Intracytoplasmic Sperm Injection Fertilization Research Articles

P-066 A multi-bioassay approach to identify male gamete fertilization dysfunction

Abstract Study question Can a multi-bioassay approach identify a specific male gamete dysfunction often overlooked by a standard semen analysis? Summary answer In cases of persistent fertilization failure, additional testing may assist in pinpointing underlying factors contributing to compromised sperm embryo developmental competence and tailor treatment. What is known already Conventional semen analysis, while informative, ineffectively captures the complete spectrum of male gamete dysfunction. Despite adequate sperm concentration and motility, some couples may still experience fertilization failure with Intracytoplasmic Sperm Injection (ICSI). This highlights the complexity of male infertility, that extends beyond standard parameters. In addressing sperm-related fertilization challenges, various assisted oocyte activation (AOA) methods have been proposed to trigger intracellular Ca2+ release, leading to calcium spikes and meiosis initiation. Study design, size, duration In the past 10 months, we identified couples (n = 50) with a history of poor ICSI fertilization, exhibiting suboptimal head morphology. Screening for acrosomal dysfunction involved assessing the presence of PLCxand related gene mutations. We measured the degree of chromatin compaction by histone-to-protamine ratio and evaluated sperm chromatin fragmentation (SCF). To correct for eventual acrosomal dysfunction, couples were counseled to undergo subsequent ICSI cycles with AOA. Clinical outcome was compared between historical and treatment cycles. Participants/materials, setting, methods Fifty men with a history of poor ICSI fertilization were included. A standard semen analysis was conducted according to WHO guidelines. The presence of PLCx was measured through immunofluorescent staining (normal threshold, ≥ 30%), corroborated by gene mutation detection though DNA-seq. Sperm histone-to-protamine ratio was evaluated by Aniline Blue (normal threshold, <15% histone retention). SCF was assessed utilizing the TUNEL assay (normal threshold, ≤15%). AOA was performed using ionomycin or recombinant PLCx protein. Main results and the role of chance Fifty men experienced a poor clinical outcome following ICSI with a fertilization of 7.4% (48/647), embryo implantation of 6.6% (1/15), clinical pregnancy of 6.6% (1/15), and pregnancy loss rate of 100% (1/1). Their semen analysis, interestingly, revealed the following semen parameters: 3.0±1 mL volume, 50.0±20 x106/mL concentration, 41.4±21% motility, and 1.9±1% normal morphology. Nonetheless, PLCx content was diminished, with values ranging from 2-27%. Genomic assessment confirmed mutations in PLCx, DPY19L2, SPATA16 and PICK1 genes, all implicated in sperm nuclear remodeling and acrosome formation. An elevated histone content at 29.7% was positively correlated with an increased SCF at 18.0% (R2=0.6, P<0.001). In subsequent cycles employing AOA, the normal fertilization increased to 42.3% (305/721), the embryo implantation reached 13.0% (9/69), the clinical pregnancy rose to 21.1% (8/38), and the delivery rate was 21.1% (8/38), without any pregnancy loss. (P<0.001). Limitations, reasons for caution While this study affirms the effectiveness of a multifaceted assessment in pinpointing specific sperm dysfunctions and customizing treatment, it is still a retrospective observation requiring independent validation in a larger population. Wider implications of the findings Semen analysis alone fails to address specific aspects of male gamete dysfunction. Incorporating additional bioassays to assess chromatin structure, integrity, and acrosomal function offers valuable insights for fertility management in tailoring personalized treatments. This comprehensive approach enhances the understanding of male reproductive health, maximizing the potential for successful clinical outcome. Trial registration number not applicable

Successful pregnancy with intracytoplasmic sperm injection after bacterial contamination of embryo culture in in vitro fertilization: a case report

BackgroundBacterial infection of embryo culture medium is rare but may be detrimental. The main source of embryo culture contamination is semen. Assisted reproduction centers currently lack consensus regarding the methods for preventing and managing embryo culture infection. In our recent case, a successful pregnancy was achieved with intracytoplasmic sperm injection after failed conventional in vitro fertilization owing to bacterial contamination.Case presentationWe present a case report of two consecutive in vitro fertilization–intracytoplasmic sperm injection cycles with photo and video documentation of the bacterial growth. A 36-year-old Hungarian woman and her 37-year-old Hungarian partner came to our department. They had two normal births followed by 2 years of infertility. The major causes of infertility were a closed fallopian tube and asthenozoospermia. Bacterial infection of the embryo culture medium was observed during in vitro fertilization and all oocytes degenerated. The source was found to be the semen. To prevent contamination, intracytoplasmic sperm injection was used for fertilization in the subsequent cycle. Intracytoplasmic bacterial proliferation was observed in one of the three fertilized eggs, but two good-quality embryos were successfully obtained. The transfer of one embryo resulted in a successful pregnancy and a healthy newborn was delivered.ConclusionIntracytoplasmic sperm injection may be offered to couples who fail conventional in vitro fertilization treatment owing to bacteriospermia, as it seems to prevent infection of the embryo culture. Even if bacterial contamination appears, our case encourages us to continue treatment. Nevertheless, the development of new management guidelines for the prevention and management of bacterial contamination is essential.

Metabolomic profiling of embryo culture media in patients with repeated implantation failure during assisted reproductive technology cycles.

This study investigated the metabolic status of the spent culture media from embryos of patients with repeated implantation failure (RIF) undergoing in vitro fertilization-intracytoplasmic sperm injection cycles in comparison with the embryos from healthy fertile women. Metabolite levels in spent culture media were assessed and compared between embryos from RIF patients (n=35) and oocyte donors as controls (n=15). Protein levels of insulin-like growth factor 1 (IGF-1) were determined using Western blotting. Concentrations of glucose, pyruvate, and lactate were measured using spectrophotometry. Ionic colorimetric assay kits were utilized to analyze the concentrations of sodium, chloride, calcium, and magnesium ions. High-performance liquid chromatography was employed to measure the concentrations of glutamic acid, aspartic acid, methionine, phenylalanine, and histidine. Glucose consumption and lactate secretion were higher in the control group than in the RIF group. The magnesium concentration was significantly higher in the control group than in the RIF group, but glutamic acid and aspartic acid concentrations were lower in the control group than in the RIF patients (p<0.05). The levels of IGF-1, sodium, calcium, chloride, methionine, histidine, and phenylalanine did not show statistically significant differences between the two groups. The metabolic profile of the culture medium of the embryos in the RIF group differed from that of the control group. These findings suggest potential factors that may affect implantation capacity in RIF patients and provide a new perspective on embryo selection.

Aggressive sperm immobilization improves reproductive outcomes in patients with suboptimal semen parameters and previous ICSI fertilization failure

Intracytoplasmic sperm injection (ICSI) is the most effective procedure to resolve male infertility, enhancing overall fertilization and pregnancy outcomes. However, it is important to note that fertilization failure (FF) can still occur in a few cases after ICSI. This study aims to introduce a specialized technique of aggressive sperm immobilization for ICSI and evaluate its impact on reproductive outcomes in cases involving prior fertilization failure. All infertile couples with male partners having suboptimal semen samples and previous ICSI fertilization failure were evaluated using retrospective data from National Taiwan tertiary university hospital (NTUH) between January 2016 and February 2022. Fertilization failure in our study was defined as less than 30% fertilization rate (FR, the number of normally fertilized oocytes divided by the total number of injected mature oocytes). Data involving both standard (routine procedure) and aggressive sperm immobilization (SI) techniques during different ICSI cycles were included in this study. Standard and aggressive SI methods were performed by compressing the distal half tail of the spermatozoa ≦ 5 and 15 times prior to ICSI respectively. Generalized estimating equations analysis were applied to compare the clinical outcomes between two procedures. Overall, data from 23 infertile couples who had undergone 65 ICSI cycles (31 standard SI with low fertilization rate and 34 aggressive SI) were included in the study. The average FR in the ICSI cycles with standard SI and aggressive SI were 23.6 ± 23.1% and 49.5 ± 31.8 respectively (P = 0.0002). The majority of embryos were transferred at the day 3 stage, with an average number transferred of 2.6 ± 0.9 in the aggressive SI group and 1.9 ± 0.9 in the standard group. The number of embryos transferred per transfer cycle was higher in the aggressive SI (P = 0.015), whereas the number of good-quality embryos was similar between the two procedures (P = 0.44). There were one and seven live births from the standard SI cycles and aggressive SI cycles respectively. In conclusion, aggressive SI was associated with a significantly higher FR, resulting in more available embryos for transfer without compromising embryo quality. Therefore, this specialized technique improved pregnancy outcome among infertile couples with a previous ICSI–FF. It can be a safe, economic, and effective method to improve the assisted reproductive technologies outcomes for infertile patients affected by previous ICSI–FF.

Ex-vivo microscopic oncotesticular sperm extraction: step-by-step surgical technique at time of radical orchiectomy

To demonstrate the intraoperative surgical techniques required for simultaneous radical orchiectomy and microscopic oncotesticular sperm extraction (m-OncoTESE) in a step-by-step fashion. Video presentation. University Hospital (University of Chicago). A 37-year-old man (status after right orchiectomy at another institution for stage II-C testicular seminoma with positive preoperative tumor markers) was referred for contralateral orchiectomy of multifocal left testis mass and fertility preservation. Semen analysis before, microscopic testicular sperm extraction during, and semen or testicular specimen analysis after the first orchiectomy were unable to identify any sperm. A postoperative analysis of the m-OncoTESE performed on the left testis resulted in the cryopreservation of 200,000 motile sperm for future assisted reproductive technology (i.e., invitro fertilization or invitro fertilization-intracytoplasmic sperm injection). Left radical orchiectomy and left m-OncoTESE. A comprehensive visual documentation of m-OncoTESE surgical techniques with concurrent commentary detailing the reasons behind each surgical step. A brief discussion on the background of m-OncoTESE and alternative fertility preservation methods accompanies the procedure. This video provides a step-by-step guide to performing an m-OncoTESE (proceeding a radical orchiectomy in a patient with testicular cancer) as a means of fertility preservation in an azoospermic patient. Successful extraction and cryopreservation of testicular spermatozoa were achieved after targeted ex-vivo testicular microdissection. Sperm extraction via m-OncoTESE is a viable option for azoospermic patients with testicular cancer undergoing radical orchiectomies. The use of preoperative imaging and microsurgical techniques facilitates and optimizes surgical dissection and sperm recovery.

Involvement of CATSPER 2 mutation in a familial context of unexplained infertility and fertilization failure associated with hearing loss: a case report

To explore the functional implications of a homozygous CATSPER 2 (cation channel for sperm) deletion within the acrosome reaction pathway during fertilization in 2 brothers, who have unexplained infertility and hearing loss. Case report. Two twin brothers aged 30 years with hearing loss and unexplained infertility. Molecular genetic diagnosis of deafness. Evaluation of the acrosome reaction and calcium mobilization assays after induction by progesterone and ionomycin on spermatozoa of the CATSPER 2-mutated patient and on fertile controls. Fertilization rate during conventional in vitro fertilization. Molecular genetic test. Percentage of acrosome-reacted spermatozoa with peanut agglutinin lectin staining. Recording of progesterone and ionomycin-induced intracellular calcium signals with a fluorescent probe. Mr. S and his brother have normal, conventional sperm parameters. Both brothers have had repeated intrauterine insemination failures and one fertilization failure after conventional in vitro fertilization. Mr. S obtained 2 healthy babies after intracytoplasmic sperm injection. Genetic analysis found a homozygote deletion of the STRC (stereocilin) gene (NM 153700: c.1-? 5328+?del) that removes the CATSPER 2 gene. Mutation of the STRC gene is known to be associated with hearing loss. Sperm functional tests revealed an inability of progesterone to activate intracellular calcium signaling and to induce acrosome reaction. We demonstrate the absence of a calcium signal and acrosome reaction after progesterone in our patient with a CATSPER 2 mutation. We emphasize the importance of the male medical interview and of the genetic investigation of hearing loss. We show that in vitro fertilization-intracytoplasmic sperm injection is necessary, even where normal sperm parameters are present.

Molecular mechanisms of cellular dysfunction in testes from men with non-obstructive azoospermia.

Male factor infertility affects 50% of infertile couples worldwide; the most severe form, non-obstructive azoospermia (NOA), affects 10-15% of infertile males. Treatment for individuals with NOA is limited to microsurgical sperm extraction paired with in vitro fertilization intracytoplasmic sperm injection. Unfortunately, spermatozoa are only retrieved in ~50% of patients, resulting in live birth rates of 21-46%. Regenerative therapies could provide a solution; however, understanding the cell-type-specific mechanisms of cellular dysfunction is a fundamental necessity to develop precision medicine strategies that could overcome these abnormalities and promote regeneration of spermatogenesis. A number of mechanisms of cellular dysfunction have been elucidated in NOA testicular cells. These mechanisms include abnormalities in both somatic cells and germ cells in NOA testes, such as somatic cell immaturity, aberrant growth factor signalling, increased inflammation, increased apoptosis and abnormal extracellular matrix regulation. Future cell-type-specific investigations in identifying modulators of cellular transcription and translation will be key to understanding upstream dysregulation, and these studies will require development of in vitro models to functionally interrogate spermatogenic niche dysfunction in both somatic and germ cells.

Defects in phospholipase C zeta cause polyspermy and low fertilization after conventional IVF: not just ICSI failure.

Phospholipase C zeta (PLCζ) is a key sperm-borne oocyte-activating factor that triggers Ca 2+ oscillations and the subsequent block to polyspermy following gamete fusion. Mutations in PLCZ1 , the gene encoding PLCζ, cause male infertility and intracytoplasmic sperm injection (ICSI) fertilization failure; and PLCζ expression and localization patterns are significantly correlated with ICSI fertilization rate (FR). However, in conventional in vitro fertilization (cIVF), whether and how sperm PLCζ affects fertilization remain unclear. Herein, we identified one previously reported and two novel PLCZ1 mutations associated with polyspermy in vitro that are characterized by excessive sperm-zona binding and a delay in pronuclei (PN) formation. Immunofluorescence staining and oocyte activation testing revealed that virtually all spermatozoa from patients lacked functional PLCζ and were thus unable to evoke Ca 2+ oscillations. ICSI with an artificial oocyte activation treatment successfully rescued the polyspermic phenotype and resulted in a live birth. Furthermore, we analyzed PLCζ in an additional 58 males after cIVF treatment in the Reproductive and Genetic Hospital of CITIC-Xiangya (Changsha, China) between February 2019 and January 2022. We found that the proportion of spermatozoa that expressed PLCζ was positively correlated with both 2PN rate and total FR. The optimal cutoff value below which males were likely to experience low FR (total FR ≤30%) after cIVF was 56.7% for the proportion of spermatozoa expressing PLCζ. Our study expands the mutation and the phenotypic spectrum of PLCZ1 and further suggests that PLCζ constitutes a promising biomarker for identifying low FRs cases in cIVF due to sperm-related oocyte activation deficiency and that sperm PLCζ analysis may benefit the wider male population and not only men with ICSI failure.

Microdissection Testicular Sperm Extraction.

Nonobstructive azoospermia (NOA) is among the most common causes of male infertility. For men with NOA seeking fertility treatment, microdissection testicular sperm extraction (microTESE) is the best option for retrieving sperm, which can be used with in vitro fertilization-intracytoplasmic sperm injection to achieve pregnancy in their partner. With the aid of the operating microscope, microTESE allows for thorough evaluation of the testis tissue and selection of seminiferous tubules that appear most capable of sperm production. Rates of success with microTESE vary depending on the underlying cause of NOA and the center at which the procedure is performed. Not all patients are candidates for microTESE, and those who are candidates should be counseled on the likelihood of sperm retrieval and the potential for changes in postoperative testis function.

Benefits of Blastocyst Transfer With at Least Three Good-Quality Cleavage-stage Embryos in Women of Advanced Maternal Age: A Retrospective Analysis

ObjectivesLimited studies on the benefits of blastocyst transfer in advanced maternal age (AMA) (≥40 years) have been reported. Our objective was to find whether blastocyst-stage embryo transfer improves pregnancy and live birth rates in women ≥40 years who have 3 or more good-quality cleavage-stage embryos. MethodsAll fresh in vitro fertilization-intracytoplasmic sperm injection cycles performed from January 2020 to December 2021 in AMA women that progressed to transfer were considered for analysis. We compared fresh and cumulative ongoing pregnancy rates in AMA women of those who had a cleavage-stage transfer, while meeting the criteria for extended culture (≥3 high-quality embryos, group 1), and those who underwent blastocyst transfer (group 2). Demographic parameters, stimulation, embryology, fresh and cumulative ongoing pregnancy rates, and clinical miscarriage rates were compared. ResultsDuring the study period, 255 cycles were analyzed including group 1 (n = 99) and group 2 (n = 156). Group 1 participants were older and had a greater number of embryos for transfer. Fresh and cumulative ongoing pregnancy rates per transfer were higher in group 2 compared to group 1 (23.4% vs. 13.1%, P = 0.04; 25.5% vs. 14.1%, P = 0.03), while overall miscarriage rates were higher in group 1 than group 2 (51.7% vs. 25%, P = 0.01). ConclusionsBlastocyst culture provides a benefit to AMA women who have at least 3 good-quality embryos on day 3 resulting in significantly higher fresh and cumulative ongoing pregnancy rates and lower miscarriage compared to cleavage-stage transfers.

Artificial oocyte activation with Ca2+ ionophore improves reproductive outcomes in patients with fertilization failure and poor embryo development in previous ICSI cycles.

Does artificial oocyte activation (AOA) by a calcium ionophore (ionomycin) improve the previous fertilization failure or poor embryo development of intracytoplasmic sperm injection (ICSI) account for male factor infertility or other infertility causes? This retrospective study involved 114 patients receiving ICSI-AOA in Shanghai First Maternity and Infant Hospital with previous ICSI fertilization failure or poor embryo development. The previous ICSI cycles of the same patients without AOA served as the control group. The fertilization rates, cleavage rates, transferable embryo rates and blastocyst formation rates of the two groups were compared. Additionally, the clinical pregnancy, implantation rate and live birth rates were also compared to assess the efficiency and safety of AOA. Furthermore, two subgroup analyses were performed in this study based on the cause of infertility and the reason for AOA. The fertilization rate, embryonic development potential and clinical outcome were compared among groups. Among 114 ICSI-AOA cycles, the fertilization rate, top-quality embryo rate, implantation rate, clinical pregnancy per patient and live birth rate per patient were improved significantly compared with previous ICSI cycles (p<0.05 to P< 0.001), and the miscarriage rate in the AOA group was significantly lower than that of the control group (p<0.001). In the AOA subgroups based on the cause of infertility, the fertilization rates of each subgroup were significantly improved compared with previous control cycles except for the mixed factor infertility subgroup (p<0.05 to p<0.001). In the AOA subgroups based on the reason for AOA, the fertilization rates of each subgroup were significantly increased compared with those in their previous ICSI cycle without AOA (p<0.001); however, there was no significant difference in the top-quality embryo rate. No significant improvement was found in the implantation rates and the clinical pregnancy rate in each subgroup except for the poor embryo development subgroup. In the 114 AOA cycles, 35 healthy infants (21 singletons and 7 twins) were delivered without major congenital birth defects or malformations. This study showed that AOA with the calcium ionophore ionomycin can improve the reproductive outcomes of patients with previous fertilization failure and poor embryo development after ICSI.

P-278 What are the effects of ICSI procedure time intervals on in vitro fertilization outcomes?

Abstract Study question Do time intervals between oocyte pick-up (OPU), oocyte denudation (OD), and Intra-cytoplasmic Sperm Injection (ICSI) impact ICSI outcomes? Summary answer ICSI time intervals impact oocyte maturation, blastulation and pregnancy rates. There is no effect on fertilization and cleavage rates. What is known already Cytoplasmic and nuclear oocyte maturation are crucial for the success of in vitro fertilization and embryo development. Cumulus cells enhance oocyte maturation. Therefore, prolonged oocyte culture prior to denudation may induce oocyte apoptosis. There is no consensus for optimal time intervals between different ICSI procedures and their effects on fertilization, embryo and blastocyst, development and pregnancy rates. Study design, size, duration A retrospective analysis of 794 ICSI fresh cycles was performed at the Alyssa Fertility Group Center, between Juin 2021 and December 2022. Participants/materials, setting, methods ICSI cycles were performed on women aged ≤40 years, with at least 3 mature oocytes. Semen partner concentrations greater than 1million/mL were included. Time intervals between OPU, OD, and ICSI were recorded. We evaluate the effects of these timing intervals on oocyte maturation, fertilization, embryo development and pregnancy rates. Statistical analysis was performed using SPSS 22.0. Kolmogorov–Smirnov, Student t, Pearson tests, and ROC curve analysis were used. Level of signification p-value &amp;lt;0.05. Main results and the role of chance The mean women’s age was 33,2 years (SD 4,4). Mean times in hours for OPU/OD, OD/ICSI, and OPU/ ICSI were respectively: 01:28(SD 00:37); 02:35(SD 01:01); 04:03(SD 01:08). Positive correlations were seen between maturation rate and OPU/OD; OPU/ICSI intervals respectively (r = 0,217, p = 0,002; r = 0,193, p = 0,005).There were no significant correlation between ICSI time intervals, fertilization and cleavage rates. Top quality embryos rates were positively associated with OPU/OD time interval (r = 0,136: p = 0,049). The blastulation rate was positively associated with OPU/OD, OD/ICSI, and OPU/ICSI time intervals respectively(r = 0,183: p = 0,008; r = 0,211: p = 0,002; r = 0,279: p &amp;lt; 0,001).Pregnancy rates were positively correlated to OD/ICSI and OPU/ICSI respectively: (r = 0,158: p = 0,022; r = 0,188: p = 0,006). The ROC curves analysis revealed that the cut off value of time intervals between: OPU/OD was 01:10 (AUC = 0,554; IC 95% 0,476-0,632), p = 0,185; OD/ICSI was 02:20 (AUC = 0,600; IC 95% 0,516-0,669), p = 0,023 and for OPU/ICSI: 03:50, (AUC = 0,600; IC 95% 0,524-0,676), p = 0,014. Limitations, reasons for caution Trigger time is an important factor for oocyte maturation that should be noted. This study is limited by its retrospective nature. Potential unmeasured confounding factor cannot be excluded Wider implications of the findings Time intervals between all the ICSI procedures affect reproductive outcomes. According to our findings, excessive oocyte incubation time before denudation should be avoided. ICSI should be performed 2 hours after OD. Time intervals should be adjusted according to laboratory practice to improve ICSI outcomes. Trial registration number Not applicable

P-206 An evaluation of the incidence of total failed fertilization (TFF) following conventional IVF (cIVF) and the outcome of subsequent intracytoplasmic sperm injection (ICSI) cycles

Abstract Study question What is the incidence of TFF following conventional IVF (cIVF) and the chance of live birth in subsequent ICSI treatment cycles after TFF? Summary answer The incidence of TFF might be higher than previously reported and establishment of in-house benchmarks are recommended. Treatment continuation results in acceptable live birth rates. What is known already TFF following a cIVF is a devastating outcome for patients and physicians. The fear of this unfortunate event is assumable one of the main drivers for overutilizing ICSI. The incidence of TFF after cIVF should be less than 5 % of stimulated cycles based on ESHREs recommended performance indicators. Data on TFF of an unselected population is indeed however only available from an era when ICSI was not in use and showed a higher rate. We investigated the incidence of TFF and evaluated the chance of live birth following of such event at the first cycle. Study design, size, duration A single center, retrospective chart review for the time period of 2010-2021 was done to identify all cycles where cIVF did not result in at least one oocyte with ≥ 2 pronuclei. Also, patients who had more than 2 oocytes retrieved in their first cycle and continued their treatment using ICSI as a fertilization method in their upcoming cycles were selected. Participants/materials, setting, methods A total of 5772 initiated IVF cycles were identified in the electronic database for the examined time period. Criterion for cIVF was normozoospermia and applied as fertilization method in 1001 (17.3%) cycles. All the cases where TFF was noted following cIVF were retrieved and the number of oocytes, the rank of the cycle, female age and the result of upcoming cycles if any were recorded. Data was analyzed using Chi-Square test, p &amp;lt; 0.05 was considered significant. Main results and the role of chance A total of 125 TFF cycles were identified out of the 1001 cIVF cycles (12.49% failed fertilization rate per cIVF). A significantly higher frequency of TFF was seen in cycles where less than 3 oocytes were retrieved compared to those with more than two oocytes (38/99 [38.38%] vs. 87/902 [9.65%], p &amp;lt; 0.001). No statistical difference was detected between the risk of TFF occurring at the first, second or the third rank cIVF cycle (87/664 [13.10%], 30/224 [13.39%], 5/75 [6.67%] respectively; p &amp;gt; 0.05). The differences remained non-significant when only cycles with more than 2 oocytes were analyzed (67/608 [11.02%], 16/199 [8.04%], 4/68 [5.88%] respectively; p &amp;gt; 0.05). Also, different age groups showed similar incidence of TFF (16/157 [10.19%], 41/362 [11.33%], 54/381 [14.17%] and 14/101 [13.86%] in &amp;lt; 30, 31-35, 36-40 and &amp;gt;40 years of age respectively; p &amp;gt; 0.05). A total number of 56 patients were identified who continued their treatment using ICSI as fertilization method in their upcoming cycles. These patients completed 101 cycles resulting in 3 TFF cases (3%) and a fertilization rate of 75.1% (524/698) which was significantly higher to our baseline ICSI fertilization rate (70.9% [25231/35568]; p &amp;lt; 0.05). Nineteen of these 56 patients (33.9%) ended up giving at least one live birth following treatment continuation. Limitations, reasons for caution Selection criteria, e.g. patient characteristics and treatment type such as PGT-A, for choice of fertilization method are probably a source of variation in TFF incidence between centers. It is recommended that laboratories establish their own benchmark values based on their own data for the incidence of TFF. Wider implications of the findings Previous studies identified serum luteinizing hormone and progesterone levels at trigger and the number of oocytes as critical predictors for TFF. The similar chance of TFF after previous successful fertilization suggests more involvement of female factors in TFF. Acceptable live birth rates can be achieved following treatment continuation. Trial registration number not applicable

Sperm deoxyribonucleic acid fragmentation index at the time of intracytoplasmic sperm injection and standard in vitro fertilization is correlated with lower fertilization but not with blastocyst genetic diagnosis

To determine the effects of sperm deoxyribonucleic acid (DNA) fragmentation at the time of fertilization on invitro fertilization (IVF) outcomes and genetic diagnosis using next generation sequencing. Prospective double-blinded study. Private Clinic. Couples (n = 150). Invitro fertilization with preimplantation genetic testing for aneuploidy and sperm DNA fragmentation assay, as in sperm chromatin structure assay the day of retrieval. Laboratory outcomes are listed in the results section. Statistical analysis was performed using JMP, XYLSTAT, and STATA version15. The sperm DNA fragmentation index (DFI) in the neat ejaculate did not predict fertilization rate, quality, blastulation, or genetic diagnosis. No statistically significant results were obtained comparing <15% with >15%, <20% with >20%, <30% with >30% except for DFI. No statistically significant differences in oocyte source age or male age were observed. No statistically significant differences comparing <15% with >15%, <20% with >20%, <30% with >30% DFI at the time of standard IVF or intracytoplasmic sperm injection (ICSI) were observed for % euploid, aneuploid, mosaic, blastulation, biopsied, or D5/total biopsied. The DFI of >15% had more good quality D3 embryos than the <15% group, as did the >20% group compared with the <20% group. The ICSI fertilization was significantly higher in all 3 lower percentage groups compared with the higher counterpart. Standard IVF had significantly more blastocysts/fertilized suitable for biopsy and more D5/total number biopsied than ICSI embryos despite no difference in DFI. The DFI at fertilization is correlated with decreased fertilization for ICSI and IVF.

The Association of Sperm DNA Fragment and Assisted Reproductive Outcomes: A Meta-Analysis.

Objective To analyze the effect of sperm DNA fragmentation index (DFI) on the outcomes of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Methods Literature search was conducted on English databases PubMed, Cochrane, and Embase to obtain eligible studies. Results A total of 11 cohort studies were included and analyzed using the random effects model. The results illustrated that the IVF fertilization rate (RR = 0.94, 95% CI: 0.77-1.14, P = 0.61), pregnancy rate (RR = 0.83, 95% CI: 0.57-1.21, P = 0.32), and live birth rate (RR = 0.53, 95% CI: 0.16-1.80, P = 0.31) in the high DFI group were statistically insignificant with those in the low FI group. The correlations between DFI and ICSI fertilization rate (RR = 0.79, 95% CI: 0.52-1.18, P = 0.25), pregnancy rate (RR = 0.89, 95% CI: 0.74-1.06, P = 0.18), and live birth rate (RR = 0.89, 95% CI: 0.70-1.14, P = 0.36) were also not statistically significant. Conclusion This study has observed no significant correlation between sperm DFI and assisted reproductive outcomes. Multicenter large-sample clinical trials are required to conclusively determine the impact of DNA damage on the clinical outcomes of assisted reproduction.

Debate: Are We Limiting the Potential of Oocytes by Not Using ICSI for All? - ICSI for Some

Intracytoplasmic sperm injection (ICSI) was invented nearly thirty years ago and the first pregnancies in Australia were obtained at the Queen Elizabeth Hospital in the University of Adelaide’s Reproductive Medicine Program. Since then, ICSI has become the predominant method of fertilisation for IVF in cycles in Australia and in many parts of the world used for 100% of all cycles. While its value for male factor infertility, IVM egg fertilisation and injection into previously frozen eggs have been validated, the use of ICSI for non-male-factor infertility remains controversial and unproven. Given the significant concerns about the potential effects of ICSI on the health of children conceived by this method, a debate about its use in an excessive number of IVF cycles is warranted. The first randomised control trial of ICSI versus standard IVF fertilisation for unexplained infertility occurred more than twenty years ago and showed a lower ongoing pregnancy rate for those who received the ICSI form of fertilisation. Despite this evidence, ICSI continued to grow in prevalence across Europe, North America, the Middle East, Asia, and Australia for non-male factor infertility. Significant cohort studies started to emerge from the US and from Europe that showed either no benefit from ICSI for unexplained infertility, or indeed harm, in terms of ongoing pregnancy rates. This year we published in The Lancet, a randomised control trial where couples with unexplained infertility and a normal total sperm count and motility were randomised to receive either ICSI or standard IVF as a fertilisation method. No significant benefit of using ICSI was found with respect to ongoing pregnancy or live birth. The potential health benefits to the child and the cost effectiveness of the treatment weigh against the routine use of ICSI in the presence of normal sperm. Other cohort studies show that there is no benefit of ICSI for low egg numbers, advanced maternal age, or preimplantation genetic diagnosis. Examination of the use of ICSI across Australia, shows enormous variations between states and within the same state. It is clear, that members of the Society have very heterogeneous view of the use of ICSI and my intention in this talk is to encourage everyone to be more critical of its use when there is no sperm abnormality. I would encourage clinicians to use the normal natural processes of sperm selection which occur in routine IVF without sperm injection.

Effects of donor sperm on perinatal and neonatal outcomes resulting from in vitro fertilization-intracytoplasmic sperm injection and embryo transfer cycles: a retrospective cohort study.

BackgroundThe impact of donor sperm on pregnancy outcomes is controversial. The aim of this study was to investigate whether donor sperm in in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment could reduce the rate of live births or increase the incidence of adverse pregnancy outcomes and birth defects in neonates.MethodsThis single-centre, retrospective cohort study included 1,559 patients with infertility who received donor sperm at our hospital from 2015 to 2019. All the patients received fresh embryos and underwent first-cycle transfer. After propensity score matching, 4,677 controls who received their partners’ sperm were matched at 1:3. Clinical pregnancy, perinatal, and neonatal outcomes were compared between the donor sperm and partner sperm groups.ResultsThe embryo development was better in the donor sperm group than in the partner sperm group. The high-quality embryo and available embryo rates were significantly higher in the donor sperm group (P<0.05 for both groups). The rate of high-quality embryos transferred from the donor sperm group was higher than that from the partner sperm group (P<0.05). The clinical pregnancy (62.99% vs. 59.65%; P=0.02) and live birth (54.65% vs. 51.59%; P=0.036) rates were higher in the donor sperm group. After adjusting for confounding factors, no significant difference in live birth rates was observed between the two groups (adjusted P=0.057). The low birthweight (18.21% vs. 21.39%; P=0.023) and small for gestational age (SGA) (7.60% vs. 11.97%; P<0.001) rates were lower in the donor sperm group. To exclude the effect of multiple pregnancies, we evaluated neonatal outcomes of singleton pregnancies. No significant differences were noted in preterm and very preterm birth, SGA, mean birthweight, high birthweight, and low birth weight (LBW) and very low birth weight (VLBW) rates (P>0.05 for both groups). Further, no significant between group differences were observed in the ectopic pregnancy rate, early and late spontaneous abortion rates, gestational age, rate of large for gestational age (LGA), and neonatal defects.ConclusionsCompared with partner sperm, donor sperm did not reduce live birth rate and did not increase neonatal LBW or low birth defects.

Recombinant Human Chorionic Gonadotropin Versus Purified Human Chorionic Gonadotropin Trigger for In vitro Fertilization Intracytoplasmic Sperm Injection Cycle

The recombinant human luteinizing hormone or human chorionic gonadotropin (hCG) has substantially taken over the product which was formulated from the urine of pregnant women. In addition to this, a number of randomized controlled trials have been conducted comparing the significance of recombinant hCG (rHCG) with urinary hCG in assisted reproduction. Nevertheless, the present study has collected secondary information based on the nature of the chosen research area. The acquired data and information have been analyzed using content analysis technique to review the methodology and findings of the selected research articles. With the use of rHCG as well as purified hCG, the serum level was equivalent at day 5 among women underwent intracytoplasmic sperm injection (ICSI) but this level abnormally increased among those injected with rHCG thus, minimizing the rate of pregnancy. It has been concluded that purified hCG trigger plays a more efficient role in inducing oocyte maturation and increasing the rate of pregnancy among women undergoing in vitro fertilization ICSI cycle in comparison to rHCG trigger.

Embryo selection through non-invasive preimplantation genetic testing with cell-free DNA in spent culture media: a protocol for a multicentre, double-blind, randomised controlled trial

IntroductionMorphological evaluation is used to select embryos for in vitro fertilisation. However, it does not fully reflect the implantation potential. Preimplantation genetic testing for aneuploidies (PGT-A) can detect embryonic aneuploidy,...

Triggering with 1,500 IU of human chorionic gonadotropin plus follicle-stimulating hormone compared to a standard human chorionic gonadotropin trigger dose for oocyte competence in in vitro fertilization cycles: a randomized, double-blinded, controlled noninferiority trial

To assess if triggering with 1,500 IU of human chorionic gonadotropin (hCG) with 450 IU of follicle-stimulating hormone (FSH) induces noninferior oocyte competence to a standard dose of hCG trigger used in invitro fertilization (IVF). The alternative trigger will be considered noninferior if it is at least 80% effective in promoting oocyte competence. Randomized, double-blinded, controlled noninferiority trial. Academic infertility practice. Women aged 18-41 undergoing IVF with antral follicle count ≥8, body mass index ≤30 kg/m2, and no history of ≥2 IVF cycles canceled for poor response were enrolled. Participants with a serum estradiol >5,000 pg/mL on the day of trigger were excluded because of high risk of ovarian hyperstimulation syndrome. Participants were randomized to receive an alternative trigger of 1,500 IU of hCG plus 450 IU of FSH or a standard trigger dose of hCG (5,000 or 10,000 IU) for final oocyte maturation. The primary outcome was total competent proportion, defined as the probability of 2 pronuclei from an oocyte retrieved. The alternative trigger will be considered noninferior to the standard trigger if a 1-sided 95% confidence interval (CI) of the relative risk (RR) is not <0.8. Secondary outcomes included oocyte recovery and maturity, intracytoplasmic sperm injection fertilization, embryo quality, pregnancy rates, as well as serum and follicular hormones. Secondary outcomes were compared using a 2-sided superiority test. Outcomes were analyzed by intention-to-treat and per-protocol. A total of 105 women undergoing IVF were randomized from May 2015 to June 2018. The probability of the primary outcome was 0.59 with the alternative trigger and 0.65 with the standard trigger, with a RR of 0.91 and a 1-sided 95% CI of 0.83. Noninferiority of the alternative trigger was demonstrated. Live birthrate from all fresh transfers in the alternative trigger group vs. standard trigger was 46.9 vs. 46.4% (RR, 1.01; 95% CI, 0.62-1.62), respectively. Live birthrate per randomized participant was 48.1% in the alternative trigger group vs. 62.7% with the standard trigger (RR, 0.73; 95% CI, 0.48-1.11). No participants had a failed retrieval. Triggering with 1,500 IU of hCG plus 450 IU of FSH promoted noninferior oocyte competence compared to a standard hCG trigger dose. NCT02310919.