Intracytoplasmic Sperm Injection Group Research Articles

Should ICSI be implemented on patients with poor-quality embryos in the previous IVF cycle?

This study was to evaluate whether Intracytoplasmic sperm injection (ICSI) can improve the quality of embryo in patients with poor-quality embryos in the previous In-vitro fertilization (IVF) cycle, which was cancelled before transfer. This was a retrospective cohort study of 178 IVF and 158 ICSI cycles for patients with poor-quality embryos in the previous IVF cycle in the Center for Reproductive Medicine, Women and Children's Hospital of Chongqing Medical University from March 2016 to June 2022. The 2 PN rate, oocyte utilization rate , high-quality embryo rate and clinical pregnancy rate were compared between the two groups. Furthermore, the implantation rate, miscarriage rate and cycle cancelation rate were measured and compared. ICSI resulted in a comparable 2 PN rate, oocyte utilization rate and cycle cancelation rate with IVF. The high-quality embryo rate of ICSI group was significantly higher than that of IVF group (5.56% vs. 2.60%, P < 0.05). Eventually, a total of 239 patients performed embryo transfer. ICSI resulted in a significantly higher clinical pregnancy rate (55.56% vs. 40.98%, P < 0.05) compared with IVF, however, there were no notable differences in miscarriage rate and implantation rate. The present study suggested that ICSI significantly improved the high-quality embryo rate and clinical pregnancy of the patients with poor-quality embryos in the previous IVF cycle. Prospective randomized controlled trials are needed to further verify.

Comparison of clinical and preimplantation genetic testing for aneuploidy outcomes between in vitro fertilization and intracytoplasmic sperm injection in sibling mature oocytes from high-risk patients: A retrospective study.

To evaluate the influence of insemination methods on clinical outcomes by assessing preimplantation genetic testing for aneuploidy (PGT-A) outcomes in embryos obtained using in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in sibling mature oocytes from high-risk patients. This retrospective study involved 108 couples with nonmale or mild male factor infertility who underwent split insemination cycles from January 2018 to December 2021. PGT-A was performed using trophectoderm biopsy, array comparative genome hybridization, or next-generation sequencing with 24-chromosome screening. Mature oocytes were divided into IVF (n = 660) and ICSI (n = 1028) groups. The normal fertilization incidence was similar between the groups (81.1% vs. 84.6%). The total number of blastocysts biopsied was significantly higher in the IVF group than in the ICSI group (59.3% vs. 52.6%; p = 0.018). However, euploidy (34.4% vs. 31.9%) and aneuploidy (63.4% vs. 66.2%) rates per biopsy and clinical pregnancy rates (60.0% vs. 58.8%) were similar between the groups. Implantation (45.6% vs. 50.8%) and live birth or ongoing pregnancy (52.0% vs 58.8%) rates were slightly higher in the ICSI group than in the IVF group and miscarriage rate per transfer was slightly higher in the IVF group than in the ICSI group (12.0% vs 5.9%); however no significant difference was observed. IVF and ICSI using sibling mature oocytes had similar clinical outcomes, and euploidy and aneuploidy rates in couples with nonmale and mild male factor infertility. These results suggest that IVF is a useful option, along with ICSI, as an insemination method in PGT-A cycles, especially in high-risk patients.

P-245 Birth of twenty-seven healthy babies following transfer of fresh and frozen-thawed embryos derived from monopronuclear zygotes: a retrospective study

Abstract Study question What are the neonatal outcomes of fresh/frozen-thawed embryos derived from monopronuclear zygotes (1PN)? Is there any influence of the method of fertilization? Summary answer A total of 231 monoembryonic transfers (TF) of fresh or frozen/thawed embryos derived from 1PN zygotes allows to the birth of twenty-seven healthy babies. What is known already Zygotes were classified 16 to 18 hours post insemination according to the extrusion of the second polar body and the number of pronuclei (PN). The «normally fertilized» zygotes display 2 PN and those with 0, 1 or more than 2 PN (&amp;gt; 2PN) are considered as abnormal. If some zygotes are always discarded (&amp;gt;2PN) due to the increased risk of aneuploidy, the transfer of embryos derived from 1PN zygotes remains more controversial. Study design, size, duration Our retrospective study included all conventional in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and frozen embryo transfer (FET) cycles performed between january 2018 and december 2022. Embryo and pregnancy outcomes were analyzed in a first group of 781 fresh cycles (731 patients) and in a second group of 167 FET (150 patients) with embryos derived from 1PN zygote. Only monoembryonic transfers were analyzed for live birth rate (LBR). Participants/materials, setting, methods Only embryos derived from 1PN zygotes were analyzed in our study. For fresh cycles (n = 2762), embryo outcome was analyzed for 1234 (1PN) zygotes (IVF = 648 and ICSI = 586). Fresh embryo TF was performed in 64 cycles. For frozen cycles, TF was achieved in 167 cases. For a total of 231 TF on day 2, 3 or 5, pregnancy and live birth rates were analyzed. Main results and the role of chance At first, 46% of 1PN zygotes from fresh IVF cycles gave rise to embryos of sufficient quality to be transferred or frozen (day 3 or 5/6). This rate decreased to 33% in the fresh ICSI cycles. Blastulation rate was also more important in IVF group (44%) in comparison to ICSI group (20%) without signification (p = 0.1). Fresh TF of 64 patients (32 in each fertilization group) allowed 7 pregnancies in the IVF group (PR = 22%) as compared to 4 pregnancies in the ICSI group (PR = 12%). The 7 IVF pregnancies ended in 4 deliveries of healthy newborns, 2 miscarriages and one pregnancy still ongoing. In the ICSI group, 1 birth of a healthy newborn and 3 miscarriages were observed. Secondary, thirty-six pregnancies were obtained in the 167 FET cycles. A non significative difference was observed between embryos derived from IVF cycles (PR = 26%) and ICSI cycles (PR = 16%) with respectively 15 and 6 healthy babies born. Two pregnancies in each group are still ongoing. Limitations, reasons for caution The main limitation of this study is that it was a retrospective, single-center study with a relatively small number of births. Wider implications of the findings In conclusion, we observed better outcomes in IVF cycles in comparison to ICSI cycles and our center policy to transfer a good quality embryo developed from 1PN zygotes allowed 26 deliveries of 27 healthy newborns during the period analyzed. Trial registration number not applicable

Pregnancy and neonatal outcomes of monozygotic twins resulting from assisted reproductive technology: a 10-year retrospective study

BackgroundMonozygotic twins (MZTs) are associated with high risks of maternal and fetal complications. Even with the widely used elective single embryo transfer (SET), the risk of MZTs following assisted reproductive technology (ART) treatments remains. However, most studies of MZTs focused on the relevant etiology, with few studies describing pregnancy and neonatal outcomes.MethodsThis retrospective cohort study included 19,081 SET cycles resulting from in-vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), preimplantation genetic testing (PGT) and testicular sperm aspiration (TESA) performed between January 2010 and July 2020 in a single university-based center. A total of 187 MZTs were included in this investigation. The main outcome measures were the incidence, pregnancy and neonatal outcomes of MZTs. Multivariate logistic regression analysis was performed to figure out the risk factors for pregnancy loss.ResultsThe overall rate of MZTs from ART treatment in SET cycles was 0.98%. No significant difference was found in the incidence of MZTs among the four groups (p = 0.259). The live birth rate of MZTs in the ICSI group (88.5%) was significantly more favorable than in the IVF, PGT and TESA groups (60.5%, 77.2% and 80%, respectively). IVF resulted in a significantly increased risk of pregnancy loss (39.4%) and early miscarriage (29.5%) in MZT pregnancies compared to ICSI (11.4%, 8.5%), PGT (22.7%, 16.6%) and TESA (20%, 13.3%). The total rate of twin-to-twin transfusion syndrome (TTTS) in MZTs was 2.7% (5/187); however, the TESA group had the highest rate at 20% and was significantly higher than the PGT group (p = 0.005). The four ART groups had no significant effect on the occurrence of congenital abnormalities or other neonatal outcomes in newborns from MZT pregnancies. Multivariate logistic regression analysis revealed that infertility duration, cause of infertility, the total dose of Gn used, history of miscarriages, and the number of miscarriages were not related to the risk of pregnancy loss (p > 0.05).ConclusionsThe rate of MZTs was similar among the four ART groups. The pregnancy loss and the early miscarriage rate of MZTs was increased in IVF patients. Neither the cause of infertility nor the history of miscarriage was correlated with the risk of pregnancy loss. MZTs in the TESA group had a higher risk of TTTS, placental effects influenced by sperm and paternally expressed genes may play a role. However, due to the small total number, studies with larger sample sizes are still needed to validate these result. Pregnancy and neonatal outcomes of MZTs after PGT treatment seem to be reassuring but the duration of the study was short, and long-term follow-up of the children is needed.

Compared with conventional insemination, intracytoplasmic sperm injection provides no benefit in cases of nonmale factor infertility as evidenced by comparable euploidy rate

To evaluate if differences in euploidy rates exist between intracytoplasmic sperm injection (ICSI) and conventional insemination (CI) in non-male factor infertility cases. Retrospective cohort study. Single, academically affiliated infertility center in the U.S. A total of 3,554 patients who underwent in-vitro fertilization (IVF) cycles from January 2014 to December 2021. All cycles that had pre-implantation testing for aneuploidy (PGT-A) performed by trophectoderm biopsy and had a post preparation sperm concentration >4 million total motile sperm per milliliter were included. The primary outcome was embryo euploidy rate per embryo biopsied in the ICSI versus CI group. Secondary outcomes included fertilization rate and number of embryos biopsied. Generalized estimating equations with a Poisson distribution was used to estimate the euploid rate ratio (with total embryos biopsied as an offset), while accounting for multiple retrievals per patient. To adjust for confounding, a propensity score model was fit for ICSI using 14 baseline female and male characteristics. Oocytes retrieved and number of embryos biopsied were similar in both groups, while fertilization rate per oocyte retrieved was significantly lower with ICSI (0.64 vs 0.66, p=0.04). The proportion of euploid embryos in the ICSI group was significantly lower when compared to CI (0.47 vs 0.52), with a euploid rate ratio of 0.89 (p<0.001). Interestingly, when accounting for variation in PGT reference laboratories over the study time period, adjusting for date of procedure did not change the relationship between ICSI and euploid rate (RR=0.89, p<0.001); however, after adjusting for PGT reference laboratory, the relationship between ICSI and euploid rate was no longer significant (RR=0.97, p=0.07). In the setting of non-male factor infertility, ICSI resulted in a lower fertilization rate and an 11% lower embryo euploid rate compared to CI. Although the data is not statistically significant when adjusted for PGT reference laboratory, we can still conclude that ICSI does not provide any benefit. This data supports the recommendation that CI should be the preferred methodology of fertilization in non-male factor infertility cases.

Safety of early cumulus cell removal combined with early rescue ICSI in the prevention of fertilization failure

Research questionWhat are the clinical outcomes and safety implications of early cumulus cell removal after short-term insemination combined with early rescue intracytoplasmic sperm injection (ICSI) in preventing fertilization failure? DesignIn this retrospective study, a total of 14,360 cycles were divided into four groups based on insemination method and fertilization ability: conventional IVF group (n = 5519); early cumulus cell removal group (n = 4107); conventional ICSI group (n = 4215); and early rescue ICSI group (where failed or low fertilization was predicted, n = 519). Fertilization outcomes, pregnancy outcomes, neonatal outcomes and birth defects were analysed by comparing the early cumulus cell removal group with the conventional IVF group, and the early rescue ICSI group with the conventional ICSI group. ResultsThere were no significant differences in the outcomes of fertilization, pregnancy, neonates or birth defects between the conventional IVF group and the early cumulus cell removal group (P > 0.05). When compared with the conventional ICSI group, the early rescue ICSI group had similar rates of two pronuclei (2PN) at fertilization, clinical pregnancy, miscarriage, ectopic pregnancy, live birth, sex, mean gestational age, very low birthweight, macrosomia and birth defects (P > 0.05) but a higher polyploidy rate, lower high-quality embryo rate (both P < 0.001), lower twin pregnancy rate (P < 0.01), lower rate of low birthweight, and a higher rate of normal birthweight (both P = 0.024). ConclusionsEarly cumulus cell removal combined with early rescue ICSI led to good pregnancy and neonatal outcomes without an increase in birth defects. This approach could therefore be an effective and safe method for patients with fertilization failure in conventional IVF.

Correlation analysis of sperm DNA fragmentation index with semen parameters and the effect of sperm DFI on outcomes of ART

Routine semen analysis provides limited information about a man’s male reproductive potential and can not always fully explain male infertility. Many male infertilities are caused by sperm DNA defects that routine semen quality analyses fail to detect. In this study, we analyzed the association of sperm DNA fragmentation index (DFI) with the semen routine, sperm morphology, in vitro fertilization embryo transfer (IVF-ET)/intracytoplasmic sperm injection (ICSI). Further, we explored the predictive value of sperm DFI in evaluating male fertility and the outcome of IVF-ET/ICSI. Data on sperm DFI, sperm routine, and sperm morphology were collected from 1462 males with infertility. According to DFI levels, there were 468 cases in group I (DFI ≤ 15%), 518 cases in group II (15% < DFI < 30%), and 476 cases in group III (DFI ≥ 30%). The correlations of sperm DFI with semen routine and malformation rate were analyzed. Seminal plasma malondialdehyde (MDA), and total antioxidant capacity (TAC) were assessed. Sperm DFI, semen routine, and sperm morphology were detected in male patients of 101 pairs of IVF-ET/ICSI infertile couples and subdivided into IVF-I group (DFI ≤ 15%), IVF-II group (15% < DFI < 30%), IVF-III group (DFI ≥ 30%), ICSI-I group (DFI ≤ 15%), ICSI-II group (15% < DFI < 30%) and ICSI-III group (DFI ≥ 30%) according to DFI value. The effect of sperm DFI on the outcome of IVF-ET/ICSI was analyzed. There were significant differences in sperm survival rate, sperm concentration, and PR% between groupIII and group II (P < 0.01). There were significant differences in sperm survival rate, sperm concentration and PR% between group III and group I (P < 0.01). There was no significant difference in semen volume, age, abstinence days, or percentage of normal sperm between the three groups (P > 0.05). DFI was positively correlated with MDA content ( P < 0.01) and negatively correlated with TAC (P < 0.01). Sperm DFI was negatively correlated with sperm survival rate, sperm concentration, and PR% (P < 0.01). There was no correlation with age, abstinence days, semen volume, or percentage of normal-form sperm (r = 0.16, 0.05, 0.04, -0.18, p > 0.05). Compared with IVF-I group, the sperm concentration and PR were decreased in IVF-III group. The sperm malformation rate was higher in IVF-III group than that in IVF-II group. Comparatively, the PR was decreased in ICSI-III group. The sperm malformation rate was higher in ICSI-III group than that of the ICSI-I group (P < 0.05). There were no statistically significant differences in fertilization rate, cleavage rate, embryo rate, and clinical pregnancy rate between IVF group or ICSI group, and between all subgroups (P > 0.05). Sperm DFI is negatively associated with sperm survival rate, sperm concentration, and PR%. Antioxidants can decrease the rate of DNA fragmentation. Sperm DFI has proven to be very valuable in the male fertility evaluation, but its significance as a predictor of pregnancy outcomes following assisted reproductive technology. (ART) requires further investigation.

Comparison of clinical outcomes between conventional in vitro fertilization and intracytoplasmic sperm injection in poor responders with only single oocyte retrieved

ObjectiveTo compare the clinical outcomes between conventional insemination (IVF) and intracytoplasmic sperm injection (ICSI) in poor responders with only a single oocyte retrieved. Materials and methodsThis is a retrospective case–control study. Couples who were treated with assisted reproductive technology (ART) with a single oocyte retrieved in Mackay Memorial Hospital from 1996 to 2016 were recruited. All data were categorized into three groups, according to their fertilization method and semen quality: group A, conventional insemination with non-male factor (IVF-NMF, n = 115), group B, ICSI with male factor (ICSI-MF, n = 30), and group C, ICSI with non-male factor (ICSI-NMF, n = 49). ResultsNo statistically significant difference was observed between IVF and ICSI groups in pregnancy outcomes, including the chemical or clinical pregnancy rate, miscarriage rate, and live birth rate. Similar fertilization rates per oocyte obtained were observed in IVF and ICSI patients, but significantly lower per mature oocyte in the ICSI group (IVF: 91.5%, ICSI-MF: 75.0%, ICSI-NMF: 77.8%). Although there is no statistical significance, the lower live birth rate is observed in group C than others (A:11.5%, B:25%, C:5%, p = 0.187). ConclusionIn this study, pregnancy outcomes of conventional in vitro fertilization and ICSI in poor responders with only a single oocyte retrieved were similar. However, the fertilization rate of matured oocytes in ICSI groups is significantly lower than that in the IVF group, indicating that ICSI procedures might cause oocyte damage. Therefore, the choice of fertilization method should be based on semen quality. A randomized controlled trial should be performed to confirm our findings.

P-223 Women aged 40 or older and non-male factor have the same reproductive outcome using intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (cIVF)

Abstract Study question Do women aged 40 or older have the same reproductive outcome using ICSI or cIVF when male factor infertility is not present? Summary answer Women aged 40 or older with non-male factor partners achieved similar reproductive outcomes either with ICSI or cIVF. What is known already Although initially proposed to treat couples with severe male factor infertility, ICSI has become the preferred tool to treat patient populations others than initially intended. Many studies and international forums, however, have alerted that its overuse is not free due to its safety, cost and time-consuming concerns. Moreover, it was noted that it does not increase clinical outcomes even in cases of compromised oocyte factor such as poor responder patients or few oocytes retrieved. However, there is still no information regarding its use in the group of patients of advanced reproductive age. Study design, size, duration Retrospective cohort study including 412 women aged 40 or older with non-male factor infertility performing ICSI or cIVF between May 2015 and October 2017. Participants/materials, setting, methods Patients underwent ICSI (n = 109) or cIVF (n = 303) according to medical indication. The decision to use ICSI in non-male factor patients was merely operational, with the objective of evaluating oocyte maturity.Those cycles with own oocytes, non-male factor and fresh embryo transfers were included. Semen samples were considered normal following the World Health Organization fifth edition sperm parameters values, and prepared by density gradient selection. Main results and the role of chance The primary outcome was clinical pregnancy rate. Secondary outcomes included ongoing pregnancy, miscarriage and implantation rates. Statistical significance was denoted by P &amp;lt; 0.05. The mean age of the women were 41.4 in the ICSI group and 41.6 years in the cIVF group, while the mean age for male couple was 43.6 and 41.6 respectively. No differences between ICSI and cIVF groups were observed regarding clinical pregnancy [16.5% (18/109) vs. 20.8% (63/303)], ongoing pregnancy [9.2% (10/109) vs. 7.9% (24/303)], miscarriage [41.2% (7/17) vs. 46,5% (20/43)], and implantation rates [11.3% (17/151) vs. 9.9% (50/506)]. Limitations, reasons for caution These results need confirmation with a bigger population size. As with any retrospective study, the potential for residual confounding exists. Wider implications of the findings These data suggest that ICSI offers no clinical benefit for women aged 40 and older with non-male factor infertility, beyond oocyte maturation classification. Trial registration number not applicable

P-121 Azoospermia: are there neonatal differences regarding the source of sperm for ICSI?

Abstract Study question In couples with azoospermia undergoing intracytoplasmic sperm injection (ICSI), are neonatal outcomes different depending on sperm origin (epididymal-PESA, testicular-TESA or donor sperm)? Summary answer Newborns from PESA or TESA had similar neonatal outcomes. Comparing to donor cases, PESA/TESA cycles didn’t present increased levels of malformation or stillbirth. What is known already Since the introduction of ICSI and the demonstration that the spermatozoa derived either from the epididymis or the testis were capable of normal fertilization and pregnancy, it was possible for azoospermic men to father their own children. However, it raised concerns that the quality of spermatozoa in terms of DNA damage or maturation when collected from non-ejaculated semen could differ from that collected from ejaculated and whether sperm of different origins will affect the neonatal outcome and safety of ICSI. Study design, size, duration Retrospective cohort study performed at a reproductive medicine center, using data from 359 deliveries from 945 cycles of ICSI of couples with infertility due to azoospermia, performed between 1995 and 2021. Data were collected from electronic records. Participants/materials, setting, methods A total of 359 deliveries were divided in 3 groups, according to the sperm source: PESA (n = 138), TESA (n = 139) and DONOR (n = 82). A total of 402 newborns were evaluated: PESA (n = 162); TESA (n = 156); DONOR (n = 84). Neonatal outcomes and congenital malformations were analyzed for singletons, twins and triplets separately. Statistical analysis: Anova, chi-square and Fischer test, considering p &amp;lt; 0.05. Main results and the role of chance Comparing the 3 groups (PESA, TESA, and donor sperm), the following results were found: The mean birth weight (BW), length and Apgar score &amp;gt; 7 for singletons and twins did not differ between groups; neither did prematurity or extreme prematurity. According to birth percentile, there was a lower rate of small for gestational age (SGA) newborn in the epididymal group (PESA, 1.5% vs. TESA, 8.5% vs. sperm donor, 5.5%, p = 0.004). Also, in 158 twin births, the number of newborns with low birth weight (&amp;lt;2500g) was significantly smaller in the epididymal group (PESA, 32.9% vs. TESA, 62.5%; donor sperm, 73.1%, p = 0.001). There were three stillbirths in triplets (PESA group). We also found six cases of neonatal mortality (one in singleton pregnancy in TESA group, 3 in twins (two in PESA group and one donor sperm group), and two in triplets (one in TESA and one in PESA groups). The total number of congenital malformations was 8 (2.48%) (PESA n = 1, 0.62%; TESA, n = 2, 1.28%, and donor sperm, n = 5, 5.95%, p = 0.02). Limitations, reasons for caution The possibility of incomplete medical records is an important limitation to be considered in retrospective studies. Larger studies are important to better assess the risk of malformation in this population. Wider implications of the findings In this study, there were no differences on neonatal outcomes regarding the source of sperm used for ICSI, with PESA, TESA and donor groups, as well as congenital malformation. It showed equally safe options with testis, epididymal or ejaculated sperm for azoospermia treatments. Trial registration number not applicable

P-190 The first cleavage plane in human embryos is dictated by the topology of pronuclei, not by the polar body position

Abstract Study question Is the first cleavage plane pre-determined in human embryos? Summary answer Unlike in mice, the human zygotes tend to divide at the right angle to the longitudinal pronuclear axis, irrespective of polar body position. What is known already In lower species, the orientation of the first cleavage plane defines the development of individual blastomeres and the three-dimensional (3D) body plan. The existence of embryonic pre-patterning in mammals remains controversial. Mouse studies suggested that the juxtaposed pronuclei (PN) are aligned with the polar body (PB) and cleavage furrow bisects the zygote along the polar axis thus ensuring symmetrical distribution of maternal resources to arising blastomeres. However, whether the same spatial control applies to humans is unclear. Study design, size, duration Retrospective observational analysis of high-resolution footage from 300 human embryos that gave rise to live births after a single embryo transfer. Participants/materials, setting, methods Embryo monitoring has been performed by the GERI time-lapse incubator. Image acquisition of individual embryo development lasted 6 days, starting within 10 minutes after intracytoplasmic sperm injection (ICSI). Transmitted light images have been automatically captured in 5-minute intervals and 11 focal planes covering the whole embryo volume. Analysis of 3D image data was performed at the time frame corresponding to the onset of the first mitosis. Main results and the role of chance The systematic analysis of the zygotic division pattern showed that the PN axis (defined by apposed pronuclei) was not stereotypically aligned with the animal-vegetal (AV) axis (with animal pole defined by the PB). Instead, PN/AV arrangement categories were equally represented in our cohort (PN/AV angle 0-30° - 34.7% vs. 30-60° - 36.0% and 60-90° - 29.3%; chi-square test p=.91351). Remarkably, we observed that the direction of the first cleavage was preferentially perpendicular to the PN axis (65.3%), while only 2.7% of embryos divided along the longitudinal PN axis. The remaining 32% embryos showed intermediated phenotype with cleavage plane/PN axis angle 30-60o. The results of this human embryo analysis contrast with the previous reports from mice. We hypothesize that this discrepancy might be attributed to the difference in centrosome inheritance and configuration of the zygotic spindle apparatus. Limitations, reasons for caution All analyzed embryos were produced by ICSI. Deposition of spermatozoa might affect the pronuclei positioning. Nevertheless, preliminary data from conventional IVF cycles are in agreement with the ICSI group results. Wider implications of the findings Our observations challenge the textbook view of the first division’s geometry, which has been derived from experiments in mice. The analysis of clinical time-lapse records improves our understanding of symmetry breaking during normal human development and creates scientific ground for evidence-based (de)selection of embryos for infertility treatment. Trial registration number not applicable

Perinatal outcome of babies born after using a simplified IVF culture system versus ICSI with sibling oocytes: a prospective cohort study

Research questionIs there a difference in perinatal outcome in the same patient cohort for babies conceived following randomization of sibling oocytes allocated to a simplified IVF culture system (SCS) or intracytoplasmic sperm injection (ICSI) followed by conventional culturing? DesignThe study compared the perinatal outcomes of 367 babies born from 1 January 2013 until 31 December 2020 after using split SCS and ICSI insemination of sibling oocytes in a selected group of normo-responsive women, excluding cases of severe male infertility. Primary outcome measures were preterm birth (PTB; <37 weeks’ gestation), low birthweight (LBW; <2.5 kg) and small for gestational age (SGA) as a primary outcome parameter while secondary outcome measures included mean birthweight, mean gestational age, extreme prematurity (<32 weeks), very low birthweight (<1.5 kg), perinatal mortality, multiple pregnancy and Caesarean section rate. ResultsA total of 105 and 103 singleton babies were born after fresh embryo transfer (FRET) and 71 and 50 singletons after frozen embryo transfer (FET) in the SCS and ICSI groups, respectively. For babies born after FRET, the LBW rate was 2.9% (3/105) for SCS and 7.8% (8/103) for ICSI (P = 0.10). LBW occurred in 4.2% (3/71) and 0% (0/50) of babies born after the transfer of cryopreserved-thawed SCS and ICSI embryos, respectively (P = 0.14). The rate of PTB was 3.8% and 6.8% for SCS and ICSI in FRET cycles (P = 0.33), and 8.5% and 6.0% for SCS and ICSI in FET cycles (P = 0.62). One congenital malformation was found in the SCS FET group. ConclusionThere was no difference in perinatal outcome for singleton and twin babies born after SCS and ICSI.

Clinical outcome of microsurgical vasoepididymostomy versus epididymal or testicular sperm retrieval combined with intracytoplasmic sperm injection in obstructive azoospermia males.

This purpose of the retrospective study was to compare the clinical outcome of microsurgical vasoepididymostomy (MVE) and sperm retrieval combined with intracytoplasmic sperm injection (ICSI) in the treatment of males with obstructive azoospermia. From February 2018 to December 2019, a total of 69 males with obstructive azoospermia underwent MVE while 351 males with obstructive azoospermia in their first ICSI cycles were enrolled. Patients in the MVE group achieved higher pregnancy rate and births delivered rate than those in ICSI group (pregnancy rate, 44.6% vs. 27.6% for MVE and ICSI, respectively; delivery rate, 38.5% versus 25.8% for MVE and ICSI, respectively). Besides, lower female age (<35 vs. ≥35 years old, OR 1.15, 95% CI=1.08-1.21), higher sperm concentration and forward motility after MVE((sperm concentration, OR (95% CI)=1.12 (1.08-1.23), sperm forward motility, OR (95% CI)=1.10 (1.06-1.20)) were associated with higher probability of pregnancy. Based on these data, it has been found that MVE is the prior option for males with obstructive azoospermia suitable for the surgery, other than direct subjected to ICSI. More high quality studies are needed in the future due to the non-randomized design and the relative small size of this study.

Relationship between sperm quality and total fertilization failure in intracytoplasmic sperm injection and in vitro fertilization cycles: A cross-sectional study.

BackgroundTotal fertilization failure (TFF) is associated with essential mechanistic and cellular events.ObjectiveThe present study is a comprehensive examination of detrimental effects with well-known assays for predicting TFF in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles.Materials and MethodsSemen parameters of 90 men, including 60 cases who had experienced IVF/ICSI failure and a control group of 30 individuals, were evaluated. Sperm chromatin/DNA quality assessments were done by aniline blue, toluidine blue, chromomycin A3, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays. A lipid hydroperoxide (LPO) kit was used to measure the LPO, and JC1 staining was used to evaluate mitochondrial membrane potential (MMP).ResultsThere were statistically significant differences found between the IVF, ICSI and control groups by the toluidine blue (p = 0.01), TUNEL (p = 0.02), and chromomycin A3 (p 0.001) tests, but not by the aniline blue staining. Furthermore, there was a significant difference regarding LPO concentration and high MMP in cases of IVF fertilization failure compared to the control group (p = 0.04, p = 0.02, respectively). The logistic regression model showed that sperm viability was predictive for fertilization failure in the ICSI group. Sperm chromatin and DNA quality assays were not predictors for TFF in either group.ConclusionCellular events such as high DNA fragmentation damage, high levels of reactive oxygen species, and low MMP levels can cause TFF in IVF and ICSI programs. Diagnostic tests, especially in cases with previous fertilization failure, showed significant differences in sperm chromatin and DNA quality between groups but could not predict the risk of TFF.

In vitro production of ICSI derived horse blastocysts

In vitro production of horse embryos via intracytoplasmic sperm injection (ICSI) is a useful clinical and research technique. However, current rates of blastocyst production are still sub-optimal. Here, we compared results from our commercial ICSI program to evaluate whether slight modifications in protocols can result in improved blastocyst rates. Firstly, we evaluated the time cumulus oocyte complexes (COCs) can be maintained in holding medium before starting in vitro maturation (IVM) (Group A: 4-10 hours and Group B: overnight). Secondly, we evaluated the effects of using a microfluidic device (Zymöt) for the preparation of sperm samples for ICSI (Group C). This device comprises a microporous membrane that allows to select spermatozoa in base of their motility, avoiding samples centrifugation. This could eventually reduce DNA fragmentation and oxidative stress, which have been associated with conventional sperm preparation protocols. Frozen-thawed cut straws with semen from different stallions were used and prepared either with a gradient and swim-up protocol or with the Zymöt microfluidic device. COCs were recovered from immature follicles using transvaginal ultrasound-guided follicle aspiration (TVA) procedures. After collection, COCs were transferred to holding medium for 4 to 10 hours or maintained in holding medium overnight at room temperature (22–24°C) before starting IVM. In vitro maturation was performed in T-199 Earle's medium supplemented with 5 mU/ml FSH, 50 ng/ml EGF and 10% FBS. After IVM, oocytes were denuded from cumulus and those showing a polar body were subjected to piezo-ICSI (PiezoXpert, Eppendorf). Spermatozoa were first immobilized with strong piezo pulses and then injected individually in the matured oocytes after penetrating the zona pellucida and oolemma with low intensity pulses.The injected oocytes were then cultured in a mix of KSOM-DMEM/F12 medium supplemented with 6% FBS, which was refreshed on Day 5 of development. Embryo culture was performed in humidified benchtop incubators at 6.5% CO2 and 5%O2 at 38.2°C. A Chi-square test was used to compare results.

Fertilization and neonatal outcomes after early rescue intracytoplasmic sperm injection: a retrospective analysis of 16,769 patients

PurposeTo evaluate the efficacy and safety of short-term insemination and early-rescue intracytoplasmic sperm injection (ICSI), an approach that rescued oocytes with unclear second polar body 6 h after initial insemination by ICSI (early R-ICSI) to avoid total or near-total fertilization failure in conventional in vitro fertilization (IVF).MethodsWe performed a retrospective study in 16,769 patients (short-term IVF, n = 12,094; ICSI, n = 3452; early R-ICSI, n = 1223) who received IVF/ICSI treatment in our hospital from January 2009 to October 2018. Fertilization and clinical outcomes were compared among those three groups.ResultsWhen considering the R-ICSI embryos in the early R-ICSI group independently, the rates of fertilization and day-3 cleaved embryos in 2PN oocytes were comparable, the rates of fertilization (2PN) and high-quality embryos were lower, whereas the multi-PN fertilization rate (3.27%) was significantly higher than the ICSI group (1.26%). The difference of clinical pregnancy rate between the part of transferred R-ICSI embryos (40.81%) and the ICSI group (44.73%) remained nonsignificant. Furthermore, the rate of congenital birth defects in the early R-ICSI group (0.99%) was not significantly different from those in the short-term IVF (0.76%) and ICSI groups (1.07%).ConclusionDespite the multi-PN fertilization rate, our study highlights early R-ICSI as a safe and effective alternative in assisted reproduction to decrease complete IVF fertilization failure and reduce ICSI utilization. Additional large amount and long-term follow-up studies are needed to further validate the use of early R-ICSI.

Placental histology and pregnancy complications following intracytoplasmic sperm injection for non-male factor infertility.

To assess obstetric outcomes and placental histology following intracytoplasmic sperm injection (ICSI), for non-male infertility. This was a retrospective cohort of live born singleton deliveries after in vitro fertilization (IVF) at a single university affiliated medical center between 2009 and 2017. Excluded were IVF cycles with male infertility and oocyte recipients. We compared obstetric outcomes and placental histology in cases ICSI was performed (ICSI group) and cases with no ICSI (IVF group). A total of 400 deliveries following ICSI were compared to 218 in the IVF group. Maternal age was similar between the groups, while diminished ovarian reserve was more common among ICSI patients and tubal disease less common (p < 0.001). The rate of blastocyte transfer was also significantly lower in the ICSI group-67.5% vs. 77%, p = 0.01. Pregnancies following ICSI were characterized by similar rates of preeclampsia, preterm birth, and small for gestational age neonates. Although cesarean delivery rate was significantly higher in the group, this did no attain significance after adjustment for confounders. Placentas in the ICSI group were notable for a lower rate of villitis of unknown etiology (1% vs. 4.5%, p = 0.007) and a higher rate of maternal surface calcifications (33% vs. 23.8%, p = 0.01) after adjustment for confounders. The employment of ICSI with no male indication is associated with similar obstetric outcomes. Despite isolated placental differences among many investigated, placental histology seems overall comparable as well. These results are reassuring to clinicians and patients.

Analysis of Clinical Outcomes of Different Fertilization Methods in Patients with ≤3 Eggs Retrieved.

Objective To explore the intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) method on the clinical outcomes of infertile women with ≤3 eggs retrieved. Study Design. We retrospectively analyzed a cohort of female patients who received IVF/ICSI to assist pregnancy with retrieved eggs ≤3. The general conditions, i.e., two pronuclei (2PN) fertilization rate, abnormal fertilization rate, high-quality embryo rate, cycle cancellation rate, pregnancy rate of fresh embryo transfer, cumulative pregnancy rate, and miscarriage were compared between the two groups. Results When the number of retrieved eggs was one, the fertilization rate of 2PN was higher and the cycle cancellation rate was lower in the ICSI group than in the IVF group (P < 0.05). The pregnancy rates of fresh embryo transfer, frozen-thawed embryo transfer, and the cumulative pregnancy rate were all higher in the ICSI group than in the IVF group (P < 0.05). When the number of retrieved eggs was two, the pregnancy rate of frozen-thawed embryo transfer and cumulative pregnancy rate in the ICSI group were higher than those in the IVF group (P < 0.05). When the number of retrieved eggs was three, the fertilization rate of 2PN and the pregnancy rate of frozen-thawed embryo transfer were higher in the ICSI group than those in the IVF group (all (P < 0.05)). Conclusions For patients with one egg retrieved, ICSI fertilization can reduce abnormal fertilization rate and cycle cancellation rate and improve cumulative pregnancy rate significantly enhancing patients' benefits. However, increasing the number of eggs retrieved decreases the advantages of ICSI fertilization.

Effects of leukocytospermia on the outcomes of assisted reproductive technology.

Leukocytospermia is one of the common causes of male infertility, and its effects on the clinical outcomes of assisted reproduction are controversial. There are no recommendations for the management of leukocytospermia in cases of assisted reproductive technology (ART). To investigate the impact of leukocytospermia on ART, we retrospectively compared the clinical outcomes in ART couples with or without leukocytospermia and further analysed the impact of the insemination method itself by split insemination treatment in ART couples with leukocytospermia. In this study, leukocytospermia was detected in 133 patients, namely 63 in the conventional in vitro fertilization (IVF) group, 38 in the intracytoplasmic sperm injection (ICSI) group and 32 in the split insemination group. Leukocytospermia has a negative influence on the parameters of semen samples; however, leukocytospermia did not affect the clinical outcomes of IVF or ICSI. Different insemination methods did not affect the fertilization, clinical pregnancy or live birth rates. In the split insemination study, no significant differences in clinical pregnancy and live birth rates between the IVF and ICSI groups were found; however, the numbers of two pronuclei (2PN), available embryos and good‐quality embryos in the ICSI group were higher than those in the IVF group. Leukocytospermia may be a risk factor affecting semen parameters, and more attention should be given to IVF insemination. Leukocytospermia has no significant negative effect on the outcomes of ART. ICSI may obtain better embryos than IVF, but it cannot improve the clinical pregnancy and live birth rates.

45 Does ovarian endometriosis increase oocyte sensitivity to icsi-induced mechanical damage?

Introduction and aims of the study: Some studies on sibling oocytes have shown that intracytoplasmic sperm injection (ICSI) obtains poorer results than conventional in vitro fertilization (cIVF) in women with ovarian endometriosis, suggesting that their oocytes could be particularly sensitive to some ICSI-induced mechanical damage. Aims of this study were to clarify (a) whether ovarian endometriosis could induce peculiar fragility in the oocytes of women with endometriosis, so that ICSI would finally result harmful, and (b) whether endometrioma removal before IVF could be advisable in order to avoid any hypothetical detrimental effect on oocytes. Methods: In the present case-control study, data of women who underwent their first IVF cycle between January 2010 and December 2018 at the IVF Unit of S. Anna Hospital, Turin (Italy), were retrospectively reviewed. We included 368 women: 203 with in situ endometrioma at the time of IVF (128 of which underwent ICSI and 75 cIVF), and 165 who received laparoscopic stripping of endometrioma before undergoing ICSI. Results: For women with in situ endometrioma at the time of treatment, ICSI and cIVF outcome was comparable for all studied parameters, including the clinical pregnancy rate per embryo transfer (PR/ET: 31.8% (95% confidence interval (CI), 20.8-44.4%) in cIVF group vs. 39.5% (95%CI, 30.7-48.9%) in ICSI group, p=0.15) and the cumulative live birth rate per ovum pick-up (CLBR/OPU: 24.4% (95%CI, 15.6-35.1%) in cIVF group vs. 27.7% (95%CI,20.7-35.7%) in ICSI group, p=0.29). ICSI outcome was also similar comparing women with in situ endometrioma (CLPR/OPU: 27.7% (95%CI, 20.7-35.7%)) and women previously submitted to laparoscopic stripping of the ovarian cyst (CLPR/OPU: 25.1% (95%CI, 19.1-32.0%)) (p=0.30). Conclusions: Our findings suggest that (a) in women with in situ endometrioma ICSI may be performed, when needed, without harming oocytes and compromising the outcome, and (b) there is no advantage in removing endometrioma before performing ICSI.