Assisted Reproductive Procedures Research Articles

Contribution of sperm DNA fragmentation testing by flow cytometry in medically assisted reproduction procedures

Contribution of sperm DNA fragmentation testing by flow cytometry in medically assisted reproduction procedures

Microfluidics as an emerging paradigm for assisted reproductive technology: A sperm separation perspective.

Millions of people are subject to infertility worldwide and one in every six people, regardless of gender, experiences infertility at some period in their life, according to the World Health Organization. Assisted reproductive technologies are defined as a set of procedures that can address the infertility issue among couples, culminating in the alleviation of the condition. However, the costly conventional procedures of assisted reproduction and the inherent vagaries of the processes involved represent a setback for its successful implementation. Microfluidics, an emerging tool for processing low-volume samples, have recently started to play a role in infertility diagnosis and treatment. Given its host of benefits, including manipulating cells at the microscale, repeatability, automation, and superior biocompatibility, microfluidics have been adopted for various procedures in assisted reproduction, ranging from sperm sorting and analysis to more advanced processes such as IVF-on-a-chip. In this review, we try to adopt a more holistic approach and cover different uses of microfluidics for a variety of applications, specifically aimed at sperm separation and analysis. We present various sperm separation microfluidic techniques, categorized as natural and non-natural methods. A few of the recent developments in on-chip fertilization are also discussed.

Correlation between standard sperm parameters and sperm DNA fragmentation from 11,339 samples

Conventional semen parameters have long been considered fundamental in male fertility analyses. However, doubts have been raised regarding the clinical utility of the assessment of spermatozoa (sperm) DNA damage. In this retrospective study, we investigated the potential correlation between conventional semen parameters and semen DNA fragmentation (SDF) assessed as sperm DNA damage, in 11,339 semen samples collected between January 2019 and June 2022. We observed significant negative correlations between the DNA fragmentation index (DFI) and sperm viability (correlation coefficient [r] = −0.514) as well as progressive sperm motility (r = −0.512, p < 0.05). Samples were categorized into three groups according to DFI levels (Groups A, B, and C: ≤15%, 15 < DFI ≤30%, and >30%, respectively). Furthermore, the percentage of semen samples with normal sperm conventional parameters in Groups A, B, and C was 76.7% (4369/5697), 61.4% (2351/3827), and 39.7% (721/1815), respectively. Moreover, according to the reference values of conventional sperm parameters, the samples were divided into Groups F, G, and H with all normal, only one abnormal, and > two abnormal parameters, respectively. In addition, the proportions of samples with abnormal DFI values (>30) in Groups F, G, and H were 9.7% (721/7441), 23.1% (618/2676), and 39.0% (476/1222), respectively. Multivariate logistic regression models demonstrated that sperm vitality, progressive sperm motility, normal sperm form, total sperm count, semen volume, age, and some sperm kinematics collectively improved the area under the receiver operating characteristic curve (AUROC) to 0.861, surpassing the predictive value of a single predictor of pathologically damaged sperm DNA. Our study suggests that samples with abnormal sperm parameters may have a higher likelihood of high DNA fragmentation. Furthermore, certain semen parameters could be potential indicators of sperm DNA fragmentation, aiding sperm selection in assisted reproductive procedures.

Between choices and decisions. Genetics in homoparental families through surrogacy in Argentina.

Comprehensive access to medically assisted reproduction procedures and techniques in Argentina has been assured by National Law No. 26,862 since 2013. This Law does not include surrogacy procedures, and the lack of specific regulation shifts practices to a paralegal setting. In this context, planned parenthood by male couples through surrogacy is performed through actions that convey demands for access rights and active State policies. For these couples, the argument is that surrogacy is the only option to have a child with a genetic bond with at least one of the two parents and recognize both filiatory bonds. This work results from field work in progress with parents from the Province of Buenos Aires running this practice in Argentina. Based on in-depth interviews, we attempted to rebuild personal experiences and analyze the meanings that the narratives construct regarding their parenting, the biological connections in establishing or defining family relationships, and the importance of genetics in constructing and maintaining affiliations.

The Role of Male Consent in Assisted Reproductive Technology Procedures: an Examination of Japanese Court Cases.

The online version contains supplementary material available at 10.1007/s41649-023-00274-1.

Debate 4: Morphological Assessment of Embryos is Outdated

Motion: For The Outdated Significance of Morphological Assessment in Embryo Selection and the Rise of Advanced Technologies in Reproductive Medicine This symposium lecture presents a compelling debate, shedding light on the diminishing relevance of traditional static morphological assessment as a primary criterion for embryo. The speaker aims to challenge the long-standing reliance on morphological evaluation and advocate for the integration of cutting-edge technologies as a more reliable and objective means of embryo quality assessment. During the debate, the speaker will examine the limitations of morphological assessment, acknowledging its subjective nature, potential bias, and lack of comprehensive insight into embryonic development. The traditional morphological assessment based solely on visual appearance may not adequately predict an embryo’s true potential for successful implantation and healthy pregnancy. A variety of available new technologies will be highlighted during the lecture, showcasing alternatives to morphological embryo assessment. Attendees will gain insight into how these innovative methods can offer more objective and precise measures of embryo quality, enhancing the success rates of assisted reproductive procedures. The symposium will foster an engaging environment, encouraging active participation and thoughtful discussions among attendees from the reproductive medicine community. Participants will have the opportunity to challenge prevailing notions and explore the implications of adopting new technologies in clinical practice. In summary, this debate aims to catalyze a paradigm shift in embryo evaluation, emphasizing the need to move beyond traditional morphological assessment towards embracing advanced technologies, to optimize embryo selection and lead to improved outcomes in IVF. Motion: Against Morphological Assessment of Embryos is Not Outdated In the era of new technologies and artificial intelligence, looking good has never been more important! On-time progression through specific events is requisite for human preimplantation development and provides important visual insights into the health and physiology of the embryo. The formation of 2 distinct pronuclei, the times and patterns of cleavage, and how the cells compact are all key markers of development. Following compaction, the formation of the blastocyst is not only a temporally regulated event, but also an energetically demanding process, one that reflects the metabolic activity (a biomarker of viability) of the embryo. Further, at the blastocyst stage it is possible to visualise and quantitate the formation of both the inner cell mass (ICM) and trophectoderm, with all parameters of blastocyst development linked to transfer outcomes. The development of time-lapse incubation, which can be described as embryo morphology over time, has led to the incorporation of several key morphological features to support embryo selection, further reflecting the significance of embryo morphology. Interestingly, AI algorithms used for embryo selection reflect predominantly those morphological parameters that are known to affect transfer outcomes, such as blastocyst quality. While only specific molecular tests can accurately determine the precise chromosomal composition of an embryo, key morphological features and events can also be used to tag those embryos at greatest risk of being genetically abnormal. In conclusion, key morphological developmental events indirectly inform us of the physiology of the embryo (degree of blastocyst expansion reflects metabolic activity for example), and hence this is why morphological grading systems (such as the Gardner Grade for blastocysts) have been shown to be effective in embryo selection for over the past 25 years, and further have proven to be as useful as AI or biomarker analysis for the majority of patients.

The Impact of Growth Hormone Co-Treatment Duration on Outcomes in IVF/ICSI Cycles Among Poor Ovarian Responders.

The efficiency of in vitro fertilization is improved by growth hormone (GH) during ovarian stimulation. Additionally, patients with diabetes experience impaired insulin resistance and compromised glucose tolerance, which further exacerbate their condition. Due to these side effects, in this study, the duration of GH treatment was compared in IVF/ICSI cycles among poor ovarian responders. In this study, POSEIDON criteria were used to choose patients. Subcutaneous administration of gonadotropin-releasing hormone (GnRH) antagonist was done beginning on the sixth day of the cycle and continuing through the day of human chorionic gonadotropin (hCG) injection. In one group, GH was administered 4 units/day from the 2nd day of the cycle until hCG injection, and in another group, the first dose was administered on the 6th day of the cycle. Following the administration of hCG, which lasted from 24 to 36 hr, oocytes were retrieved with the support of B-mode sonography. In our analysis, no significant differences were observed between the two groups in terms of the number of retrieved oocytes, metaphase II oocytes, and quality of grade A and B embryos. The results show that the treatment or conditions did not have a significant impact on the outcomes among the studied groups. Our findings indicate that a shorter duration of GH administration can yield similar outcomes compared to a longer duration in IVF/ICSI cycles involving poor ovarian responders. This result holds the potential for a more cost-effective and patient-friendly approach in managing assisted reproductive technology procedures. It may lead to reduced side effects and improved adherence to medication regimens in patients.

Effect of L. crispatus M247 Administration on Pregnancy Outcomes in Women Undergoing IVF: A Controlled, Retrospective, Observational, and Open-Label Study.

The aim of our study was to retrospectively evaluate whether the oral administration of L. crispatus (M247) could increase pregnancy and live birth rates in women undergoing assisted reproductive technology procedures. Enrolled women (N = 160) were divided into two groups: treated (N = 80) or untreated (N = 80) with the probiotic strain. The odds ratio (OR) for a treated woman to have a clinical pregnancy (CP) was 1.56. In women aged 30-40 years, M247 increased the probability of a CP in correlation with the progressive rise in BMI, reaching 47% (35% in controls) with a BMI of 35 (OR: 2.00). The CAID statistics showed that in a woman of the blastocyst subgroup, below 43 years, with a BMI over 18.6, treatment with M247 increased the chance of a CP from 28.4% to 44.5% (OR: 2.08; p < 0.05). Considering live births, the rate of the probiotic group was 12.5% versus 7.5% (OR: 1.76). Considering only the blastocyst subgroup, the treatment increased the number of live births by 200% (OR: 3.64; p = 0.05). As confirmed also by statistical indices NNT, NNH, and LHH, the use of M247 demonstrated a risk-benefit ratio to the full advantage of the benefits.

Chronic endometritis and infertility — in vitro fertilization outcomes: systematic review and meta-analysis

Background. The relevance of the problem is related to the high prevalence of chronic endometritis (CE); its role in female infertility, implantation failures during assisted reproductive technology procedures, and recurrent miscarriage; as well as the lack of a unified strategy in the diagnosis and treatment of this pathology. The present systematic review with a meta-analysis focuses on evaluating the impact of CE and its therapy on the outcome of in vitro fertilization. In addition, the effect of CE of various severity on the outcomes of assisted reproductive technologies is analyzed. Objective. To analyze the effect of CE of varying severity and its treatment on the outcomes of in vitro fertilization. Methods. Using PubMed, Medline, Scopus, Embase, ELibrary, Cochrane Central Register of Controlled Trials (CENTRAL), WHO International Clinical Trials Registry, and Russian Science Citation Index, a systematic search was conducted for articles published over the past 12 years that met the following criteria: randomized controlled trial examining the effect of CE of varying severity on fertility and ways to treat it. The following indicators were calculated: ongoing pregnancy/live birth, clinical pregnancy, and miscarriage rates. A total of 4145 patients (from ten studies) were included. A meta-analysis was performed using Stata 11.0 software (The Cochrane Collaboration, Oxford, UK). The heterogeneity was considered low at I2 &lt;30%, moderate at 30–50%, and high at &gt;50%. Results. Women with CE exhibited lower ongoing pregnancy/live birth (OR 1.97; p = 0.02) and clinical pregnancy rates (OR 2.28; p = 0.002) as compared to women without it. CE treatment increased the ongoing pregnancy/live birth (OR 5.33; p &lt; 0.0001) and clinical pregnancy rates (OR 3.64; p = 0.0001). In vitro fertilization outcomes were comparable in women treated for CE and women without CE (ongoing pregnancy/live birth rate, clinical pregnancy rate, and miscarriage rate: p = ns). Women with severe CE exhibited lower ongoing pregnancy/live birth (OR 0.43; p = 0.003) and clinical pregnancy rates (OR 0.40; p = 0.0007). Mild CE showed no significant effect on in vitro fertilization outcomes (ongoing pregnancy/ live birth rate, clinical pregnancy rate, and miscarriage rate: p = ns). Conclusion. The conducted meta-analysis showed that CE significantly reduces the ongoing pregnancy/live birth and clinical pregnancy rates in infertile women undergoing in vitro fertilization. Noteworthy is that antimicrobial therapy in such patients improves the results of assisted reproductive technologies, which are comparable to those of patients without CE. The negative impact of this pathology on the implantation capacity of the endometrium is most often observed in the severe form, while its mild form has virtually no effect on the in vitro fertilization outcome.

Differential sperm histone retention in normozoospermic ejaculates of infertile men negatively affects sperm functional competence and embryo quality.

The unique epigenetic architecture that sperm cells acquire during spermiogenesis by retaining <15% of either canonical or variant histone proteins in their genome is essential for normal embryogenesis. Whilst heterogeneous levels of retained histones are found in morphologically normal spermatozoa, their effect on reproductive outcomes is not fully understood. Processed spermatozoa (n=62) were tested for DNA integrity by sperm chromatin dispersion assay, and retained histones were extracted and subjected to dot-blot analysis. The impact of retained histone modifications in normozoospermic patients on sperm functional characteristics, embryo quality, metabolic signature in embryo spent culture medium and pregnancy outcome was studied. Dot-blot analysis showed heterogeneous levels of retained histones in the genome of normozoospermic ejaculates. Post-wash sperm yield was affected by an increase in H3K27Me3 and H4K20Me3 levels in the sperm chromatin (p<0.05). Also, spermatozoa with higher histone H3 retention had increased DNA damage (p<0.05). Spermatozoa from these cohorts, when injected into donor oocytes, correlated to a significant decrease in the fertilisation rate with an increase in sperm histone H3 (p<0.05) and H3K27Me3 (p<0.01). An increase in histone H3 negatively affected embryo quality (p<0.01) and clinical pregnancy outcome post-embryo transfer (p<0.05). On the other hand, spent culture medium metabolites assessed by high-resolution (800MHz) nuclear magnetic resonance showed an increased intensity of the amino acid methionine in the non-pregnant group than in the pregnant group (p<0.05) and a negative correlation with sperm histone H3 in the pregnant group (p<0.05). Histone retention in spermatozoa can be one of the factors behind the development of idiopathic male infertility. Such spermatozoa may influence embryonic behaviour and thereby affect the success rate of assisted reproductive technology procedures. These results, although descriptive in nature, warrant further research to address the underlying mechanisms behind these clinically important observations.

DNA strand displacement and TdT-Mediated DNA extension for swift, convenient, and quantitative evaluation of sperm DNA integrity and its clinical implications

DNA integrity is crucial for the clinical pregnancy outcome and offspring health, while detection methods currently used (comet assay, TUNNEL assay, SCSA, etc.) can only provide the ratio of positive sperms at the cellular level and are unable to quantitatively detect the breakpoints at the DNA molecular level. Herein, we developed a detection system based on terminal deoxynucleotidyl transferase and DNA strand displacement fluorescent probe, which could efficiently and conveniently measure the number of 3′-OH (equivalent to the number of breakpoints). We further investigated the use of this technique in assisted reproduction after completing the principle verification, system optimization, and research on analytical performance. The detection system was shown to have a good linear range from 0.01 nM to 4 nM, using single-stranded DNA with 3′-OH end as the calibrator. The system underwent thorough optimization for stability and accuracy. In comparison to the widely accepted index DFI detected by SCSA, the new system demonstrated reasonable correlation and better prediction efficiency. Its applicability was also proven through its use in assisted reproductive technology procedures.

Hormonal Pretreatment for Ovarian Stimulation: A Narrative Review

Introduction: Assisted reproductive technologies are necessary and important for the management of infertility and can be applied with and without the use of strategies such as hormonal pretreatment. However, there is a discussion in the literature on whether hormonal pretreatment in ovarian stimulation protocols can improve the outcome of oocytes and embryos. This study aimed to conduct an integrative review of hormonal pretreatment and its impact on women undergoing assisted reproduction procedures. Methods: We made searches in the PubMed and the Cochrane Library databases. The inclusion criteria were articles published in peer-reviewed journals in English, Portuguese, or Spanish from January 1, 2010 to December 31, 2021 and available as free full texts. We excluded gray literature works, including term papers, theses, and dissertations. We used a narrative synthesis for data analysis. Results: The 189 articles retrieved from the two databases (2010-2021) were narrowed down to eight articles. In most studies, pretreatment seems to play a role beyond stimulation that includes an early inhibition of the FSH peak, a more homogeneous cohort, and better results. Conclusions: Using a pretreatment with combined oral contraceptives (COC) produces conflicting results in assisted reproduction. The COC improves the quantity and quality of eggs, but it neither benefits nor harms reproductive outcomes. Also, the use of COC has contradictory results regarding clinical pregnancy

Outcomes of assisted reproductive technology procedures performed on weekdays versus weekends: a retrospective cohort study.

To evaluate if assisted reproductive technology (ART) outcomes are different based on whether procedures - oocyte retrieval, insemination, embryo biopsy, or embryo transfer - are performed on a weekday versus weekend/holiday. Retrospective cohort study of all patients ≥ 18years old who underwent oocyte retrieval for in vitro fertilization or oocyte banking (n = 3,197 cycles), fresh or natural-cycle frozen embryo transfers (n = 1,739 transfers), or had embryos biopsied for pre-implantation genetic testing (n = 4,568 embryos) in a large academic practice from 2015-2020. The primary outcomes were as follows: oocyte maturity for oocyte retrievals; fertilization rate for insemination; rate of no result on pre-implantation genetic testing for embryo biopsy; and live birth rate for embryo transfers. The average number of procedures performed per embryologist per day was higher on weekends/holidays than weekdays. For oocyte retrievals performed on weekdays vs. weekends/holidays, there was no difference in oocyte maturity rate (88% vs 88%). There was no difference in the fertilization rate (82% vs 80%) in cycles that had intracytoplasmic sperm injection performed on weekdays vs. weekends/holidays. No difference was found in the no result rate for embryos biopsied on weekdays vs. weekends/holidays (2.5% vs 1.8%). Finally, there was no difference by weekday vs. weekend/holiday in the live birth rate per transfer among all transfers (39.6% vs 36.1%), or when stratified by fresh (35.1% vs 34.9%) or frozen embryo transfer (49.7% vs. 39.6%). We found no differences in ART outcomes among women who had their oocyte retrievals, inseminations, embryo biopsies, or embryo transfers performed on weekdays versus weekends/holidays.

P-059 Sperm DNA fragmentation (SDF) after cryopreservation and sperm selection: implications for clinical pregnancies and live births after intrauterine insemination with donor sperm

Abstract Study question Can SDF assessment pre-cryopreservation, post-thaw and after post-thaw sperm selection predict clinical outcome in a heterologous intra-uterine donor insemination program (IUI)? Summary answer SDF increased post-cryopreservation in donor sperm and after post-thaw density gradient without affecting clinical pregnancy, live birth and miscarriage rates. What is known already Sperm cryopreservation is effective for preservation of male fertility and facilitates the storage of donor semen, while infectious disease screening can be completed and confirmed negative. Sperm viability and motility are the most vulnerable parameters during the freeze-thaw process. Inherent reactive oxygen species may induce SDF via oxidative stress. Controversial results are reported regarding induction of SDF after cryopreservation due to differences in cryo tolerance, methods of cryopreservation and methods used to evaluate SDF. Sperm selection post-thaw is indispensable to select a normal motile fraction for insemination. Again, results are controversial regarding influence on SDF levels after sperm selection post-thaw. Study design, size, duration This was a prospective, observational study. Between March 2015 and March 2019, eighteen potential sperm donors were screened via a three step plan – semen assessment, medical assessment and serological blood tests. In accordance with the Belgian legislation (2007), a sperm donor was matched to obtain a maximum of 6 pregnancies in acceptor women/couples, allowing for more than one offspring/woman or couple. A total of 106 acceptors were matched for heterologous intra-uterine insemination. Participants/materials, setting, methods Semen samples were collected after 2-7 days abstinence and standard semen parameters analyzed within 60 mins after ejaculation. Within the same time frame SDF test was performed using TUNEL assay both before and after cryopreservation and after a two-step discontinuous density gradient centrifugation post-thaw. Samples were cryopreserved by the conventional slow freezing method using sperm freeze solution. A yield of ≥ 2 M progressive spermatozoa post-thaw after selection was sufficient to contemplate IUI. Main results and the role of chance Mean female age at first cycle was 33.9 ± 4.1 years and donor age 28.5 ± 5.6 years. Out of 429 cycles, 100 (23.3%) resulted in clinical pregnancy. Excluding three ongoing pregnancies, we counted 75 live births (17.6% of cycles or 77.3% of pregnancies), while 21 pregnancies ended in miscarriage (4.9% of cycles or 21.6% of pregnancies) and 1 resulted in stillbirth. Progressive motility (59.3 ± 12.5%) decreased significantly after cryopreservation (30.0 ± 13.2%; p &amp;lt; 0.001), but increased post-thaw after density gradient centrifugation (61.6 ± 16.0%; p &amp;lt; 0.001). On the contrary, SDF (12.0 ± 5.9%) increased post-cryopreservation (26.3 ± 14.5%; p &amp;lt; 0.001) and further increased after sperm selection (34.9 ± 22.1%; p = 0.04). Using multilevel mixed-effects logistic regression (Odds ratio [95%CI]), we found that female age significantly influenced clinical pregnancy (0.911 [0.847-0.981]; p = 0.01), live birth (0.894 [0.834-0.959]; p = 0.002) and miscarriage rates (1.180 [1.033-1.347]; p = 0.015). There was no significant age-adjusted effect of SDF post-thaw after density gradient selection on clinical pregnancy (1.007 [0.994-1.021]; p = 0.30), live birth (1.001 [0.988-1.014]; p = 0.84) or miscarriage (1.021 [0.997-1.046]; p = 0.08). Median time-to-live birth was 4 cycles. Analyses of time-to-live birth with mixed-effects Cox models revealed no significant association with progressive motility; SDF before or after cryopreservation and after sperm selection post-thaw. Limitations, reasons for caution Results concern a small donor population, with high normal semen parameters and should therefore be extrapolated cautiously to subfertile men with normal and subnormal semen parameters. Wider implications of the findings In a donor IUI program with strict selection on conventional sperm parameters, differences in SDF before and after cryopreservation and gradient centrifugation do not change IUI outcomes, although cryopreservation and centrifugation increase SDF. Findings could be useful for the optimization of sperm freezing and selection in different assisted reproductive procedures. Trial registration number not applicable

P-181 The impact of high proportion of immature oocyte in a cohort on the reproductive outcome following icsi

Abstract Study question Whether high proportion of immature oocyte in a cohort impacts outcome of sibling mature oocyte in ICSI cycles. Summary answer Study demonstrate that if immature oocyte retrieved was more than 50% in a cycle then it reduces reproductive outcome with the remaining mature sibling oocytes. What is known already The selection of competent oocyte is crucial to assisted reproductive technique procedure outcome. In ICSI procedure where only metaphase II oocytes (MII) are injected correlates with number of MII oocytes available, embryo development and pregnancy outcome. Percentage of immature oocyte {Metaphase I (MI) and Germinal Vessicle (GV) } retrieved in a cycle is always a concern for stimulation protocol, patient response which affects IVF laboratory outcome. Study design, size, duration A prospective study was conducted from 1st January 2020 to 1st December 2022 in the Department of Reproductive medicine at a tertiary infertility centre in India. Participants/materials, setting, methods Patients were divided into 3 groups on the basis of percentage of immature oocyte retrieved in a single cycle. Group A (n = 100), where percentage of immature oocyte (MI and GV) was between 0 to 20%, while Group B (n = 60) where percentage of immature oocyte was between 21 to 50% and Group C (n = 20) where percentage of immature oocyte was between 51 to 100%. Main results and the role of chance This study included all normal responder patients and cycles using in-vitro fertilisation (IVF) and poor responders were excluded. All blastocyst formed were vitrified and transferred in frozen embryo replacement cycle. All the 3 groups were compared on the basis of fertilisation rate, blastocyst formation rate, clinical pregnancy rate and miscarriage rate. Group A and group B had no significant difference in fertilisation rate (82% vs. 78%, p &amp;gt; 0.5), blastocyst formation rate (60% vs. 58%, p &amp;gt; 0.5), clinical pregnancy rate (58% vs. 55%, p &amp;gt; 0.5) and miscarriage rate (12% vs. 11%, p &amp;gt; 0.5). While significant reduction was observed in group C ( &amp;gt; 50% MI &amp; GV oocyte) in fertilisation rate (40%, p &amp;lt; 0.1), blastocyst formation rate (28%, p &amp;lt; 0.1), clinical pregnancy rate (15%, p &amp;lt; 0.1). Limitations, reasons for caution Larger randomised control studies are needed to strengthen these results. Wider implications of the findings Percentage of immature oocyte can help in predicting the success of assisted reproductive technique. This will help in counselling patient about outcome. Trial registration number NA

Chronic kidney disease, female infertility, and medically assisted reproduction: a best practice position statement by the Kidney and Pregnancy Group of the Italian Society of Nephrology

Fertility is known to be impaired more frequently in patients with chronic kidney disease than in the general population. A significant proportion of chronic kidney disease patients may therefore need Medically Assisted Reproduction. The paucity of information about medically assisted reproduction for chronic kidney disease patients complicates counselling for both nephrologists and gynaecologists, specifically for patients with advanced chronic kidney disease and those on dialysis or with a transplanted kidney. It is in this context that the Project Group on Kidney and Pregnancy of the Italian Society of Nephrology has drawn up these best practice guidelines, merging a literature review, nephrology expertise and the experience of obstetricians and gynaecologists involved in medically assisted reproduction. Although all medically assisted reproduction techniques can be used for chronic kidney disease patients, caution is warranted. Inducing a twin pregnancy should be avoided; the risk of bleeding, thrombosis and infection should be considered, especially in some categories of patients. In most cases, controlled ovarian stimulation is needed to obtain an adequate number of oocytes for medically assisted reproduction. Women with chronic kidney disease are at high risk of kidney damage in case of severe ovarian hyperstimulation syndrome, and great caution should be exercised so that it is avoided. The higher risks associated with the hypertensive disorders of pregnancy, and the consequent risk of chronic kidney disease progression, should likewise be considered if egg donation is chosen. Oocyte cryopreservation should be considered for patients with autoimmune diseases who need cytotoxic treatment. In summary, medically assisted reproduction is an option for chronic kidney disease patients, but the study group strongly advises extensive personalised counselling with a multidisciplinary healthcare team and close monitoring during the chosen medically assisted reproduction procedure and throughout the subsequent pregnancy.Graphical abstract

How to select healthy sperm for intracytoplasmic sperm injection in samples with high sperm DNA fragmentation?

The body of evidence supports the negative impact of increased sperm DNA fragmentation (SDF) on natural fertility as well as assisted reproduction conditions. High SDF has been correlated with low pregnancy and delivery rates following intrauterine insemination. Also, high SDF is accused of reducing the rates of fertilization, implantation, pregnancy, and live birth following in-vitro fertilization (IVF). Despite no impact of high SDF on fertilization or pregnancy rates following intracytoplasmic sperm injection (ICSI), it has been correlated with poor embryo quality and a higher risk of miscarriage. Several methods have been introduced to help select sperm with the best DNA quality to be used in assisted reproductive technology procedures. These include magnetic-activated cell sorting, intracytoplasmic morphologically selected sperm injection, physiologic ICSI, and microfluidic sperm sorters, among others. This article aimed to discuss the impact of high SDF in infertile men on the reproductive outcome of couples undergoing IVF/ICSI. Additionally, this review highlights the principles, advantages, and limitations of different techniques that are currently used for the selection of sperm with intact DNA to be utilized for ICSI.

The influence of concomitant extragenital pathology on the success of assisted reproductive technology procedures in women (according to clinics of the Volgograd region)

At the moment, the problem of infertility does not lose its relevance. In the structure of this disease, the violation of reproductive function in women is dominant. The present study is devoted to the study of the influence of concomitant extragenital pathology on the success of assisted reproductive technology procedures. The data of 184 women who underwent infertility treatment by ART methods in clinics of the Volgograd region in the period from 2015 to 2020 were analyzed. The following parameters of the patients were studied: age, gynecological history, the main cause of infertility, transferred diseases of non-productive organs, the protocol of ovarian stimulation with gonadotropins, the number of mature follicles by the end of stimulation, the number of received, fertilized eggs, the number of embryos by day 3, 5. It has been established that concomitant extragenital pathology affects the processes of folliculogenesis and qualitatively affects oogenesis.

Female infertility from oocyte maturation arrest: assembling the genetic puzzle.

Assisted reproduction procedures often encounter an issue called oocyte maturation arrest (OMA), which is manifested as failed IVF/ICSI attempts using oocytes from some infertile women. In this issue of EMBO Molecular Medicine, Wang etal identify infertile women bearing novel DNA sequence variants in a gene called PABPC1L, which is essential for translation of maternal mRNAs. By conducting a series of in vitro and in vivo experiments, they demonstrated certain variants as being causal for OMA, confirming a conserved requirement for PABPC1L in human oocyte maturation. This study offers a promising therapeutic target for treating OMA patients.

Equine Spermatozoa Selection by Magnetic Activation for Use in Assisted Reproduction.

This study aimed to select high-quality spermatozoa by sperm separation by magnetic activation of the fresh equine semen, compared to density gradient centrifugation and evaluating cell quality after selection. The semen of 10 stallions was collected by the artificial vagina technique. The samples analyzed were: (1) fresh semen; (2) density gradient centrifugation (DGC); (3) separation by magnetic activation (MASS) (nonapoptotic portion NAP); (4) separation by MASS (apoptotic portion-APT). Was analyzed: motility (light microscopy), concentration (Neubauer chamber), semen morphology (humid chamber in phase contrast), and supravital test (eosin/nigrosine). In DGC, 20×106 spermatozoa were used in the gradient of Percoll at 90% and 45% (400 μL each), centrifugation at 900 G/5 min, the pellet was diluted in HEPES. In MASS, 10×106 spermatozoa were diluted in 1.5 mL of HEPES, centrifugation at 300 G/10 min, pellet was resuspended in 150 μL of HEPES with 20 μL of nanoparticles bound to annexin V, incubation for 15 minutes and filtered in the magnetic separation column. The nonapoptotic fraction was collected directly and the apoptotic fraction after removal the column from the magnet and adding 300 μL of HEPES. The total abnormalities were 43.2% ± 2.78%, with the DGC and MASS being effective in reducing sperm abnormality by 15.6% ± 2.10% and 24.30% ± 1.63%, respectively, like the observed for the number of cells with intact membranes (50% lower in the APT portion). This nanotechnological method is efficient in producing high-quality semen samples for assisted reproduction procedures.