Intracytoplasmic Sperm Injection Procedure Research Articles

Protocols for Cryopreservation and Rederivation of Rat Gametes.

New genome-editing tools, such as ZFNs, TALEN, and CRISPR/Cas9, have enabled the generation of gene-modified models effectively in mammals. These technologies are a powerful tool for studying gene function and creating animal models for human diseases. On the other hand, such gene-modified animals are raised in numerous experimental animal facilities, which puts pressure on breeding space and maintenance costs. Embryo and sperm cryopreservation is not only the most simple and cost-effective method available for most gene-modified strains but also the most reliable method to preserve strains to avoid breeding problems and contamination. We have established a reliable, high quality embryo and sperm cryopreservation system for rat strains, ensuring the longevity of these valuable resources for the scientific community. These cryopreserved resources have been successfully used to rederive next generation pups using embryo transfer and intracytoplasmic sperm injection (ICSI). In this chapter, we describe in detail protocols for rat embryo vitrification and sperm cryopreservation followed by pup rederivation using the ICSI procedure and embryo transfer.

Effect of endometrial scratch using a Pipelle catheter on recurrent implantation failure with intracytoplasmic sperm injection

Back ground In the last two decades, assisted reproductive technology procedures for culturing and transferring embryos have been continually improved, but the clinical pregnancy rate has not improved substantially over the last 10 years. The most important cause for implantation failure is embryo quality or uterine receptivity, but remains unexplained in many cases. About 75% of conceptus are lost before or at the time of implantation. Aim This work aims to assess the effect of endometrial scratch (ES) using a Pipelle catheter on the outcome of intracytoplasmic sperm injection (ICSI) in patients with recurrent implantation failure. Patients and methods The current study included 130 patients who were divided into two equal groups of 65 patients each. In the scratch group, endometrial local scratch using a Pipelle biopsy catheter on the mid luteal phase of spontaneous menstrual cycle before the ICSI procedure was performed, whereas the control group did not undergo ES. ICSI failure was recorded two or more times in each group. Results In the current study, there were no statistically significant differences between the two groups in age, gravidity and parity, type, duration of infertility, and the number of previous ICSI trials. However, the BMI was higher in the control group. In the current study, there were no statistically significant differences between the two groups in basal hormonal profile follicle-stimulating hormone, luteinizing hormone, thyroid-stimulating hormone, prolactin, estradiol, and anti-Mullerian hormone. In this study, there was highly significant increase in the pregnancy rate in the scratch group than the nonscratch group. The number of pregnant cases in the scratch group was 36 (55.4%), whereas in the control group, it was 21 (32.3%), with a highly significant P value of 0.008. In this study, there was no statistically significant difference between positive and negative pregnancy in the basal hormonal profile, except anti-Mullerian hormone and follicle-stimulating hormone levels, P values of 0.040 and 0.000. Conclusion ES in the cycle before the ICSI cycle improved the chemical and clinical pregnancy rates compared with ICSI performed without ES in cases of recurrent implantation failure. It seems that increasing pregnancy rates could be because of the local inflammatory reaction elicited by the endometrial sampling procedure.

Assessment of Sub-Endometrial Junction Zone by 3-Dimensional Transvaginal Ultrasound in Unexplained Recurrent Implantation Failure and Its Effect on ICSI Outcomes

BACKGROUND: Sub-endometrial junction zone (JZ) plays an important role in most of reproductive functions. OBJECTIVE: To find out the effect of the sub-endometrial JZ thickness assessment by 3-D trans-vaginal ultrasound (TVUS) on intracytoplasmic sperm injection (ICSI) outcomes in patients with unexplained recurrent implantation failure (RIF). SETTING: ART Unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Qena, Egypt. DURATION: From April 2016 to October 2018. STUDY DESIGN: A prospective observational study. METHDS: Fifty couples with history of unexplained RIF in previous ICSI cycles and prepared for another ICSI cycle (group I) and fifty couples with unexplained infertility prepared for ICSI for the first time (group II) had been included in this study. At time of ovum pick up, 3-DTVUS was done for all cases in both groups for assessment of sub endometrial junction zone thickness and correlated with ICSI outcome. RESULTS: There were statistically significant differences between group I and group II in JZ thickness in the 3 uterine regions (fundus, anterior and posterior walls) with p < 0.001, but there were mildly statistically significant differences between both groups in chemical and clinical pregnancy rates with p < 0.01. CONCLUSIONS: The thickness of JZ in patients with history of unexplained RIF was higher than those with unexplained infertility scheduled for ICSI. The JZ thickness was inversely correlated with increased embryo implantation rates in ICSI procedures, the thinner the JZ thickness was associated with higher pregnancy rates (both chemical and clinical pregnancy rates).

Secondary sex ratio of assisted reproductive technology babies

Objective: To assess the secondary sex ratio (SSR) of assisted reproduction technology babies within a fertility clinic in Melbourne, Australia.Methods: A retrospective cohort study was conducted on 3 369 babies who were born following single embryo transfer between 2011 and 2016. Variables examined included embryo creation date, maternal and paternal ages, maternal body mass index (BMI), type of infertility, sperm parameters, fertilization method, type of embryo and stage of transfer. Multivariate regression analysis was performed on the data set using STRATA V9.2.Results: More males were found to be born to embryos created in summer (adjusted odds ratio=1.46, 95% confidence interval: 1.08-1.95; P=0.01). Lower BMI of mother (≤30 kg/m2) and younger age of parents (≤35 years) were associated with an increase in SSR (50.1% vs. 47.1%; 50.6% vs. 48.8%; 50.0% vs. 48.9%) respectively. Decreased SSR value was associated with primary subfertility, using in-vitro fertilization procedure and fresh embryo compared to those with secondary subfertility, using intracytoplasmic sperm injection procedure and thawed embryo (49.3% vs. 50.6%; 47.1% vs. 50.8%; 49.4% vs. 50.1%) respectively.Conclusions: The fertilization methods, type of embryo, stage of embryo transfer, parameters of the sperm and status of subfertility do not significantly affect the SSR. SSR is affected by the seasonality, maternal age, and BMI.

Genes for anti-Müllerian hormone and androgen receptor are underexpressed in human cumulus cells surrounding morphologically highly graded oocytes.

Objectives:The aim of this study was to investigate the expression of genes crucial for the quality of the oocyte and whether expression levels of these genes in cumulus cells can be biological markers for the quality of the oocyte, zygote or embryo, or even for achievement of pregnancy after the assisted reproductive technology procedure. We examined the expression profile of the anti-Müllerian hormone (AMH) gene and its respective receptors: anti-Müllerian hormone receptor type 2 (AMHR2), follicle-stimulating hormone receptor (FSHR) and androgen receptor (AR) in cumulus cells (CCs) surrounding the oocyte, as well as AMH concentrations in follicular fluid of the associated follicle. The obtained gene expression levels were correlated with the morphological quality of the associated oocyte, zygote and embryo as well as with assisted reproductive technology outcome following the intracytoplasmic sperm injection procedure.Methods:This study involved 129 cumulus cells and 35 follicular fluid samples, taken from 58 patients undergoing the intracytoplasmic sperm injection procedure. Oocytes, zygotes and embryos were assessed for morphological quality. The relative gene expression of AMH, AMHR2, FSHR and AR was calculated using the delta–delta Ct method. Anti-Müllerian hormone concentrations in follicular fluids were measured by enzyme-linked immunosorbent assay.Results:The results yielded suggest a relationship between AMH, AR and oocyte morphology: AMH and AR gene expression levels in CCs surrounding morphologically optimal oocytes were significantly lower than in CCs surrounding oocytes with suboptimal morphology (p = 0.011 and p = 0.008, respectively). Statistically significant positive correlation was found between mRNA expression levels of AMH and FSHR (p < 0.001), AMH and AR (p = 0.001), AMHR2 and FSHR (p < 0.001), AMHR2 and AR (p < 0.001), as well as between FSHR and AR (p < 0.001).Conclusion:Assessed results point to AMH and AR relation with oocyte maturity, but not with its fertilization potential, or with embryo quality.

Effect of Follicular Fluid Lactoferrin Level on Oocytes Quality and Pregnancy Rate in Intracytoplasmic Sperm Injection Cycles

Assessment of oocyte quality to avoid overproduction of embryos is now considered an important goal in ICSI cycles. Although in classic Intra Cytoplasmic Sperm Injection (ICSI) cycles selection is mainly done for embryos and not for follicles. Follicular fluid and its contents representing the oocyte environment are now gaining more attention because not only of its crucial influence on oocyte developments but also due to easy isolation of follicular fluid with every case of ICSI. By its aspiration during ovum pick up (OPU). substances isolated from follicular fluid to assess oocyte quality were in the form of: hormones like, Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), Anti-mullerian Hormon (AMH), Growth Hormone (GH) etc., growth factors like Insulin Like Growth Factor (ILGF), proteins and amino acids like Lactoferrin (LF) which is an iron-binding glycoprotein that was detected in follicular fluid and was thought to be related to good oocyte quality when present in high concentration. Objectives: The aim of this study was to detect the possible effect of follicular fluid lactoferrin on oocyte quality and hence pregnancy rate in ICSI cycles. Methods: Follicular fluid was obtained from 64 patients undergoing Intra-cytoplasmic sperm injection (ICSI) procedure in Ain Shams University Maternity Hospital. Follicular fluid (FF) was collected at the time of oocyte harvesting. The lactoferrin concentration in FF was assayed by ELISA. Results: The mean LF concentration in follicular fluid of the positive biochemical pregnancy group (0.63 ± 0.17 ng/mL) was not significantly higher than that in the negative biochemical pregnancy (0.61 ± 0.16 ng/mL). A positive correlation between a number of mature oocytes and lactoferrin concentration was not found (r.101). Conclusion: Lactoferrin level in follicular fluid does not correlate with oocytes quality or pregnancy rate.

A Meta-Analysis of Oocyte Quality and Clinical Outcome of Intracytoplasmic Sperm Injection Cycles in Patients with Polycystic Ovary Syndrome

Whether treatment outcomes in intracytoplasmic sperm injection (ICSI) cycles differ between polycystic ovary syndrome (PCOS) and non-PCOS patients remains inconclusive. The aim of this meta-analysis is to compare oocyte quality and clinical outcomes in PCOS patients and non-PCOS patients undergoing ICSI procedure. Methods: Databases including Medline, Embase and the Cochrane Library were searched for studies published up to 18 January 2018. All articles searched were independently reviewed and selected by two evaluators according to predetermined inclusion and exclusion criteria. Results: Eight studies were selected for inclusion, and a total of 474 PCOS patients (627 cycles) were compared to age-matched controls (962 patients, 992 cycles). The total gonadotropin usage was significant less in PCOS group, and the MD was –481.72 IU [95% CI:– 603.76–359.68, P &lt; 0.00001]. A significantly higher mean number of oocytes, metaphase II (MII) oocytes and 2-pronucleated (2PN) oocytes per cycle were observed in the PCOS group than in the non-PCOS group (P &lt; 0.005), the total MDs were 4.20 [95% CI: 2.50–5.90], 1.52 [95% CI: 0.84–2.20] and 1.71 [95% CI: 0.73–2.69], respectively. The fertilization rate was comparable. The clinical pregnancy rate per transfer cycle was higher in the PCOS group, the odd ratio (OR) was 1.59 [95% CI: 1.23–2.05, P &lt; 0.005] and the live birth rate was comparable. A higher miscarriage rate was observed in the PCOS group, but this did not reach statistical significance. The risk of ovarian hyperstimulation syndrome (OHSS) was significantly higher in the PCOS group [OR: 3.01, 95% CI: 1.16–7.82, P &lt; 0.05]. Conclusions: This meta-analysis demonstrated that PCOS patients may have more favourable ICSI outcomes than non-PCOS patients. Our findings should be evaluated in further prospective cohort studies.

Intra-Technician Comparison of Sperm Morphology Evaluation

Objective To identify intra-technician subjective variability in sperm morphology assessment at a single site. Design Intra-technician evaluation of sperm morphology results compared to reported outcome on initial semen analysis using Kruger (Tygerburg) Strict criteria. Materials and Methods This study was a retrospective analysis of 35 semen sample slides stained using a Romanowski stain (Diff-Quick). Slides were randomly selected for semen samples analyzed January 1, 2018 – February 28, 2018 and numbered (1-35). Morphology assessment was performed by 3 andrologists, blinded to initial reported morphology results. The median percentage of normal forms (normal, ≥ 4%) were compared between 3 technicians to the initial reported values. Statistical analysis was performed using One-Way Analysis of Variance (ANOVA) and ANOVA on Ranks using Kruskan-Wallis test, with p < 0.05 considered significant. Results Median percentage of normal forms was 4% for the initial semen analysis report. Whereas, technician 1: 3%, technician 2: 2% and technician 3: 3%. The median values among the technicians were significant (P = 0.021). There were no differences between reported values compared to each technician. However, the median results for technician 3 and technician 2 were different (P < 0.05). Conclusions The findings suggest that there is variability in assessment of sperm morphology between technicians. Inaccurate assessment reporting high percentage of abnormal forms for sperm morphology can result in misrepresented sperm data by clinicians. As a result, since low morphology is an indication for intra-cytoplasmic sperm injection (ICSI) during in-vitro fertilization (IVF) procedures, ICSI can be unnecessarily ordered. The additional financial cost associated with an ICSI procedure may result in financial and emotional burden for the patient. Furthermore, an incorrect abnormal morphology result leads to emotional strain on the male patient leading to the feelings of inadequacy. An increased frequency of quality assurance training of andrology technicians in groups reviewing Kruger (Tygerburg) Strict Criteria is recommended to limit assessment variability. Randomly selected images of sperm morphology from the Kruger Strict Atlas can be used to assess technician sperm morphology evaluations. Disclosures Nothing to disclose. Funding None. To identify intra-technician subjective variability in sperm morphology assessment at a single site. Intra-technician evaluation of sperm morphology results compared to reported outcome on initial semen analysis using Kruger (Tygerburg) Strict criteria. This study was a retrospective analysis of 35 semen sample slides stained using a Romanowski stain (Diff-Quick). Slides were randomly selected for semen samples analyzed January 1, 2018 – February 28, 2018 and numbered (1-35). Morphology assessment was performed by 3 andrologists, blinded to initial reported morphology results. The median percentage of normal forms (normal, ≥ 4%) were compared between 3 technicians to the initial reported values. Statistical analysis was performed using One-Way Analysis of Variance (ANOVA) and ANOVA on Ranks using Kruskan-Wallis test, with p < 0.05 considered significant. Median percentage of normal forms was 4% for the initial semen analysis report. Whereas, technician 1: 3%, technician 2: 2% and technician 3: 3%. The median values among the technicians were significant (P = 0.021). There were no differences between reported values compared to each technician. However, the median results for technician 3 and technician 2 were different (P < 0.05). The findings suggest that there is variability in assessment of sperm morphology between technicians. Inaccurate assessment reporting high percentage of abnormal forms for sperm morphology can result in misrepresented sperm data by clinicians. As a result, since low morphology is an indication for intra-cytoplasmic sperm injection (ICSI) during in-vitro fertilization (IVF) procedures, ICSI can be unnecessarily ordered. The additional financial cost associated with an ICSI procedure may result in financial and emotional burden for the patient. Furthermore, an incorrect abnormal morphology result leads to emotional strain on the male patient leading to the feelings of inadequacy. An increased frequency of quality assurance training of andrology technicians in groups reviewing Kruger (Tygerburg) Strict Criteria is recommended to limit assessment variability. Randomly selected images of sperm morphology from the Kruger Strict Atlas can be used to assess technician sperm morphology evaluations.

Confocal Raman spectroscopy and multivariate data analysis for evaluation of spermatozoa with normal and abnormal morphology. A feasibility study

This paper investigates a feasibility of using confocal Raman spectroscopy (CRS) and multivariate analysis for classification of sperm cells. The spectral based classification is compared with the morphological analysis, which is the main criterion for sperm selection in intracytoplasmic sperm injection procedure. The spectral analysis is conducted using the data driven soft independent modeling of class analogies method. The supervised classification reveals numerous outliers that pass from the 'normal' class to the 'abnormal' class, and vice versa. The ultimate result shows that the initial morphological discrimination overlaps with the spectral classification only partly. It is shown that CRS provides additional information regarding the nuclear DNA stability and helps to reveal spermatozoa with fragmented and defective DNA. This can be a promising direction for future evaluation of spectra from live, unfixed cells.

In-vitro equine embryo production: an adventure in multi-unit collaboration

The accessibility of assisted reproductive techniques related to oocyte collection and in vitro fertilization in the mare is increasingly becoming more widespread for private equine practitioners and their clients. The necessity of hauling a valuable mare long distances to the closest facility for transvaginal aspiration (TVA) of oocytes and on site intracytoplasmic sperm injection (ICSI) over a period of potential weeks before obtaining a pregnancy no longer exists. Instead, a mare’s primary veterinarian can manage the patient’s cycle and briefly transport her to a nearby facility where TVA can be performed. With TVA services being offered with increasing frequency in densely equine populated areas it is increasingly common for TVA to be offered as an outpatient procedure even if ICSI laboratories are not present at that location. The majority of reports associated with TVA and ICSI are associated with benchtop research and typically do not continue beyond blastocyst development to report pregnancy rates. A cumulative pregnancy rate after TVA and ICSI were performed on immature oocytes, with shipment of blastocysts to an off-site recipient herd has been reported at 72% with a 52% foaling rate.1 In 2017, in collaboration with an exclusively equine reproduction veterinary practice, we worked together to provide TVA services for client-owned mares. Mares were referred from the veterinary practice to our university setting where outpatient TVA procedures were performed; 330 follicles were aspirated and 212 oocytes were recovered (mare age range from 4-26 years) and shipped overnight at room temperature in an Equitainer® to an ICSI laboratory via FedEx™. A total of 43 (26.1%) blastocysts resulting from 165 ICSI procedures were obtained; 29 embryos were vitrified for future use and 14 embryos were transported to the referring reproductive veterinary practice via courier for fresh transfer to recipient mares on day 7 or 8 after ICSI. A 14 day pregnancy rate of 71.4% (10/14) and a 50% (7/14) pregnancy rate at heartbeat check were recorded. Three recipient mares have already successfully foaled and four are due to foal later in the spring of 2018. In light of these results sending a mare to a local practice specializing in TVA, as opposed to a distant, self-contained facility that has both TVA and ICSI capabilities has benefits for referring equine practitioners, their clients and patients. Being a key element in the management of the mare’s cycle prior to TVA has allowed the primary veterinarian to remain an integral partner in providing these specialty services. Shorter duration stays at referral facilities translates into less expense for the mare owner. Finally, as collaborators in this working partnership we have found working as a team to offer such services has enhanced the working relationship between practitioners and specialists.

An efficiency comparison of different in vitro fertilization methods: IVF, ICSI, and PICSI for embryo development to the blastocyst stage from vitrified porcine immature oocytes

BackgroundMost studies carried out to evaluate recovery and development after porcine oocyte vitrification, reported better rates when cryopreserved in embryonic development stages or zygotes, but not in immature oocytes. For this reason, many studies are performed to improve immature oocyte vitrification protocols testing the use of different cryoprotectant concentrations, cooling devices, incubation times; but only a few of them have evaluated which fertilization procedure enhances blastocyst rates in vitrified oocytes. Therefore, this study was aimed to evaluate: 1) if the sperm selection with hyaluronic acid (HA) or polyvinylpyrrolidone (PVP) before injection could play a key role in increasing fertilization and blastocyst formation and 2) the embryo developmental ability and blastocyst production of porcine immature oocytes retrieved after vitrification-warming and co-cultured with granulosa cells during IVM, using different fertilization techniques: in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and conventional ICSI with hyaluronic acid (HA) sperm selection, known as physiological intracytoplasmic sperm injection (PICSI) and.ResultsSperm selected with HA-PICSI displayed a higher percentage of live/acrosome reacted status compared to those in control and exposed to PVP. Higher dead/acrosome reacted rates were obtained after PVP exposure compared to control and HA. In oocytes, viability significantly decreased after IVM in vitrified oocytes. Besides, IVM rates were not different between control denuded oocytes cultured with granulosa cells (DO-GC) and vitrified oocytes. Regarding fertilization parameters, IVF showed higher percentages of total fertilization rate than those obtained by ICSI and PICSI. However, results demonstrate that PICSI fertilization increased the blastocysts formation rate in control DO-GC and vitrified oocytes compared to IVF and ICSI.ConclusionsTo achieve high blastocyst formation rates from vitrified GV oocytes, it is recommended that sperm should be selected with HA instead of PVP before injection since high viability and acrosome reaction rates were obtained. Also, PICSI fertilization was the best method to produce higher blastocyst rates compared to the IVF and ICSI procedures.

Delayed childbearing and female ageing impair assisted reproductive technology outcome in survivors of male haematological cancers.

To analyse the impact of female characteristics on assisted reproductive technology outcome among male haematological cancer survivors. A retrospective analysis of 93 haematological cancer survivors attending our tertiary referral fertility centre between June 1998 and June 2017 for achieving fatherhood with assisted reproductive technology treatments. A progressive increase in the median female age was observed during the study period (32.2years until the year 2007 and 36.9years from the year 2012). Fifty-five out of 93 patients were treated with intracytoplasmic sperm injection (ICSI) (113 ovarian stimulations, 108 ICSI procedures). Cryopreserved ejaculated sperm was used in 28 couples, fresh sperm in 19, and thawed testicular sperm in 8 couples. Mean female age at ovarian stimulation was 37.0±4.7years. Twenty-six pregnancies resulted in a full-term birth (23% per started ovarian stimulation; 43.6% per couple) and 33 children were born. No significant differences were observed according to source of sperm (fresh, frozen, testicular) and multivariate analysis confirmed that maternal age was the only variable inversely related to the cumulative delivery rate, being five times lower (15.7%) when the female partner was ≥ 40years (OR = 0.22, 95% CI 0.06-0.77) vs. 58.3% with younger women (p = 0.0037). Delayed childbearing and female ageing affect ICSI outcome in couples where the male is a survivor of haematological cancer. This topic should be discussed when counselling male cancer patients about fertility preservation.

Mitochondrial activity and cytoskeleton organization in three pronuclei oocytes after intracytoplasmic sperm injection.

SummaryDigyny, the presence of a third pronucleus due to the failure of second polar body extrusion, is problematic after intracytoplasmic sperm injection (ICSI) practices. Mitochondria have critical roles such as production of adenosine triphosphate (ATP) and regulation of Ca2+ homeostasis during oocyte maturation, fertilization and the following development, while the regulation of meiotic spindle formation, chromosome segregation, pronuclear apposition and cytokinesis is closely associated with the cytoskeleton. In this study, mitochondrial membrane potential, distribution of F-actin and γ-tubulin, and the ultrastructure of three pronuclear (3PN) oocytes were investigated. 3PN oocytes after ICSI procedure were taken from patients who were enrolled in assisted reproduction programmes. For mitochondrial membrane potential analysis, fresh oocytes stained with the mitochondrial membrane potential probe JC-1, were evaluated under fluorescence microscopy. The mitochondrial membrane potential of three pronuclear oocytes showed similar results to normal zygotes. γ-Tubulin was stained strongly at the subplasmalemmal domain and microfilaments were localized at the cortical, but not the perinuclear, area. Cytoplasmic halos were moderately or not detected by electron microscopy; lipofuscin granules, degenerated mitochondria, and multilamellated bodies were seen in the ooplasm. Immunohistochemistry and electron microscopic findings suggested that mitochondrial membrane potential has no direct effect on second polar body extrusion. This abnormality can be associated with an altered cytoskeleton due to poor oocyte quality.

Non-invasive metabolomic profiling of culture media of ICSI- and IVF-derived early developmental cattle embryos via Raman spectroscopy

The aim of the present study was to compare differences in composition between in vitro cultured early developmental embryos resulting from either in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Non-invasive metabolomic profiling of culture media was conducted with laser tweezer Raman spectroscopy (LTRS), providing molecular information that was used to aid the diagnosis or treatment of embryos that were adversely affected by ICSI treatment, ultimately improving the ICSI embryonic developmental potential. Cattle embryos were generated via ICSI and IVF with development to the 2-, 4-, 8-, 16-,32-cell, and blastocyst stages with individual in vitro culturing occurring for 4 h. The culture media for embryos in different developmental stages were separately analyzed using LTRS. The resulting composition of culture media used for culturing IVF- and ICSI-derived embryos was mainly altered in contents of carbohydrates, lipids, DNA, and proteins. Bands at 1004 cm−1 (phenylalanine) and 1529 cm−1 (-C = C-carotenoid) had specific patterns related to the metabolicactivity of embryos; using LTRS, and these may be considered as biomarkers for embryonic development. Furthermore, the vibrations of lipids at different stages increased more with assessment of ICSI culture media than in IVF media. Discriminant function analysis can be utilized for the classification of culture media used for culture of ICSI- and IVF-derived embryos. In conclusion, LTRS can be used for development of an independent assay to assess embryo status during both ICSI and IVF procedures, which provides novel insights into differences in structure and components of single cells.

Three Dimentional Transvaginal U/S Measurements Of Uterine Junctional Zone Thickness And Its Relation To Implantation Success Rate In ICSI Cycles

Background: Sub-endometrial junction zone (JZ) plays an important role in most of reproductivefunctions. Objective: To find out the effect of the sub-endometrial JZ thickness assessment by 3-D transvaginalultrasound (TVUS) on intracytoplasmic sperm injection (ICSI) outcomes in patients withunexplained recurrent implantation failure (RIF). Setting: ART Unit of Obstetrics and GynecologyDepartment, Faculty of Medicine, South Valley University, Qena, Egypt. Duration: From April 2016 toOctober 2018. Study Design: A prospective observational study. Methds: Fifty couples with history ofunexplained RIF in previous ICSI cycles and prepared for another ICSI cycle (group I) and fifty coupleswith unexplained infertility prepared for ICSI for the first time (group II) had been included in this study.At time of ovum pick up, 3-DTVUS was done for all cases in both groups for assessment of subendometrial junction zone thickness and correlated with ICSI outcome. Results:There were statisticallysignificant differences be- tween group I and group II in JZ thickness in the 3 uterine regions (fundus,anterior and posterior walls) with p < 0.001, but there were mildly statistically significant differencesbetween both groups in chemical and clinical preg- nancy rates with p < 0.01. Conclusions: Thethickness of JZ in patients with history of unexplained RIF was higher than those with unexplained infertilityscheduled for ICSI. The JZ thickness was inversely correlated with increased embryoimplantation rates in ICSI procedures, the thinner the JZ thickness was associated with higher pregnancyrates (both chemical and clinical pregnancy rates).

657 Nomogram to predict the outcomes of intracytoplasmic sperm injection (ICSI)

Although many studies have shown that the success of ICSI depends on a number of male factors, female factors and laboratory procedures, no one has analyzed these variables collectively. We aimed to create a nomogram capable of predicting the outcome of ICSI based on all important available clinical and laboratory prognostic variables. We prospectively analyzed 1112 ICSI cycles for 1112 infertile couples performed for male or female infertility.We used multiple factors to build the first nomogram to predict the probability of clinical pregnancy of ICSI. All ICSI procedures were performed in the same institute using the same technique and the same operators. 11 variables (male factors, female factor, and laboratory factors) were included and the final nomogram was internally validated with bootstrap analysis. Multiple logistic regression analysis was conducted. The final nomogram was built from the logistic model. The model discrimination was quantified by a concordance index (CI), which is equal to the non-parametric area under the Receiver operating characteristic (ROC) curve.

Electroejaculation combined with assisted reproductive technology in psychogenic anejaculation patients refractory to penile vibratory stimulation.

BackgroundPsychogenic anejaculation (PAE) is a rare disease in clinic seriously affecting quality of life, especially fertility of patients. However, there is not an effective treatment for these patients. The aim of the study was to evaluate the results of treatment with electroejaculation (EEJ), intrauterine insemination (IUI), and intracytoplasmic sperm injection (ICSI) in patients with PAE who were refractory to penile vibratory stimulation (PVS).MethodsTwenty PVS-refractory patients with PAE were included in the retrospective study. Semen was retrieved through EEJ procedures. According to the semen quality and patient preference, IUI or ICSI was selected.ResultsA total of 23 procedures of EEJ were performed in 20 patients. Sperm was successfully retrieved in all of the cases. Yielded semen was used in a total of 16 ICSI procedures in 6 couples. The fertilization rate was 54.0%. The ICSI cycles resulted in three pregnancies; of these one resulted in a live birth. Fourteen couples were treated with 26 IUI cycles, which resulted in live births in five pregnancies.ConclusionsEEJ offers an encouraging pregnancy opportunity for men with PAE who were refractory to PVS. The combined use of EEJ and assisted reproductive techniques is excellent management for PAE infertility. In our study, EEJ resulted in pregnancies and the birth of six healthy children.

Human oocyte-like germ cells derived in vivo from testicular tissue of a Klinefelters’ patient: A case report

The combination of testicular biopsy and intracytoplasmic sperm injection procedures has assisted potentially azoospermic men with Klinefelters’ syndrome to have children. Approximately two-thirds of the time, a testicular sperm extraction procedure performed on a nonmosaic Klinefelter 47,XXY patient fails to isolate sperm. In the case report described herein, both wet preparation and histological examinations confirmed typical cellular morphologies associated with Klinefelter testicular tissue, including no spermatozoa. It did however, observe the unusual presence of oocyte-like cells representing a unique, theoretical in vivo transformation of pluripotent stem cells, probably spermatogonial, into primordial germ cells and gametes of the opposite sex, as previously documented only in the experimental mouse model.

Obstetric and neonatal outcome following ICSI with assisted oocyte activation by calcium ionophore treatment.

Calcium ionophore treatment is being used in assisted reproductive technology (ART) for cases with previous low fertilization rate or total absence of fertilization after insemination by intracytoplasmic sperm injection or when a specific indication such as globozoospermia is present. As this technique is more invasive and differs from the physiological process of fertilization, a thorough investigation of the health of the children born following this procedure is required. We intent to report the medical outcome of all children conceived following calcium ionophore treatment in our IVF center. One-armed descriptive study is performed to report the obstetrical and neonatal outcome of children born after using calcium ionophore treatment during the intracytoplasmic sperm injection procedure in our center. A number of 237cycles were included in this study, with 74 pregnancies reported, from which 47 children (31 singletons and 16 twin children) were born. No major malformations were detected in singletons. In twins, three children were diagnosed with major malformations. Minor malformations were present in seven singletons and in one twin. In conclusion, our results regarding birth characteristics and congenital malformations are within the expected range but, although reassuring, should be interpreted with caution due to the small number of children included.

Effect of Vitamin D Supplementation on Intracytoplasmic Sperm Injection Outcomes: A Randomized Double-Blinded Placebo-Controlled Trial

OBJECTIVE: To assess the effect of vitamin D supplementation on enhancement of embryos implantation rates in intracytoplasmic sperm injection procedures. STUDY DESIGN: A randomized double-blinded controlled trial. SETTING: Assisted Reproduction Unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Egypt. DURATION: From December 1st 2017 to the end of October 2018. SUBJECTS and METHDS: Eighty infertile couples that underwent ICSI procedure were entrapped in this study. Patients were divided into two groups; Group I (Vitamin D group) = 40 patients (received vitamin D supplementation in the form of Vidrop oral drops 600 IU/day starting after ovum pickup) and Group II (placebo group) = 40 patients (received a placebo “normal saline oral drop”). RESULTS: There was a significantly increased embryo implantation rates (53.2% in vitamin D group versus 46.7% in placebo group), there was a mild statistically significant difference in the clinical pregnancy rates (45.9% in vitamin D group versus 39.4% in placebo group) with p-value CONCLUSIONS: Vitamin D supplementation could improve embryos implantation rates and rate of ongoing pregnancy by decline in the first trimester miscarriage rate.