Isoform-resolved transcriptome of the human preimplantation embryo.

Human preimplantation development involves extensive remodeling of RNA expression and splicing. However, its transcriptome has been compiled using short-read sequencing data, which fails to capture most full-length mRNAs. Here, we generate an isoform-resolved transcriptome of early human development by performing long- and short-read RNA sequencing on 73 embryos spanning the zygote to blastocyst stages. We identify 110,212 unannotated isoforms transcribed from known genes, including highly conserved protein-coding loci and key developmental regulators. We further identify 17,964 isoforms from 5,239 unannotated genes, which are largely non-coding, primate-specific, and highly associated with transposable elements. These isoforms are widely supported by the integration of published multi-omics datasets, including single-cell 8CLC and blastoid studies. Alternative splicing and gene co-expression network analyses further reveal that embryonic genome activation is associated with splicing disruption and transient upregulation of gene modules. Together, these findings show that the human embryo transcriptome is far more complex than currently known, and will act as a valuable resource to empower future studies exploring development.

High Incidence of Obstetric Anal Sphincter Injuries among Immigrant Women of Asian Ethnicity.

(1) Background: Obstetric anal sphincter injuries (OASI) may complicate vaginal deliveries. The aim of the present study was to explore the incidence and clinical characteristics of OASI among Asian women living in a Western country compared to local Caucasian women. (2) Methods: A retrospective cohort study of 380 women diagnosed with OASI, following singleton vaginal deliveries, during a 10-year period (January 2011 to December 2020). Exclusion criteria: age < 18 years, stillbirth, and breech presentation. Demographic, clinical, and obstetrical data were obtained, and a comparison between Asian and Caucasian women was performed. (3) Results: There were 35 cases of OASI among 997 women of Asian ethnicity compared to 345 cases of OASI among 86,250 Caucasian women (3.5% vs. 0.4%, respectively, p < 0.001). Asian women endured a significantly higher rate of fourth-degree OASI (17.1%) even though they bore smaller newborns (3318 g vs. 3501 g, p = 0.004), and birth weights rarely exceeded 3800 g (2.8% vs. 25.8%, p < 0.001). Asian ethnicity was also associated with a significantly higher risk for blood transfusion following OASI and a lower tendency for postpartum follow up. (4) Conclusions: Immigrant women of Asian ethnicity had a nine-fold higher rate of OASI, much higher than previously reported. Furthermore, Asian women had higher rates of fourth-degree OASI.

Sperm parameters in Israeli transgender women before and after cryopreservation.

BACKGROUND: The application of fertility preservation, initially intended for oncological patients prior to gonadotoxic treatment, has extended in recent years to transgender and gender-non-conforming individuals undergoing therapy for gender compatibility. OBJECTIVES: To examine semen quality and survival in transgender women pursuing semen cryopreservation in the presence or absence of gender-affirming hormonal medication. MATERIALS AND METHODS: In this retrospective cohort study, we reviewed data of 74 consecutive transgender women presenting for semen cryopreservation at a single center between 2000 and 2019. Semen parameters before and after cryopreservation were compared to a control group composed of 100 consecutive sperm bank donor candidates. A subgroup analysis of subjects who had used gender-affirming hormonal treatment was also performed. RESULTS: Compared to the control group, transgender women had lower total sperm count (144.0 vs. 54.5 million, respectively, p < 0.001), lower sperm motility percentage (65.0% vs. 51.0%, respectively, p < 0.001), and lower total motile sperm count (94.0 vs. 27.0 million, respectively, p < 0.001). Values were further decreased in transgender women who had received hormonal treatment before sperm cryopreservation. Post-thawing motility rate remained lower in the transgender group compared to the control group (20.0% vs. 45.0%, respectively, p < 0.001), and the total motile count remained lower as well (2.7 vs. 9.0 million, respectively, p < 0.001). Following sperm cryopreservation, the post-thaw decreases in total motile sperm count were higher in the transgender group compared with the control group (91.5% vs. 90.0%). Further subdivision in the transgender group showed that the decrease in total motile sperm count was lower for transgender women who did not use gender-affirming hormonal treatment compared to those who did (-89.7% vs. -92.6%, respectively, p < 0.01). DISCUSSION AND CONCLUSION: Sperm parameters in transgender women are poor compared to candidates for sperm donation representing the general population. Specimens collected after discontinuation of gender-affirming hormone treatments were further impaired. Moreover, post-thawing sperm total motile count, motility, and overall sperm survival were reduced in transgender women.

Cleavage stage at compaction-a good predictor for IVF outcome.

OBJECTIVE: To analyze whether cleavage stage at compaction, and not only kinetics, can serve as a reliable predictor for clinical outcome. METHODS: A retrospective cohort study including 1194 embryos, classified by compaction initiation stage (Group 1: compaction at fewer than eight cells, Group 2: compaction at eight cells, Group 3: compaction at more than eight cells). Of these, 815 embryos were evaluated for morphokinetic preimplantation parameters, and 379 embryos were analyzed for clinical implantation following thawing and transfer of single blastocysts during the same period. RESULTS: In total, 1194 embryos were analyzed. Embryos that underwent compaction from more than eight cells (Group 3) exhibited more synchronous cleavage compared with Groups 1 and 2 (at both S2 and S3; P < 0.001), and displayed a significantly lower fragmentation rate. The likelihood of obtaining top-quality blastocysts decreased by 73% and 44% when comparing Group 3 embryos with those of Groups 1 and 2, respectively, (P < 0.03). Clinical validation of the results shows that while compaction from fewer than eight cells barely produced blastocysts for transfer, compaction at eight or more cells is crucial for implantation and birth (birth rates 11.1% and 18.5% for Groups 2 and 3, respectively). CONCLUSION: Cleavage stage at compaction has a direct effect on blastocyst quality and subsequent pregnancy, so can be included in newly developed deep learning models for embryo selection.

Substantial Obstetric Anal Sphincter Injury during Vacuum Assisted Delivery: An Obstetrical Issue or Device Related?

BACKGROUND: Obstetric anal sphincter injuries (OASIS) might be associated with long-term urinary and anorectal morbidities. The aim of the study was to investigate the risk factors and clinical implications of OASIS associated with vacuum-assisted deliveries versus normal vaginal deliveries. METHODS: A series of 413 consecutive OASIS cases were retrospectively analyzed. A comparison was made between OASIS cases diagnosed following vacuum-assisted deliveries versus OASIS cases diagnosed following normal vaginal deliveries. Multivariable analysis was used to study the association between vacuum-assisted deliveries and superficial (3A and 3B) versus deep (3C and 4) perineal tears. RESULTS: The study population comprised 88,123 singleton vaginal deliveries. Diagnosis of OASIS was made in 413 women (0.47% of the total cohort), 379 (91.8%) of whom had third-degree tears and 34 (8.2%) of whom had fourth-degree tears. Among the 7410 vacuum-assisted deliveries, 102 (1.37%) had OASIS, whereas, among the 80,713 normal vaginal deliveries, only 311 (0.39%) had OASIS. In a multivariate analysis, only vacuum-assisted delivery was found to be associated with a significant risk of deeper (3C or 4) perineal tears (OR = 1.72; 95% CI 1.02-2.91; p = 0.043). CONCLUSIONS: Vacuum-assisted instrumental intervention is a significant risk factor for OASIS and especially for deeper tears, independent of other maternal and obstetric risk factors.

Stabilization of hESCs in two distinct substates along the continuum of pluripotency.

A detailed understanding of the developmental substates of human pluripotent stem cells (hPSCs) is needed to optimize their use in cell therapy and for modeling early development. Genetic instability and risk of tumorigenicity of primed hPSCs are well documented, but a systematic isogenic comparison between substates has not been performed. We derived four hESC lines in naive human stem cell medium (NHSM) and generated isogenic pairs of NHSM and primed cultures. Through phenotypic, transcriptomic, and methylation profiling, we identified changes that arose during the transition to a primed substate. Although early NHSM cultures displayed naive characteristics, including greater proliferation and clonogenic potential compared with primed cultures, they drifted toward a more primed-like substate over time, including accumulation of genetic abnormalities. Overall, we show that transcriptomic and epigenomic profiling can be used to place human pluripotent cultures along a developmental continuum and may inform their utility for clinical and research applications.

Similar fertilization rates and preimplantation embryo development among testosterone-treated transgender men and cisgender women.

RESEARCH QUESTION: What are the effects of testosterone treatment on oocyte fertilization and preimplantation embryo development among transgender men who have undergone fertility preservation? DESIGN: A retrospective study was undertaken in a university-affiliated tertiary hospital between April 2016 and November 2021. Embryos were divided into three groups by source: 210 embryos from 7 testosterone-exposed transgender men, 135 from 10 cisgender women who cryopreserved embryos, and 276 from 24 cisgender women who underwent fertility treatment. Statistical analyses compared assisted reproductive technology outcomes between the group of transgender men and both groups of cisgender women. Morphokinetic and morphological parameters were compared between the embryos derived from these three groups. RESULTS: The transgender men (30.2 ± 3.5 years of age) were significantly younger than the cisgender women who cryopreserved embryos (35.1 ± 1.8 years; P = 0.005) and the cisgender women who underwent fertility treatment (33.8 ± 3.2 years; P = 0.017). After adjusting for participant age, the fertilization rate was comparable between the transgender men and both groups of cisgender women (P = 0.391 and 0.659). There were no significant differences between the transgender men and the cisgender women who preserved fertility in terms of number of cryopreserved embryos (7.2 ± 5.1 and 3.5 ± 2.6; P = 0.473) or the distribution of embryo age at cryopreservation (P = 0.576). All morphokinetic parameters evaluated by time-lapse imaging, as well as the morphological characteristics, were comparable for the embryos in all three groups. CONCLUSIONS: Testosterone exposure among transgender men has no adverse impact upon fertilization rates or preimplantation embryo development and quality.

Maternal underweight does not adversely affect the outcomes of IVF/ICSI and frozen embryo transfer cycles or early embryo development.

OBJECTIVE: To compare assisted reproductive technology (ART) outcomes and preimplantation embryo development between underweight and normal-weight women. METHODS: This retrospective cohort study included 26 underweight women (body mass index [BMI] < 18.50 kg/m2) and 104 normal-weight women (BMI >20 and <24.9 kg/m2) who underwent a total of 204 in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles and 358 fresh/frozen embryo transfers (ET) in our institution between January 2016 and December 2018. Statistical analyses compared selected ART outcomes (ovarian stimulation, fertilization, and pregnancy) between both weight groups. Morphokinetic and morphological parameters were also compared between 346 and 1467 embryos of underweight and normal-weight women, respectively. RESULTS: The mean ± standard deviation age of the underweight and normal-weight women was similar (31.6 ± 4.17 vs 32.4 ± 3.59 years; p = .323). There were no differences in the peak estradiol levels, the number of retrieved oocytes, the number of metaphase II oocytes, and the oocyte maturity rates between the two groups. The IVF/ICSI fertilization rates and the number of embryos suitable for transfer or cryopreservation were similar for both groups. All morphokinetic parameters that were evaluated by means of time-lapse imaging as well as the morphological characteristics were comparable between low and normal BMI categories. There were no significant differences in pregnancy achievement, clinical pregnancy, live births, and miscarriage rates between the suboptimal and optimal weight women. CONCLUSION: Underweight status has no adverse impacts on the outcomes of IVF/ICSI with either fresh or frozen ET or on preimplantation embryo development and quality.

Why fertility preservation rates of transgender men are much lower than those of transgender women.

RESEARCH QUESTION: What are the fertility preservation rates of transgender women and transgender men, and what are the factors that affect their decision-making? DESIGN: This prospective study included 97 transgender women and 91 transgender men referred to the Gender Clinic of the study medical centre's Endocrinology Institute and to the Gan Meir Community Health Care Centre. The responders completed a 28-item questionnaire during 2018. RESULTS: Most of the transgender women and transgender men wished to parent a child (67.4% and 61.9%, respectively, P = 0.447), but only 40.4% of the transgender women and 5.8% of the transgender men used fertility preservation (P < 0.001). The main reasons for not pursuing fertility preservation were unwillingness to postpone gender-affirming treatment (58.8% and 74.0%, respectively, P = 0.076), preference to adopt a child (58.8% and 60.9%, respectively, P = 0.818) and cost (44.9% and 60.9%, respectively, P = 0.086). Factors related to the fertility preservation process itself were specifically chosen by transgender men compared with transgender women as the reason for not pursuing this treatment, including distress caused by the fertility preservation technique (60.3% versus 29.3%, respectively, P = 0.006), fear of gender dysphoria caused by hormonal treatment (63.5% versus 28.3%, P = 0.002) and concern over the attitude of medical staff (44% versus 19%, P = 0.027). CONCLUSIONS: Fertility preservation rates were considerably lower among transgender men than transgender women, strongly related to the fertility preservation process itself. Finding ways to overcome the obstacles confronted by transgender individuals, especially transgender men, will enhance their future biological parenting.

Mental Health, Work Presenteeism, and Exercise in Inflammatory Bowel Disease.

BACKGROUND: Chronic diseases, such as IBD, can lead to anxiety and depression which can have a significant impact on productivity at work [presenteeism]. The aim of this study was to assess the prevalence of depression/anxiety, presenteeism and exercise levels among IBD patients. METHODS: This was a multicentre study whereby adult IBD patients, in clinical remission, were asked to answer a questionnaire anonymously. Hospital Anxiety and Depression Score [HADS], Stanford Presenteeism Scale [SPS-6] and Godin Exercise Score were also collected. RESULTS: A total of 585 patients were recruited. The majority had Crohn's disease [CD, 62.2%] and were male [53.0%], with a median age of 39 years [IQR 30-49]. A psychiatric diagnosis was present in 10.8% of patients prior to their IBD diagnosis. A further 14.2% of patients were psychiatrically diagnosed after IBD diagnosis, this being commoner in CD patients [41.6% of CD, p <0.01]. A raised HADS-Anxiety or a HADS-Depression score ≥8 was present in 46.1% of patients, with 27.4% having a score ≥11. Low presenteeism at work was present in 34.0%. Patients diagnosed with depression/anxiety had a more sedentary lifestyle [p <0.01], lower presenteeism at work [p <0.01] and a higher rate of unemployment [p <0.01]. CONCLUSIONS: A significant percentage of IBD patients in remission suffer from anxiety and/or depression. Risk factors for these are CD, female gender, use of biologic medications, long-standing and/or perianal disease. Depression/anxiety was associated with a sedentary lifestyle, lower presenteeism at work and unemployment. Validated screening tools and appropriate referrals to psychologists and/or psychiatrists should be employed within IBD clinics.

Transgender men's preferences when choosing obstetricians and gynecologists.

BACKGROUND: Transgender men are a marginalized population with unique health care needs. However, their usage of health services is low because of considerable discrimination. A major factor in their avoidance is patient-provider interactions. METHODS: This cross-sectional study included 102 transgender men who anonymously completed a 55-item questionnaire in clinic, between 10/2017 and 01/2019. In addition, 92 transgender women filled out the part about family physician's preferences. We examined which characteristics transgender men prefer in their obstetricians/gynecologists in order to promote their usage of healthcare services. RESULTS: A small majority of the transgender men (54.1%) had no gender preference for their obstetrician/gynecologist, while 42.9% preferred a female obstetrician/gynecologist and 3.1% preferred a male obstetrician/gynecologist. Most transgender men with a same-gender preference preferred female obstetricians/gynecologists for both invasive procedures (e.g., pelvic examination, 97.4%) and non-invasive procedures (e.g., cesarean section, 60%). The reasons for preferences regarding invasive procedures were feeling comfortable, embarrassment and feeling that female obstetricians/gynecologists are gentler. Transgender men who preferred female obstetricians/gynecologists ranked ability (90.5%), sexual tolerance (92.9%) and gender identity tolerance (90.5%) as the top three desirable qualities of obstetricians/gynecologists, while the responders who did not prefer female ranked ability (94.6%), experience (92.9%) and knowledge (92.9%) as the top three qualities. Transgender men with female preferences considered female obstetricians/gynecologists to be more accepting of gender identity compared to the responders that did not prefer females (47.5% vs. 9.1%, P < .001).. CONCLUSION: A small majority of the transgender men exhibited no gender preference when choosing an obstetrician/gynecologist, although 42.9% preferred females. The latter choice was associated with the assumption that female obstetricians/gynecologists are more tolerant towards their transgender men patients. Educating the medical staff about their special needs and establishing dedicated SGM centers staffed with high percentages of female healthcare providers are highly recommended.

The Manchester operation - is it time for it to return to our surgical armamentarium in the twenty-first century?

We conducted a study to assess the efficacy and outcome results of Manchester operation for women with symptomatic elongation of uterine cervix between 2010 and 2020. Forty-five women were enrolled. Mean age was 54.2 ± 10.5 years, 57.5% were premenopausal. Mean follow up was 4.53 years. Most patients were pleased from the surgery: 29 (76.3%) very pleased and three (7.9%) quite pleased. There were four cases of late postoperative complications: hematometra, pyometra, vesicovaginal fistula and small bowel evisceration through the posterior vaginal fornix. All four cases required surgical interventions with complete recovery. Manchester operation is an effective procedure for symptomatic elongation of uterine cervix. The surgery is short, minimal blood loss and without significant intraoperative complications. However, it is not free of late postoperative complications and it is therefore important that the surgery will be carried out with a strict technique and by skilled surgeons who are capable to manage unique postoperative complications.Impact StatementWhat is already known on this subject? Elongation of the uterine cervix is a condition that has not been sufficiently studied. Up-to-date data regarding the efficacy and safety of the Manchester operation for patients with cervical elongation and normal pelvic support are scarce, as most previously published studies included mainly patients with uterine prolapse.What do the results of this study add? The present study presents the efficacy, complications and clinical outcomes of Manchester operation for women with elongation of the uterine cervix. Our results show that Manchester operation is safe, effective method for symptomatic uterine cervix elongation.What are the implications of these findings for clinical practice and/or further research? This suggests Manchester operation is an effective procedure for symptomatic uterine cervix elongation should be implemented more widely. It must be carried out with a strict technique by skilled surgeons capable to manage unique postoperative complications.

Adolescent Transgender Females Present Impaired Semen Quality That Is Suitable for Intracytoplasmic Sperm Injection Even Before Initiating Gender-Affirming Hormone Treatment.

The present study aimed to determine the semen quality and cryopreservation outcomes among adolescent transgender females at the time of fertility preservation (FP) before initiating gender-affirming hormone (GAH) treatment. This retrospective cohort study included 26 adolescent transgender females who underwent FP in our Fertility Institute between 06/2013 and 10/2020. Pre-freezing semen parameters were compared to WHO 2010 reference values. Post-thaw semen parameters were used to determine the adequate assisted reproductive technology (ART). A multivariate linear regression analysis was performed to assess the impact of medical and lifestyle factors on semen quality. The mean age at which adolescent transgender females underwent FP was 16.2 ± 1.38 years. The median values of all semen parameters in our study group were significantly lower compared to the WHO data, including volume (1.46 mL vs 3.2 mL, respectively, P = 0.001 ), sperm concentration (28 × 106/mL vs 64 × 106/mL, P < 0.001), total sperm number (28.2 × 106 vs 196 × 106, P < 0.001), total motility (51.6% vs 62%, P < 0.001), and normal morphology (2% vs 14%, P < 0.001). The frequency of semen abnormalities was teratozoospermia 72%, hypospermia 52%, oligozoospermia 28%, and azoospermia 4%. The median post-thaw total motile count was 0.17 × 106/vial, and the quality was adequate only for ICSI in 87.7% of the thawed semen samples. No correlation was found between selected medical and lifestyle factors and poor semen parameters. Semen quality is strongly reduced among adolescent transgender females before hormone therapy and their stored sperm samples are suitable for intracytoplasmic sperm injection (ICSI) rather than conventional IVF/intrauterine insemination (IUI).

Skin exposure to UVB light induces a skin-brain-gonad axis and sexual behavior.

Ultraviolet (UV) light affects endocrinological and behavioral aspects of sexuality via an unknown mechanism. Here we discover that ultraviolet B (UVB) exposure enhances the levels of sex-steroid hormones and sexual behavior, which are mediated by the skin. In female mice, UVB exposure increases hypothalamus-pituitary-gonadal axis hormone levels, resulting in larger ovaries; extends estrus days; and increases anti-Mullerian hormone (AMH) expression. UVB exposure also enhances the sexual responsiveness and attractiveness of females and male-female interactions. Conditional knockout of p53 specifically in skin keratinocytes abolishes the effects of UVB. Thus, UVB triggers a skin-brain-gonadal axis through skin p53 activation. In humans, solar exposure enhances romantic passion in both genders and aggressiveness in men, as seen in analysis of individual questionaries, and positively correlates with testosterone level. Our findings suggest opportunities for treatment of sex-steroid-related dysfunctions.

Gonadotropin-Releasing Hormone Agonist Versus Recombinant Human Chorionic Gonadotropin Triggering in Fertility Preservation Cycles.

The purpose of this research is to study the efficacy of GnRH-a versus r-hCG triggering in patients who go through fertility preservation cycles. This retrospective cohort study was performed in a tertiary university-affiliated medical center. It includes 191 patients undergoing fertility preservation cycles between May 2013 and September 2018, in which ovulation was induced by either GnRH-a or r-hCG. Main outcome measures were number and rate of mature oocyte. Among treatment cycles with medical indication, GnRH agonist significantly increases the odds for high mature rate by 3.55 (1.30-9.66), while in treatment cycles with social indication, there is no significant effect of the triggering agent. An advantage for GnRH-a triggering was observed in medically indicated preservation cycles.

A tailored flexible vaginal pessary treatment for pelvic organ prolapse in older women.

OBJECTIVES: To present a flexible protocol of vaginal pessaries for older women with significant pelvic organ prolapse (POP). DESIGN: A prospective cohort study of 140 consecutive older women (aged ≥65 years) with significant POP treated with individually fitted vaginal pessaries. After initial insertion, each patient was invited for a first evaluation after 1 month and subsequent follow-ups at increasing intervals of 1 month. During each examination, the pessary was removed and the vagina was inspected for infection, bleeding, or erosions. All patients were advised to use a vaginal estrogen cream twice a week. SETTING: The study was conducted during 2020 in the urogynecology clinic of a tertiary medical center. PARTICIPANTS: The study cohort included 140 consecutive older women with significant and symptomatic POP treated with vaginal pessaries. MAIN OUTCOME: Primary outcomes included time intervals between follow-up visits, pessary-associated complications, need to change the pessary, need to remove the pessary temporarily, and whether the patients eventually underwent POP surgery. RESULTS: The mean age of the patients at the time of pessary fitting was 76.7 ± 9.2 years (range 65-100 years). Mean interval between follow-up examinations was 3.3 ± 1.1 months (range 1-6 months). Most patients (83.6%) used a ring pessary with support, but only a small number of patients were able to maintain the pessary by themselves. Of the 140 patients, five (3.6%) patients only eventually underwent POP surgery, and in 11 (7.9%) patients, the vaginal pessary had to be removed for 2-4 weeks because of significant vaginal discharge or superficial erosions. Additionally, 12 (8.6%) patients developed stress urinary incontinence following pessary insertion. CONCLUSIONS: Customized management with vaginal pessary for symptomatic POP in older women is effective and safe, and is a suitable therapeutic alternative for older women who are unable or unwilling to undergo reconstructive pelvic surgery.

Principles of signaling pathway modulation for enhancing human naive pluripotency induction.

Isolating human MEK/ERK signaling-independent pluripotent stem cells (PSCs) with naive pluripotency characteristics while maintaining differentiation competence and (epi)genetic integrity remains challenging. Here, we engineer reporter systems that allow the screening for defined conditions that induce molecular and functional features of human naive pluripotency. Synergistic inhibition of WNT/ß-CATENIN, protein kinase C (PKC), and SRC signaling consolidates the induction of teratoma-competent naive human PSCs, with the capacity to differentiate into trophoblast stem cells (TSCs) and extraembryonic naive endodermal (nEND) cells in vitro. Divergent signaling and transcriptional requirements for boosting naive pluripotency were found between mouse and human. P53 depletion in naive hPSCs increased their contribution to mouse-human cross-species chimeric embryos upon priming and differentiation. Finally, MEK/ERK inhibition can be substituted with the inhibition of NOTCH/RBPj, which induces alternative naive-like hPSCs with a diminished risk for deleterious global DNA hypomethylation. Our findings set a framework for defining the signaling foundations of human naive pluripotency.

Unmet communication needs and moral work in the disposition decision concerning surplus frozen embryos: The perspectives of IVF users.

The disposition decision is a frequently unresolved issue for many IVF users with surplus frozen embryos (SFEs), and this study draws attention to their experiences and moral work, locating it in the Jewish-Israeli context that legally enables the donation of SFEs to research but prohibits donation to other infertile people. To explore the (mis)understandings and (mis)communication underlying IVF users' decisions concerning the fate of their SFEs, the records of 674 IVF users with SFEs stored for more than 5 years during 1996-2011 were analyzed, and 89 IVF users with different disposition decisions were recruited for semi-structured interviews. With an average of 5.1 SFEs, after an average of 8 years of storage, no response to a written request for a disposition decision came from 60% (n = 404) of IVF users with SFEs. Payment for storage and defrosting were the two most frequent choices (13%, n = 89 and 89, respectively) followed by donation to research and transfer (7%, n = 47 and 45, respectively). Three themes emerged from the interviews: misunderstanding the consequences of not returning the disposition form, communication gaps regarding donation to research, and the unmet wish to donate embryos to infertile people. We conclude by discussing the experiences and views of IVF users as reflecting the implications of the liminality and boundary-work surrounding the frozen embryo as a moral work object, and their consequences for policy recommendations.