Embryos In IVF Cycles Research Articles

Title: Repeated implantation failure is associated with increased Th17/Treg cell ratio, during the secretory phase of the human endometrium

Repeated implantation failure (RIF) is a significant limiting factor in assisted reproduction. Chronic endometrial inflammation has been noted in RIF women, therefore we sought to investigate the potential association of endometrial Th17/Treg ratio and endometrial inflammation in these cases. Endometrial pipelle biopsies were obtained from volunteers, 29 women with RIF (failure to achieve pregnancy following at least 3 transfers of high-grade embryos in IVF-cycles) and 27 fertile women (at least one child) in total, at the secretory phase of the menstrual cycle. Using tissues from 17 fertile and 18 RIF endometrial samples, stromal and immune cells were isolated and flow cytometry analysis was performed to determine Th17 and CD4+CD25high FOXP3+ cell populations in endometrial stromal cell suspensions. Another group of tissues from 10 fertile and 11 RIF samples were used for mRNA expression levels of Treg and Th17-cell transcription factors, FOXP3 and RORγt respectively. Endometrial inflammatory mediators’ mRNA expression was also analyzed. A statistically significant increase in protein flow cytometry analysis of Th17/Treg ratio (p≤0.05) as well as a reduction in absolute Treg cells in the endometrium (p≤0.05) was noted in women with RIF. Additionally, RNA analysis on the same set of women indicated RORγt/FOXP3 significantly increased in women with RIF compared to fertile ones (p≤0.05). Finally, women with RIF exhibited significantly (p≤0.05) elevated mRNA levels of pro-inflammatory mediators (ΤΝF-a, ΙL-6, IL-8 and CCl2). Women with RIF exhibit elevated Th17/Treg ratio, mostly due to endometrial Treg depletion, as well as a pro-inflammatory state in the endometrium.

Application of the FISH Method for Analyzing Chromosome Segregation Patterns in Preimplantation Embryos from Robertsonian Translocation Carriers

Robertsonian translocations (RTs) are very common balanced structural chromosome rearrangements in humans. Due to alterations in the chromosome segregation pattern and the formation of unbalanced gametes and embryos, the carriers of RTs have a significant risk of reproductive failure. For over 30 years, fluorescent in situ hybridization (FISH) has been used for preimplantation genetic testing for chromosomal structural rearrangement (PGT-SR) in carriers of RTs. The data obtained by performing FISH for PGT-SR can be useful for analyzing segregation patterns in preimplantation embryos. We analyzed chromosome segregation patterns in 964 embryos from 100 couples who were carriers of various types of RTs by performing FISH or using the array comparative genomic hybridization (aCGH) method. When the carriers were male, the predominant segregation mode was alternate (detected in 42.4% of embryos), whereas, when the carriers were female, alternate and adjacent types of segregation were detected in most embryos (32.3% and 26.6% of embryos, respectively). About 33% of balanced/euploid embryos in IVF cycles were formed when the carriers of RT were male, and about 24% of such embryos were formed when the carriers were female. No association was found between sperm parameters and malsegregation rates in male carriers of RT. In this study, we found that female and male RT carriers have different reproductive risks associated with chromosome segregation patterns and the rate of balanced/euploid embryos.

Strictly selected Mono- and non-pronuclear blastocysts could result in appreciable clinical outcomes in IVF cycles

The aim of the study was to examine the clinical value of blastocysts derived from mono-pronuclear (1PN) or non-pronuclear (0PN) zygotes with two polar bodies (2PB), which were selected by our criteria. We retrospectively analysed 610 frozen-thawed blastocyst transfer (FET) cycles and the corresponding oocyte retrieval cycles from 2014 to 2017. Developmental potential and clinical outcomes of embryos derived from zygotes with various numbers of pronuclei were analysed. Based on more detailed pre-selection settings, blastulation rates of 1PN/2PB and 0PN/2PB-derived embryos were 70.18% and 69.17%, respectively. Blastocyst FET results were not significantly different between 2PN/2PB, 1PN/2PB and 0PN/2PB groups in terms of clinical pregnancy rates (59.79%, 47.06% and 56.25%), implantation rates (47.24%, 40.00% and 47.62%), live birth rates (49.39%, 29.41% and 43.75%) or malformation rates (0%, 0% and 0%). In conclusion, after strict morphological selection and blastocyst culture, 1PN/2PB and 0PN/2PB-derived embryos in IVF cycles can have considerable clinical value. Blastocysts derived from 1PN/2PB or 0PN/2PB zygotes are worthwhile FET option for patients who have no available 2PN-derived embryos.

Assessment of a double freezing approach in the management of surplus embryos in IVF.

What pregnancy rates are achieved after transfer of cryopreserved double slow-frozen embryos in IVF cycles? Patients in whom surplus thawed cleaved embryos (day 2 or 3) were grown to the blastocyst stage, re-frozen and then re-thawed for transfer (double freezing) were included. Data were collected on all patients who had undergone the above procedure at the IVF unit of Assuta Ramat Hachayal Hospital, Tel Aviv, during a 7-year period. For each patient in the study group, the two-consecutive, matched-by-age patients treated with frozen-thawed single blastocyst transfer were selected to form a 2:1 ratio control group. All embryos were frozen using the slow freeze protocol. A total of 54 patients had 70 embryos that were re-frozen at the blastocyst stage. Twenty-eight of these blastocysts were thawed and 27 underwent transfer to 25 patients. A single embryo was transferred to 23 patients and two embryos were transferred to two patients. The survival rate of the second thawing was 96.4% (27/28). Clinical pregnancy rate was 16% (4/25) and implantation rate was 14.8% (4/27). In the study group, pregnancies were achieved in 22 out of the 25 patients using IVF treatment, indicating good receptivity of the uterus. In the control group, the implantation/pregnancy rates were significantly higher (44.2% [23/52]; P < 0.01). The transfer of twice slow-frozen and thawed embryos does not seem to be a beneficial approach in the planned management of cryopreserved surplus embryos owing to the low pregnancy rate achieved after transfer of the re-frozen blastocyst embryos.

Metabolomics: the perspective search of methods to overcome infertility

Nowadays, the reproductive medicine has started to develop precipitously. Nevertheless, 60 to 80 millions of couples worldwide suffer from infertility. The etiology and pathogenesis of many diseases leading to infertility is still causing a lot of debates and needs to be researched further. In 25% of the cases, the cause of infertility may not be identified that forces couples to seek help in auxiliary reproductive technologies. However, even in this case the probability of a positive result makes up no more than 40%. In recent years, scientists' attention became increasingly driven to the so-called omics-disciplines, including metabolomics. Many researchers believe that metabolomics is that science which will give a possibility to improve the quality of diagnosis, will lead to a deeper understanding of the etiology and pathogenesis of diseases that lead to infertility and will improve the quality of the assessment of oocytes and embryos in IVF cycles.

Do multinucleated embryos in IVF cycles affect IVF outcome?

One of the morphological embryo selection criteria is the nuclear status of blastomeres. Human embryos with multinucleated blastomeres (MNB) are considered to be abnormal in general. The purpose of this study was to evaluate the pregnancy rates of patients whose age/basal FSH were matched having both normal and multinucleated embryos to a group of patient who had didn't have multinucleated embryos.

P52 Blastocyst formation rate for aneuploid and euploid embryos in IVF cycles

P52 Blastocyst formation rate for aneuploid and euploid embryos in IVF cycles

P5 Correlation between preimplantation genetic screening for chromosomal aneuploidies and amount of embryos in IVF cycles

P5 Correlation between preimplantation genetic screening for chromosomal aneuploidies and amount of embryos in IVF cycles

Corifollitropin alfa in a long GnRH agonist protocol: proof of concept trial

Fifty healthy women, aged 18-39 years with no known risk of ovarian hyperstimulation were treated with 100 or 150 μg corifollitropin alfa (dependent on body weight) in a long GnRH agonist protocol. At these doses, corifollitropin alfa initiated and supported growth of a large cohort of follicles during the first week of ovarian stimulation.

Optimal follicle and oocyte numbers for cryopreservation of all embryos in IVF cycles at risk of OHSS

Ovarian hyperstimulation syndrome (OHSS) is a rare but potentially fatal complication of IVF treatment. The risk of OHSS increases with increasing numbers of follicles aspirated and oocytes retrieved, but there is little evidence to support whether threshold values of either can be used to correctly predict OHSS. Since the most severe forms of OHSS are usually associated with pregnancy, cryopreservation of all embryos may prevent this. The authors attempted to find thresholds of follicle and oocyte numbers that would optimally predict OHSS, through a retrospective analysis of 2253 consecutive cycles of IVF/intracytoplasmic sperm injection treatment reaching oocyte retrieval, between 1 January 2003 and 31 March 2006. Receiver operator characteristic (ROC) curves were calculated for both parameters, to determine threshold values that might predict OHSS in women with ≥20 oocytes. For the prediction of early onset OHSS, ROC curves showed that an optimal balance between sensitivity and specificity was achieved using thresholds of 24 oocytes (79%, 60%) and 29 follicles (82%, 65%) respectively. Using these thresholds, cryopreservation of all embryos may be offered as an alternative to cancellation of a treatment cycle due to excessive ovarian response, thus minimizing the number of unnecessary interventions while still correctly predicting most cases of early onset OHSS.

Ketotifen improves sperm motility and sperm morphology in male patients with leukocytospermia and unexplained infertility

In an open, uncontrolled study, the effect of 12 weeks of daily administration of ketotifen, an antihistamine-like drug with a mast cell stabilizing effect, on the semen quality of 55 men with leukocytospermia and unexplained infertility was examined. After 4 weeks of treatment, white blood cell count dramatically diminished and was accompanied by a significant improvement in sperm motility. A significant increase of morphologically normal sperm cells was observed at 8 weeks of treatment, and these changes remained until at least 4 weeks after the end of treatment.

The CryoTip Method: Aseptic Vitrification of Oocytes and Embryos

The CryoTip Method: Aseptic Vitrification of Oocytes and Embryos

Early Cleavage Improves Ongoing Pregnancy Rate When Transferring Non-Top Quality Embryos in IVF Cycles

To evaluate the development potential of embryos with early cleavage into top quality embryos and the impact of early cleavage on ongoing pregnancy rates in women undergoing IVF treatment. Analysis of a large, randomized, open-label, assessor-blind, multicenter, multinational trial (MERIT) comparing two gonadotropin preparations in IVF cycles with the long GnRH agonist protocol. Embryo quality and ongoing pregnancy data were collected from 731 ovulatory women with tubal factor, mild male factor, endometriosis stage I/II or unexplained infertility. All were downregulated with triptorelin 0.1 mg SC daily from the mid-luteal phase and randomised to either HP-hMG (MENOPUR, Ferring Pharmaceuticals) (n=363) or recombinant FSH (GONAL-F, Serono) (n=368). The starting dose was 225 IU SC for the first five days and thereafter dose adjustments of 75 IU were allowed not more frequently than every four days. Recombinant hCG 250 μg SC was administered when 3 or more follicles of 17 mm or greater were observed. A total of 7535 oocytes were obtained. Embryo quality was evaluated at 28h, 44h and 68h (±1h) after standard IVF insemination locally at site and subsequently centrally by three independent embryologists based on digital images. All embryologists were blinded to treatment allocation and each others results. A top quality embryo was pre-defined as having 4-5 cells on Day 2, ≥ 7 cells of equal size and ≤ 20% fragmentation on Day 3 and no multinucleation. Transfer of 1-2 embryos of pre-defined quality criteria (minimum: ≥ 4 cells with no cleavage arrest and ≤ 20% fragmentation) was done on Day 3. Ongoing pregnancy was defined as at least one viable fetus 10-11 weeks after embryo transfer. The analysis was based on the central evaluation of embryos and included embryos for which individual outcome was determined, meaning all women with 0% or 100% ongoing implantation rate after single or double embryo transfer. A total of 1123 embryos were 2-cells at 28h based on the central evaluation. Embryos with 2-cells at 28h had a significantly (p<0.001) higher chance of developing into a top quality embryo (32%) compared to those embryos that were not 2-cells at 28h (5%). The ongoing pregnancy rate for women with transfer of top quality embryos was 39% with early cleaved embryos (2-cell at 28h) and 40% with not early cleaved embryos. Concerning non-top quality embryos, the ongoing pregnancy rate was 35% for women with transfer of early cleaved embryos which was significantly (p<0.05) higher than the 19% observed for embryos with no early cleavage. Among single embryo transfers, the ongoing pregnancy rate/implantation rate was 17% for embryos that were 1-cell at 28h compared to 36% for 2-cell embryos (p<0.05). For women with double-embryo transfer, the ongoing pregnancy rate was 36% when both embryos transferred were 2-cells at 28h and 29% when both embryos were 1-cells. Embryos that reach the 2-cells stage at 28h after insemination have a significantly higher chance of developing into top quality embryos. Information on early cleavage will not increase pregnancy rates when transferring top quality embryos on Day 3, but will dramatically increase the pregnancy rates if the embryo transferred is of non-top quality.

R-089. Cryopreservation of all embryos in IVF cycles results in satisfactory birth rates

R-089. Cryopreservation of all embryos in IVF cycles results in satisfactory birth rates