To evaluate whether endometrial compaction using sequential transvaginal ultrasound is associated with improved live birth rates in medicated single euploid frozen embryo transfer (FET) cycles. Prospective observational cohort study at a private fertility clinic. Patients who underwent FETs between January and December 2018 were assessed for inclusion. The change in endometrial thickness between the end of the estrogen phase and the day before embryo transfer, measured by sequential transvaginal ultrasound, was used to categorize cycles with compaction (≥ 5%), no change, or expansion (≥ 5%). FET cycle outcomes were then compared between groups. The primary outcome was live birth. Secondary outcomes include clinical pregnancy rate and rate of spontaneous abortion. Of the 259 single euploid medicated FETs performed during the study period, only 43/259 (16.6%) of the cycles demonstrated ≥ 5% compaction, whereas 152/259 (58.7%) expanded and 64/259 (24.7%) were unchanged. Live birth rates did not differ between cycles with compaction (58.1%), no change (54.7%), or expansion (58.6%), p = 0.96. Clinical pregnancy and spontaneous abortion rates were also similar between groups. The vast majority of cycles did not demonstrate endometrial compaction. Endometrial compaction is not associated with live birth rate or spontaneous abortion rate in medicated single euploid FETs in this cohort.

To evaluate whether endometrial compaction using sequential transvaginal ultrasound is associated with improved ongoing pregnancy rate in programmed single euploid FETs. Retrospective observational study. All single euploid FET cycles between January and December 2018 were assessed for eligibility. Natural cycle and controlled ovarian stimulation FETs as well as gestational carrier cycles were excluded. Patients were also excluded if their endometrial thickness was <7 mm at the initiation of progesterone. All patients received the same exogenous hormones for endometrial preparation per clinic protocol. Endometrial thickness was measured with transvaginal ultrasound after an average of 9-10 days of estrogen. Progesterone in oil 50mg daily and endometrin 100 mg TID were started within two days of the final lining measurement if the endometrium was ≥7 mm and trilaminar in appearance. Patients returned the day prior to embryo transfer for reassessment of the endometrial lining using transvaginal ultrasound. All FETs were performed on the sixth day of progesterone. The primary outcome was ongoing pregnancy rate defined as the presence of fetal cardiac activity at 10 weeks. We grouped patients by percentage of endometrial compaction, defined as the difference in endometrial thickness at the end of the estrogen-only phase and the day before embryo transfer, divided by the thickness at the end of estrogen-only phase. Cycles in which the percent compaction was less than +/-5% were considered unchanged. A total of 294 single euploid FETs were included in the analysis. Only 17.3% (51/294) of the cycles demonstrated ≥5% compaction, whereas 82.7% (243/294) either expanded or were unchanged. Of those without any compaction, 30.0% (73/243) had no change in compaction and 70.0% (170/243) expanded ≥5%. Ongoing pregnancy rate did not significantly differ between cycles with compaction, no change, or expansion (69.8% vs 57.5% vs 58.8%, p=0.94) (Table 1). When considering only those cycles in which the endometrium compacted, no difference was seen in ongoing pregnancy rates at different levels of compaction (p=0.72) (Table 2). Endometrial compaction is not associated with ongoing pregnancy rate in single euploid FETs in our cohort.Table 1Ongoing pregnancy rate (any compaction vs no change vs any expansion)Any CompactionNo ChangeAny Expansionp-valueaOngoing Pregnancy Rate31/51 (69.8%)42/73 (57.5%)100/170 (58.8%)0.94achi-squared Open table in a new tab Table 2Ongoing pregnancy rate by percent compactionNo Change5-9%10-14%≥15%p-valueaOngoing Pregnancy Rate42/73 (57.5%)12/17 (70.6%)9/15 (60%)10/19 (52.6%)0.72achi-squared Open table in a new tab

To determine whether the presence of abnormal cell cleavage, cell fusion or both seen on morphokinetic analysis of preimplantation embryos using time-lapse monitoring (TLM) is predictive of chromosomal aneuploidy when compared to embryos with normal development. Retrospective cohort study at a large, private fertility center. All patients who utilized both the TLM device (Embryoscope®, Vitrolife, Sweden) and preimplantation genetic testing for aneuploidy (PGT-A) at a single IVF center from 1/2018 to 12/2019 were included in the study. Patients using donor oocytes were excluded. Evaluation of embryo development was performed on all embryos from the time of first cell division until 72 hours. Abnormal cell cleavage (AC) was defined as irregular first cleavage of the zygote resulting in more than two daughter cells and/or irregular second cleavage that formed five or more cells instead of four. Cell fusion (CF) was defined as a decrease in number of cells at any time point up until 72 hours. Normal development (ND) was defined as the absence of AC or CF. Trophoectoderm biopsy was performed on day 5, 6, or 7 in blastocysts of good or fair quality. PGT-A was performed using next generation sequencing. All embryos that were sufficient quality for blastocyst biopsy were included in the study. Statistical analyses were conducted using logistic regression with Huber-White standard errors to correct for multiple embryos within a patient. A total 6,171 embryos were analyzed with TLM and of those, 1,083 embryos from 311 patients developed into blastocysts of sufficient quality to biopsy and were included in the analysis. Of the biopsied blastocysts, 8.1% (87/1,083) had AC, 1.2% (14/1,083) had CF and 0.4% (4/1,083) displayed both AC and CF. The vast majority of embryos, 90.2% (969/1,083), did not have AC or CF. The presence of AC, CF or both did not significantly increase with older age (p=0.77). Abnormal development (AC, CF or both) was significantly more likely to be seen in fair-quality compared to good-quality embryos (17.5% v. 5%, p<0.001). The aneuploidy rate was not increased in embryos displaying AC (49%), CF (48%) or both (48%) compared to embryos with ND (49%), after controlling for age (p=0.99). While the presence of early abnormal cell cleavage or cell fusion on TLM is associated with lower quality morphological grade at the blastocyst stage, these markers of abnormal development are not associated with increased aneuploidy. Additional research is needed to assess ongoing pregnancy rates from these embryos to evaluate the clinical significance of AC and CF in early pre-implantation embryo development.

INTRODUCTION: The objective of this study was to compare characteristics of donor in vitro fertilization (IVF) cycles that did and did not result in pregnancy. Additionally, we sought to identify the optimal number of retrieved oocytes and good-quality embryos needed to maximize pregnancy rates in fresh donor IVF cycles. METHODS: We conducted a retrospective analysis of donor oocyte cycles at a single large private fertility clinic between 2010 and 2013. Parametric and nonparametric statistical analyses were used to compare cycles that did and did not result in pregnancy. Receiver operating characteristic curves were used to evaluate the optimal number of retrieved oocytes and good-quality embryos predictive of pregnancy. RESULTS: An overall clinical pregnancy rate of 58.5% was found among the 318 fresh donor cycles analyzed. There was no difference in donor age, number of total or mature oocytes retrieved, fertilization rate, or number of embryos transferred or frozen between the two groups. There was a trend toward higher number of good-quality embryos in the pregnant group (6.3 compared with 7.2, P=.06). The optimal number of retrieved oocytes and good-quality embryos predictive of pregnancy was 18 oocytes and 5.5 embryos, respectively (area under the curve [confidence interval] 0.55 [0.49–0.61] and 0.57 [0.50–0.63]). CONCLUSION AND IMPLICATION: Donor egg banks should consider avoiding cycles that result in extremely high numbers of retrieved oocytes because this does not improve pregnancy outcomes. Larger studies are needed to validate our findings and to demonstrate the optimal use of a limited quantity of viable, pregnancy-producing oocytes.

Removal of cumulus cells surrounding oocytes, denudation, is a necessary step prior to some Assisted Reproductive Technology procedures. An important aspect during the chemical/physical removal of cumulus cells is to minimize the risk of fractured zona pellucida (FZ) damage. In our labratory, two denudation medias have been used to facilitate this action: Irvine Scientific's Hyaluronidase Solution (HYAL) and Origio's ICSI Cumulase® (CUMU). Over a 12 month time period we initially used HYAL, then transitioned to CUMU and finally returned back to HYAL. The HYAL is made up of serum supplemented HEPES buffered HTF media containing a variable concentration of at least 80 IU/mL of bovine derived hyaluronidase enzyme. There is/maybe a risk of animal pathogen contamination and oocyte toxicity when using HYAL. Cumulase® is also made up of serum supplemented HEPES buffered media, however, it contains a verified 80 IU/mL of recombinant human hyaluronidase enzyme (rHuPH20). This recombinant human derived enzyme may make the CUMU less toxic to oocytes and eliminates the risk of animal contamination. Procedurally, these two medias recommend differing denudation exposure times, with HYAL incubation time of <30 seconds while the exposure time for CUMU is >1 minute. These two denudation media are different in composition and procedure, so our study analyzed their effects on the FZ rate of oocytes between transitioning their uses. Collection of oocytes during retrieval was performed in a laminar flow hood with a heated surface at 37°C in a prepared HEPES buffered and HSA supplemented media. After the cumulus masses surrounding the retrieved oocytes were trimmed, oocytes were transferred using a Pasteur pipette to a prepared and warmed 35mm dish for denudation. The dish contained a ∼100 μL drop of either HYAL or CUMU, with ∼12 drops of 50 μL of gamete wash media with an oil overlay. The oocytes were first placed in the large drop of HYAL or CUMU for the recommended time given by their respective manufacturers, while gently pipetting the oocytes up and down (no more than 30 seconds for HYAL and no less than one minute for CUMU). Following this exposure, the oocytes were moved by means of a Stripper and a 300μM stripper tip into the first of the wash drops to rinse off any remaining denudation media while continuing to remove as many cumulus cells as possible. Continual gentle aspiration of the oocytes between wash drops occurred while decreasing the size of stripper tips from to 170μM, to as low as 140μM. The denuding continued until the oocytes' maturity could be determined by the presence or absence of the first polar body or germinal vesicle. Any oocytes that had FZ were removed and discarded as damaged. The cumulative totals of FZ for 2 months of HYAL use before the transition to CUMU, for the middle 2 months of CUMU use and for the first 2 months following the return to HYAL were statistically analyzed using a two-tailed Chi-square with Yates correction. The number of patients, of oocyte denuded and of FZ were calculated and analyzed for statistical differences before, during and after transitioning denudation media (Table). No difference in FZ rates were found between the three different time periods.TableDenudation Media# Patients# Oocytes Denuded# Fractured Zona (%)Hyaluronidase Solution –Before Transition4757010 (1.7)a,cICSI Cumulase®6566517 (2.5)a,bHyaluronidase Solution –After Return Transition3852619 (3.6)b,ca,b,c p>0.05. Open table in a new tab a,b,c p>0.05. When using HYAL, oocytes are out of the incubator for a shorter period of time, due to its <30 second exposure, and the amount/intensity of physical/mechanical denudation is conceivably lessened. Congruently, when using CUMU, oocytes are exposed for a longer amount of time with potentially more manipulation. However, when FZ rates with HYAL were compared to the CUMU, the data did not demonstrate a difference in FZ rates before or after transitioning between media. Despite the fact that these two medias are chemically different (bovine vs. recombinant human derived), they both perform the task of denudation of oocytes safely and at an equivalent level of risk for damage to the oocytes.

Morphokinetic analysis of developing embryos (MADE) using the Embryoscope® (Unisense, Denmark) allows time-lapse video review of embryo development. The process of growth from a single cell to a blastocyst can be visually analyzed 24 hours a day without disruption to the culture environment using this technology. The non-invasive, 72 per embryo daily observations with 7 individual planes allow for over 3000 images containing morphokinetic information about each embryo's development to the blastocyst stage. Our study set out to compare multiple single stage embryo culture media to determine the developmental processes that could only be observed when using this new technology. Mouse 1-cell embryos (n=187) were thawed and placed individually into the Embryoscope® culture slides containing equilibrated media. The media studied included our standard 2-stage formulation along with 3 other commercially produced single stage culture media (A, B and C). The slides were inserted into an Embryoscope®, exposing all embryos to the same culture conditions of 37C, 6% CO2 and 5% O2. Continuous culture with video observation in 7 planes (slices) every 20 minutes for up to 110 hours was carried out. Video analysis of embryo development was performed to isolate specific developmental stages reached and the time points that corresponded. Compilation and comparison of the expanded blastocyst rates for each media were analyzed. The percentage of mouse embryos reaching the expanded blastocyst (or more advanced stages) at 110 hours of culture was compiled along with the time point of the first observed cavitation for each medium studied (Table).Tabled 1MEDIAExpanded blastocysts at 110 hours1st cavitation observed2-Stage88.80%78.2 hoursSingle Stage-A74.30%84.4 hoursSingle Stage-B93.60%85.4 hoursSingle Stage-C75.80%87.5 hours Open table in a new tab The use of an Embryoscope® allows for the non invasive observation of embryo development using time-lapse technology. The comparison of the 4 different media show that both the time to initiation of a blastocyst cavity and the rate of reaching the expanded blastocyst stage by a set time point vary dramatically. This continuous observation using MADE, can produce images that demonstrate development with accuracy never before possible. These findings are crucial in selecting an optimal media and for quality control analysis in any laboratory and have been demonstrated only through the use of this new non-invasive technology.

Morphokinetic analysis of developing embryos (MADE) using the Embryoscope® (Unisense, Denmark) allows time-lapse video review of embryo development. The process of growth from a single cell to a blastocyst can be visually analyzed 24 hours a day without disruption to the culture environment using this technology. The non-invasive 72 per embryo daily observations with 7 individual planes allow for over 3000 images containing morphokinetic information about each embryo’s development to the blastocyst stage. Our study set out to discover the processes of blastocyst recovery and reexpansion after trophectoderm biopsy that could only be observed when using this new technology. Mouse 1-cell embryos were placed individually into an Embryoscope® culture slide containing equilibrated media. Slides were inserted into an Embryoscope® for continuous culture with video observation every 20 minutes for 160 hours. Video review of embryo development was performed to isolate time points associated with post-trophectoderm biopsy recovery and reexpansion that would not have been observed using only the standard static single observations each day. Embryos were biopsied a second time on the following day and placed back in the Embryoscope® to judge the recovery and reexpansion after rebiopsy. Overall 87.5% of trophectoderm biopsy embryos begin to reexpand within 20 minutes after completion of biopsy. Hatching blastocysts begin to recover within 20 minutes 100% of the time, with expanded (but not hatching) and fully hatched blastocysts recovering within 20 minutes 66.7% and 50% respectively. After the first biopsy, recovery was within 20 minutes 87.5% of the time and after the second biopsy the following day embryos recovered 77.8% within 20 minutes. All embryos biopsied recovered from both the first and second biopsies, with 80 minutes being the longest time before reexpansion. Static single observations of embryo development the day after a trophectoderm biopsy does not allow for the determination of how an embryo responds and recovers. The use of an Embryoscope® allows for the non-invasive observation of embryo survival and reexpansion using time-lapse technology. Our results demonstrate that the technique of trophectoderm biopsy has a 100% recovery rate, has a very short recovery time of less than 20 minutes in 82% of embryos and that the optimal timing of biopsy is at the hatching blastocyst stage. Continuous observation of embryos using MADE, can produce images that demonstrate developmental processes with accuracy that can improve trophectoderm biopsy techniques and potentially aid in embryo selection for transfer.

This study retrospectively analysed array comparative genomic hybridization (CGH) results of 7753 embryos from 990 patients to determine the frequency of embryonic euploidy and its relationship with the cohort size (i.e. the number of embryos available for biopsy and array CGH analysis). Linear regression analysis was performed to assess the effect of cohort size on euploidy rate adjusted for the effect of female age. While increasing female age was associated with a significant decrease in euploidy rate of day-3 and day-5 embryos (P<0.001 for both groups), cohort size was not significantly associated with euploidy rate. Logistic regression analysis was performed to assess the effect of cohort size, adjusted for maternal age, on the likelihood of having at least one euploid embryo available for transfer. The odds of having at least one euploid embryo in an assisted cycle was significantly decreased by increasing female age (P<0.01 for both day-3 and day-5 embryos) and was significantly increased by every additional embryo available for analysis (P<0.001 for both day-3 and day-5 embryos).

BACKGROUND AND SIGNIFICANCE: The reduction of clinical outcomes in cycles where high numbers of oocytes are retrieved has been debated in the literature. The hypothesis that extremely super physiological estrogen concentrations may impair implantation has been discussed as has the direct effect on embryo viability. In order to be able to study the effect of high numbers of oocytes retrieved independent of the many variables infertility type brings, this study was performed on anonymous oocyte donors only. OBJECTIVE(S): This prospective case cohort study investigated the embryo viability rate which is defined as the number of embryos transferred and cryopreserved divided by the total number of oocytes retrieved in donor oocyte cycles in a private ART clinic. MATERIALS AND METHOD(S): Donor oocyte cycles between January 2008 and December 2009 were studied for the number of embryos transferred plus the number of embryos cryopreserved and the total number of oocytes retrieved. Cycles with male factor infertility were excluded from this study. Cycles that used preimplantation genetic diagnosis (PGD) or preimplantation genetic screening (PGS) were excluded from the study. The non-parametric Chi Sq test was used to determine statistical significance. RESULT(S):Tabled 1Total Number of oocytes retrievedNumber of cyclesTotal number of oocytesTotal number transferred and cryopreserved% viability% immature oocytes% Live Birth Rate / cycle< 1014903640.119.668.310–155160622837.723.167.916–203971828439.621.270.1> 2050136841430.2 = statistically significant where P<0.001.22.868.5∗∗ = statistically significant where P<0.001. Open table in a new tab CONCLUSION: The percent viability or useable embryos per oocyte retrieved was significantly reduced in cycles where more than twenty oocytes were retrieved. This suggests that overly aggressive ovarian hyperstimulation may lead to a direct compromise in oocyte quality.

BACKGROUND AND SIGNIFICANCE: The considerable medical, financial and sociological burden caused by the complications of multiple pregnancies is one of the main criticisms of reproductive medicine around the world. Despite ASRM urging physicians in the USA to reduce the number of embryos for transfer in 1998, this country continues to fall behind most western nations in implementing a single embryo transfer policy for high risk patients. OBJECTIVE(S): This ongoing prospective case cohort study presents four years implementation of e-SET in couples presenting for infertility treatment who fall into the high risk category for multiple pregnancy in a private ART clinic. MATERIALS AND METHOD(S): Patients that were undergoing assisted reproductive treatment cycles between January 2006 and December 2009 who had at least one grade 5AA blastocyst on day five of culture were offered eSET and if the patient declined they were offered elective double embryo transfer (eDET). The clinical pregnancy rate, the singleton pregnancy rate, the twinning pregnancy rate, the triplet pregnancy rate and the quadruplet pregnancy rates were compared between eSET and eDET. The patient characteristic of the number of previous failed treatment cycles was also compared between the two groups. The non-parametric Mann Whitney test was used to determine statistical significance. RESULT(S):Tabled 1Number of embryo transfers# of previous failed cyclesClinical pregnancy rateSingleton rateTwin rateTriplet rateQuad rateLive Birth rateeSET1682 (±2)136/168 (81.5%)120/136 (88.2%)13/136 (9.5%)3/136 (2.2%)0131/136 (96.3%)eDET6452 (±2)510/645 (79.1%)149/510 (29.2%)= statistically significant where P<0.001.321/510 (62.9%)= statistically significant where P<0.001.35/510 (6.8%)= statistically significant where P<0.001.5/510 (0.9%)= statistically significant where P<0.001.496/510 (97.3%)∗∗ = statistically significant where P<0.001. Open table in a new tab CONCLUSION(S): e-SET leads to high pregnancy rates with low risk of multiple pregnancies in patients with high risk of multiple pregnancies. Less than one third of the patients in this study choose to transfer one embryo compared to two. We need more than just physician based education to convince patients at risk of multiple pregnancy to accept single embryo transfer.

Objective: The purpose of this Study was to determine the positive predictive value (PPV) and negative predictive value (NPV) of FISH analysis and to determine which chromosomal abnormalities are most frequently confirmed.Design: Prospective observational.Setting: IVF laboratory.Patient(s): Two hundred forty-one embryos were analyzed from 98 patients.Intervention(s): FISH reanalysis.Main Outcome Measure(s): Embryos that would have been discarded in patients undergoing preimplantation genetic diagnosis (PGD) were fixed and FISH reanalysis was performed. Results of reanalysis were compared with the day 3 diagnosis while PPV and NPV were calculated.Result(s): Among the 241 embryos, 198 embryos were abnormal and 43 were normal by day 3 FISH analysis. The PPV was 83% and the NPV was 81%. PPV was also determined for specific categories of aneuploidy, and certain abnormalities such as monosomies, trisomies, tetrasomies, and polyploidies were frequently confirmed on reanalysis (PPV >80%), whereas Turner syndrome diagnosis was not (PPV = 17%).Conclusion(s): FISH analysis offers a PPV of 83% and NPV of 81% when evaluating a single blastomere in conjunction with PGD. FISH errors and mosaicism are primarily responsible for the errors associated with FISH analysis in PGD.

The actual effectiveness rates of natural and barrier methods of family planning are lower than the theoretical ones. If couples accurately defined the limits of the fertile phase and used barriers at that time, then actual effectiveness might increase. A randomized, controlled clinical trial was initiated to determine the effectiveness of the contraceptive sponge used only during the fertile time and to compare this with sponge use at every intercourse. Recruitment problems and discontinuation forced the early termination of this study, but qualitative information about compliance and acceptability was collected. Common sponge problems were reported as were misuses of the sponge, but problems and misuse were not related. Determination of the fertile phase was reportedly easy, but complaints of and discontinuation for inconvenience occurred. For unplanned pregnancies, contraceptive behaviors around the time of conception are presented.