ObjectiveTo investigate the effects of in vitro myo-inositol (Myo-Ins) supplementation of cryopreserved human semen on the cryo-survival rate (CSR).MethodsSemen samples were obtained from 41 infertile men. Following routine semen analysis, each sample was divided into two equal aliquots (0.5 mL each). One aliquot was treated with 1 mg of Myo-Ins dissolved in 10 µL of sperm preparation medium. The second aliquot was treated with 10 µL of the same medium (control). Both aliquots were incubated for 20 minutes prior to freezing to slow the freezing process. The frozen samples were examined for post-thaw percentages of total motility (TM), progressive motility (PM), and the CSR, defined as the percentage of post-thaw TM divided by the percentage of pre-freeze TM and multiplied in 100. The results were expressed as median and interquartile range (25th and 75th percentiles).ResultsThe pre-freeze TM (50% [30%–50%]) and PM (35% [20%–35%]) were significantly higher than the post-thaw TM and PM in the Myo-Ins group (15% [10%–35%] and 10% [5%–20%]; p<0.001 and p<0.001, respectively) and the control group (10% [6%–30%] and 5% [3%–15%]; p<0.001 and p<0.001, respectively). The CSR of the 41 semen aliquots supplemented with Myo-Ins (40% [25%–70%]) was significantly higher than that of the control samples (30% [13%–58%], p=0.041). The CSR of the 26 abnormal semen samples that were supplemented with Myo-Ins (38% [20%–50%]) was significantly higher than that of the control samples (23% [12%–30%], p=0.031).ConclusionIn vitro Myo-Ins supplementation of ejaculated human sperm from infertile men resulted in a significant increase in the CSR in samples with abnormal pre-freeze sperm parameters.

Objective: To evaluate the effects of cytokine enrichment of culture medium on embryological and clinical outcomes after intracytoplasmic sperm injection (ICSI). Design: A randomized clinical trial. Setting: In vitro fertilization centers. Patient(s): This trial included 443 ICSI cycles randomized into two groups. Intervention(s): This study evaluated the influence of integration of granulocyte-macrophage colony-stimulating factor, heparinbinding epidermal growth factor-like growth factor, and leukemia inhibitory factor into culture media on human embryo development after ICSI. Main Outcome Measure(s): Ongoing pregnancy rate per a randomized participant. Result(s): Cytokine enrichment of culture medium showed improvement in ongoing pregnancy rate compared with no cytokines (106/ 224 [47%] vs. 78/219 [36%]; absolute rate difference [ARD] ¼ 12; 95% confidence interval [CI], 2.5-21). This integration of cytokines also showed better rates of live birth (101/224 [45%] vs. 71/219 [33%]; ARD ¼ 13; 95% CI, 4-21) and cumulative live birth (132/224 [60%] vs. 97/219 [44%]; ARD ¼ 12; 95% CI, 4-20) and lower rate of pregnancy loss (27/124 [22%] vs. 37/103 [36%]; ARD ¼ -14; 95% CI, -26 to -2) than conventional medium. Embryos developed in the cytokine-supplemented medium showed better blastocyst formation, quality, cryopreservation, and use than control medium. Conclusion(s):Integration of cytokines into human embryo culture media showed improvement in embryological and clinical outcomes after ICSI. However, the long-term effect of cytokine enrichment of a medium is still unclear and warrants further studies with longitudinal follow-up.

This study examined the relationship between oxidation–reduction potential (ORP) in frozen‐thawed semen and the post‐thaw sperm parameters. Levels of ORP were measured in 25 samples from men presenting for routine infertility work‐up and were expressed as millivolt (mV)/106 sperm/ml. Frozen‐thawed samples were examined for post‐thaw total motility (TM%), progressive motility (PM%), total sperm count (TSC) and ORP. The cryo‐survival rate (CSR) was calculated as post‐thaw TM/pre‐freeze TM × 100. Data are provided as median and interquartile range (25th and 75th percentiles). The post‐thaw TM% (10.0% [4.00%, 15.1%]), PM% (5.88% [2.97%, 9.33%]) and TSC (12.5 [10.0, 17.5] × 106 sperm) were significantly lower than the pre‐freeze TM% (45.9% [32.9%, 59.1%], PM% (31.5% [24.4%, 40.0%] and TSC (120 [90, 250] ×106 sperm) (p < .001). Post‐thaw ORP (2.62 [2.52, 3.13] mV/106 sperm/ml) was significantly higher than pre‐freeze ORP (0.73 [0.54, 1.21] mV/106 sperm/ml; p < .001). The CSR was 21.7% (11.3%, 31.9%). The post‐thaw seminal ORP was negatively correlated with post‐thaw TM% (r = −.5; p = .02), PM% (r = −.41; p = .03), TSC (r = −.60; p = .03) and CSR (r = −.52; p = .01). Increased levels of ORP are significantly correlated with poor post‐thaw sperm quality and CSR.

OBJECTIVE: Myo-inositol is a naturally occurring component of vitamin B complex that is involved in cellular signaling pathways. There is a growing evidence on the potential beneficial effects of Myo-inositol supplementation on both natural and assisted fertility. The objective of this study was to investigate the effects of in vitro Myo-inositol supplementation of cryopreserved human semen on the post-thaw sperm quality.DESIGN: A randomized controlled trial (RCT). MATERIALS AND METHODS: The study had been approved by the institutional review board, and conducted in an infertility center between September 2016 and March 2017. The study included semen samples obtained from 25 infertile men during routine infertility work-up. Myo-inositol was supplied in a powder form (Sigma Aldrich Chemical Co, USA). Under complete aseptic condition, Myo-inositol was dissolved in sperm nourishment media (AllGrad washÒ, Life global, Canada) to prepare aliquots of 10 ml stock solutions, each containing 1 mg Myo-inositol. Standard semen analysis was performed according to the WHO guidelines (fifth edition, 2010). Azoospermic and leukocytospermic samples were excluded. Each semen sample was divided into two equal aliquots (0.5 ml each) for cryopreservation using slow freezing technique. The two identical aliquots of each semen sample were randomized into two groups (A and B). Group A was treated with cryo-protectant plus 10 ml Myo-inositol solution, while group B was treated with cryo-protectant alone (control). Frozen samples were thawed at 37 C, and examined for post-thaw total motility, progressive motility and progressive recovery rate (PRR). PRR ¼ post-thaw progressive motility/pre-freeze progressive motility X100. Data were presented as median (25 th and 75 th percentiles). Paired sample t test was used for comparison of the pre and post-thaw results. P value < 0.05 was considered significant.RESULTS: The median (25 th , 75 th ) of pre-freeze sperm concentration was 25 (18, 41) X10 6 sperm, and percentage of normal sperm forms was 4 (2, 5). Comparisons of pre-freeze and post-thaw percentages of total motility, progressive motility and PRR between samples in group A and group B are shown in Table 1.CONCLUSIONS: In vitro Myo-inositol supplementation of cryopreserved ejaculate sperm, from infertile men, resulted in a significant enhancement of post-thaw sperm quality. Such finding is interesting, and may have important implications on the future outcome of assisted reproductive techniques using cryopreserved sperm.

STUDY QUESTION Does the use of a laser to open the zona pellucida during ICSI (laser assisted or LA-ICSI) improve oocyte survival, embryo development and clinical outcomes? SUMMARY ANSWER Compared to conventional ICSI, LA-ICSI increased rates of oocyte survival and some aspects of embryo development but it did not alter the ongoing pregnancy rate; after adjusting for oocyte survival, there was no beneficial effect of LA-ICSI on embryo development and utilization. WHAT IS KNOWN ALREADY Oocyte degeneration occurs in a 10th of mature oocytes after ICSI. Pilot studies suggest that LA-ICSI may improve oocyte survivability. STUDY DESIGN, SIZE, DURATION In a randomized controlled trial, 966 couples (16 122 metaphase II oocytes) were allocated to receive LA-ICSI (intervention) or conventional ICSI (control) between 17 September 2018 and 5 August 2019. Oocyte survival (primary endpoint), embryo development and ongoing pregnancy rates were compared. PARTICIPANTS/MATERIALS, SETTING, METHODS Couples included in this study were recommended for ICSI due to female or male factor, unexplained infertility or a combination of factors. Patients were ineligible to participate in the study if they had uterine abnormality including thin endometrium, recurrent pregnancy loss, endometriosis or a severe medical condition. Concealed randomization to LA-ICSI or conventional ICSI, allocated in a 1:1 ratio, took place on stimulation Day 1 with replacement of blastocysts on only Day 5. The primary endpoint was oocyte survival with membrane integrity 24 h after the ICSI procedure. The sample size was estimated to detect a 3% increase in oocyte survival after LA-ICSI with 99% power at a 1% significance level. This also permitted the detection of 10% increase in ongoing pregnancy rate after LA-ICSI with 85% power at 5% alpha level. We used Poisson regression with zero-inflation for count data to estimate relative risk (RR) with 95% CI and logistic regression for clinical outcomes to estimate odds ratio (OR) with 95% CI. Both models adjusted for age as a covariate. MAIN RESULTS AND THE ROLE OF CHANCE Compared with conventional ICSI, LA-ICSI resulted in a higher number of surviving oocytes (RR 1.08, 95% CI 1.05–1.12, P &lt; 0.001), as well as a higher number of fertilized oocytes (RR 1.08, 95% CI 1.04–1.13, P &lt; 0.001) and utilizable blastocysts (RR 1.09, 95% CI 1.04–1.15, P &lt; 0.001). Sensitivity analyses adjusted for oocyte survival showed no between-group difference in utilizable blastocysts (OR 1.01, 95% CI 0.95–1.08, P = 0.73) and by calculating the mean rate, a reduction in utilizable blastocysts was shown (RR 0.95, 95% CI 0.94–0.97, P &lt; 0.001). Ongoing pregnancy showed no between-group difference (LA-ICSI 179/489 (37%) vs ICSI 201/477 (42%), OR 0.79, 95% CI 0.61–1.03, P = 0.09). LIMITATIONS, REASONS FOR CAUTION It was not possible to blind the embryologists involved in the ICSI procedure. However, there was concealment of randomization and blinding of outcome assessments reducing the risk of selection and measurement bias. WIDER IMPLICATIONS OF THE FINDINGS A beneficial effect of LA-ICSI on oocyte survival should be shown to improve clinical outcomes, before its use in clinical practice is justified. STUDY FUNDING/COMPETING INTEREST(S) The study received no funding, and the authors declare that there are no conflicts of interest. TRIAL REGISTRATION NUMBER NCT03665103 TRIAL REGISTRATION DATE 11 September 2018 DATE OF FIRST PATIENT'S ENROLMENT 17 September 2018