The impact of microfluidics sperm processing on blastocyst euploidy rates compared with density gradient centrifugation: a sibling oocyte double-blinded prospective randomized clinical trial.

OBJECTIVE: To compare the euploidy rates among blastocysts created from sibling oocytes injected with sperm and processed using microfluidics or density gradient centrifugation. DESIGN: Sibling oocyte randomized controlled trial. SETTING: Single university-affiliated infertility practice. PATIENTS: A total of 106 patients aged 18-42 years undergoing fresh in vitro fertilization treatment cycles with preimplantation genetic testing between January 2021 and April 2022 contributed 1,442 mature oocytes, which were injected with sperm and processed using microfluidics or density gradient centrifugation. INTERVENTION(S): The sperm sample is divided and processed using a microfluidics device and density gradient centrifugation for injection into sibling oocytes. MAIN OUTCOME MEASURE(S): The primary outcome was the embryo euploidy rate. Secondary outcomes included fertilization, high-quality blastulation, and ongoing pregnancy rates. RESULT(S): The blastocyst euploidy rate per mature oocyte was not significantly different in the study group compared with the control group (22.9% vs. 20.5%). The blastocyst euploidy rate per biopsied embryo was also similar between the 2 groups (53.0% vs. 45.7%). However, the fertilization rate per mature oocyte injected was found to be significantly higher in the study group compared with the control group (76.0% vs. 69.9%). The high-quality blastulation rate per mature oocyte injected was similar between the 2 groups, as was the total number of embryos frozen. There were no differences in the number of participants with no blastocysts for biopsy or the number of participants with no euploid embryos between the 2 groups. Among the male factor infertility and recurrent pregnancy loss subgroups, there were no differences in euploidy rates, fertilization rates, blastulation rates, or total numbers of blastocysts frozen, although the study was underpowered to detect these differences. Seventy-seven patients underwent frozen embryo transfer; there were no significant differences in pregnancy outcomes between the 2 groups. CONCLUSION(S): Microfluidics processing did not improve embryo euploidy rates compared with density gradient centrifugation in this sibling oocyte study, although fertilization rates were significantly higher. CLINICAL TRIAL REGISTRATION NUMBER: NCT04744025.

Attitudes towards disposition of cryopreserved sperm in the event of death.

INTRODUCTION: To evaluate patient preference for sperm disposition in case of death based on demographic factors and infertility etiology. MATERIALS AND METHODS: This retrospective cohort study was performed at a university hospital-affiliated fertility center. Charts of 550 men undergoing cryopreservation for assisted reproductive technologies (ART) between 2016-2019 were reviewed to create a descriptive dataset. Patients previously signed consent forms stating their preference for sperm transfer to their partner or disposal in the event of their subsequent death. Patients undergoing sperm cryopreservation for the purpose of ART were analyzed to assess associations between demographic characteristics and etiology of infertility and their choice to either transfer sperm to their partner or discard. RESULTS: A total of 84.9% (342/403) of patients included in final analyses elected to transfer their sperm to their partner in the event of their death. Factors associated with a significantly increased likelihood to transfer versus discard included a male-factor infertility diagnosis compared to female-factor infertility diagnosis (transfer rate 89.3% vs. 79.9%; p = .022) and commercial insurance coverage versus non-commercial/no insurance coverage (transfer rate 86.3% vs. 75.0%, p = .029). No significant differences relating to age, race/ethnicity, occupation classification, marital status or duration of marriage, or prior paternity were found. CONCLUSION: A majority of male patients seeking sperm cryopreservation for ART elected to transfer their sperm to their partner if future death should occur. There does not appear to be a clear factor that would impact this decision based on demographic characteristics.

Rationale and study design of a double-blinded prospective randomized clinical trial comparing euploidy rates among embryos created from sibling oocytes injected with sperm processed by microfluidics or by density gradient centrifugation.

BACKGROUND: During the in vitro fertilization (IVF) process, sperm must be processed prior to insemination. While the most common method, density gradient centrifugation, can potentially damage sperm during centrifugation, a recent advancement in sperm processing uses a microfluidics system which selects for the most highly motile sperm. In selecting for these sperm which may be of higher quality, the euploidy rates of embryos created as a result may also be improved. The primary aim of this study is to compare the euploidy rates per mature oocyte between embryos created from sibling oocytes injected with sperm processed by microfluidics sorting or by density gradient centrifugation. METHODS: This is a double-blinded prospective randomized sibling oocyte study including patients undergoing treatment with IVF with intracytoplasmic sperm injection (ICSI) and preimplantation genetic testing (PGT). After controlled ovarian hyperstimulation, oocytes from each patient will be separated into two groups. Each group will be randomized to sperm processed using either microfluidics or density gradient centrifugation and embryos biopsied for PGT to assess euploidy rates. A sample size of 686 oocytes in each group for a total of 1372 oocytes will provide 80% power to detect a significant difference in the euploidy rates per mature oocyte between the two groups. An ancillary study examining the relationship between sperm processing method and sperm DNA fragmentation will be assessed. CONCLUSION: This study will offer insight into the sperm's contribution to embryo euploidy, and has the potential to provide an alternative method of improving euploidy rates in clinical practice.

Independent serum markers of corpora lutea function after gonadotropin-releasing hormone agonist trigger and adjuvant low dose human chorionic gonadotropin in in vitro fertilization.

OBJECTIVE: To characterize corpora lutea (CL) function after gonadotropin-releasing hormone agonist (GnRHa) trigger with the use of adjuvant human chorionic gonadotropin (hCG). DESIGN: Secondary analysis of serum from prospective randomized clinical trial. SETTING: University-based fertility center. PATIENT(S): Women under 40 years of age at risk of ovarian hyperstimulation syndrome (OHSS) with serum E2 level <4,000 pg/mL. INTERVENTIONS(S): All subjects underwent ovarian stimulation with the use of a GnRH antagonist protocol. Within a larger study, subjects were randomized to receive 1,000 IU hCG at the time of GnRHa trigger and placebo at the time of vaginal oocyte retrieval (VOR) or placebo at the time of GnRHa trigger and 1,500 IU hCG at the time of VOR. MAIN OUTCOME MEASURE(S): Luteal phase and early pregnancy curves of serum prorenin and 17α-hydroxyprogesterone (17OH-P). RESULT(S): Thirty subjects enrolled in this secondary analysis. Serum 17OH-P peaked in the early luteal phase, 5 days after GnRHa trigger, with a nadir in the mid-luteal phase 9 days after trigger. Serum prorenin peaked in the luteal phase 2 days after GnRHa trigger, independently from adjuvant hCG timing, and reached a nadir at 9 days after trigger. CL function appears higher when adjuvant hCG is given at VOR compared with adjuvant hCG given at the time of trigger. CONCLUSION(S): CL function, as interpreted by proxy measures of serum prorenin and 17OH-P with pregnancy, continues despite GnRHa trigger. Both options for adjuvant hCG timing are sufficient for CL rescue and successful pregnancy, so the potential for OHSS risk with increased CL activity after hCG at VOR should be considered. CLINICAL TRIAL REGISTRATION NUMBER: NCT01815138.

The National Physicians Cooperative: transforming fertility management in the cancer setting and beyond.

Once unimaginable, fertility management is now a nationally established part of cancer care in institutions, from academic centers to community hospitals to private practices. Over the last two decades, advances in medicine and reproductive science have made it possible for men, women and children to be connected with an oncofertility specialist or offered fertility preservation soon after a cancer diagnosis. The Oncofertility Consortium's National Physicians Cooperative is a large-scale effort to engage physicians across disciplines - oncology, urology, obstetrics and gynecology, reproductive endocrinology, and behavioral health - in clinical and research activities to enable significant progress in providing fertility preservation options to children and adults. Here, we review the structure and function of the National Physicians Cooperative and identify next steps.

Sequential embryo assessment outperforms investigator-driven morphological assessment at selecting a good quality blastocyst.

Two selection methods (morphology-only and a sequential embryo assessment algorithm) were compared within the same IVF clinic to determine which method best identifies the embryos on day 3 that will develop into the highest quality on day 5. The sequential embryo assessment algorithm was significantly better at selecting the best embryo and selecting a blastocyst compared with the morphology-only method.

A soluble molecule secreted by human blastocysts modulates regulation of HOXA10 expression in an epithelial endometrial cell line.

OBJECTIVE: To determine the existence of a soluble signal, secreted from the human blastocyst embryo, that induces HOXA10 gene expression before cell-cell contact. DESIGN: To analyze, by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), cell-free media that had contained human embryos cultured to the blastocyst stage for a soluble molecule that induces HOXA10 expression in an endometrial epithelial cell line (Ishikawa). SETTING: Assisted reproduction technology program of Yale University, New Haven, Connecticut. PATIENT(S): Patients undergoing intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF) cycles. Treatment of Ishikawa cells with blastocyst-conditioned media. MAIN OUTCOME MEASURE(S): Determination of HOXA10 gene expression. RESULT(S): We demonstrate that cell-free media that had contained human embryos cultured to the blastocyst stage contain a soluble molecule that induces HOXA10 expression in an endometrial epithelial cell line (Ishikawa). We found that hCG does not induce HOXA10 in Ishikawa cells. CONCLUSION(S): Soluble molecules induce a well-characterized marker of endometrial receptivity in endometrial cells without blastocyst apposition. Additionally, HOXA10 induction can serve as a means of evaluating human embryos cultured for IVF and ET. High quality embryos may induce local endometrial receptivity before trophectoderm-endometrial contact.

Ubiquitin-dependent sperm quality control mechanism recognizes spermatozoa with DNA defects as revealed by dual ubiquitin-TUNEL assay.

Defective mammalian spermatozoa become ubiquitinated during epididymal passage, a mechanism that may mark the abnormal spermatozoa for proteolytic destruction (Sutovsky et al., 2001a: J Cell Sci 114:1665-1675). It is not known how such spermatozoa are recognized by the epididymal ubiquitination pathway and whether there is a selection against certain types of sperm defects. We examined the relationship between sperm ubiqutination, lifelong sperm morphology and sperm DNA defects using a single chanel, ubiquitin-activated flow cytometric assay, and a dual, ubiquitin-TUNEL assay. Semen samples from nine service sires of good-to-average fertility were screened. A positive correlation was found between sperm ubiquitination and the average frequency of morphological semen abnormalities from field evaluations performed throughout the reproductive life of individual sires. Sample correlation coefficients were r=0.65 for primary (head and tail) and r=0.60 for total semen abnormalities in the single channel assay. In a dual assay, we found a high, positive correlation (r=0.93) between the ubiquitin-positive sperm and the TUNEL positive sperm. Substantial correlations (r=0.47-0.64) were observed when the measurements from these two respective assays were compared for individual sires. While anti-ubiquitin antibodies recognized most of the TUNEL-positive sperm cells, the TUNEL-positive spermatozoa represented only a subset (approximately 20-40%) of all ubiquitin-positive cells. It appears that the ubiquitin-dependent sperm quality control, residing in the epididymal epithelium, has the ability to detect spermatozoa with apoptotic or necrotic DNA, while spermatozoa with defects other than DNA fragmentation are also recognized and ubiquitinated.