The percentage of reproductive age women classified as obese has increased over the past decades. Obesity has been associated with several adverse pregnancy outcomes including lower implantation rates and pregnancy loss. Various theories have been proposed to identify a cause including poor embryo quality and endometrial receptivity. We attempted to assess an association between maternal BMI and live birth in frozen embryo transfers with known euploid embryos. Retrospective cohort. Autologous cycles with embryos undergoing preimplantation genetic screening (PGS) between March 2014 and July 2015 were available for analysis. Live birth outcomes from the first FET of a PGS-normal blastocyst(s) after retrieval were included in this analysis. PGD for single gene disorders or known parental translocations were excluded from the analysis. The association of BMI and live birth were assessed using GEE models, accounting for multiple cycles within the same patients and adjusting for patient age and the number of embryos transferred. Outcomes were also analyzed using adjusted GEE models according to BMI categories per WHO guidelines; underweight (<18.5), normal (18.5-24.9), overweight (25-29.9), and obese (>30). 376 FET cycles from 361 unique patients were available for analysis. The average age in this patient population was 36.1 years. The number of embryos transferred and clinical pregnancy rates did not differ significantly in any BMI category. There was no association between BMI and live birth when BMI was analyzed as a continuous variable (OR 1.02, 95% CI 0.98-1.06, P=0.31). Patients with normal BMI had no significant difference in live birth rate when compared to overweight (OR 0.81, 95% CI 0.47-1.40, P=0.81) and obese patients (OR 1.18, 95% CI 0.67-2.07, P=0.55). BMI was not associated with clinical pregnancy or live birth in patients undergoing euploid FET in this cohort when analyzed as both a continuous and dichotomous variable. These results suggest that BMI may not have a significant impact in FET cycle outcomes when known euploid embryos are transferred.Tabled 1Underweight (n=8)Normal (n=206)Overweight (n=87)Obese (n=75)Age (years)37.735.236.636.1Average Embryos Transferred11.21.21.2Clinical Pregnancy Rate56%56%53%60%Live Birth Rate44%46%41%51% Open table in a new tab

Several studies have demonstrated that day 5 blastocysts have higher implantation rates than day 6 blastocysts in fresh embryo transfers. It is unclear if this is due to poor embryo quality with delayed blastulation, or dyssynchronous endometrium. We aimed to assess embryo quality through preimplantation genetic screening (PGS) and determine if day 6 blastocysts had higher rates of aneuploidy. Retrospective cohort. Fresh autologous IVF cycles from March 2014 through July 2015 with embryos undergoing PGS were available for analysis. Patients who elected for PGD for single gene disorders or known parental translocations were excluded from the analysis. Mixed-effects logistic regression models were used to assess for an association between day of embryo biopsy and aneuploidy. Analysis was performed adjusting for maternal age using piece-wise model. Unadjusted analysis was also performed. Available for analysis were 707 cycles from 544 unique patients. 3,055 embryos were biopsied and stratified according to blastulation rate. An average of 4.3 embryos were biopsied per completed cycle, with similar numbers blastulating and biopsied on days 5 and 6. The average age in this population was 37 years. Prior to adjusting for maternal age, embryos biopsied on day 6 were more likely to be aneuploid than those reaching the blastocyst stage on day 5 (OR 1.246 (1.064-1.460), p =0.006). Aneuploidy was significantly associated with maternal age (≤37 years old OR 1.05 (1.02-1.09) p=0.002; >37 years old OR 1.26 (1.13-1.41) p<0.001). After adjusting for maternal age, embryos biopsied on day 6 continued to have higher aneuploidy rates (OR 1.169 (1.003-1.363), p=0.046). Our data demonstrated embryos with delayed blastulation are more likely to be aneuploid. To our knowledge this is the largest cohort with adequate power to address this question. Though this difference is statistically significant, the absolute difference is small, therefore factors other than aneuploidy may also contribute to lower pregnancy rates in day 6 fresh transfers.Tabled 1Day 5 Blastocyst (n=1538)Day 6 Blastocyst (n=1517)No. Biopsied/Cycle2.22.2Unadjusted Aneuploidy Rate53%(50%-56%)58%(55%-61%)Adjusted Aneuploidy Rate56%(53%-59%)60%(57%-62%) Open table in a new tab

Prior to ovulation, oocytes are arrested in meiosis I (MI), and the luteinizing hormone (LH) surge triggers ovulation and completion of MI. Maternal nondisjunction in MI is the most common cause of aneuploidy. In patients undergoing ART, the ovulation sequence has historically been initiated with human chorionic gonadotropin (hCG) acting directly on LH receptors in the ovary. Alternatively, ovulation can be initiated with a GnRH agonist acting on the level of the pituitary, utilizing the body's endogenous store of LH, which has a significantly shorter half-life. We aimed to determine if aneuploidy rates in embryos differed by the mechanism of IVF trigger. Retrospective cohort study Fresh autologous IVF cycles from March 2014 through July 2015 with embryos undergoing PGS were available for analysis. The associations between trigger type and aneuploidy were analyzed using linear mixed models with random intercepts to account for repeated measurements. Rates were adjusted for maternal age, BMI, and ovarian reserve. 707 cycles from 543 unique patients were available for analysis. 3049 embryos were biopsied. DNA was unable to be amplified from 108 embryo biopsies; thus, 2941 embryos were available for analysis. The average patient age was 37 years. Higher rates of diminished ovarian reserve (DOR) were seen in patients receiving hCG compared to GnRH agonist and dual trigger. Patients receiving hCG trigger had fewer oocytes retrieved, 2PNs, and blastocysts compared to those with GnRH agonist and dual triggers. No significant difference in percentage of embryos with aneuploidy was observed with GnRH agonist when compared to hCG trigger (Beta=-0.003, 95%CI: -0.05, 0.05, p=0.89). Further dual trigger had similar aneuploidy percentages (Beta =-0.004, 95%CI: -0.10, 0.10, p=0.93) relative to hCG trigger. IVF trigger type does not affect aneuploidy rates in patients undergoing ART. The specific trigger type should be determined based on the patient's clinical presentation, and patients should be counseled that there is no increased risk of aneuploidy with either method. This provides reassurance that GnRH trigger, which may be selected to optimize patient safety, does not compromise egg quality.Tabled 1hCG Trigger (n=446)GnRH AgonistTrigger (n=230)Dual Trigger(n=27)Maternal age (years)37.535.837.1Patients with DOR19%10%7%Average oocytes retrieved13.220.420.3Average No. 2PN812.813Average No. blastocysts3.86.36.9No. embryos biopsied15161266159Adjusted aneuploidy percentage60.7%60.4%60.3% Open table in a new tab

OBJECTIVE: To develop a mathematical formula resulting in an accurate determination of mitochondrial DNA (mtDNA) levels in human blastocysts substratified by ploidy, age, and implantation potential.DESIGN: Retrospective analysis of mtDNA content in human blastocysts used in preimplantation genetic diagnosis for IVF selection.MATERIALS AND METHODS: 833 embryos derived from 181 patients were tested for mtDNA content by next generation sequencing (NGS), and 150 embryos derived from 96 patients were tested by quantitative polymerase chain reaction (qPCR).For each embryo, the level of mtDNA was determined from a trophectoderm biopsy by whole genome amplification followed by NGS and/or qPCR.The value was subjected to mathematical analysis tailored to the genomic DNA composition of said embryo.Grouped values were compared by Welch's two-tailed upaired t-test.RESULTS: On average our quantitation method changed the conventionally determined mtDNA level of a given embryo via NGS by 1.35% +/-1.58%, with changes ranging up to 17.42%, and via qPCR by 1.33% +/-8.08%, with changes ranging up to 50.00%.Levels of mtDNA in euploid and aneuploid embryos showed a statistically insignificant difference of P¼0.102 by NGS (euploid N¼494, aneuploid N¼339) and P¼0.642 by qPCR (euploid N¼100, aneuploid N¼50).Blastocysts derived from younger or older patients had comparable mtDNA values, with P¼0.293 by NGS (20-37 age group N¼559, 38-46 age group N¼274) and P¼0.101 by qPCR (20-37 age group N¼92, 38-46 age group N¼58).Blastocysts that upon transfer resulted in implantation did not contain significantly different mtDNA levels compared to blastocysts that failed to implant, with P¼0.813 by NGS (implanted N¼51, non-implanted N¼69) and P¼0.103 by qPCR (implanted N¼49, non-implanted N¼51).CONCLUSIONS: We recommend the implementation of our correction factor to all laboratories evaluating mtDNA levels in their embryos by NGS or qPCR.Applied to our in-house data, our quantitation method reveals that overall levels of mtDNA are largely equal between human blastocysts regardless of embryo ploidy, age, or implantation potential.

The prevalence of obesity has increased, and several studies have associated increased BMI with poor outcomes in patients undergoing IVF. The mechanism is speculated to be secondary to oocyte quality or uterine environment. We aimed to assess embryo quality using PGS to determine if aneuploidy rates were higher in patients with elevated BMI. Retrospective cohort study. Fresh autologous IVF cycles from February through July 2015 with embryos undergoing PGS were available for analysis. Patients who elected for PGD for single gene disorders or known parental translocations were excluded from the analysis. BMI was classified according to World Health Organization guidelines; underweight (<18.5), normal (18.5-24.9), overweight (25-29.9), and obese (>30). The associations between BMI and aneuploidy were analyzed using generalized estimating equations to account for patients with multiple cycles, and were adjusted for female age. 309 cycles from 267 unique patients were available for analysis. 1,237 embryos were biopsied and stratified according to BMI (n=28 underweight, n=713 normal, n=285 overweight, n=211 obese). The average age in this cohort was 37 years. Age was a significant predictor of aneuploidy (OR 1.02, 95% CI 1.003-1.016, p=0.001). No significant difference in aneuploidy rate per patient was observed in obese vs normal BMI patients (OR 0.96 (0.88-1.04), p=0.37). No significant difference in aneuploidy rates was found in overweight vs normal BMI patients (OR 0.98 (0.89-1.08), p=0.72). There were also no differences in number of embryos transferred or clinical pregnancy rate in obese vs normal BMI patients. Elevated BMI was not associated with aneuploidy in patients undergoing PGS in this large cohort. In our study population, the transfer of only euploid embryos resulted in similar clinical pregnancy rates in obese vs normal BMI patients. Further studies are needed to determine the etiology and extent to which obesity impacts IVF outcomes.Tabled 1Cycle Outcomes by BMI# CyclesUnderweight (n=6)Normal (n=175)Overweight (n=72)Obese (n=56)Age (yrs)35.337.137.436.7No. MII1012.111.712.2No. 2PN910.39.69.4No. Biopsied4.74.14.03.8No. Transferred11.21.11.2ClinicalPregnancy Rate/Transfer50%57%41%60%Aneuploidy Rate60.7%59.1%55.8%60.3% Open table in a new tab