Standard Intracytoplasmic Sperm Injection Research Articles

Sperm selection by zona adhesion improves assisted reproductive treatment outcomes.

Zona pellucida is one of the main selective barriers for the spermatozoa before reaching the oocyte. Using native zona in the sperm selection prior to Intracytoplasmic Sperm Injection (ICSI) has been proven effective but inconvenient approach in In Vitro Fertilisation (IVF) laboratory. The application of autologous solubilised zonae pellucidae in the sperm selection prior to ICSI has not been studied yet. To compare the assisted reproductive treatment (ART) outcomes (implantation, pregnancy, live birth, and miscarriage rates) after ICSI performed with spermatozoa selected on their ability to adhere to immobilised solubilised zonae pellucidae and conventionally selected spermatozoa. In total, 500 couples fulfilled the inclusion criteria and 368 of them were included in the study. After random allocation, 192 couples had spermatozoa selected by sperm-zona adhesion for ICSI (study group) and 176 patients underwent standard ICSI (control group). In the study group, patients' own zonae were acid solubilised and immobilised on petri dishes. The partner's motile spermatozoa were placed in the dishes and the adhered spermatozoa were used for ICSI. For the control group, the conventional sperm selection by morphological criteria was applied prior ICSI. All women underwent frozen ET with euploid embryos. Chi square test was used to compare the data. The sperm selection by zona adhesion resulted in significantly higher implantation rate (50.4%vs. 37.0%, p=0.003), clinical pregnancy rate (43.8%vs. 33.3%, p=0.018) and live birth rate (38.0%vs. 25.9%, p=0.004) and significantly lower incidence of miscarriage (11.3%vs. 22.2%, p=0.044) in comparison to the conventional method of the sperm selection. The application of solubilised zonae pellucidae in the sperm selection for ICSI benefits ART outcomes in couples with unexplained infertility. Moreover, sperm-zona selection significantly reduces the risk of miscarriages.

The importance of sperm DNA fragmentation in the choice of a method for the treatment of male infertility using assisted reproductive technologies

Background. Sperm DNA fragmentation is an important marker of male fertility and may influence the outcome of the infertility treatment based on assisted reproductive technologies. Numerous studies have been done to assess the effect of sperm DNA fragmentation on reproductive outcomes, however the extent of this effect with various assisted reproductive technologies remains a matter of debate.Materials and methods. Search, analysis and systematization of publications in the PubMed and eLIBRARY databases using the keywords: male infertility, sperm DNA fragmentation, intrauterine insemination, in vitro fertilization, intracytoplasmic sperm injection. We have selected 49 sources that are included in this literature review.Results and discussion. The presence of a high level of sperm DNA fragmentation is associated with a low probability of natural pregnancy, as well as a low frequency of successful intrauterine insemination. The high level of sperm DNA fragmentation appears to have a negative impact on the reproductive outcomes of in vitro fertilization and intracytoplasmic sperm injection (ICSI), but the degree of this effect on the results of ICSI is less pronounced. ICSI with testicular spermatozoa can be used in treatment of male infertility associated with high sperm DNA fragmentation, however, the data on the advantage of this approach over standard ICSI are contradictory.Conclusion. High level of sperm DNA fragmentation associated with the low likelihood of natural pregnancy and success of intrauterine insemination and in vitro fertilization. More large, well-designed studies are needed to establish the role of ICSI with testicular sperm in the treatment of patients with high levels of sperm DNA fragmentation.

Influence of a hyaluronan-binding system for sperm selection in intracytoplasmic sperm injection cycles on embryo morphokinetic parameters and in vitro fertilization cycle outcomes.

Although the impact of the paternal contribution to embryo quality and blastocyst formation is a well-known phenomenon, the current literature provides insufficient evidence that hyaluronan-binding sperm selection methods improve assisted reproductive treatment outcomes. Thus, we compared the cycle outcomes of morphologically selected intracytoplasmic sperm injection (ICSI) with hyaluronan binding physiological intracytoplasmic sperm injection (PICSI) cycles. A total of 2415 ICSI and 400 PICSI procedures of 1630 patients who underwent in vitro fertilization cycles using a time-lapse monitoring system between 2014 and 2018 were analyzed retrospectively. Fertilization rate, embryo quality, clinical pregnancy rate, biochemical pregnancy rate and miscarriage rate were evaluated, differences in morphokinetic parameters and cycle outcomes were compared. In total, 85.8 and 14.2% of the whole cohort were fertilized with standard ICSI and PICSI, respectively. The proportion of fertilized oocytes did not significantly differ between groups (74.53 ± 1.33 vs. 72.92 ± 2.64, p > 0.05). Similarly, the proportion of good-quality embryos according to the time-lapse parameters and the clinical pregnancy rate did not significantly differ between groups (71.93 ± 4.21 vs. 71.33 ± 2.64, p > 0.05 and 45.55 ± 2.91 vs. 44.96 ± 1.25, p > 0.05). No statistically significant differences were found between groups in clinical pregnancy rates (45.55 ± 2.91 vs. 44.96 ± 1.25, p > 0.05). Biochemical pregnancy rates (11.24 ± 2.12 vs. 10.85 ± 1.83, p > 0.05) and miscarriage rates (24.89 ± 3.74 vs. 27.91 ± 4.91, p > 0.05) were not significantly different between groups. The effects of the PICSI procedure on fertilization rate, biochemical pregnancy rate, miscarriage rate, embryo quality, and clinical pregnancy outcomes were not superior. The effect of the PICSI procedure on embryo morphokinetics was not apparent when all parameters were considered.

P-045 Sperm chromatin structure resilience is related to fertilization and blastocyst rates in ICSI cycles involving both oocyte and sperm donors

Abstract Study question Does the structural maintenance of sperm chromatin have an implication in assisted reproduction outcomes? Summary answer Sperm deprotamination is related to fertilizing capacity. Also, the time needed for the reduction of protamine disulphide bridges is a parameter underlying early embryo development. What is known already How male factor affects embryo development has been a controversial topic for decades, with opposite results. A meta-analysis recently showed that whereas sperm genotoxic damage has an impact on IVF success, its repercussion on ICSI remains unclear. Revealing the factors that may predict embryo development is much warranted to understand the sperm contribution in the ICSI era. Sperm DNA is preferentially protamine-condensed and necessarily needs to be switched to histone-condensed chromatin upon fertilization. This study aims to determine the degree of protamination and the disulphide bond integrity to provide new insights into the influence of chromatin resilience on embryo development. Study design, size, duration The study setting included a public university and a private sperm bank. The design was a blinded retrospective study, including samples from 51 sperm donors that were used for a total of 55 cycles conducted with donated oocytes in the fertility clinic. Sperm quality parameters, including those focused on chromatin structure, were evaluated after 0 min and 240 min post-thawing and compared to embryo outcomes. Participants/materials, setting, methods Sperm samples from donors selected by the germplasm bank were collected and used to fertilize oocytes in double-donation cycles (oocyte donation) with standard intracytoplasmic sperm injection. Cryopreserved samples were used to assess sperm viability (SYBR14/PI), incidence of thiol reduction (Dibromobimane, DBB) and chromatin deprotamination (Chromomycin A3, CMA3) through flow cytometry, at 0 min and 240 min after thawing. Positive controls after incubation with Triton-X100, DTT and DTT+NaCl were used to establish the flow cytometry thresholds. Main results and the role of chance In fresh semen samples, sperm concentration was 34.12±18.13 sperm/mL and progressive motility was 37.49±13.22% (mean±standard deviation). At 0 min and 240 min post-thawing, sperm viability was 24.80±12.12% and 18.65±10.37%, respectively, and the death rate was -1.54±1.24%/hour. Percentages of sperm with reduced thiols were 17.37±11.26% and 11.53±9.16% after 0 min and 240 min post-thawing, the reduction rate being -1.46±2.31%. Percentages of sperm with chromatin deprotamination were 21.12±9.03% and 35.34±14.10% after 0 min and 240 min post-thawing, and the deprotamination rate was 14.22±11.90%/hour. ICSI was conducted with an average of 11.86±7.89 oocytes per sperm sample, of which 8.78±5.95 were fertilized; the mean fertilization rate was 75.77±17.61%. For each sperm sample, 5.98±4.93 blastocysts at day 5 were obtained, leading to an average blastocyst rate of 65.17±22.22%. Interestingly, the deprotamination status at 0 min post-thaw was correlated to fertilization rates (Rs= -0.388; P = 0.005) and the decreasing rate of sperm with reduced thiols was correlated to blastocyst rate (Rs= -0.351; P = 0.012). Reduced thiols and deprotamination were also correlated with each other (Rs= -0.320; P = 0.022). Limitations, reasons for caution The main limitation of the present study was the sample size. More donors are thus needed in order for the promising results found in this study to be confirmed. Wider implications of the findings The research conducted herein has evidenced that the resilience of the sperm chromatin structure is related to embryo development outcomes from ICSI cycles conducted with oocyte donors. Trial registration number N/A

Hyaluronic acid-sperm selection significantly improves the clinical outcome of couples with previous ICSI cycles failure.

Beneficial effects of hyaluronic acid (HA)-based selection of spermatozoa for intracytoplasmic sperm injection (ICSI) are still controversial, and further studies are needed to categorize patients that might benefit from such a method. We investigated whether HA sperm selection improved ICSI outcome of couples with previous ICSI cycle failure. In this retrospective study, we prospectively collected data of (i) Group 1: 96 couples who performed one failed ICSI cycle ("1st procedure," n=96) followed by another ICSI cycle ("2nd procedure," n=101); ii) Group 2: 87 couples who performed one failed ICSI cycle (n=87) followed by an HA-ICSI cycle (n=104). Differences between procedures and groups were measured by paired and independent statistical tests, respectively. A generalized linear mixed model analyzed the effect of procedure on the outcomes and the interaction between procedures and groups. Injection of HA-bound sperm significantly improved cleavage rate with respect to standard ICSI (p=0.026). No evolutive pregnancies were obtained in the 1st ICSI attempts. The 2nd ICSI cycles resulted in successfully seven pregnancies. In HA-ICSI cycles, the better quality of embryos with respect to ICSI (p=0.034) increased the choice of day 5 embryo transfer (p=0.030), which resulted in successfully 28 pregnancies. No differences were observed in clinical outcomes of the two ICSI procedures in Group 1, while pregnancy and implantation rates were significantly higher in HA-ICSI with respect to ICSI cycles (p=0.001, p<0.0001, respectively). No negative perinatal outcomes were recorded. In couples where previous 1st ICSI failed, selection of HA-bound spermatozoa significantly improved clinical outcomes with respect to further standard ICSI. This study identified couples with previous ICSI cycles failure as a category of infertile patients that really may benefit from HA sperm selection before ICSI.

Conventional ICSI vs. physiological selection of spermatozoa for ICSI (picsi) in sibling oocytes.

Physiological selection of spermatozoa for ICSI (PICSI) is a sperm selection method based on sperm binding to hyaluronic acid. Previous studies on the effect of hyaluronic acid binding assays on fertilization and embryo quality have shown inconsistent results. Previous sibling oocyte studies have not found a significant improvement in fertilization or embryo development with hyaluronic acid binding assays. To compare fertilization and embryo development between standard intracytoplasmic sperm injection (ICSI) and PICSI in sibling oocytes. This is a retrospective analysis of all in vitro fertilization (IVF) cycles between January 2017 and April 2020 in which sibling oocytes were randomly fertilized by both ICSI and PICSI. Fertilization rate and the rate of embryos eligible for transfer were compared. Forty-five IVF cycles, in which 257 oocytes were fertilized with PICSI and 294 with standard ICSI, were compared. Most of the patients included in the study had previous failures of fertilization, poor embryonic development, implantation failure, or miscarriage. All but two of the patients had at least one previous unsuccessful IVF cycle. Both fertilization rates (71% vs. 83%) and transfer eligible embryo rates (38% vs. 51%) were significantly higher in PICSI fertilized oocytes (p=0.008 and p=0.01 respectively). Our study is the largest sibling oocyte study comparing ICSI and PICSI, and the first to find a significant improvement in fertilization and embryo quality with PICSI using sibling oocytes. The fact our cohort included almost exclusively couples with previous unsuccessful IVF cycles might suggest that PICSI should be used in selected cases. PICSI improves fertilization rates and transfer eligible embryo rates in sibling oocytes in a selected study group.

The prognostic value of seminal anti-sperm antibodies screening in men prepared for ICSI: a call to change the current antibody-directed viewpoint of sperm autoimmunity testing.

Aims:We aimed to prospectively evaluate the prognostic value of seminal anti-sperm antibodies (ASA) screening in couples prepared for intracytoplasmic sperm injection (ICSI).Methods:A prospective comparative study was conducted, including 184 non-azoospermic infertile men scheduled for ICSI. For each patient, detection of ASA in seminal plasma using the ELISA technique was done and its relation to standard semen parameters and ICSI outcomes, including fertilization, embryo development, and pregnancy rates, was analyzed.Results:Sperm count, motility, and morphology were negatively affected by existence of seminal ASA (p = 0.012, 0.006, and 0.011, respectively). However, no statistically significant difference was detected between patients with positive and negative seminal ASA regarding the median values of fertilization (56.3% versus 66.7%, p = 0.091), percentage of couples with grade A embryo development (91.4% versus 89.9%, p = 0.520), and pregnancy rates (31.4% versus 32.2%, p = 0.98) after ICSI.Conclusion:ICSI seems able to overcome the problem of ASA in semen. The routine screening of ASA in men prepared for ICSI has no additional prognostic value and cannot be recommended for the time being, until more specific antigen-concerned testing can be developed.

Embryo morphokinetics analysis and reproductive outcomes with assisted oocyte activation by a calcium ionophore after prior fertilization failure. A multicentric retrospective study

BackgroundFertilization failure or low fertilization rate after ICSI is around 1–3% in IVF treatments. Several strategies have been studied in order to bypass the lack of activation. The aim of this study is to evaluate embryo morphokinetics and reproductive outcomes after intracytoplasmic sperm injection (ICSI) with assisted oocyte activation (AOA) using a calcium ionophore (CaI) in patients with previous fertilization failure or low fertilization rate (under 30%) and severe male factor. MethodsMulticentric retrospective cohort study including 70 patients with fertilization failure or low fertilization rate and severe male factor (sperm concentration under 1million/mL) who underwent ICSI with CaI (756 oocytes), and 76 patients with severe male factor without previous fertilization failure who had standard ICSI (748 oocytes) between January 2011 and December 2016. ResultsCaI Exposed and non-exposed groups differed significantly for normal fertilization rates, pronuclear disappearance timing (tPNf), time to 4cells stage, multinucleation at the 2- and 4-cell stages, and direct cleavage events. Implantation rate was higher in the non-exposed group (p=0.023). Other morphokinetic variables were similar between groups. Pregnancy (higher in the non exposed group), abortion, and live birth rates, were also not statistically different among groups. ConclusionsAlthough the fertilization and implantation rates were higher in the non-exposed group, ICSI-CaI was associated with an increased fertilization rate compared to patient previous attempts and thus with increased pregnancy chances. Because of the specific patient populations involved (patients with severe male factor with or without previous fertilization failure), the results might not generalize for patients with different etiologies.Trial Registration: 1506-VLC-045-MM.

Physiological, hyaluronan-selected intracytoplasmic sperm injection for infertility treatment (HABSelect): a parallel, two-group, randomised trial

SummaryBackgroundSperm selection strategies aimed at improving success rates of intracytoplasmic sperm injection (ICSI) include binding to hyaluronic acid (herein termed hyaluronan). Hyaluronan-selected sperm have reduced levels of DNA damage and aneuploidy. Use of hyaluronan-based sperm selection for ICSI (so-called physiological ICSI [PICSI]) is reported to reduce the proportion of pregnancies that end in miscarriage. However, the effect of PICSI on livebirth rates is uncertain. We aimed to investigate the efficacy of PICSI versus standard ICSI for improving livebirth rates among couples undergoing fertility treatment.MethodsThis parallel, two-group, randomised trial included couples undergoing an ICSI procedure with fresh embryo transfer at 16 assisted conception units in the UK. Eligible women (aged 18–43 years) had a body-mass index of 19–35 kg/m2 and a follicle-stimulating hormone (FSH) concentration of 3·0–20·0 mIU/mL or, if no FSH measurement was available, an anti-müllerian hormone concentration of at least 1·5 pmol/L. Eligible men (aged 18–55 years) had not had a vasovasostomy or been treated for cancer in the 24 months before recruitment and were able, after at least 3 days of sexual abstinence, to produce freshly ejaculated sperm for the treatment cycle. Couples were randomly assigned (1:1) with an online system to receive either PICSI or a standard ICSI procedure. The primary outcome was full-term (≥37 weeks' gestational age) livebirth, which was assessed in all eligible couples who completed follow-up. This trial is registered, number ISRCTN99214271.FindingsBetween Feb 1, 2014, and Aug 31, 2016, 2772 couples were randomly assigned to receive PICSI (n=1387) or ICSI (n=1385), of whom 2752 (1381 in the PICSI group and 1371 in the ICSI group) were included in the primary analysis. The term livebirth rate did not differ significantly between PICSI (27·4% [379/1381]) and ICSI (25·2% [346/1371]) groups (odds ratio 1·12, 95% CI 0·95–1·34; p=0·18). There were 56 serious adverse events in total, including 31 in the PICSI group and 25 in the ICSI group; most were congenital abnormalities and none were attributed to treatment.InterpretationCompared with ICSI, PICSI does not significantly improve term livebirth rates. The wider use of PICSI, therefore, is not recommended at present.FundingNational Institute for Health Research Efficacy and Mechanism Evaluation Programme.

Do immature and mature sibling oocytes recovered from stimulated cycles have the same reproductive potential?

How can laboratory and clinical outcomes of spontaneously, early maturing germinal-vesicle oocytes and sibling in-vivo-matured (metaphase II [MII]) oocytes be quantified and compared? A prospective, non-randomized intra-cohort study of oocytes from women aged 38 years or younger, with six or fewer MII oocytes and four or more germinal vesicles retrieved. No indication was identified for genetic tests or oocyte or embryo cryopreservation. The study was carried out at IVIRMA-Valencia. Early maturing germinal vesicles were selected for reproductive purposes. In vitro- and in-vivo MII oocytes were fertilized. After time-lapse culture, hatching blastocysts from germinal vesicles were biopsied for aneuploidy screening and vitrified. Laboratory and clinical outcomes were compared according to oocyte origin. Almost 70% of germinal vesicles had matured early and spontaneously, and had comparable in vitro-outcomes and morphokinetics to sibling in vivo-matured oocytes. Fifty per cent of biopsied blastocysts were euploid. Germinal-vesicle rescue increased the number of MII oocytes per cycle to 3.9, finally adding one extra-blastocyst per cycle. A live birth confirmed the feasibility of this approach. Further data, however, are needed to quantify its real contribution to standard intracytoplasmic sperm injection cycles. Nevertheless, 40% of patients obtained either an immediate advantage (reduction of cancellation rate) or long-term benefit (availability of extra blastocysts of attempts). Germinal-vesicle rescue can be considered as a complementary approach when folliculometry (expected) and number of MII (observed) are unequal.

The clinical and neonatal outcomes after stimulation of immotile spermatozoa using SperMagic medium.

To evaluate the efficiency and safety of SperMagic medium on stimulating the immotile spermatozoa in testicular sperm extraction (TESE) and absolute asthenozoospermia, 96 patients with TESE and 106 patients with absolute asthenozoospermia were enrolled in this study. The motile spermatozoa were detected in 47 TESE patients and 68 absolute asthenozoospermia and these patients were assigned to control group. The immotile spermatozoa in 49 TESE patients and 34 absolute asthenozoospermia were stimulated with SperMagic medium. Patients were treated by standard intracytoplasmic sperm injection (ICSI). There were no significant differences in fertilisation, cleavage, implantation, pregnancy, live birth and neonatal outcomes. SperMagic medium does not increase incidence of adverse neonatal outcomes and is a reliable tool for selection of viable spermatozoa in ICSI.

Claims for fertility interventions: a systematic assessment of statements on UK fertility centre websites

ObjectivesFertility services in the UK are offered by over 200 Human Fertilisation and Embryology Authority (HFEA)-registered NHS and private clinics. While in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI)...

Delayed intracytoplasmic sperm injection (ICSI) with trophectoderm biopsy and preimplantation genetic screening (PGS) show increased aneuploidy rates but can lead to live births with single thawed euploid embryo transfer (STEET).

The aim of this study was to report the results of IVF with trophectoderm biopsy and preimplantation genetic screening (PGS) following delayed intracytoplasmic sperm injection (ICSI). Patients undergoing IVF with PGS and delayed ICSI were included in the study. Indications for delayed ICSI included absent or poor fertilization via standard insemination or more than 50% immature oocytes, noted post-cumulus stripping for standard ICSI procedure. Delayed ICSI was performed the day after retrieval due to absent or poor fertilization. The immature oocytes were kept in extended culture, and if demonstrated maturity, ICSI was performed. Primary outcome included fertilization rate and blastocyst stage formation, defined by the number of blastocysts for biopsy. Secondary outcome included aneuploidy rate and pregnancy outcomes following single thawed euploid embryo transfers (STEET). Sixteen patients with delayed ICSI were included in the study. Twelve were due to poor fertilization and four secondary to immature oocytes. A total of 219 oocytes were retrieved; ten were frozen upon patient request, 168 had standard insemination, and 13 had routine ICSI on the day of retrieval. A total of 129 oocytes underwent delayed ICSI. Sixty-three (49%) fertilized, 19 (14.7%) reached blastocysts for biopsy; fivw of which were chromosomally normal (26.3%). Three patients underwent STEET of a delayed ICSI embryo; all three resulted in live births, including one embryo biopsied on day 8 of development. Fertilization failure or an excessive proportion of immature oocytes in an IVF cycle, necessitating delayed ICSI, showed equivalent fertilization and blast formation rates. With the implementation of trophectoderm biopsy and PGS, these embryos can lead to healthy live born babies.

Commentary: Physiological Intracytoplasmic Sperm Injection (PICSI), An Alternative to the Standard ICSI Procedure

The rationale of this study is to uncover sperm selection by hyaluronic acid (HA) binding which could advance fertilization pace and embryo quality in Intracytoplasmic sperm injection (ICSI) cycles. HA a sperm binding nonsulfated glycosaminoglycan required as a part of normal fertilisation. Physiological Intracytoplasmic Sperm Injection employs sperm binding to Hyaluronic acid to opt for which sperm to use for Intracytoplasmic sperm injection. PICSI and their latent benefit are still in tranquil being explored. We expect more detailed facts in the coming few years in relation to for whom these modus operandi are most useful apart from cases discussed. Usually if the sperm binding is less than 65% the PICSI dish is used to select the sperm. The medley of sperm selection for ICSI is essential and directly authorise the male input to the embryo therefore visual evaluation alone should certainly not be the only aspect involved when selecting a sperm for ICSI failed cycle twice. But unfit sperm with DNA fragmentation and compromised maturity is able to appear normal and healthy, but if selected for injection/insemination can go ahead to deprived embryo quality and likely a pregnancy loss.

Does embryo ploidy status differ in patients whose eggs were fertilized by standard insemination versus intra cytoplasmic sperm injection (ICSI)?

To determine if standard insemination technique versus ICSI leads to measurable differences in embryonic chromosomal status. Retrospective analysis. 26 patients undergoing ovarian stimulation/pre-gestational screening protocols at our center from 2013-2014 were included. Patients were only included if they underwent both standard insemination and ICSI within a single egg retrieval cycle, thus patients served as their own controls. Primary outcomes included fertilization rates, number of embryos that reached an appropriate stage (2Bc or greater) for trophoectoderm biopsy (TEBx) and number of euploid embryos. T-tests of means and paired t-tests were used for statistical analysis. Average age of the patients in this study was 35.6 years +/- 5.2. Average day 2 estradiol (pg/ml) and follicle stimulating hormone (mIU/ml) values were 55.1 +/-56.5 and 6.75 +/- 2.87. Average total gonadotropin dosage (IU) and estradiol at trigger were 3648.6 +/- 144.6 and 2789 +/- 1418.3, respectively. Approximately 19.8 +/- 9.3 oocytes on average were retrieved with a maturation percentage of 81%. Insemination fertilization rates were 69% +/- 23.8 and 75% +/- 13.8 (p-value of 0.28) for the ICSI cohort. The percentage of embryos available for TEBx for the insemination cohort was 32% and 38% for the ICSI cohort, with a p-value of 0.15 and RR 0.84 (0.652-1.07). 52.8% of embryos biopsied resulting from ICSI and 47% of embryos from standard insemination were euploid, and paired t-test analysis showed no significance between the two groups, p = 0.07. Standard insemination techniques and ICSI showed no difference in fertilization rates, and no difference in percent of chromosomally competent embryos. In the absence of standard indications for ICSI, there does not appear to be any benefit or harm in the use of ICSI for increased development of chromosomally competent embryos.

Influence of different oocyte insemination techniques on early and late morphokinetic parameters: retrospective analysis of 500 time-lapse monitored blastocysts

To determine how standard IVF vs. intracytoplasmic sperm injection (ICSI) fertilization influences early and late morphokinetic parameters during prolonged embryo culture. Five-hundred expanded blastocysts that were monitored in a time-lapse monitoring incubator were analysed retrospectively. Early (pronuclear fading [PNf], t2-t9) and late (start of blastulation, expanded blastocyst) morphokinetic variables were scored according to published consensus criteria. Private infertility clinic. A total of 209 consecutive infertile patients (mean ± SD age, 38.4 ± 4 years; range, 28-47 years) undergoing 238 natural IVF/minimal ovarian stimulation cycles during 2012-2014. Minimal ovarian stimulation, oocyte retrieval, fertilization with standard IVF or ICSI, prolonged embryo culture in a time-lapse monitoring incubator. Differences in morphokinetic parameters according to insemination techniques. In total, 29% and 71% of the whole cohort was fertilized with standard IVF and ICSI, respectively. During early cleavage stages (PNf to t4) there was a statistically significant delay (+1.5 to +1.1 hours) among IVF-fertilized embryos. By contrast, at the expanded blastocyst stage IVF-fertilized embryos showed faster development (-3.3 to -4.1 hours). After normalizing to the time point of PNf, differences in cleavage-stage parameters disappeared, but those at all blastocyst stages increased even further in favor of IVF-fertilized embryos (-3.2 to -5.7 hours). The observed 1.5-hour time difference between standard IVF- and ICSI-fertilized embryos is an artificial phenomenon. At the blastocyst stages, however, genuine timing differences arise between IVF- and ICSI-fertilized embryos, possibly related to their different quality. Normalization to a common time point permits the joint analysis of IVF- and ICSI-fertilized embryos, thus increasing the size of studied cohorts.

Advanced sperm selection techniques for assisted reproduction.

Assisted reproductive technologies (ART) such as in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) bring together gametes outside of the body to enhance the probability of fertilisation and pregnancy. Advanced sperm selection techniques are increasingly being employed in ART, most commonly in cycles utilising ICSI. Advanced sperm selection techniques are thought to improve the chance that structurally intact and mature sperm with high DNA integrity are selected for fertilisation. Advanced sperm selection strategies include selection according to surface charge; sperm apoptosis; sperm birefringence; ability to bind to hyaluronic acid; and sperm morphology under ultra-high magnification. These techniques theoretically improve ART outcomes. To evaluate the impact of advanced sperm selection techniques on ART outcomes. Systematic search of electronic databases (Cochrane Menstrual Disorders and Subfertility Group Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, PsycINFO, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Latin American and Caribbean Health Science Information Database (LILACS)), trials registers (ClinicalTrials.gov, Current Controlled Trials, World Health Organization International Clinical Trials Registry Platform), conference abstracts (Web of Knowledge) and grey literature (OpenGrey) for relevant randomised controlled trials. We handsearched the reference lists of included studies and similar reviews. The search was conducted in May 2014. We included randomised controlled trials (RCTs) comparing an advanced sperm selection technique versus standard IVF or ICSI or versus another advanced sperm selection technique. We excluded studies of sperm selection using ultra-high magnification (intracytoplasmic morphologically selected sperm injection, or IMSI), as they are the subject of a separate Cochrane review. Quasi-randomised and pseudo-randomised trials were excluded. Our primary outcome measure was live birth rate per woman randomly assigned. Secondary outcome measures included clinical pregnancy per woman randomly assigned, miscarriage per clinical pregnancy and fetal abnormality per clinical pregnancy. Two review authors independently assessed eligibility of studies and risk of bias, and performed data extraction. Disagreements were resolved by consultation with a third review author. Study investigators were consulted to resolve other queries that arose. Risk ratios (RRs) were calculated with 95% confidence intervals (CIs). We planned to combine studies using a fixed-effect model, if sufficient data were available. The quality of the evidence was evaluated using Grades of Recommendation, Assessment, Development and Evaluation (GRADE) methods. Two RCTs were included in the review. Both evaluated sperm selection by hyaluronanic acid binding for ICSI, but only one reported live births. No studies were identified that were related to surface charge selection, sperm apoptosis or sperm birefringence.One RCT compared hyaluronanic acid binding versus conventional ICSI. Live birth was not reported. Evidence was insufficient to show whether there was a difference between groups in clinical pregnancy rates (RR 1.01, 95% CI 0.84 to 1.22, one RCT, 482 women). This evidence was deemed to be of low quality, mainly as the result of poor reporting of methods and findings. Miscarriage data were unclear, and fetal abnormality rates were not reported.The other RCT compared two different hyaluronanic acid binding techniques, SpermSlow and physiological intracytoplasmic sperm injection (PISCI). Evidence was insufficient to indicate whether there was a difference between groups in rates of live birth (RR 1.16, 95% CI 0.65 to 2.05, one RCT, 99 women), clinical pregnancy (RR 1.07, 95% CI 0.67 to 1.71, one RCT, 99 women) or miscarriage (RR 0.76, 95% CI 0.24 to 2.44, one RCT, 41 women). The evidence for these comparisons was deemed to be of low quality, as it was limited by imprecision and poor reporting of study methods. Fetal abnormality rates were not reported. Evidence was insufficient to allow review authors to determine whether sperm selected by hyaluronanic acid binding improve live birth or pregnancy outcomes in ART, and no clear data on adverse effects were available. Evidence was also insufficient to show whether there is a difference in efficacy between the hyaluronic acid binding methods SpermSlow and PICSI. No randomised evidence evaluating sperm selection by sperm apoptosis, sperm birefringence or surface charge was found.Further studies of suitable quality are required to evaluate whether any of these advanced sperm selection techniques can be recommended for use in clinical practice.

Epigenetic status in the offspring of spontaneous and assisted conception.

Is DNA methylation in buccal cell DNA from children born following IVF (in vitro fertilization) and ICSI (intra-cytoplasmic sperm injection) different from that of spontaneously conceived children? DNA methylation in the imprinted gene, small nuclear ribonucleoprotein polypeptide N (SNRPN), was higher in children conceived by ICSI and in those born to women with the longest duration of infertility regardless of the method of conception. Fertility treatment is associated with a small but significant increase in the risk of a range of adverse obstetric outcomes, birth defects and longer term sequelae, but the biological basis for this is unknown. A growing evidence base suggests that epigenetics may play a role in subfertility and the link between fertility and health. In this retrospective cohort study of children born between 2002 and 2008, we measured DNA methylation in paternally expressed gene 3 (PEG3), insulin-like growth factor II (IGF2), SNRPN, long interspersed nuclear element 1 (LINE1) and the insulin gene (INS) in buccal cell DNA from children born following IVF (n = 49) and ICSI (n = 20) and compared them with a matched spontaneous conception group (n = 86). Participants were identified from the Aberdeen Maternity and Neonatal Databank and IVF and ICSI pregnancies were matched to spontaneous conception pregnancies on year of birth and maternal age at delivery. Only singleton pregnancies following fresh embryo transfer were included. DNA methylation was determined by pyrosequencing. Regression with adjustment for covariates was used to determine the effect of infertility on offspring DNA methylation. SNRPN methylation in the offspring was linked to fertility treatment in the parents. This effect was specific to children conceived using ICSI and was apparent in the comparison of ICSI versus spontaneous conception (1.03%; 95% CI 0.10, 1.97; P = 0.031), ICSI versus standard IVF (1.13%; 95% CI 0.04, 2.23; P = 0.043) and ICSI versus standard IVF and spontaneous conception (1.05; 95% CI 0.15, 1.94; P = 0.023). In all comparisons, the use of ICSI was associated with a higher level of SNRPN methylation in the offspring. A higher level of SNRPN methylation in the offspring was also associated with a longer duration of infertility in the parents. This was observed in all cases of infertility (0.18% per year of infertility; 95% CI 0.02, 0.33; P = 0.026) and after excluding ICSI cases (0.21% per year of infertility; 95% CI 0.04, 0.37; P = 0.017). There was a significant increase in the level of LINE1 methylation with age between birth and 7 years (0.77% per year; 95% CI 0.49, 1.05; P < 0.001). Methylation in the INS gene decreased significantly over the same period (-0.46% per year; 95% CI -0.89, -0.03; P = 0.035). There was no evidence from this cross-sectional data that methylation within the imprinted genes changed over the first 7 years of life. The ICSI sample size was limited but the groups were carefully selected and well matched and the SNRPN findings were consistent across different outcomes. The results of this study provide support for a role for epigenetics, and imprinting in particular, in fertility. The specific changes point to possible long-term consequences of fertility treatment for the health and fertility of future generations. The authors report no conflict of interest in relation to this work. Funding was provided by the University of Aberdeen and the Scottish Government. Not applicable.

V7-02 ROBOTIC LEVEL 3 CAVA THROMBECTOMY

increased DNA fragmentation and clinical outcome from IMSI, although results are conflicting. The aim of this study was to compare pregnancy rate (PR) from standard intra-cytoplasmic sperm injection (ICSI) and IMSI, in couples who had failed previous ART attempts in whom the male partner had been demonstrated to have a raised DFI. METHODS: Data was collected prospectively between July 2011 and October 2012 at a fertility clinic. Couples who had undergone at least one previous failed attempt of ART at the clinic were advised DNA fragmentation testing using the TUNEL technique. DNA fragmentation was considered raised if the index (DFI) was above 30%. Couples were offered either standard ICSI or IMSI and PR, live birth rates (LBR) and miscarriage rates were compared. Sperm for injection with the IMSI technique were selected under 6600 x magnification based on the motile sperm organelle morphology examination (MSOME) as previously described by Bartoov et. al. RESULTS: 54 couples were included in the study analysis (ICSI, n1⁄420; IMSI n1⁄434). Mean age in women (ICSI, 38; IMSI, 37) (p1⁄40.14) and menwere similar in both groups (ICSI, 40; IMSI, 42) (p1⁄40.31). There was no statistical difference in themeanDFI in both coupleswho had ICSI and IMSI (ICSI, 44.3%; IMSI, 48.3%) (p1⁄40.34). Therewas no difference in the number of oocytes collected (ICSI, 9.0; IMSI, 12.5) (p1⁄40.06) or in the number of oocytes fertilized (ICSI, 5.1; IMSI 6.7) (p1⁄40.17). Two women who had ICSI had no fertilization of their oocytes on the day following egg collection and were excluded in the final analysis. Of women who had embryo transfer (n1⁄452), there was no difference in PR (ICSI, 55.6%; IMSI, 52.9%) (p1⁄40.56) or LBR (0.61)(ICSI, 50.0%; IMSI, 50.0%) between groups. Miscarriage rate was 5.6% in the ICSI group and 8.8% in the IMSI group. CONCLUSIONS: This study would suggest that IMSI may not improve the pregnancy outcome from ART in couples where men are found to have high DNA fragmentation.

Analysis of clinical outcomes with respect to spermatozoan origin after artificial oocyte activation with a calcium ionophore

Fertilization failures have occurred repeatedly in reproductive centers after intracytoplasmic sperm injection (ICSI) and artificial oocyte activation (AOA) has been used to prevent it. This study was performed to investigate whether spermatozoan origin influences clinical outcomes of AOA with a calcium ionophore. A total of 185 ICSI cycles with a history of no or low fertilization was included in this retrospective study. The outcomes of AOA after ICSI were compared with ejaculated-normal, ejaculated-oligo-astheno-terato or extracted-testicular spermatozoa. There were significant differences between the previous standard ICSI cycles and AOA cycles in the rate of fertilization and clinical outcomes among cases with different sperm origins. Thirty-eight healthy babies (20 singles and 18 twins, 29cycles) were successfully delivered, and no congenital birth defects were observed. Most patients with a no or low fertilization history obtained an increased fertilization rate and a positive clinical outcome with AOA regardless of the origin of spermatozoa.