The exact synchronization timing between the cleavage embryo stage and duration of progesterone therapy-improved pregnancy rates in frozen embryo transfer cycles: A cross-sectional study.

BACKGROUND: Synchronization between the embryonic stage and the uterine endometrial lining is important in the outcomes of the vitrified-warmed embryo transfer (ET) cycles. OBJECTIVE: The aim was to investigate the effect of the exact synchronization between the cleavage stage of embryos and the duration of progesterone administration on the improvement of clinical outcomes in frozen embryo transfer (FET) cycles. MATERIALS AND METHODS: 312 FET cycles were categorized into two groups: (A) day-3 ET after three days of progesterone administration (n = 177) and (B) day-2 or -4 ET after three days of progesterone administration (n = 135). Group B was further divided into two subgroups: B1: day-2 ET cycles, that the stage of embryos were less than the administrated progesterone and B2: day-4 ET cycles, that the stage of embryos were more than the administrated progesterone. The clinical outcome measures were compared between the groups. RESULTS: The pregnancy outcomes between groups A and B showed a significant differences in the chemical (40.1% vs 27.4%; p = 0.010) and clinical pregnancies (32.8% vs 22.2%; p = 0.040), respectively. The rate of miscarriage tended to be higher and live birth rate tended to be lower in group B than in group A. Also, significantly higher rates were noted in chemical pregnancy, clinical pregnancy, and live birth in group A when compared with subgroup B2. CONCLUSION: Higher rates of pregnancy and live birth were achieved in day-3 ET after three days of progesterone administration in FET cycles.

Efficacy of the in vitro splitting of human preimplantation embryos from ART programs

Background/aim: The aim of this study was to evaluate the efficiency of in vitro embryo splitting (IES) procedures. We also assessed the quality of the blastocysts developed from embryos obtained from different sources. Materials and methods: Good quality embryos at 6­8-cell stages were categorized according to their fertilization sources: 1) frozenwarmed donated embryos, 2) chromosomally abnormal embryos, 3) parthenogenetic embryos, and 4) embryos derived from fertilization of in vitro matured oocytes (rescue IVM). After IES, splitting and developmental efficiency was assessed. Furthermre, the quality of the developed blastocysts was evaluated by Hoechst and propidium iodide (PI) staining. Results: The data showed a high rate of both splitting and developmental efficiency in the frozen-warmed embryos after IES (140% and 71.7%, respectively), followed by chromosomally abnormal embryos (96.8% and 52.5%, respectively). Results of the Hoechst and PI staining showed that the mean ± SD cell numbers of the control group were higher (113.11 ± 16.01) than that of twins A (donor blastomeres embryos, 58 ± 12.2) and B (recipient blastomeres embryos, 50.4 ± 8.5), respectively. Conclusion: Chromosomally normal embryos enrolled in IES are more potent to develop into viable blastocysts. For research purposes, 1PN and 3PN embryos are the best options for splitting procedures, regardless of the poor quality of developed blastocysts.

Endometrial preparation using gonadotropin-releasing hormone agonist prior to frozen-thawed embryo transfer in women with repeated implantation failure: An RCT.

BACKGROUND: Preparation of endometrial thickness in frozen-thawed embryo transfer (FET) is extremely important, particularly in repeated implantation failure (RIF) patients. OBJECTIVE: This study aimed to investigate the clinical outcomes of FET cycles among RIF women, based on the effects of administering gonadotropin-releasing hormone (GnRH) agonist prior to estrogen-progesterone preparation of the endometrium. MATERIALS AND METHODS: In this randomized clinical trial, 67 infertile women who were candidates for FET were divided into two groups: A) case group (n = 34), treated with GnRH agonist prior to endometrial preparation and B) control group (n = 33), which received the routine protocol. (6 mg daily estradiol started from second day) The clinical outcomes) including chemical and clinical pregnancy, in addition to implantation rates, were compared between the two groups. RESULTS: The results showed no significant differences in women's age (p = 0.558), duration (p = 0.540), type (p = 0.562), and cause of infertility (p = 0.699). Regarding pregnancy and implantation rates, there was a trend toward an increase in the case group; however, differences were not statistically significant. CONCLUSION: Although our results showed no significant differences between groups. Because there are trends to better results in case group larger sample size may show significant difference.

The Determination of Estradiol to Cumulus Oocyte Complex (COC) Number Ratio: Does it Predict the Outcomes of ART Cycles?

BACKGROUND: The aim of this study was to assess the impact of total serum E2 on the day of human chronic gonadotropin (hCG) administration and the serum E2 per oocyte ratio on the outcomes of assisted reproductive technology (ART) cycles. METHODS: A total of 205 women were categorized into 3 groups according to the serum E2 levels: 1: ≤1500 pg/ml; 2: 1500-3000 pg/ml; 3: >3000 pg/ml. Another categorization included 3 groups according to E2/oocyte ratio: A: ≤150 pg/ml per oocyte; B: 150-200 pg/ml per oocyte; and C: >200 pg/ml per oocyte. The outcome compared between groups included laboratory and clinical characteristics. One-way analysis of variance (ANOVA), chi-square and Kruskal-Wallis, and multiple logistic regression model were performed, and appropriate differences were considered significant at p<0.05. RESULTS: There was a significant difference between the groups based on the E2 levels with respect to laboratory parameters. In group C, the rates of chemical pregnancy (54.1%), clinical pregnancy (50%) and live birth (45.8%) were significantly higher, when compared to other groups. Moreover, according to E2/oocyte ratio, the rate of live birth was higher in group C compared with group A (18.3%, p=0.04), and group C (29.7%, p<0.0001). Logistic regression showed the number of good quality embryos was a positive predictor for live birth (odds ratio=2.03, 95% CI=1-4.1), but the level of E2 on day of HCG was a negative predictor (odds ratio=0.99, 95% CI=0.99-1). CONCLUSION: Supraphysiological levels of E2 had no adverse effects on the quality of the embryos in IVF cycles, but may have adverse effect on live birth in fresh transfer. Also, it is confirmed that both the pregnancy and live birth rates were elevated with E2/oocyte ratio ≥200 pg/ml.

Quality of Blastocysts Created by Embryo Splitting: A Time-Lapse Monitoring and Chromosomal Aneuploidy Study.

OBJECTIVE: The aim of this study was to screen the potential of human embryos to develop into expanding blastocysts following in vitro embryo splitting and then assess the quality of the generated blastocysts based on chromosomal characteristics and using morphokinetics. MATERIALS AND METHODS: In this experimental study, a total of 82 good quality cleavage-stage donated embryos (8- 14 cells) were used (24 embryos were cultured to the blastocyst stage as controls and 58 embryos underwent in vitro splitting). After in vitro splitting, the blastomere donor and blastomere recipient embryos were named twin A and twin B, respectively. Morphokinetics and morphological parameters were evaluated using a time-lapse system in the blastocysts developed from twin embryos. Aneuploidy of chromosomes 13, 15, 16, 18, 21, 22, X and Y were analyzed in the twin blastocysts. RESULTS: Following in vitro splitting, of the 116 resulting twin embryos, 80 (69%) developed to the expanded blastocyst (EBL) stage compared to 21 (87.5%) embryos in the control group (P>0.05). The morphokinetics analysis suggested that the developmental time-points were influenced by the in vitro splitting. Moreover, the blastocysts developed from A and B twins had impaired morphology compared to controls. Regarding chromosome abnormalities, there was no significant difference in the rate of aneuploidy or mosaicism between the different groups. CONCLUSION: This study showed that while no chromosomal abnormalities were seen, in vitro embryo splitting may affect the embryo morphokinetics.

Does Overnight Culture of Cleaved Embryos Improve Pregnancy Rate in Vitrified-Warmed Embryo Transfer Programme?

BACKGROUND: Vitrification is a routine procedure in assisted reproductive technique (ART) lab. However, there is widespread variability between protocols of different centres. The aim of this study was to compare the chemical pregnancy, clinical pregnancy and live birth rates between one-day embryo culture and immediate transfer for frozen-thawed embryo transfer (FET) cycles. METHODS: In this cohort retrospective study, 366 FET cycles were divided into two groups: Group A, the embryos were warmed one day before transfer, and were cultured overnight; Group B, the embryos were warmed on the same day of transfer, at least were cultured 1 h before embryo transfer (ET). Chemical and clinical pregnancy and live birth rates were compared between two groups. RESULTS: The chemical pregnancy was higher in group A than B (37.9% versus 28.9%), but this difference was not significant (P = 0.07). Clinical pregnancy (30.8% versus 24.1%) and live birth (19.8% versus 22.05%) were similar in group A and B, (P = 0.15), and (P = 0.8). Conclusion: In conclusion, overnight culture and confirmation of mitosis resumption was not essential for FET cycles in vitrification method.

Attempts for Generation of Embryonic Stem Cells from Human Embryos Following In Vitro Embryo Twinning.

In vitro embryo twinning can be used to increase the number of the human embryos available for production of human embryonic stem cell (hESC) lines. The aim of this study was to generate hESCs following the production of the twin embryos by in vitro embryo splitting procedures. In total 21 chromosomally abnormal (three pronuclei) embryos underwent in vitro embryo twinning and were allowed to develop to the blastocyst stage. As a result, 42 twin embryos were obtained, of which 24 developed to blastocyst stage. Using micromanipulation technique, the zona-free blastocysts were recovered and plated onto mitotically inactivated Yazd human foreskin fibroblast (Batch18; YhFF#18) feeder layers in microdrops. After 3 to 5 days of blastocyst culture onto human foreskin fibroblast feeder layers, the hESC-like outgrowths were passaged onto new feeders in microdrops. The initial outgrowths of hESC-like cells were generated, and cells were proliferated, passaged, and some of them expressed hESC and trophoblastic markers; however, no cell lines were established. This might be due to the low cell number and poor quality of inner cell mass within these twin blastocysts. In vitro embryo twinning by increasing the number of the human embryos could be useful in the future for the generation of new pluripotent stem cell lines. However, the challenge remains to optimize the methods.

Morphometric analysis of human oocytes using time lapse: does it predict embryo developmental outcomes?

The aim of this prospective study was to evaluate the relationship between morphometric parameters of metaphase II (MII) oocytes and the morphokinetic behaviour of subsequent embryos derived by intra-cytoplasmic sperm injection (ICSI). The association between oocyte morphometry: (whole oocyte), ooplasm, width of zona pellucida (ZP) and perivitelline space (PVS) and first polar body (PB) with embryo morphokinetic variables, including time of second PB extrusion (tPB2), pronuclei appearance (tPN), pronuclei fading (tPNf), formation of two to eight cells (t2 to t8) and irregular cleavage events [uneven at two cells stage, cell fusion (Fu) and trichomonas mitoses (TM)] were assessed. tPB2, t5 and t8 timings were related to the ooplasm diameter (p = 0.003, r = -0.12; p = 0.001, r = -0.16; p < 0.001 r = -0.36, respectively); otherwise, there were no significant relationships apart from an association between the oocyte morphometry and other morphokinetic parameters, irregular cleavage embryos as well as embryo arrest which approached significance (p > 0.05). Overall, the data showed that morphometric parameters of oocytes did not provide a tool for the prediction of embryo morphokinetic or embryo selection in ICSI cycles. However, ooplasm diameter might be useful as a marker for predicting the timing of embryo cleavage.

Evaluation of sheep ovarian tissue cryopreservation with slow freezing or vitrification after chick embryo chorioallantoic membrane transplantation.

The aim of our investigations was to compare the effectiveness of two methods for cryopreservation of sheep ovarian tissue, slow freezing and vitrification. The quality of cryopreserved tissues was evaluated after 5 days of thawing and chorioallantoic membrane (CAM) transplantation. Follicular structure, stromal integrity and neovascularization were assessed. The areas of fibrosis and necrosis were measured using MICROVISIBLE software, and proliferation was assessed with Ki-67 immunostaning. After 5 days of culture, the proportion of primordial follicles decreased, whereas the primary and intermediary follicles increased insignificantly (p > .05). Only necrosis in the vitrified culture group increased significantly (p < .05). It was established also that 5 days CAM culture was not suitable methodology for detection of folliculogenesis. Follicular quality decreased after culture, but was better in fresh and slow frozen tissues than after vitrification (p < .05). Cellular proliferative activity fell, but it preserved to some extent in all groups. In conclusion, follicles was preserved better in grafted tissue after slow freezing than vitrification and stroma was more susceptible to ischemia in vitrified rather than conventional freezing in this view. Vitrification may not be a suitable alternative to the slow freezing.

Pronuclear pattern does not predict morphokinetics behavior in human embryos.

The purpose was to investigate the correlation between pronuclei (PN) morphology and morphokinetic behaviors of derived embryos with time lapse monitoring (TLM) in assisted reproduction setting. Over time, PN morphology from PN appearance (PNA) to PN fading (PNF), PNF according to size, contact, number and position of nuclear precursor bodies (NPBs) within each PN and morphokinetics variables, including absolute time points, relative timing parameters, cleavage patterns and arrest rate, were evaluated using TLM. There were insignificant relationship between morphokinetics variables including tBP2, tPNA, tPNF, t2, t3, t4, t5, t6, t7, t8, S1, CC2, S2 and Z scoring according Z1 to Z4 (p > .05). Also, an insignificant relationship was noticed between uneven blastomeres, reverse cleavage embryos and Z scoring (p > .05). However, there were significant correlations between the rates of direct and arbitrary cleavage as well as arrested embryos and Z scores. Combined PN morphology and embryo kinetic evaluation were suggested in assisted reproduction programs.

Administration of high dose of methamphetamine has detrimental effects on sperm parameters and DNA integrity in mice.

BACKGROUND: Methamphetamine (MA) was shown to have harmful effects on male reproductive system. OBJECTIVE: To investigate probable effects of daily administration of MA on sperm parameters and chromatin/DNA integrity in mouse. MATERIAL AND METHODS: Thirty-five NMRI male mice were divided into five groups including low, medium, and high dosage groups which were injected intraperitoneally with 4, 8 and 15 mg/kg/day for 35 days, respectively. Normal saline was injected in sham group and no medications were used in control group. Then, the mice were killed and caudal epididymis of each animal was cut and placed in Ham's F10 medium for sperm retrieval. To evaluate sperm chromatin abnormalities, the aniline blue, toluidine blue and chromomycine A3 were used. For sperm DNA integrity and apoptosis, the acridine orange, sperm chromatin dispersion, and TUNEL assay were applied. For sperm morphology, Papanicolaou staining was done. RESULTS: Normal morphology and progressive motility of spermatozoa decreased in medium and high dosage groups in comparison with the control group (p=0.035). There was a significant increase in rate of aniline blue, toluidine blue, and chromomycine A3 positive spermatozoa in high dosage group. In a similar manner, there was an increase in rates of acridine orange, TUNEL and sperm chromatin dispersion positive sperm cells in high dosage group with respect to others. CONCLUSION: MA abuse in a dose-dependent manner could have detrimental effects on male reproductive indices including sperm parameters and sperm chromatin/DNA integrity in mice.

Is there any correlation between oocyte polarization microscopy findings with embryo time lapse monitoring in ICSI program?

PURPOSE: The aim was to investigate the relationship between the presence of the meiotic spindle (MS) and zona pellucida (ZP) birefringence of MII oocytes with morphokinetics variables of derived embryos in ICSI setting. METHODS: Using a polarization imaging system, the ZP birefringence and presence of MS were evaluated pre ICSI. Also, morphokinetics variables including time of second PB extrusion (tPB2), time of pronuclei appearance (tPNa), time of pronuclei fading (tPNf), time of two to eight discrete cells (t2-t8) ECC1 (t2-tPB2), cc2a (t3-t2), S2 (t4-t3) and S3 (t8-t5) as well as irregular cleavage events of 368 embryos were analyzed with time lapse monitoring (TLM). RESULTS: t5 occurred earlier in high birefringent ZP (HB-ZP) compared with low birefringent oocytes (LB-ZP; p = 0.001). In addition, t2 happened later in invisible MS compared to visible MS oocytes (p = 0.013). There were significantly lower rates of cell fusion (Fu) in oocytes with HB-ZP and also the Fu and trichotomous mitoses (TM) together in visible MS oocytes (p = 0.005, p = 0.001 and p = 0.001, respectively). CONCLUSIONS: Both t2 and t5 timings and irregular cleavage events of embryos were correlated with ZP birefringence and MS status, respectively. So, combining the information from both oocyte polarization microscopy imaging and embryo TLM can be a useful tool for single embryo transfer (SET) program.

Noninvasive imaging systems for gametes and embryo selection in IVF programs: a review.

Optimizing the efficiency of the in vitro fertilization procedure by improving pregnancy rates and reducing the risks of multiple pregnancies simultaneously are the primary goals of the current assisted reproductive technology program. With the move to single embryo transfers, the need for more cost-effective and noninvasive methods for embryo selection prior to transfer is paramount. These aims require advancement in a more acquire gametes/embryo testing and selection procedures using high-tech devices. Therefore, the aim of the present review is to evaluate the efficacy of noninvasive imaging systems in the current literatures, focusing on the potential clinical application in infertile patients undergoing assisted reproductive technology treatments. In this regards, three advanced imaging systems of motile sperm organelle morphology examination, polarization microscopy and time-lapse monitoring for the best selection of the gametes and preimplantation embryos are introduced in full.

Analysis of meiotic spindle and zona pellucida birefringenceof IVM oocytes in PCOS patients.

BACKGROUND/AIM: Polycystic ovarian syndrome (PCOS) is one of the most common causes of infertility. One of the best therapeutic options for PCOS patients is intracytoplasmic sperm injection (ICSI). In vitro maturation (IVM) can also be a useful technique for these women. The goal of this study was to evaluate both the zona pellucida (ZP) birefringence and meiotic spindle (MS) of in vivo- and in vitro-matured oocytes from PCOS patients using the PolScope system. MATERIALS AND METHODS: Immature oocytes undergoing IVM and MII oocytes were obtained from PCOS patients in an ICSI program. Using PolScope, the presence of MS and ZP birefringence was assessed in both in vivo-matured oocytes (n = 32) and IVM oocytes (n = 24). Oocytes were classified as having highly birefringent (HB) ZP and lowly birefringent (LB) ZP. Furthermore, the rates of fertilization after ICSI were evaluated. RESULTS: The maturation rate was 68.5% after IVM. The percentage of a HB-ZP was significantly higher in the IVM oocytes than in vivo-matured ones (58.3% vs. 31.2%, respectively; P = 0.04). There were similar outcomes for the fertilization rates and MS detection between the two groups (P = 0.80 and P = 0.53, respectively). CONCLUSION: Clinical IVM is a safe technology for the maturation and maintenance of oocyte integrity in PCOS patients. The use of the noninvasive PolScope is recommended for detection of healthy oocytes in ICSI.

Vitrification of mouse MII oocytes: Developmental competency using paclitaxel.

OBJECTIVE: Oocyte cryopreservation provides an important alternative for fertility preservation for women who will be treated with cytotoxic drugs. However, it can cause spindle disorganization of microtubules, putting the zygote at risk for aneuploidy. Paclitaxel is known to stabilize the microtubules that constitute the spindle. The aim of this study was to investigate the suitable concentration of paclitaxel for adding to the vitrification media to improve the developmental potential of post-thawed mature oocytes to blastocyst formation in mice. MATERIALS AND METHODS: A total of 300 MII oocytes were retrieved from superovulated mice, and were divided into three groups of control, Experimental I, and Experimental II. Oocytes in Experimental I and Experimental II were cryopreserved in the presence of 0.5µM or 1µM of paclitaxel in vitrification media, respectively. After thawing, all oocytes were incubated in G-IVF medium for 1 hour. From each group,12 oocytes were selected for viability evaluation by Hoechst/propidium iodide nuclear staining. Standard in vitro fertilization was performed on the rest of the oocytes and embryo development was followed to the blastocyst stage. RESULTS: Fertilization rate was not significantly different between the three groups. However, the cleavage rate (55%) in Experimental II group was significantly lower compared to Experimental I (88%) and control groups (83%). There was a detectable difference between the three groups at the blastocyst rate (Experimental I and control groups, p = 0.004; Experimental II vs. control and Experimental I, p < 0.001). The highest rates of parthenogenesis and arrest were in Experimental II (16% and 21%, respectively) compared with control (6% and 5%, respectively) and Experimental I (5% and 3%, respectively). There was also a significant decrease in viability rate of oocytes in Experimental II compared to the other groups. CONCLUSION: A high concentration of paclitaxel, an anticancer drug, interrupted the mouse oocyte competency when supplemented to vitrification media. Consequently, the optimal concentration of this cytoskeleton stabilizer may improve the post-thawed developmental abilities of oocytes.

Developmental competence of immature oocytes aspirated from antral follicles in patients with gynecological diseases.

BACKGROUND: In vitro maturation (IVM) of immature oocytes collected from ovary has been proposed for fertility preservation. In addition, quality of oocytes post IVM is one of the factors determining its developmental competence. By using the non-invasive Polscope system, both meiotic spindle (MS) and zona pellucida (ZP) can be assessed in living oocytes. OBJECTIVE: The aim was to investigate the developmental potential of immature oocytes retrieved from ovarian tissue after IVM, as a method for fertility preservation, in patients with gynecological diseases. MATERIALS AND METHODS: The ovarian cortex from 26 patients with malignant and benign diseases (21-45 years old), were obtained directly from collaborating hospitals, and transported to the IVF center on ice. In total 61 immature oocytes were aspirated, of which 18 (29.5%) were degenerated and discarded. The remaining 43 (70.5%) healthy oocytes were cultured in IVM culture media for 48 hr. The rate of maturity was assessed, and the ZP birefringence and MS were imaged with Polscope technology. RESULTS: Overall 43 immature oocytes underwent IVM technology, of which 30.2% reached viable metaphase II (MII) oocytes. The ovarian tissues of 9 (34.6%) women were lacking oocytes at any stage. During polarized light microscopy examination, MS could be visualized only in one of the MII oocytes, but high ZP birefringence's were observed in the majority of the oocytes post IVM (61.5%). CONCLUSION: Oocytes maturation post IVM from unstimulated ovaries showed a good developmental competence in gynecologic patients. Further studies should be performed to advance the oocyte maturation program, such as co-culture system, for fertility preservation.

Noninvasive assays of in vitro matured human oocytes showed insignificant correlation with fertilization and embryo development.

PURPOSE: Recently, the upgrading of in vitro maturation (IVM) of human oocytes as a promising strategy has emerged in assisted reproductive technology (ART). The goal was to evaluate the correlation of the in vitro matured oocytes selected on the basis of the zona pellucida (ZP) birefringence and meiotic spindles (MS) detection with fertilization and subsequent embryo development in ICSI program. METHODS: A total of 168 immature oocytes [germinal vesicle (n = 140) and metaphase I (n = 28)] obtained from patients undergoing oocytes retrieval for ICSI. After in vitro culture for 24-40 h, 112 (67 %) oocytes reached to MII stage. Using a polarized microscopy, the presence of MS and ZP birefringence were assessed in matured oocytes, followed by ICSI performance. RESULTS: The rates of fertilization in oocytes with spindles (51.3 %) were similar to that of the oocytes without spindles (50.7 %; P = 1.00). Moreover, the fertilization rates in high birefringence (HB) oocytes was not statistically different than oocytes with low birefringence (LB) (P = 0.44). The findings also showed that 64.9 % of the fertilized oocytes developed to embryos, in which 33.3 % were derived from spindle-detected oocytes. Regarding the ZP birefringence, 35.5 % of the embryos were derived from HB oocytes. CONCLUSIONS: There were insignificant relationships between the MS detection and ZP birefringence score with the rates of fertilization and embryo development in IVM oocytes.

The effect of embryo catheter loading technique on the live birth rate.

OBJECTIVE: Embryo loading (EL) is a major step in embryo transfer (ET) and affect on the success of in vitro fertilization (IVF). This study aimed to compare the effect of two different EL techniques on the rates of pregnancy and delivery in IVF/ET cycles. METHODS: 207 fresh ET and 194 Frozen-thawed ET (FET) cycles were included in this retrospective study. Two groups (A and B) were defined based on the EL technique used. In group A, the entire catheter was flushed with Ham's F-10 medium. The embryos were then drawn into the catheter using one air bracket. In group B, 70 µL of air was aspirated into the syringe and the catheter was flushed using Ham's F10 medium. The medium, air, embryos, air, and finally another layer of medium were then sequentially drawn into the catheter. The main outcome measures were the pregnancy and delivery rates. RESULTS: The groups did not differ with respect to the etiology of infertility, the source of spermatozoa, the quality of the embryos, the type of EL catheter, and the ease of transfer. The pregnancy rate was similar between two groups. In fresh ET cycles, a higher delivery rate was observed in group B than it group A (78.1% vs. 60%, p=0.1). In FET cycles, the rate of delivery was significantly higher in group B than in group A to a nonsignificant extent (88.9% vs. 58.8%, p=0.06). CONCLUSION: EL techniques did not have a significant impact on the delivery rate in either fresh or FET cycles.

Saccharin consumption increases sperm DNA fragmentation and apoptosis in mice.

BACKGROUND: Saccharin is an artificial non-caloric sweetener that used to sweeten products such as drinks, candies, medicines, and toothpaste, but our bodies cannot metabolize it. Sodium saccharin is considered as an important factor in tumor promotion in male rats but not in humans. OBJECTIVE: The objective of this study was to investigate the effect of saccharin consumption on sperm parameters and apoptosis in adult mice. MATERIALS AND METHODS: Totally 14 adult male mice were divided into 2 groups. Group 1 served as control fed on basal diet and group 2 or experimental animals received distilled water containing saccharin (0.2% w/v) for 35 days. After that, the left cauda epididymis of each mouse was cut and placed in Ham's F10. Swimmed-out spermatozoa were used to analyze count, motility, morphology (Pap-staining) and viability (eosin-Y staining). Sperm DNA integrity, as an indicator of apoptosis, was assessed by SCD (sperm chromatin dispersion) and terminal deoxynucleotidyl transferase (TUNEL) assay. RESULTS: Following saccharin consumption, we had a reduction in sperm motility with respect to control animals (p=0.000). In addition, the sperm count diminished (17.70±1.11 in controls vs. 12.80±2.79 in case group, p=0.003) and the rate of sperm normal morphology decreased from 77.00±6.40 in control animals into 63.85±6.81 in saccharin-treated mice (p=0.001). Also, we saw a statistically significant increase in rates of sperm DNA damage and apoptosis in experimental group when compared to control one (p=0.001, p=0.002 respectively). CONCLUSION: Saccharin consumption may have negative effects on sperm parameters, and increases the rate of sperm DNA fragmentation and apoptosis in mice.

Zona pellucida birefringence and meiotic spindle visualisation of human oocytes are not influenced by IVM technology.

The aim of the present study was to investigate the relationship between the presence of the meiotic spindle and zona pellucida (ZP) birefringence with morphology of in vivo- and in vitro-matured human oocytes. Germinal vesicles (n=47) and MI (n=38) oocytes obtained from stimulated ovaries of patients undergoing intracytoplasmic sperm injection (ICSI) underwent IVM. Using a PolScope (OCTAX PolarAID; Octax, Herbon, Germany), the presence of spindles and ZP birefringence was assessed in both in vivo-matured (n=56) and IVM (n=56) oocytes. In addition, the morphology of each matured oocyte was evaluated microscopically. There were insignificant differences for ZP birefringence and meiotic spindle between the in vivo-matured and IVM MII oocytes. Subanalysis revealed that the rates of morphologically abnormal oocytes did not differ significantly between the two groups, except in the case of irregular shape (P=0.001), refractile body (P=0.001) and fragmented polar body (P=0.03), which were higher in IVM oocytes. In the case of in vivo-matured oocytes, a significantly higher percentage of oocytes with intracytoplasmic and both intra- and extracytoplasmic abnormalities have a low birefringent ZP (P=0.007 and P=0.02, respectively). There was no relationship between morphological abnormalities and spindle detection. The findings suggest that clinical IVM is a safe technology that maintains the high maturation rate and integrity of oocytes. In addition, the use of the non-invasive PolScope is recommended for the detection of oocytes most suitable for ICSI.