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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. Key words: Endometrium, Embryo transfer, Pregnancy, Live birth, Progesterone.

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 6–8-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.

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. Key words: Implantation failure, Gonadotropin-releasing hormone, Embryo transfer, Pregnancy, Implantation.

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.

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

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.

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.

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. 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). 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). 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.

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.

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.

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.

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.

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 (

Objective Chronic ethanol abuse causes reproductive organ failure and infertility in both humans and laboratory animals. Since sperm has a critical role in reproductive function, the objective of this unique study was to evaluate the effects of different doses of ethanol on sperm parameters, chromatin structure and apoptosis in adult mice. Study design A total of 36 adult male mice were equally divided into four groups. Group 1 received ethanol (10%, v/v) containing saccharin (0.2%, w/v), group 2 received ethanol (5%, v/v) containing saccharin (0.1%, w/v), group 3 was treated with saccharin (0.2%, w/v) and group 4 served as control and fed on basal diet for 35 days. Finally, the left cauda epididymis of each animal was cut and placed in Ham's F10 medium. Retrieved spermatozoa were used to analyze count, motility, morphology and viability. Sperm chromatin condensation and DNA integrity were assessed by five different tests including chromomycin A3 (CMA3), toluidine blue (TB), sodium dodecyl sulfate (SDS), and SCD (sperm chromatin dispersion) and sperm apoptosis was assessed by TUNEL. Results Following ethanol consumption, the sperm count diminished in the ethanol-treated groups. A decrease in sperm motility and an increase in the rate of morphological abnormalities (coiled and broken tails) were seen in the experimental and saccharin groups in comparison with controls. We showed that ethanol consumption can disturb sperm DNA integrity and chromatin remodeling and it may also induce sperm apoptosis. The rates of sperm apoptosis were 51.57 ± 7.45 and 42.85 ± 6.76 in the high ethanol dose and low ethanol dose groups, respectively. Conclusion The results showed that alcohol has negative effects on sperm parameters, chromatin/DNA integrity and apoptosis in mice. These alcohol-induced sperm anomalies may be dose-dependent.

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.

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.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.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.There were insignificant relationships between the MS detection and ZP birefringence score with the rates of fertilization and embryo development in IVM oocytes.

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.

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.