azam agha-rahimi

The cases with unexplained infertility may have an abnormality in their sperm chromatin structure. Sperm selection methods can be used to separate sperm with low DNA fragmentation. The purpose of this study was to compare the efficacy of physiological intracytoplasmic sperm injection (PICSI) with magnetic- activated cell sorting (MACS) in assisted reproductive techniques in cases with un- explained infertility.

The semen samples were collected from couples with unexplained infer- tility. After semen analysis and sperm DNA fragmentation (SDF) evaluations, sam- ples were prepared with swim-up method. The rates of SDF in different fractions in- cluding raw semen (n=20), swim-up (n=20), only motile sperm after swim-up (swim-up selection) (n=20), MACS sperm selection (n=20), only motile sperm after MACS (MACS selection) (n=20), and PICSI sperm selection (n=16) were evaluated. Also, the main sperm characteristics and fine morphology of sperm suspension after MACS were assessed. Statistical analysis was performed using GraphPad Prism. The p<0.05 was considered statistically significant.

DNA fragmentation index (DFI) values in PICSI and MACS groups were significantly reduced as compared to the swim-up group. The rate of this reduction was more pronounced in MACS (58.20±13.02) than PICSI (36.57±15.52) group. Al- so, our results showed that MACS resulted in decreased sperm motility, with no al- teration in their fine morphology.

MACS was found to be more efficient in reduction of SDF rates than PICSI. However, none of the sperm selection techniques can not totally eliminated the spermatozoa with DNA fragmentation in the final sperm sample.

Synchronization between the embryonic stage and the uterine endometrial lining is important in the outcomes of the vitrified-warmed embryo transfer (ET) cycles.

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.

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.

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.

Higher rates of pregnancy and live birth were achieved in day-3 ET after three days of progesterone administration in FET cycles.

Sperm quality is an important factor in assisted reproductive technology (ART) that affects the success rate of infertile couples treatment. In vitro incubation of sperm can influence its parameters and DNA integrity. The present study focused on the effect of different incubation temperatures sperm parameters on asthenoteratozoospermia semen prepared with density gradient centrifugation at different times.

Twenty-seven samples were collected and prepared. Then, the suspension was divided into two parts. One part was incubated at room temperature (RT), and another was incubated at 37C. Immediately and after 2 hr (2H) and 4 hr (4H), spermatozoa were evaluated regarding motility, viability, morphology, sperm protamine deficiency, chromatin and DNA fragmentation. Statistical analysis was performed using paired t-test and repeated measures. The p<0.05 was considered statistically significant.

Our results showed that following 2 and 4 hr of incubation at RT, sperm progressive motility and viability decreased significantly. Sperm DNA fragmentation increased significantly following 2 and 4 hr of incubation at RT and 37C. The Trend analysis confirmed that there were no significant differences between sperm parameters and DNA fragmentation after different times at RT and 37C.

Incubation of sperm at RT in comparison to 37C didn’t preserve sperm parameters and DNA efficiently. Therefore, IVF, ICSI and IUI procedure should be performed in the soonest possible time after sperm preparation.

Cumulus cells, as oocyte nurse cells, provide a suitable microenvironment with growth factors and cellular interactions required for oocyte maturation. Thus, these cells may serve as a natural niche for in vitro studies of female germ cell development. Cumulus cells may help attain a better understanding of the causes of infertility in women and eventually improve the outcomes of cases that respond poorly to standard infertility treatment.

The aim of this study was to isolate, culture, and investigate the biological characteristics of human cumulus cells.

In this experimental study, cumulus cells were isolated, cultured, and characterized using reverse transcription-polymerase chain reaction analyses of specific genes including FOXL2, CYP19A1, FSHR, AMHR, and LHR. The presence of vimentin, a structural protein, was examined via immunofluorescent staining. Moreover, levels of anti-mullerian hormone (AMH) and progesterone secretion by cumulus cells were measured with ELISA after 2, 4, 12, 24, and 48 hr of culture.

In adherent culture, human cumulus cells expressed specific genes and markers as well as secreted AMH and progesterone into the medium.

Cumulus cells secrete AMH and progesterone in an adherent culture and might be applicable for in vitro maturation (IVM) and in vitro gametogenesis (IVG) studies.

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.

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.

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).

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.

High implantation success following in vitro fertilization cycles are achieved via the transfer of embryos with the highest developmental competence. Multiple pregnancies as a result of the transfer of several embryos per cycle accompany with various complication. Thus, single-embryo transfer (SET) is the preferred practice in assisted reproductive technique (ART) treatment. In order to improve the pregnancy rate for SET, embryologists need reliable biomarkers to aid their selection of embryos with the highest developmental potential. Time-lapse technology is a noninvasive alternative conventional microscopic assessment. It provides uninterrupted and continues the survey of embryo development to transfer day. Today, there are four time-lapse systems that are commercially available for ART centers. In world and Iran, the first time lapse babies were born in 2010 and 2015, respectively, conceived by SET. Here, we review the use of time-lapse monitoring in the observation of embryogenesis as well as its role in SET. Although, the findings from our review support common use of time-lapse monitoring in ART centers; but, future large studies assessing this system in well-designed trials are necessary.

Intrauterine insemination (IUI) is one of the therapeutic approaches for infertility. The objective of this study was to evaluate DNA integrity and apoptosis role in success of IUI in both mild male and female factor infertility. Patients were divided into two groups: M (mild male factor; n=29) and F (female factor; n=31) undergoing single IUI. Ejaculates were analyzed and chromatin quality was assessed using chromomycin A3 (CMA3) staining. In addition, spermatozoal apoptosis was recognized using TUNEL assay. Statistical analyses were done using t-test and Mann Whitney test for sperm apoptosis and sperm chromatin by SPSS. Data were expressed in mean ± SD for variables. P<0.05 was considered statistically significant. Sperm concentration and progressive motility were higher in F than M group. Sperm with normal morphology were statistically similar in M and F infertile patients (32.7±15.6% vs. 35.5±9.07%, p=0.39). Sperm chromatin immaturity was higher in patients with mild male infertility, when compared with the other group (p<0.01). Also, 32.0±5.6% and 30.8±6.1% of the spermatozoa showed signs of apoptosis in groups M and F, respectively (p=0.49). Very low (3.4%) clinical pregnancy rates were noticed in patients with mild male factor infertility Defect in sperm motility as well as high rates of DNA damage and apoptosis may be involved with very low rate of pregnancy outcomes in patients with mild male factor infertility. Therefore, it seems the application of IUI may have better outcomes in patients with female infertility compared to mild male factor infertility.

ART laboratories are quality controlled to make sure that disposable objects used for the culture of gametes and embryos are toxin-free. To maintain a high standard, all disposable objects in our ART laboratory were tested by human sperm motility assay (HuSMA). HuSMA was used as a measure for QC at the intended ART laboratory. Eighteen objects that are commonly used in IVF laboratories were tested by HuSMA. The objects included gloves, syringes, culture dishes, pipettes, tips and semen collection dishes. HuSMA was conducted at 10 and 30 minutes and also at 1, 2, 4, and 24 hours of incubation at room temperature. Sperm motility index (SMI) was calculated by dividing the percentage of progressive motile sperms of the test by that of the control at the specified intervals. An SMI value < 0.85 was defined to indicate sperm toxicity. The tests were repeated for three times. QC by HuSMA confirmed the toxicity of three objects, including embryo transfer (ET) gloves A and B, and puncture gloves A. ET gloves A (SMI=0.0) and puncture gloves A (SMI=0.0) were toxic after 10 minutes, but ET gloves B (SMI=0.63) were shown to be toxic after 24 hours (46% progressive motile sperm compared with 68% in the control group). Moreover, two other objects including culture dish (SMI=0.42) and semen collection dish (SMI=0.67) had borderline values after 24 hours; different results in four repeats after 24 hours (twice toxic and twice nontoxic). Some objects which are routinely used in ART laboratories may be toxic and their use should be discontinued as part of QC programs. To increase the efficiency of HuSMA, it seems necessary to do this test more than once for each object.

The purpose was to investigate the effect of the duration of gamete incubation on fertilization rate, embryo cleavage, and embryo quality before and after freezing in mice. Ovulated oocytes collected from superovulated mice after ip injection of PMSG and hCG were divided randomly into control and experimental groups. Oocytes from the control group were inseminated for six hours and the experimental group were inseminated for one hour, respectively. The differences in fertilization rates, embryo cleavage and percent of good quality embryos in four grades (A, B, C, D) were analyzed. Finally, two cell embryos were frozen; and after thawing, the quality of embryos from the two groups were compared. There was no difference between the two groups in regards to fertilization and cleavage rates. However, the proportion of grade A embryos was significantly higher among the experimental group (41.7%) when compared to the control group (19%). Also the proportion of grade D embryos was significantly (p=0.04) lower in the experimental group (8.3%) as compared to the control group (23.8%). In addition, percentage of good quality embryos in the experimental group did not decrease after freezing (p=0.3), however the percentage of good quality embryos were significantly decreased after freezing in the control group (p=0.01). Insemination of oocytes for a short period produced embryos of superior quality than insemination for a longer period in the experimental group. Also, the effect of freezing on embryos produced from short insemination was less than the long insemination period. After freezing, a higher percentage of good quality embryos survived post thawing in mice.