Faculty Bibliography

OBJECTIVE: Many preimplantation genetic testing (PGT) labs require intracytoplasmic sperm injection (ICSI) for PGT for aneuploidy (PGT-A) due to concern for paternal cell contamination. We sought to determine if sperm lysis occurs during PGT-A and assess the rate of paternal cell contamination in trophectoderm (TE) biopsies in embryos from insemination. MATERIALS AND METHODS: Sixty-two tripronuclear (3PN) embryos donated to research were collected from IVF with either insemination or ICSI from January - April, 2021. Embryos were cultured and assessed for development to blastocyst stage on days 5, 6 and 7 of culture. Embryos that developed into blastocysts underwent two separate TE biopsies. Biopsy procedure consisted of zona ablation on day 4 followed by TE biopsy using 2-3 pulses of laser beam at the cell junction. Biopsy samples were washed with drops of buffer 2-3 times and placed in a PCR tube. Arrested embryos were collected and assessed for approximate cell number. One group of arrested embryos was collected without washing (unwashed) and a second group was collected after removal of the zona (washed). TE biopsies, arrested embryos, and maternal and paternal samples were sent to a PGT lab to determine the genetic ploidy composition of the embryo biopsies and arrested embryos including the parent of origin. Testing included PGT-A using the PGTseq platform and SNP allele sharing that can detect parental origin of abnormalities and contamination.
RESULT(S): Of the 62 3PN embryos cultured, 17 developed into blastocysts with 4 from ICSI and 13 from insemination. There were 45 arrested embryos with 6 from ICSI (2 washed, 4 unwashed) and 39 from insemination (14 washed, 25 unwashed). PGT analysis showed varying degrees of paternal cell contamination in unwashed arrested embryos from insemination, and no paternal cell contamination in washed arrested embryos (ICSI or insemination) or unwashed ICSI embryos. Two washed arrested embryos from insemination showed no amplification. There was no paternal cell contamination in TE biopsies from either ICSI or insemination.
CONCLUSION(S): Analysis of unwashed arrested embryos from insemination demonstrates that excess sperm can lyse and cause paternal cell contamination during PGT-A. However, TE biopsies of embryos from insemination showed no evidence of paternal cell contamination, indicating that when properly washed and processed, paternal cell contamination is unlikely in inseminated embryos undergoing PGT-A. While this study was not powered to draw definitive conclusions or assess levels of contamination that interfere with PGT-A, preliminary results indicate that ICSI is not necessary for PGT-A. It should be noted that these findings are specific to the PGTseq platform, and may not translate to other methods. IMPACT STATEMENT: This study demonstrates that sperm have the ability to lyse and are a potential source of paternal cell contamination in PGT-A. However, this study also showed a 0% rate of paternal cell contamination in inseminated embryos when embryo biopsies were washed and processed as described, suggesting that ICSI is not necessary for patients desiring PGT-A

OBJECTIVE: Oocyte cryo is widely used for fertility preservation, but the value of M1 cryo remains unclear. We evaluated the utility and efficiency of M1 compared to M2 cryo. MATERIALS AND METHODS: Patients (pts) who thawed autologous oocytes at our academic center from 2004-2020 were reviewed. Pts were excluded if cryo was performed for a medical indication, as research, due to no sperm or a natural disaster, in combination with embryos or for use with a gestational carrier. At our center, all M1s retrieved from 2004-2015 were cryopreserved; after 2015, M1s were only cryopreserved if <15 M2s were retrieved during the same cryo cycle. Outcomes included survival rate, useable embryo rate and embryo transfer (ET) results.Auseable embryo was defined as an embryo that was transferred, biopsied or cryopreserved for future use. Statistics included Fisher's exact test.
RESULT(S): 543 pts (median age at 1st cryo 38y, interquartile range 37-40y) underwent 800 cryo, 605 thaw and 416 ET cycles. Cryo was performed with vitrification for 72%, slow freezing for 4% and both technologies for 24% of pts. In total, 8511 oocytes (1019M1s + 7492 M2s)were thawed.All pts thawed >=1 M2, and 60% (n=327) thawed >=1 M1. See table for thaw outcomes of M1s vs. M2s. For 30 pts, >=1 M1 led to a useable embryo (n=32 useable embryos). Vitrification was used for 69% of these M1s (n=22) and slow freezing was used for 31% (n=10). Of the 32 useable embryos from M1s, 69% (n=22) underwent PGTand 4were euploid (17 aneuploid, 1 mosaic). Therewere 3 single ETs of euploid embryos from M1s, which led to 1 spontaneous abortion (SAB) and 2 biochemical pregnancies. Therewere 3 single ETs of untested embryos from M1s, which led to 1 negative result, 1 SAB and 1 singleton ongoing pregnancy. The ongoing pregnancy is from an ETof a day 5 morula and is now in the third trimester. There were 6 ETs in which untested embryos from M1s were transferred alongwith untested embryos fromM2s, resulting in 3 negative results, 1 SAB, 1 singleton live birth and 1 unknown outcome (ongoing singleton pregnancy at last contact).
CONCLUSION(S): Cryopreserved M1s can result in useable embryos and pregnancies, but are less likely to survive or form useable embryos than cryopreserved M2s. To our knowledge, this is the first report of an ongoing third trimester pregnancy from a cryopreserved M1. This information may be helpful for pt counselling and designing oocyte cryo protocols for embryology labs. IMPACT STATEMENT: Cryopreserved M1s may be a viable option for pts with a low M2 yield. (Table Presented)

OBJECTIVE: The use of a GnRH trigger in COH cycles has increased due to an improved safety profile but not all patients have adequate response1.We sought to investigate the utility of using serum GN levels to predict response to GnRH trigger. MATERIALS AND METHODS: We performed a retrospective cohort study of all GnRH-antagonist COH cycles at an urban university affiliated fertility center from 2017-2020. Cycles that utilized GnRH-agonist (GnRH-a) alone or in combination with human chorionic GN (hCG) for trigger were included. Patient and cycle characteristics were collected from the electronic medical record, including day 2 baseline follicle stimulating hormone (B-FSH) and earliest in-cycle luteinizing hormone (EIC LH). An optimal response to GnRH-a trigger was defined as a LH R40 mIU/mL on the morning after trigger. Descriptive statistics (median +/- range for continuous variables; frequencies and percentages for categorical variables) were calculated by GnRH-a response. Statistical analyses were performed on SAS (v9.4) and included the chi-square test, Fisher's exact test, or Mann Whitney U test, as appropriate with a p<0.05 considered significant.
RESULT(S): A total of 3,865 COH antagonist cycles were included. Ninetyone percent of patients had an optimal response to GnRH-a trigger. Optimal responders had higher B-FSH levels than those with poor response (6.52 mIU/ml vs 4.36 mIU/ml, p<0.001). Similarly, the EIC LH was higher for optimal responders (4.66 mIU/ml vs 2.16 mIU/ml, p<0.001). Optimal response had a positive association with older age (p<0.00001), lower BMI (p<0.0001), less days of stimulation (p<0.001), lower starting serum estradiol (p<0.0007), and lower total gonadotropin dose (p<0.001). Optimal response was also associated with B-FSH >5 mIU/ml (p<0.0001), EIC LH >1 (p<0.0001), and Clomiphene citrate use (p< 0.009). Asian race was associated with poor response (p<0.006). There was no difference in oocyte maturity rate (p=0.6) or fertilization rate (p=0.5) for optimal or poor response. Cutoffs for B-FSH (>5 mIU/mL) and EIC LH ( >1 mIU/mL) were chosen to be reasonable clinical cutoffs to create a tool or aid to predict patient response to GnRH-a trigger. The incidence of patients with B-FSH >5 IU/ ml who had a poor response was 4.9% compared to 16.0% in patients with B-FSH <5 (p<0.0001). Twenty-four percent of patients with an EIC LH <1 had a poor response, compared to 4% of patients with EIC LH >1 (p<0.0001). The combination of B-FSH >5 IU/ml and EIC LH >1 IU/ml had a 71% sensitivity and 96% PPV in predicting an optimal response. When individually compared to a B-FSH >5 mIU/ml, an EIC LH>1 mIU/ ml had a higher sensitivity (91% vs 76%) and higher PPV (96% vs 95%) in predicting optimal response.
CONCLUSION(S): A B-FSH>5 and EIC LH>1 may be an appropriate threshold and helpful guide for physicians when determining trigger medicine for GnRH-antagonist COH cycles. Further studies are needed to understand predictors of poor response above these thresholds. IMPACT STATEMENT: In an era of personalized medicine, cycle and patient characteristics, such as GN levels, may improve cycle outcomes and provide further individualized care

OBJECTIVE: Morphologic evaluation of embryos after fertilization is the first step in embryo assessment, with two pronuclei (2PN) indicating normal fertilization. Deviations from 2PN are considered consistent with abnormal fertilization and genetic ploidy, and may be discarded instead of cultured. The aim of this study was to determine the genetic ploidy of three pronuclei (3PN) embryos diagnosed by morphology. MATERIALS AND METHODS: Sixty-two 3PN embryos donated to research that underwent IVF with either insemination or ICSI were collected from January - April, 2021. 3PN embryos were identified at time of fertilization check and vitrified. Batched 3PN embryos were subsequently warmed and cultured. Embryos were assessed for development to the blastocyst stage on days 5, 6 and 7 of culture, and embryos that developed into blastocysts underwent two separate trophectoderm (TE) biopsies. TE biopsies, along with maternal and paternal samples were sent to a pre-implantation genetic testing (PGT) lab to determine the genetic ploidy composition of the morphological based 3PN embryos. Testing included PGT for aneuploidy (PGT-A) using the PGTseq platform that routinely includes triploidy detection via single nucleotide polymorphism (SNP) B allele ratio. Testing was also performed using a second method, SNP allele sharing, with the maternal and paternal DNA samples. This method can detect both triploidy and parental of origin of abnormalities.
RESULT(S): Of the 62 3PN embryos cultured, 17 (27%) developed into blastocysts that underwent TE biopsy. In all cases paired biopsies were concordant. Three of the 17 biopsied embryos were diploid (18%) and 14 were triploid (82%). All 3 diploid embryos were the result of insemination and were aneuploid on PGT-A; no euploid embryos were identified. The overall rate of diploid tested blastocysts was 4.8% (3/62) among all 3PNs collected. Of the 14 triploid embryos, 10 were the result of IVF with traditional insemination and 4 were from ICSI. All triploid embryos from insemination were consistent with paternal origin while all triploid embryos from ICSI were consistent with maternal origin. Both methods for detecting triploidy were concordant.
CONCLUSION(S): Embryos morphologically diagnosed as 3PN are typically discarded as they are likely the result of abnormal fertilization consistent with triploidy. This study demonstrates that a small percentage of 3PN embryos have the potential to develop into blastocysts with a diploid genetic complement. While none of the diploid 3PN embryos in this study were found to be euploid, this could be due to the small sample size and it is possible that a larger number of embryos may result in 3PN euploid embryos which could impact a labs decision on what tissue to discard or culture. It should be noted that due to the inherently subjective component of morphologic assessment, these findings may not translate to other laboratories. IMPACT STATEMENT: While 3PN embryos are typically discarded in IVF after both insemination and ICSI, our study shows that a small proportion have the potential to develop into diploid blastocysts where reproductive potential remains to be seen

OBJECTIVE: The majority of patients surveyed believe that use of social media (SM) to share information benefits the patient experience (1). Our objective was to understand the SM content and activity about mosaic embryos. MATERIALS AND METHODS: This is a retrospective cohort study of the use of search terms ''mosaic embryo'' and ''mosaic embryos'' on Google (GO) and Instagram (IG). Both terms were analyzed from 1/1/2015 to 12/21/2020 on Google Trends, a publicly accessible tool that quantifies GO searches nationally. On IG, all posts that mention either search term were included. Posts were characterized by author type, content, and tone (positive, negative, neutral). Likes per post and total followers on each account were quantified. Percent of likes (PL) from total followers was calculated to assess activity. P-value < 0.05 was considered significant.
RESULT(S): Within the study time period, the term ''mosaic embryo'' was searched most in the months of 4/ 2018 and 2/2020. Both peaks occurred after mosaicism was highlighted by a major news outlet or celebrity (2,3). Use of the term increased over time with a slope of 0.64. On IG, 400 posts utilized the hashtag ''#mosaicembryo'', with 259 listed as top posts, making them most easily accessible. Of top posts, prevalence of author types parent/patient, fertility clinic and patient education (PE) platform was 233 (90%), 10 (3.9%) and 16 (6.2%) respectively. Content included personal stories (212, 81.9%), questions (10, 3.9%) and PE (37, 14.3%). 57.1% of posts were positive, 35.1% neutral and 7.7% negative. Mean number of likes, followers and PL was 104, 1571 and 14.6% respectively. PL was not associated with author type, content or tone (p=0.16, p=0.07, p=0.11). The term ''mosaic embryos'' was most searched on GO in 10/ 2020. Use remained consistent in the time window (m=-0.1). 160 posts on IG utilized the hashtag ''#mosaicembryos'', 78 being top posts. The majority of posts were by patients (58, 74.4%) compared to fertility clinics (11, 14.1%) and PE platforms (9, 11.5%). Frequency of personal story, question and PE was 52 (66.7%), 2 (2.6%), and 24 (30.8%) respectively. Distribution of tone was 55.1% positive, 39.7% neutral, and 5.1% negative. Mean number of likes, followers and PL was 84, 1515 and 8.14% respectively. PL on fertility clinic posts was significantly lower than patient and PE posts ( 2.8, 9.2, 7.3, p<0.05). PL was highest in posts with personal stories compared to questions and PE (9.7, 4.0, 5.1, p<0.05). Tone was not associated with change in PL (p=0.8).
CONCLUSION(S): Internet search of terms related to mosaic embryo has increased in recent years. These terms have a prominent SM presence where patients, clinics and PE platforms can share information. These platforms are mainly used by patients posting about their positive experiences. IMPACT STATEMENT: This is the first study to characterize the use of terms related to embryonic mosaicism on SM. This information is mainly created by non-medical professionals but is easily accessible and often used as a trusted resource. Understanding where patients may be receiving their information is important in providing adequate counseling and management. (Table Presented)

OBJECTIVE: COVID-19 has influenced family building, delayed fertility care, and affected people's decisions about where to live.We sought to understand differences in movement of cryopreserved reproductive tissue before and during the pandemic. MATERIALS AND METHODS: This was a retrospective cohort study of patients who transported tissue into or out of a single academic fertility center in New York City (NYC). Tissue transport was compared the year before (PRE, 4/1/2019-3/31/2020) and after (DUR, 4/1/2020-3/31/2021) the height of the COVID-19 pandemic in NYC, an epicenter. The primary outcome was the number of patients transporting tissue DUR compared to PRE. Secondary outcomes were the number of geographic changes, type of tissue, geographic origin/destination, and type of movement (in or out). Statistical analyses were performed using Kolmogorov-Smirnov, Wilcoxon Signed Rank Sum, Chi-Square, and Fisher's Exact tests with p<0.05 considered significant.
RESULT(S): A total of 367 tissue transports were included, with similar rates between cohorts (PRE 46.3% (170/367) vs DUR 53.7% (197/367), p=0.16). The median age at transport was the same (PRE 41 (range 29-54) vs DUR 41 (range 28-54) years, p=0.54). A similar amount of tissue was transported in (PRE 30.0% (51/170) vs DUR 35.0% (69/197)) and out (PRE 70.0% (119/170) vs DUR 65.0% (128/197), p=0.32). Patients were more likely to transport embryos pre-pandemic (37.6% (64/170) oocytes vs 61.8% (105/170) embryos, PRE) and oocytes during COVID-19 (51.8% (102/197) oocytes vs 44.2% (87/197) embryos, DUR) (p<0.01). A subgroup analysis excluding tissue moved for a gestational carrier or donor gametes found a similar number of transports were due to patient geographic relocation (PRE 50.0% (61/122) vs DUR 40.5% (60/148), p=0.12). Examination of geographic origin and destination of tissue PRE vs DUR produced no identifiable trends (p=0.38). Timing of tissue transport varied. The monthly transport rates were relatively even PRE (average 8% per month). However, during the pandemic, there were few transports in the beginning (April-May 2020, 0-1% per month) followed by a peak of transports in June-August 2020 (10-11% per month) and February-March 2021 (11-16% per month) (p<0.01). Transport activities were impacted by closure of clinics and courier service availability.
CONCLUSION(S): The rate of cryopreserved tissue movement did not differ in the year before versus during the pandemic at our center, despite being in a COVID-19 epicenter, although transport activities were concentrated into fewer days. There was peak movement of tissue three months after the pandemic onset and roughly one year from the start of the pandemic. The type of tissue transported shifted to favor oocytes during the pandemic, warranting more investigation in how COVID-19 impacted family building activities. IMPACT STATEMENT: Despite the impact of COVID-19 on reproductive and place of living choices, the pandemic did not affect the amount of cryopreserved tissue that was relocated. However, insight into the increased movement of oocytes and potential impacts on warming outcomes or timelines is necessary

OBJECTIVE: Segmental aneuploidies (SA) are generated when a piece of chromosome is gained or lost and may contribute to sub-optimal birth outcomes[ 1]. Our objective was to examine the frequency, type and association with maternal age of SA. MATERIALS AND METHODS: This is a retrospective cohort study of all embryos that underwent trophectoderm biopsy for preimplantation genetic testing for aneuploidy (PGT-A) via Next Generation Sequencing between 1/2015 and 12/2020 at a single university-based fertility center that resulted with a non-mosaic SA. SA were characterized by type of abnormality and location (p or q arm). Maternal age was stratified by SART age groups. Primary outcome was the association between age and SA. Secondary outcomes were SA location and type. Statistical analysis included ANOVA and chisquare where appropriate, with p<0.05 considered significant.
RESULT(S): There were 711 embryos with an SA included; of 30,300 embryos biopsied (2.3%). Mean maternal age was 38.8+/-4.7. The frequency of SA increased with maternal age (Table 1). The distribution of SA on p, q and both arms was 58 (8.2%), 539 (75.8%) and 114 (16%) respectively. Partial monosomy was the most common type of SA (298, 41.6%), followed by partial trisomy (154, 21.7%) and the combination of SA and whole chromosome aneuploidy (WCA; 154, 21.7%). There is a significant association between type of SA and increasing maternal age (p<0.00). SA occurred most frequently on chromosomes (CH) 1, 2 and 16 and least frequently on CHs 19, 20 and 21. For single SA only, CH 1, 2 and 4 were most common. For double SA, CH 2 and 8 occurred most. For SA with WCA, SA on CH 1 and WCA on CHs 16 and 22 was most common. Acrocentric CH have less SA than non-acrocentric. Chromosome location was not associated with maternal age across SA types (All: p=0.39, single p=0.86, SA + WCA p= 0.48)
CONCLUSION(S): Among our embryo cohort, the frequency of nonmosaic SA increased with maternal age. Since previous literature suggests that SA is mainly affected by paternal age, this association may be attributed to age similarities within romantic partnerships (2). SA were most common on chromosomes 1 and 2, and the q arm. IMPACT STATEMENT: This is the first study of the association between SA prevalence, type and location with maternal age to provide a basis for future research and understanding about SA detected by PGT-A