sperm injection was 39 twin deliveries at between 27 and 37 weeks of gestation (mean, 36 ± 3.3) and 8 successful triplet deliveries between 26 and 36 weeks of gestation (mean 32.6 ± 2.4). The outcome after regular in vitro fertilization was similar. No major malformations were observed.Conclusions:The results of this study showed that the incidence of multiple pregnancies after intracytoplasmic sperm injection was similar to that after standard, conventional in vitro fertilization. The perinatal outcome did not differ between both groups.     

Fertilization, Embryo Quality, and Cryosurvival in In Vitro Fertilization and Intracytoplasmic Sperm Injection Cycles

Purpose: Our purpose was to investigate the influence of semen quality on fertilization, embryo morphology, cleavage, and cryosurvival in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI)programs. Methods: A retrospective analysis of 513 couples undergoing IVF and 255 couples undergoing ICSI was done. Results: Semen quality influenced fertilization in IVF and abnormal fertilization in IVF and ICSI, but no effects on the development, morphology, implantation capacity, or cryosurvival of embryos were found. Fertilization, embryo quality, and cryosurvival rates were similar after IVF and ICSI. The fertilization rate of mature oocytes in IVF was lower when cytoplasmic immaturity in the oocyte population was frequent. The speed of development of embryos was 2 hr faster after ICSI than after IVF. Two-cell–stage embryos survived best after cryopreservation with propanediol and sucrose on day 2. Conclusions: After fertilization, semen parameters had no effect on the quality or cryosurvival of embryos in either IVF or ICSI.     

Intracytoplasmic Sperm Injection Increases Embryo Fragmentation Without Affecting Clinical Outcome

Purpose: To examine the effect of intracytoplasmic sperminjection (ICSI) on embryo fragmentation and implantationrates in those embryos chosen for transfer compared toconventional in vitro fertilization (IVF). Methods: We compared 253 infertility patients (71 ICSI and182 IVF) with respect to age, semen analysis, number ofembryos transferred, embryo fragmentation, implantationrate, and pregnancy rate. Embryo fragmentation wasdetermined by one observer at the same laboratory over the entirestudy period. Results: A statistically significant difference was observedin mean embryo grade between IVF (2.2 ± 0.84) and ICSI(2.5 ± 0.77), P = 0.01. Additionally, the IVF patients hadsignificantly more nonfragmented (grade I) embryoscompared to the ICSI group, P < 0.01. Conclusions: These data suggest that ICSI, irrespective ofsemen parameters, may increase embryo fragmentation andproduce fewer nonfragmented grade I embryos while maintaining implantation and pregnancy rates similar toconventional IVF.     

The Outcome of Cryopreserved Human Embryos After Intracytoplasmic Sperm Injection and Traditional IVF

Objective: Our objective was to analyze the outcome of cryopreserved embryos obtained after intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) in terms of survival rate, implantation rate (IR), total and clinical pregnancy rate (PR) in a retrospective, comparative study. Methods: Three hundred seventy-five IVF and 463 ICSI surnumerary cleaved embryos, frozen on Day 2 with 1,2-propanediol, were thawed. Results: Thirty-two percent of the thawed IVF embryos survived and 11 pregnancies (8 clinical) were obtained from 68 transfers (16.1%). Fourty-seven percent of the ICSI embryos survived, with 19 pregnancies (18 clinical) from 116 transfers (16.4%). The IR was 8.5% (8/94) in IVF cycles and 10.8% (20/185) in ICSI cycles. Conclusions: A significantly better survival rate of ICSI embryos was observed but with no difference in PR, preclinical, and clinical abortion rate, or IR.     

In Vitro Fertilization and Intracytoplasmic Sperm Injection for Couples with Unexplained Infertility After Failed Direct Intraperitoneal Insemination

Purpose: The objective was to determine the optimal insemination technique in patients undergoing in vitro fertilization (IVF) after failed direct intraperitoneal insemination (DIPI) and the outcome of intracytoplasmic sperm injection (ICSI) in such cases. Methods: In case–control studies, 53 couples with unexplained infertility who underwent IVF after four failed DIPI cycles were compared with 75 couples with tubal or endometriosis infertility as controls. Thirty couples with unexplained infertility after failing to conceive with DIPI and conventional IVF who underwent ICSI and 58 couples with male-factor infertility as controls also were compared. Fertilization cleavage, embryo quality, implantation, and pregnancy were compared after IVF and after ICSI. Results: There was a significant difference in fertilization rates after IVF between cases of unexplained infertility after failing to conceive with DIPI (40.4%) and patients with tubal or endometriosis infertility (67.9%). There also was a significant difference in total fertilization failure rates between the two groups (30.4% and 3.9%, respectively). There was a slight but significant difference in numbers of fertilized oocytes after ICSI between patients with low fertilization rate undergoing IVF after failing to conceive DIPI (85.8%) and patients with male factor (90.4%). Total fertilization failure was not observed in these cases. Conclusions: Couples with unexplained infertility after failing to conceive with DIPI show a failed fertilization or a low fertilization rate after IVF. However, they demonstrated a good chance of becoming pregnant after subsequent ICSI, even with statistically significant difference in fertilization rate as compared with male-factor cases.     

两种指征的卵胞浆内单精子注射结局的比较

目的:探讨常规IVF受精失败患者再次周期行ICSI-ET治疗时,卵子因素对胚胎结局的影响。方法:回顾分析因前次IVF-ET中受精障碍或受精率≤30%而行ICSI治疗的38个周期(A组)和因严重精液异常而行ICSI治疗的181个周期(B组)的ICSI结局。结果:A、B组的受精率(FR)、卵裂率(CR)、胚胎利用率(URE)、胚胎着床率(EIR)、临床妊娠率(CPR)和早期流产率(EAR)分别为82.5%vs78.0%(P<0.05),97.5%vs97.6%(P>0.05),76.4%vs73.1%(P>0.05),10.0%vs19.8%(P<0.05),27.0%vs31.8%(P>0.05),40.0%和16.1%(P>0.05)。结论:常规IVF-ET受精失败的患者,再次周期行ICSI-ET治疗,受精率提高,由于卵子异常对胚胎发育的不利影响,部分胚胎着床和着床后的远期发育潜能降低。     

Reproductive Performance of Patients Undergoing Intracytoplasmic Sperm Injection with 100% Implantation Rate

Purpose: To characterize the differences between twomatched groups of patients treated by ICSI: those pregnantafter all embryos transferred implanted (100% implantationrate) compared with nonpregnant patients. Methods: Twenty-one patients in whom one transferredembryo achieved a singleton pregnancy (group A) and 21pregnant patients to whom two or three embryos were transferredand achieved 11 twin and 10 triplet pregnancies(group B) compared with matched nonpregnant patients(group C and D, respectively). Results: The singleton pregnant patients were significantlyolder than the twin and triplet pregnancy patients. Althougha similar number of human menopausal gonadotropinampules were used in the singleton compared with the twinsand triplets a significantly lower number of oocytes andembryos were achieved at lower levels of estradiol on thehuman chorionic gonadotropin day in the former than inthe latter respectively. No difference was found between thepregnant women and their nonpregnant controls in any ofthe mentioned parameters. Good embryo morphology wasfound in 86% of the embryos in group A compared with62% in group C (P = 0.08) and 92% in group B comparedwith 66% in group D (P < 0.002). Conclusions: The only parameter in which pregnant patientswith 100% implantation rate differ from their nonpregnantcontrols was embryo quality.     

Assessment of Maximal Fertilization Rates with Intracytoplasmic Sperm Injection

Purpose: Maximal fertilization rates following ICSI wereassessed using two essential steps: immobilization of spermand aspiration of oocyte cytoplasm. Methods: ICSI procedure was performed for couples(N = 42) in whom the male suffered severe infertility or failureof fertilization in previous IVF cycle using different oocytecytoplasmic aspiration and sperm immobilization methods.Outcome in four patient groups was measured by oocytedamage, fertilization rate, and pregnancy rate. Results: Maximal fertilization (90%) were achieved from thegroup which used immobilization of sperm by hard-touching thetail with a pipette and optimal aspiration of oocyte cytoplasm. Conclusions: The results suggest that if the immobilizationof sperm and aspiration of oocyte cytoplasm are handledright during ICSI, this procedure can be expected to yielda 90% fertilization rate.     

Relationship Between the Sperm Motility Index Assessed by the Sperm Quality Analyzer and the Outcome of Intracytoplasmic Sperm Injection

Purpose: Intracytoplasmic sperm injection (ICSI) has been validated as a useful treatment in severe male-factor patients who could not achieve fertilization and live births by conventional in vitro fertilization treatment. To examine the impact of male factors on ICSI outcome, clinical laboratory data were retrospectively analyzed. Methods: One hundred two cycles of ICSI treatment indicated by severe male-factor infertility were entered into this study. Sperm parameters including sperm motility, sperm concentration, and sperm motility index assessed by the Sperm Quality Analyzer were evaluated. Results: Five hundred seventy-six metaphase II oocytes retrieved were manipulated. The normal fertilization (2 PN) rate per oocyte was 64.9 ± 26.0% (mean ± SD). Of the 99 transfers, 31 clinical pregnancies were obtained, yielding an average pregnancy rate of 31.3% per transfer. The mean sperm motility, sperm concentration, and sperm motility index were 20.3 ± 16.1% (range, 0 to 50%), 18.2 ± 25.1 × 10 ⁶ /ml (range, <1 to 150 × 10 ⁶ /ml), and 31.2 ± 45.0 (range, 0 to 220), respectively. Sperm concentration did not have a significant impact on fertilization rate by ICSI. In four cases, ICSI was performed using totally immotile sperm and the fertilization rate was 43.5%, which was significantly lower than that of some of the other sperm motility groups, and no pregnancy could be achieved. In 14 cases in which the sperm motility index assessed by the Sperm Quality Analyzer was 0, the fertilization rate (50.0%) was significantly lower than in most of the other sperm motility index groups. Conclusions: These findings suggest that in severe malefactor cases with totally immotile sperm or a sperm motility index of 0, the selection of good-quality sperm should be verified before injection.     

Effects of Electric Stimulation on Bovine Oocyte Activation and Embryo Development in Intracytoplasmic Sperm Injection Procedure

Purpose: This study was carried out to investigate the efficacyof electric stimulation before and/or after intracytoplasmicsperm injection (ICSI) on bovine oocyte activation andembryo development. Methods: The oocytes were treated with electric shock before(B), before and after (B&A), and after (A) sperm injection.In each group, sham ICSI (ICSI-s) was performed to excludethe effect of parthenogenesis (B ICSI-s, B&A ICSI-s, and AICSI-s). An electric pulse was applied with a single directcurrent (DC) pulse (0.8 kV/cm, 70 sec). Results: One pronucleus (PN) formation in the B&A ICSI-sgroup was slightly higher than that found in B and B&AICSI group; however, the difference was not significant. TwoPN formation in B&A ICSI group was higher than that foundin sham ICSI groups (P < 0.05). There were no differencesamong treatment groups in the cleavage rate; however, morulaeand blastocyst formation in the B&A embryos wassignificantly higher than that of other groups (P < 0.05)and got pregnant. Conclusions: Electric stimulation before and after injectionwas an effective method in inducing bovine oocyte activationand in sustaining embryo development to the morulae andblastocyst stage.     

The Status of Controlled Prospective Clinical Trials for Efficacy of Intracytoplasmic Sperm Injection in In Vitro Fertilization for Non-Male Factor Infertility

Purpose: Intracytoplasmic sperm injection (ICSI) of some sibling oocytes may have a beneficial effect in couples going through in vitro fertilization for causes of infertility not related to the male factor. Our purpose was to critically appraise the randomized controlled studies done in this area and arrive at some recommendations. Methods: The four controlled trials done so far have utilized similar methodology, i.e., they randomly allotted sibling oocytes to ICSI versus standard insemination in patients going through in vitro fertiliztion and embryo transfer. Results: In the first trial reported in 1995 there was no difference in fertilization rate, whereas the later trials reported in 1997, 1999, and 2000 showed improvement with ICSI that reached statistically significant level in the last two studies. Conclusions: Total fertilization failure of an in vitro fertilization cycle can be prevented and fertilization can be improved if half of sibling oocytes are subjected to ICSI.     

The Outcome of Sperm Retrieval and Intracytoplasmic Sperm Injection in Patients with Obstructive Azoospermia: Impact of Previous Tuberculous Epididymitis

Purpose: Our purpose was to investigate the influence of previous tuberculous epididymitis in patients with obstructive azoospermia on the outcome of sperm retrieval and intracytoplasmic sperm injection (ICSI). Methods: Eighty-eight cycles of ICSI were performed in 44 patients with obstructive azoospermia; 16 cycles (7 patients) with tuberculous obstructive azoospermia and 72 cycles (37 patients) with nontuberculous obstructive azoospermia. Results: The rates of fertilization and embryo cleavage were comparable, and there was no significant difference in the clinical pregnancy rate per fresh transfer between the two groups. The rates of embryo implantation and clinical miscarriage were also comparable. Conclusions: Embryo quality and pregnancy outcome in sperm retrieval and ICSI were comparable in both the tuberculous and the nontuberculous obstructive azoospermia patients. Although there was a preponderance of testicular sperm used in the tuberculous obstructive azoospermia group, our results suggest that previous tuberculous epididymitis in patients with obstructive azoospermia does not affect the outcome of sperm retrieval and ICSI.     

卵胞质内单精子显微注射后异常受精的影响因素分析

目的:探讨卵胞质内单精子显微注射(ICSI)后异常受精发生的影响因素。方法:回顾性分析299个ICSI周期,按照是否有异常受精分为异常受精组(n=118)和正常对照组(n=181),异常受精组至少发生1个非2原核(2PN)的受精卵。比较分析异常受精组和对照组临床和实验室资料。结果:异常受精组hCG注射日E2水平(14 097±3 066 pmol/L)高于对照组(1 2461±6 836 pmol/L),差异有统计学意义(P<0.05);异常受精组获卵数及成熟卵数分别为17.8±7.2个、15.3±6.1个,多于对照组的13.6±7.0个、10.2±5.3个,差异均有统计学意义(P<0.01);卵子成熟率(81.7%vs76.4%)、2PN受精率(78.4%vs 86.9%),组间比较差异均有统计学意义(P<0.01);而患者年龄、基础激素水平、促性腺激素(Gn)使用总量、精子来源及质量、着床率、临床妊娠率组间比较,差异均无统计学意义(P>0.05)。结论:卵巢对Gn刺激的高反应性可能与ICSI后异常受精的发生有关。异常受精卵的发生虽对临床结局无明显影响,但减少异常受精发生率,增加卵子利用率,可能会提高累积妊娠率。     

Frequency and Patterns of Premature Sperm Chromosome Condensation in Oocytes Failing to Fertilize After Intracytoplasmic Sperm Injection

Purpose: The purpose of this study was to evaluate reasonsfor fertilization failure after intracytoplasmic sperminjection as a part of internal quality control and to reviewcorresponding previous data. Methods: One hundred injected but unfertilized oocytes werefixed and examined after Giemsa staining. Results: Three oocytes (3.0%) did not show the presenceof a spermatozoon and two (2.0%) contained pronuclearstructures. An intact spermatozoon was found in 25 cases(25.0%), whereas the sperm nucleus had undergonepremature chromosome condensation (PCC) in 70 cells (70.0%).A modified classification system was established tocharacterize the different PCC patterns. Conclusions: PCC indicates a correct intracytoplasmicinjection and excludes technical problems as a major reasonfor fertilization failure in the present study. A lack of oocyteactivation due to cytoplasmic immaturity is consideredresponsible for the occurrence of PCC. A review of theliterature shows that the role of sperm chromatinabnormalities in the process of nuclear decondensation needs furtherinvestigation.     

人卵细胞冷冻保存后经显微注射授精的初步研究

建立人卵细胞冷冻保存以及融解后受精和发育颇具实用价值。本文采用慢冷、快融法，结合使用丙二醇和蔗糖作为冷冻保护剂，对２８５个临床ＩＶＦ未受精卵进行冷冻保存，３天至４０天时间不等。冷冻存活率为５４．７３％，其中未成熟卵子５１．７２％，成熟卵子５５．５０％，二者无显著性差异（Ｐ＞０．０５）。冷冻损伤的卵子中，细胞膜损坏为９６．８９％，透明带破裂为１３．１８％。选择冻融后形态结构正常的１２６个成熟卵子进行胞浆内单个精子注射（ＩＣＳＩ），受精率达４２．０６％，其中５０．９４％受精卵子进一步发育成２～８细胞胚胎。来源于常规ＩＶＦ未受精卵的再次受精率为４０．７４％，来源于ＩＣＳＩ未受精卵的再次受精率为３６．７３％。作者认为，必要时可以借助显微操作技术进行胚胎活检，了解部分染色体畸变情况，使卵子冷冻保存技术安全地应用于临床。     

Population‐Based Study of Cesarean Section After In Vitro Fertilization in Australia

Abstract: Background: Decisions about method of birth should be evidence based. In Australia, the rising rate of cesarean section has not been limited to births after spontaneous conception. This study aimed to investigate cesarean section among women giving birth after in vitro fertilization (IVF). Methods: Retrospective population‐based study was conducted using national registry data on IVF treatment. The study included 17,019 women who underwent IVF treatment during 2003 to 2005 and a national comparison population of women who gave birth in Australia. The outcome measure was cesarean section. Results: Crude rate of cesarean section was 50.1 percent versus 28.9 percent for all other births. Single embryo transfer was associated with the lowest (40.7%) rate of cesarean section. Donor status and twin gestation were associated with significantly higher rates of cesarean section (autologous, 49.0% vs donor, 74.9%; AOR: 2.20, 95% CI: 1.80, 2.69) and (singleton, 45.0% vs twin gestations, 75.7%; AOR: 3.81, 95% CI: 3.46, 4.20). The gestation‐specific rate (60.1%) of cesarean section peaked at 38 weeks for singleton term pregnancies. Compared with other women, cesarean section rates for assisted reproductive technology term singletons (27.8% vs 43.8%, OR: 2.02 [95% CI: 1.95–2.10]) and twins (62.0% vs 75.7%, OR: 1.92 [95% CI: 1.74–2.11]) were significantly higher. Conclusions: Rates for cesarean section appear to be disproportionately high in term singleton births after assisted reproductive technology. Vaginal birth should be supported and the indications for cesarean section evidence based. (BIRTH 37:3 September 2010)     

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。     

Intracytoplasmic sperm injection improving embryo quality: Comparison of the sibling oocytes of Non-Male-Factor couples

Objective: Our objective was to investigate whether the quality of embryos developed after intracytoplasmic sperm injection (ICSI) is better than that of conventional IVF embryos. Methods: Nine couples who previously achieved a normal rate of fertilization following IVF and four couples whose normal rate of fertilization was expected were involved in this study. The oocytes from those couples were randomly divided into two groups, group A by conventional insemination and group B by ICSI. The fertilization rate and quality of embryos were compared. Results: Normal fertilization was achieved in 61% of the oocytes (83/136) after conventional insemination. In group B, 69% of the oocytes (99/144) achieved normal fertilization, although only 127 metaphase II oocytes were injected using the ICSI technique. More grade A embryos were obtained when the ICSI technique was used for fertilization than by conventional IVF (35.4 and 24.3%, respectively;P=0.028). Conclusions: A similar fertilization rate can be achieved by ICSI in comparison with conventional IVF, when male factor is not involved. Embryos after ICSI have an improved quality.     

Very Low Sperm Count Affects the Result of Intracytoplasmic Sperm Injection

Purpose: The aim was to examine the influence of extremelylow sperm count on intracytoplasmic sperm injection(ICSI) outcome. Methods: Over 1000 consecutive unselected ICSI cycleswere divided into four groups according to spermconcentration of their patients: A, cryptozoospermia, 107 patients; B,sperm concentration of 1×10⁴, 146 patients; C, spermcount of 1×10⁴–1×10⁵, 135 patients; and concentration of 1×10⁵ and < 10×10⁶/ml (control group), 688 patients. Results: A significant decrease in pregnancy rate wasnoticed in the cryptozoospermic group in comparison to thecontrol group (20% vs. 31%). Fertilization rate in group Awas significantly lower in comparison to all other groups,respectively (46% vs. 52%, 54%, 61%). Embryo quality wasinferior in group A in comparison to the control group. Ahigher yet not statistically significant abortion rate wasobserved in the cryptozoospermic group (as well as in groupC) (30%, 27%) compared to the control group (15%). Conclusions: It seems that an extremely low sperm counthas a negative effect on the outcome of ICSI. Neverthelesspatients with cryptozoospermia should not be offered ICSItreatment with the ejaculated sperm before karyotype isestablished.     

Chromatin Packaging as an Indicator of Human Sperm Dysfunction

Purpose: Understanding the causes of fertilization failure is an important research field in assisted reproductive programs. The present study aimed to evaluated the possible relationship between chromatin packaging quality (CMA₃ staining) and (i) normal morphology and (ii) its ability to predict the functional integrity of spermatozoa in both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment programs. Methods: Semen of 140 men from IVF and ICSI couples were analyzed for sperm concentration, motility, morphology, and chromatin packaging (CMA₃). For CMA₃ classification, two cutoff values were used, namely, 44.5%±13 and 1 SD above the mean, i.e., 57.5% (rounded off to 60%). IVF and ICSI data were stratified using three basic cutoff values for CMA₃ staining, namely, <44%, >44–60%, and >60%. Results: Based on CMA₃ results patients were divided into four groups, namely, group A, <44% CMA₃ (n = 15, IVF); group B, 44% and <60% CMA₃ (n = 39, IVF); group C, 60% CMA₃ (n = 45 IVF); and group D, 60% CMA₃(n = 41 ICSI). During receiver operator characteristic analyses the estimated cutoff value for CMA₃ staining, to distinguish between <4% and 4% morphology groups, was 60%. The area under the curve was 0.89, sensitivity of 75%, and specificity of 100%. When IVF rates of >60% and <60% were used, the optimal CMA₃ value for prediction of fertilization success again was recorded at 60%. The area under the curve was 0.76, sensitivity of 81.5%, and specificity of 63.6%. Conclusions: Chromatin packaging assessments should be included as a complementary assay to the sequential diagnostic approach of the male-factor patients.