Machine learning for prediction of euploidy in human embryos: in search of the best-performing model and predictive features.

OBJECTIVE: To assess the best-performing machine learning (ML) model and features to predict euploidy in human embryos. DESIGN: Retrospective cohort analysis. SETTING: Department for reproductive medicine in a university hospital. PATIENT(S): One hundred twenty-eight infertile couples treated between January 2016 and December 2019. Demographic and clinical data and embryonic developmental and morphokinetic data from 539 embryos (45% euploid, 55% aneuploid) were analyzed. INTERVENTION(S): Random forest classifier (RFC), scikit-learn gradient boosting classifier, support vector machine, multivariate logistic regression, and naïve Bayes ML models were trained and used in 9 databases containing either 26 morphokinetic features (as absolute [A1] or interim [A2] times or combined [A3]) alone or plus 19 standard development features [B1, B2, and B3] with and without 40 demographic and clinical characteristics [C1, C2, and C3]. Feature selection and model retraining were executed for the best-performing combination of model and dataset. MAIN OUTCOME MEASURE(S): The main outcome measures were overall accuracy, precision, recall or sensitivity, F1 score (the weighted average of precision and recall), and area under the receiver operating characteristic curve (AUC) of ML models for each dataset. The secondary outcome measure was ranking of feature importance for the best-performing combination of model and dataset. RESULT(S): The RFC model had the highest accuracy (71%) and AUC (0.75) when trained and used on dataset C1. The precision, recall or sensitivity, F1 score, and AUC were 66%, 86%, 75%, and 0.75, respectively. The accuracy, recall or sensitivity, and F1 score increased to 72%, 88%, and 76%, respectively, after feature selection and retraining. Morphokinetic features had the highest relative predictive weight. CONCLUSION(S): The RFC model can predict euploidy with an acceptable accuracy (>70%) using a dataset including embryos' morphokinetics and standard embryonic development and subjects' demographic and clinical features.

Low feasibility of in vitro matured oocytes originating from cumulus complexes found during ovarian tissue preparation at the moment of gender confirmation surgery and during testosterone treatment for fertility preservation in transgender men.

OBJECTIVE: To study the feasibility of in vitro maturation of ovarian tissue oocytes for fertility preservation in transgender men on testosterone treatment. DESIGN: Cross-sectional study SETTING: University hospital PATIENT(S): Eighty-three transgender men enrolled from November 2015 to January 2019 INTERVENTION(S): In vitro maturation of cumulus-oocyte complexes (COCs) harvested at the time of gender confirmation surgery, and fertilization through intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S): In vitro maturation, fertilization, and blastulation rates; comparison of morphokinetics with vitrified-warmed oocytes; and analysis of the genetic profiles of embryos. SECONDARY OUTCOMES: association between serum hormone levels; COCs' morphologic characteristics, and vitrification rate. RESULT(S): All participants were on testosterone treatment for a median of 83 (64[Quartile 1]; 113.2[Quartile 2]) weeks. A total of 1,903 COCs (mean per participant, 23 ± 15.8) were collected. The in vitro maturation rate was 23.8%, vitrification rate was 21.5%, and survival rate after warming was 72.6% (n = 151). Intracytoplasmic sperm injection was performed in 139 oocytes. The rate of normal fertilized oocytes was 34.5%, and 25 (52.1%) embryos reached day 3. One blastocyst was achieved on day 5. Aberrant cleavage patterns and early embryo arrest were observed in 22 (45.8%) and 44 (91.7%) zygotes, respectively. Compared with vitrified-warmed donor oocytes, a delay was observed in pronuclei disappearance, t2 (time to reach 2 cell stage) timings, and CC1 (the duration of the 1st cell cycle) and SS3 (synchronization of cleavage pattern (calculated as t8-t5) time intervals. A normal genetic pattern was seen in 42% embryos. The proportion of vitrified oocytes was negatively associated with progesterone (odds ratio, 0.76) and positively associated with antimüllerian hormone serum levels (odds ratio, 1.23). The highest vitrification rate was achieved by the morphologic characteristic 344 at day 0 and by 433 at day 2. CONCLUSION(S): Ovarian tissue oocytes matured in vitro show low developmental capacity in transgender men, when collected under testosterone treatment.

Assisted oocyte activation significantly increases fertilization and pregnancy outcome in patients with low and total failed fertilization after intracytoplasmic sperm injection: a 17-year retrospective study.

OBJECTIVE: To investigate the extent to which assisted oocyte activation (AOA) improves clinical outcomes in patients diagnosed with oocyte activation deficiencies (OADs). DESIGN: Retrospective cohort study comparing AOA cycles and previous intracytoplasmic sperm injection (ICSI) cycles in couples experiencing low or total failed fertilization after ICSI. Importantly, the sperm-related oocyte-activating capacity was examined in all patients before AOA with the use of the mouse oocyte activation test (MOAT). SETTING: Infertility center at a university hospital. PATIENT(S): A total of 122 couples with a history of low or total failed fertilization after ICSI. INTERVENTION(S): ICSI, MOAT, AOA, and embryo transfer. MAIN OUTCOME MEASURE(S): Fertilization, pregnancy, and live birth rates. RESULT(S): MOAT revealed 19 patients with a sperm-related OAD (MOAT group 1), 56 patients with a diminished sperm-related oocyte-activating capacity (MOAT group 2), and 47 patients with a suspected oocyte-related OAD (MOAT group 3). AOA (191 cycles) significantly improved fertilization, pregnancy, and live birth rates in all MOAT groups compared with previous ICSI attempts (243 cycles). Fertilization rates after AOA were significantly different among MOAT groups 1 (70.1%), 2 (63.0%), and 3 (57.3%). Between MOAT group 1 and 3, significant differences in pregnancy (49.0% vs. 29.4%) and live birth (41.2% vs. 22.1%) rates were observed. In total, 225 embryo transfers resulted in 60 healthy live births following AOA. CONCLUSION(S): Patients undergoing diagnostic testing before AOA show a significant improvement in clinical outcomes compared with previous cycles. Our findings highlight that AOA should be reserved for patients with clear OADs.

Sperm Chromatin Dispersion Test before Sperm Preparation Is Predictive of Clinical Pregnancy in Cases of Unexplained Infertility Treated with Intrauterine Insemination and Induction with Clomiphene Citrate.

BACKGROUND/AIMS: A large proportion of men with normal sperm results as analyzed using conventional techniques have fragmented DNA in their spermatozoa. We performed a prospective study to examine the incidence of DNA fragmentation in sperm in cases of couples with previously unexplained infertility and treated with intrauterine insemination. We evaluated whether there was any predictive value of DNA fragmentation for pregnancy outcome in such couples. METHODS: The percentage of DNA fragmentation and all classical variables to evaluate sperm before and after sperm treatment were determined. We studied the probable association between these results and pregnancy outcome in terms of clinical and ongoing pregnancy rate per started first cycle. We also assessed the optimal threshold level to diagnose DNA fragmentation in our center. RESULTS: When using threshold levels of 20, 25, and 30%, the occurrence of DNA fragmentation was 42.9, 33.3, and 28.6%, respectively. Receiver operating characteristic (ROC) analysis of all cases revealed an area under the curve of 80% to predict the clinical pregnancy rate per cycle from testing the sperm motility (a + b) before treatment. We failed to generate an ROC curve to estimate pregnancy outcome from the amount of DNA fragmentation before treatment. However, when selecting only those men with a pretreatment DNA fragmentation of at least 20%, the pretreatment result was statistically different between couples who achieved a clinical pregnancy and those who did not. CONCLUSION: DNA fragmentation is often diagnosed in couples with unexplained infertility. Each center should evaluate the type of test it uses to detect DNA fragmentation in sperm and determine its own threshold values.

Slow controlled-rate freezing of human in vitro matured oocytes: effects on maturation rate and kinetics and parthenogenetic activation.

OBJECTIVE: To create a pool of frozen donated human oocytes and find the optimal stage for slow controlled-rate freezing of human in vitro matured oocytes. DESIGN: Oocytes at different developmental stages of maturation (germinal vesicle, metaphase I, or metaphase II) and oocytes that failed to fertilize after IVF or intracytoplasmic sperm injection (ICSI) were frozen using a slow controlled-rate freezing protocol. Frozen/thawed oocytes were artificially activated to verify activation potential and compared with oocytes that were not frozen. SETTING: University hospital-based fertility center. PATIENT(S): Stimulated patients undergoing IVF/ICSI treatment donated oocytes left over during their infertility treatment. INTERVENTION(S): Human oocytes were frozen at different stages of maturation. Fresh and frozen/thawed oocytes were activated by electrical pulses followed by incubation in 6-dimethylaminopurine. MAIN OUTCOME MEASURE(S): Survival rate, maturation rate and kinetics, and activation potential. RESULT(S): Human oocytes at all developmental stages have high survival rates after slow controlled-rate freezing. Frozen/thawed germinal vesicle oocytes showed decreased and delayed maturation after thawing. Activation potential was not affected. CONCLUSION(S): A pool of donated human oocytes could be established using slow controlled-rate freezing. Immature oocytes should be frozen after in vitro maturation.

Long term effects of micro-surgical testicular sperm extraction on androgen status in patients with non obstructive azoospermia.

BACKGROUND: The aim of our study was to review the results of microsurgically performed testicular sperm extraction (TESE) and to evaluate its possible long term effects on serum testosterone (T). METHODS: We operated on 48 men (35 +/- 8 years) with non-obstructive azoospermia (NOA). If no spermatozoa were found following a micro epididymal sperm extraction (Silber et al., 1994) and testicular biopsy, testicular microdissection was performed or multiple microsurgical testicular biopsies were taken. The mean follow-up of the serum T was 2.4 +/- 1.1 years. RESULTS: Sperm was retrieved in 17/48 (35%) of the men. The per couple take home baby rate if sperm was retrieved was 4/17 (24%). Serum T decreased significantly at follow-up (p < 0.05) and 5/31 (16%) de novo androgen deficiencies developed CONCLUSION: In patients with non-obstructive azoospermia in whom no spermatozoa were found following a micro epididymal sperm aspiration and a simple testicular biopsy, we were able to retrieve spermatozoa in 35% of the men. The take home baby rate was 24% among couples with spermatozoa present upon TESE. De novo androgen deficiency occurred in 16% of the male patients following TESE indicating that, in men with NOA, long term hormonal follow up is recommended after TESE.