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Sleep disordered breathing (SDB) is common in patients with neuromuscular diseases, including spinal muscular atrophy (SMA). While polysomnography (PSG) findings have been described in natural history studies of patients with SMA, reports regarding PSG in treated children are limited to nusinersen. We aim to describe the sleep characteristics in a cohort of children treated with Onasemnogene-abeparvovec.We conducted a cross-sectional cohort study of children with SMA followed at the University of Florida Center for neuromuscular and rare diseases and had a diagnostic or split night PSG after SMA treatment.Eight children were included in the cohort (four female), aged 5-250 days at diagnosis. Five children had two survival motor neuron 2 (SMN2) copies, two had three SMN2 copies and one subject had four SMN2 copies. Median age at the time of treatment was 46.5 days (range 20-257). All children received onasemnogene-abeparvovec (OA) before their PSG; in addition to OA, one received nusinersen and one received risdiplam. Apnea hypopnea index (AHI) ranged from 3.6 to 24.1/h. REM AHI was higher than NREM AHI. Median Children's Hospital of Philadelphia Infant test of neuromuscular disorders (CHOP-Intend) score at the time of PSG was 55 (range 33-64). There was no correlation between age at treatment, CHOP-Intend score and AHI.SDB is common in treated children with SMA, regardless of age at diagnosis, treatment and neuromotor scores. While AHI may not be the only indicator of SDB in this population, indications, timing of PSG in this cohort remain unknown.

Resume Objectifs Le diagnostic preimplantatoire (DPI) consiste en la recherche d’une anomalie genetique sur une ou plusieurs cellules issues d’une biopsie d’un embryon, lui-meme obtenu par fecondation in vitro. Il existe un manque certain standardisation mondiale concernant les pratiques d’un pays, voire meme d’un centre a l’autre. Dans le notre, les biopsies sont realisees preferentiellement sur des embryons clives a j3, mais aussi au stade de blastocyste a j5. La biopsie j5 demande une congelation quasi systematique de l’embryon avant l’obtention du diagnostic contrairement a la biopsie j3 qui permet un transfert intra-uterin de l’embryon sain/equilibre des reception des resultats genetiques. Nous voulions comparer les chances de reussite pour les couples pris en charge en DPI dans notre centre selon le jour de la biopsie embryonnaire. Methode Pour cela, nous avons realise une etude retrospective monocentrique incluant tous les cycles de DPI realises entre 2016 et 2019 repartis en deux groupes, biopsies j3 ou j5. Resultats Il n’en ressort aucune difference significative sur le taux de naissance vivante (p = 0,7375) concernant les transferts frais embryonnaires. Il en est de meme pour les criteres secondaires a savoir les taux de grossesse debutante, de grossesse evolutive, d’implantation et de fausse-couche. Au contraire, lorsqu’il s’agit des transferts d’embryons congeles, nous retrouvons une difference significative sur les taux de naissance vivante (p = 0,0001) comme sur tous les autres taux en faveur des biopsies j5. Nous voulions egalement evaluer quelle etait la strategie de biopsie la plus efficiente dans notre laboratoire. Nos taux d’embryons utiles sont similaires quel que soit le jour de la biopsie (34 % en j3 et 37,7 % en j5, p = 0,244). Aucune difference statistique n’est retrouvee sur le nombre d’embryons biopsies inutilement dans les deux groupes. Par contre, le pourcentage d’embryons biopsies a j5 et immediatement congeles s’eleve a 42,8 %, soit 118 embryons, alors qu’aucun embryon biopsie a j3 n’entraine ce cas de figure. Conclusion Nos resultats representent donc des arguments plutot favorables a une generalisation de la biopsie j5 comme la norme internationale. Cependant, les implications organisationnelles, financieres et logistiques qu’imposerait cette technique la rendent non systematique dans notre centre.

PURPOSE: The improvement of clinical outcome provided by time-lapse technology (TLT) in IVF over conventional incubation (CI) still remains controversial. This study aimed at evaluating whether the exclusive use of time-lapse technology (TLT) during whole IVF care improves total cumulative live birth rate (TCLBR) and shortens time to live birth (TTLB) as compared to the use of CI in couples undergoing ICSI. METHODS: This retrospective cohort study was conducted in couples with male infertility undergoing their first ICSI cycle in 2014–2015 and for whom embryo culture system remained the same during their whole IVF care, i.e., TLT or CI. Couples were followed up up to 2020, including all following frozen-embryo transfers and ICSI cycles (if any). Survival analysis was used to compare clinical outcome and time-related endpoints between both groups. RESULTS: A total of 151 and 250 couples underwent their whole IVF care with the exclusive use of TLT and CI, respectively. Survival analysis showed that TCLBR after whole IVF care was significantly higher in TLT than in CI group (66.9 vs 56.4%, p=0.02, log-rank test). Median live birth time was significantly shorter in TLT than CI group (464 vs 596 days, p=0.01). CONCLUSIONS: We found that TCLBR and TTLB were significantly improved with TLT over CI in couples undergoing ICSI for male factor. This study fuels the debate on the clinical benefit of using TLT. The use of time-related endpoints adds important information for both patients and practitioners.

Day 5 fresh blastocyst transfers results in higher clinical pregnancy and live birth rates than day 6 fresh blastocyst transfer. This study aimed to identify the strategy to adopt with slowly developing blastocysts. Should not fully expanded blastocyst on day 5 be transferred on day 5, or when expanded on day 6, or be frozen? 1093 single blastocyst transfer cycles performed between January 2016 and December 2018 were divided in 4 groups: day 5 fresh transfers of full or expanded blastocyst (≥B3), day 5 fresh transfers of slowly developing blastocysts (B1 or B2), day 6 fresh transfers of expanded blastocysts (≥B4), day 6 frozen-thawed single blastocyst transfer cycles. Clinical pregnancy rate and live birth rate were significantly higher with fresh expanded blastocyst transfer on day 5 than in any other group. No statistical difference could be found between the other 3 groups. Slowly developing day 5 blastocysts have poorer implantation potential than expanded day 5 blastocysts but can be fresh transferred on day 5 rather than being cultured until day 6 for transfer or freezing when no expanded blastocyst is available on day 5.

Study question Does a serum progesterone level higher than 1.3 ng/mL on the day of ovulation trigger have an impact on blastocyst development? Summary answer Elevated progesterone level has no significant impact on top blastocyst rate, usable blastocyst rate and on morphokinetics. What is known already Premature elevation of progesterone level on the day of ovulation trigger prior to IVF is common and causes a decrease in endometrial receptivity. A freeze all strategy is then recommended. However, cumulative live birth rates have also been described as lower in cases of high progesterone levels. Study design, size, duration This was a retrospective bicentric cohort follow-up study, including 1150 IVF/ICSI cycles performed between 2016 and 2018 with at least 1 day–5 blastocyst available for transfer or freezing. Among these cycles, 524 were performed with use of a time-lapse system (Embryoscope). Serum Progesterone level was measured on the day of ovulation trigger, and a value >1.3 ng/ml was used to identify premature progesterone elevation. Participants/materials, setting, methods The cycles were divided into 2 groups according to serum progesterone level: 1335 cycles were allocated in the normal progesterone group (P 1.3). Patient’s characteristics, ovarian stimulation characteristics, IVF cycles characteristics and embryology parameters were anonymously recorded and compared between the 2 groups. Main results and the role of chance Female age, smoking status, AFC and AMH levels were comparable between the 2 groups. Female BMI was significantly higher in the P 1.3 group (26.1 versus 24.7 kg/m² respectively). Total FSH dose, estradiol level, number of follicles >11mm and number of retrieved oocytes were significantly higher in the P > 1.3 group than in P 1.3, p = 0.04). Cumulative live birth rate per cycle was not statistically different between the two groups (23.1% for P 1.3) (p > 0.05). Limitations, reasons for caution The retrospective design of the study should lead to careful analysis of the results. The progesterone threshold refers to a specific assay, and should not be generalized to other assays. Wider implications of the findings: Premature elevation of serum progesterone level on the day of ovulation trigger does not seem to affect embryo developmental competence. This further supports the relevance of freeze all strategy in this situation. Trial registration number Not applicable

The purpose of our study was to use a time-lapse monitoring (TLM) system to determine if day 3 blastomere biopsy for preimplantation genetic testing (PGT) had an impact on subsequent morphokinetic parameters at the morula and blastocyst stages. In this retrospective monocentric study conducted between May 2013 and August 2017, we compared late morphokinetic parameters in embryos undergoing day 3 blastomere biopsy for PGT and in control non-biopsied embryos obtained in intracytoplasmic sperm injection (ICSI) cycles for male infertility. All embryos in both groups were cultured in a TLM system. The biopsy group was composed of 1691 embryos (386 PGT cycles). The control group was composed of 2578 embryos (786 ICSI cycles). Early morphokinetic parameters up to day 3 were similar in both groups. Concerning late morphokinetic parameters, the onset of compaction (tSC), fully-compacted morula stage (tM), onset of cavitation/early blastulation (tSB), and blastocyst stages (tB and tEB) appeared significantly earlier in the biopsy group than in the control group. We found that late morphokinetic events at the morula and the blastocyst stages occurred significantly earlier in biopsied embryos than in control non-biopsied-embryos. The mechanisms underlying these modifications of embryo development after biopsy should be investigated in order to determine precisely, and this phenomenon could be associated with embryo, fetal, and offspring development.

Embryo morphology assessment performs relatively poorly in predicting implantation. Embryo aneuploidy screening (PGS) has recently improved, but its clinical value is still debated, and the development of a cheap non-invasive method for the assessment of embryo ploidy status is a highly desirable goal. The growing implementation of time-lapse devices led some teams to test the effectiveness of morphokinetic parameters as predictors of embryo ploidy, with conflicting results. The aim of this study was to conduct a comprehensive review of the literature on the predictive value of morphokinetic parameters for embryo ploidy status. A systematic search on PubMed was conducted using the following key words: time-lapse, morphokinetic, aneuploidy, IVF, preimplantation genetic screening, PGS, chromosomal status. A total of 13 studies were included in the analysis. They were heterogeneous in design, patients, day of embryo biopsy, statistical approach and outcome measures. No single or combined morphokinetic parameter was consistently identified as predictive of embryo ploidy status. In conclusion, the available studies are too heterogeneous for firm conclusions to be drawn on the predictive value of time-lapse analysis for embryo aneuploidy screening. Hence, morphokinetic parameters should not be used yet as a surrogate for PGS to determine embryo ploidy in vitro.

Purpose The exploration of male infertility is mainly based on semen analysis, but its evaluation might be affected by the operator's competence and subjectivity. This led to the development of automated semen analyzing systems. Despite continuous improvement, the precision and correlation of these automated systems with manual sperm assessment performed strictly according to WHO guidelines remains variable in the literature, and their role in daily practice is debated. Methods In this double blind prospective study, we compared the results provided by 2 automated systems based on different concepts (CASA and electro-optical signal) with manual sperm assessment. Sperm concentration, motility and morphology were performed simultaneously and independently by different operators, blinded to each other. Results A total of 102 unselected men attending the andrology department for routine sperm analysis were included in the study. We found no significant difference between each automated method and manual assessment for all sperm parameters, except for sperm morphology assessment where the electro-optical system gave higher results and performed slightly poorer than CASA. Correlation was moderate to high between manual assessment and each automated methods for all sperm parameters, with randomly distributed differences. Conclusions Overall, these results show that both types of automated systems can be implemented in andrology laboratory for routine sperm analysis.

Understanding lineage specification during human pre-implantation development is a gateway to improving assisted reproductive technologies and stem cell research. Here we employ pseudotime analysis of single-cell RNA sequencing (scRNA-seq) data to reconstruct early mouse and human embryo development. Using time-lapse imaging of annotated embryos, we provide an integrated, ordered, and continuous analysis of transcriptomics changes throughout human development. We reveal that human trophectoderm/inner cell mass transcriptomes diverge at the transition from the B2 to the B3 blastocyst stage, just before blastocyst expansion. We explore the dynamics of the fate markers IFI16 and GATA4 and show that they gradually become mutually exclusive upon establishment of epiblast and primitive endoderm fates, respectively. We also provide evidence that NR2F2 marks trophectoderm maturation, initiating from the polar side, and subsequently spreads to all cells after implantation. Our study pinpoints the precise timing of lineage specification events in the human embryo and identifies transcriptomics hallmarks and cell fate markers.

STUDY QUESTION Is it possible to develop an automated annotation tool for human embryo development in time-lapse devices based on image analysis? SUMMARY ANSWER We developed and validated an automated software for the annotation of human embryo morphokinetic parameters, having a good concordance with expert manual annotation on 701 time-lapse videos. WHAT IS KNOWN ALREADY Morphokinetic parameters obtained with time-lapse devices are increasingly used for the assessment of human embryo quality. However, their annotation is time-consuming and can be slightly operator-dependent, highlighting the need to develop fully automated approaches. STUDY DESIGN, SIZE, DURATION This monocentric study was conducted on 701 videos originating from 584 couples undergoing IVF with embryo culture in a time-lapse device. The only selection criterion was that the duration of the video must be over 60 h. PARTICIPANTS/MATERIALS, SETTING, METHODS An automated morphokinetic annotation tool was developed based on gray level coefficient of variation and detection of the thickness of the zona pellucida. The detection of cellular events obtained with the automated tool was compared with those obtained manually by trained experts in clinical settings. MAIN RESULTS AND THE ROLE OF CHANCE Although some differences were found when embryos were considered individually, we found an overall concordance between automated and manual annotation of human embryo morphokinetics from fertilization to expanded blastocyst stage (r2 = 0.92). LIMITATIONS, REASONS FOR CAUTION These results should undergo multicentric external evaluation in order to test the overall performance of the annotation tool. Getting access to the export of 3D videos would enhance the quality of the correlation with the same algorithm and its extension to the 3D regions of interest. A technical limitation of our work lies within the duration of the video. The more embryo stages the video contains, the more information the script has to identify them correctly. WIDER IMPLICATIONS OF THE FINDINGS Our system paves the way for high-throughput analysis of multicentric morphokinetic databases, providing new insights into the clinical value of morphokinetics as a predictor of embryo quality and implantation. STUDY FUNDING/COMPETING INTEREST(S) This study was partly funded by Finox-Gedeon Richter Forward Grant 2016 and NeXT (ANR-16-IDEX-0007). We have no conflict of interests to declare. TRIAL REGISTRATION NUMBER N/A

Recent technological advances such as single-cell RNAseq1-3and CRISPR-CAS9-mediated knock-out4have allowed an unprecedented access into processes orchestrating human preimplantation development5. However, the sequence of events which occur during human preimplantation development are still unknown. In particular, timing of first human lineage specification, the process by which the morula cells acquire a specific fate, remains elusive. Here, we present a human preimplantation development model based on transcriptomic pseudotime modelling of scRNAseq biologically validated by spatial information and precise time-lapse staging. In contrast to mouse, we show that trophectoderm (TE) / inner cell mass (ICM) lineage specification in human is only detectable at the transcriptomic level at the blastocyst stage, just prior to expansion. We validated the expression profile of novel markers enabling precise staging of human preimplantation embryos, such as IFI16 which highlights establishment of epiblast (EPI) and NR2F2 which appears at the transition from specified to mature TE. Strikingly, mature TE cells arise from the polar side, just after specification, supporting a model of polar TE cells driving TE maturation. Altogether, our study unravels the first lineage specification event in the human embryo and provides a browsable resource for mapping spatio-temporal events underlying human lineage specification.

Recent technological advances such as single-cell (sc) RNAseq and CRISPR-CAS9-mediated knock-out have allowed an unprecedented access to processes orchestrating human preimplantation development. However, the sequence of events which occur during human preimplantation development are still unknown. In particular, the timing of the first human lineage specification, the process by which the morula cells acquire a specific fate, remains elusive. Here, we present a mouse and human preimplantation development model based on transcriptomic pseudotime modelling of scRNAseq biologically validated by spatial information and precise time-lapse staging. We show that in human, trophectoderm (TE) and inner cell mass (ICM) specification in not complete until the B3 blastocyst stage. This reveals a delay with the mouse first specification. We identified novel markers enabling precise staging of human preimplantation embryos: IFI16, associated to the developed epiblast (EPI) and NR2F2, associated with TE development. Strikingly, NR2F2 positive TE cells arise from the polar side, shortly after specification, supporting a model of faster polar TE development leading to polar implantation in the endometrium. Altogether, our study unravels preimplantation lineage specification in human embryos and provides a browsable resource for mapping spatio-temporal events orchestrating human preimplantation development.

PURPOSE: While several studies reported the association between morphokinetic parameters and implantation, few predictive models were developed to predict implantation after day 5 embryo transfer, generally without external validation. The objective of this study was to evaluate the respective performance of 2 commercially available morphokinetic-based models (KIDScore™ Day 5 versions 1 and 2) for the prediction of implantation and live birth after day 5 single blastocyst transfer. METHODS: This monocentric retrospective study was conducted on 210 ICSI cycles with single day 5 embryo transfer performed with a time-lapse imaging (TLI) system between 2013 and 2016. The association between both KIDScore™ and the observed implantation and live birth rates was calculated, as well as the agreement between embryologist’s choice for transfer and embryo ranking by the models. RESULTS: Implantation and live birth rate were both 35.7%. A significant positive correlation was found between both models and implantation rate (r = 0.96 and r = 0.90, p = 0.01) respectively. Both models had statistically significant but limited predictive power for implantation (AUC 0.60). There was a fair agreement between the embryologists’ choice and both models (78% and 61% respectively), with minor differences in case of discrepancies. CONCLUSIONS: KIDScore™ Day 5 predictive models are significantly associated with implantation rates after day 5 single blastocyst transfer. However, their predictive performance remains perfectible. The use of these predictive models holds promises as decision-making tools to help the embryologist select the best embryo, ultimately facilitating the implementation of SET policy. However, embryologists’ expertise remains absolutely necessary to make the final decision. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10815-019-01567-x) contains supplementary material, which is available to authorized users.

International audience; Purpose The purpose of our study was to use time-lapse in order to evaluate the impact of sperm origin (fresh ejaculate or surgically retrieved) on embryo morphokinetic parameters and clinical outcome in intracytoplasmic sperm injection (ICSI) cycles.Methods This retrospective monocentric study was conducted in 485 unselected couples undergoing 604 ICSI cycles with embryo culture in the Embryoscope®. Among them, 445 couples underwent ICSI cycle with fresh ejaculated sperm and 40 with surgically retrieved sperm (26 with testicular sperm and 14 with epididymal sperm). Embryo morphokinetic parameters and clinical cycle outcome were compared between fresh ejaculated sperm and surgically retrieved sperm. A subgroup analysis was also conducted between testicular and epididymal sperm ICSI cycles.Results Clinical outcome was comparable between groups according to sperm origin. Although most early morphokinetic parameters were comparable between ejaculated and surgical sperm groups, a few parameters were significantly different between both groups, but with a considerable overlap in their distribution. Late cellular events occurred significantly later in the surgical sperm group than in the ejaculated sperm group. Conclusions Morphokinetic analysis did not allow us to identify clinically relevant differences between fresh ejaculate and surgically retrieved sperm groups. Further studies are needed, especially concerning the relationship between sperm origin and late morphokinetic parameters, such as blastocyst development.

International audience; RESEARCH QUESTION: Can embryo morphokinetic parameters help identify unbalanced embryos in translocation carriers?DESIGN: This retrospective study was conducted in 67 translocation carriers undergoing 105 preimplantation genetic testing cycles for chromosomal structural rearrangements (PGT-SR) without aneuploidy screening (PGT-A). Using time-lapse imaging analysis, morphokinetic parameters of balanced and unbalanced embryos were compared, as well as the frequency of abnormal cellular events. The performance of a previously published prediction model of aneuploidy was also tested in this population.RESULTS: Significant differences were observed between balanced and unbalanced embryos for some morphokinetic parameters: t5 (P = 0.0067), t9+ (P = 0.0077), cc2 (P = 0.0144), s2 (P = 0.0003) and t5-t2 (P = 0.0028). Also, multinucleation at the two- or four-cell stages, abnormal division and cell exclusion at the morula stage were significantly (all P < 0.05) more frequent in unbalanced than in balanced embryos. None, however, could accurately predict embryo chromosomal status. A previously published morphokinetic prediction model for embryo aneuploidy did not adequately classify balanced and unbalanced embryos.CONCLUSIONS: No significant morphokinetic predictor of chromosomal status could be found. Time-lapse should not be used as a diagnostic tool for chromosomal status in translocation carriers.

Despite controversy regarding its clinical value, male fertility investigation mainly relies on semen analysis. Even though reference guidelines are available, manual sperm analysis still suffers from analytical variability, thus questioning the interest of automated sperm analysis systems. The aim of this study is to compared automated computerized semen analysis systems (SQA-V GOLD and CASA CEROS) to the conventional manual method in terms of accuracy and precision.We included 250 men in this double-blind prospective study. The SQA-V GOLD (Medical Electronic Systems) and CEROS, CASA system (Hamilton Thorne) were compared to the standard manual assessment based on the WHO 5th Edition. The main outcome measures were sperm concentration, total sperm number, total motility, progressive motility, non-progressive motility, morphology, motile sperm concentration (MSC) and progressively motile sperm concentration (PMSC) with the three methods.Statistical analysis of the test results from the automated systems and the manual method demonstrated no significant differences for most of the semen parameters. The Spearman coefficients of rank correlation (rho) for CASA and the SQA-V GOLD automated systems vs. the manual method were: Sperm concentration (0.95 and 0.95), total sperm number (0.95 and 0.95), MSC (0.94 and 0.96) and PMSC (0.94 and 0.93) correspondingly. Concerning sperm morphology, both automated systems demonstrated high specificity (Sp) and negative predictive values (NPV), despite significantly different medians (CASA: 83.7 % for Sp and 95.2 % for NPV, SQA-V: 97.9 % for Sp and 92.5 %). The highest precision (lowest 95 % confidence interval for duplicate tests) for all semen variables was found in the SQA-V GOLD.The advantages of using automated semen analysers are: Standardization, speed (lower turnaround time), precision, reduced potential for human error, automated data recording and less need for highly skilled professionals to run the systems. The disadvantages of using automated systems are: notably the problem with testing some atypical samples and the inability to perform an assessment of morphology abnormalities. Based on the results of this study, the SQA-V Gold demonstrated better agreement vs. the manual method. In conclusion, automated semen analyzers can be used for routine semen analysis providing rapid clinically acceptable results with higher precision, and positively impacting laboratory standardization.

International audience; This study aimed to describe spinal cord injured patients' semen characteristics before and after cryopreservation, and assisted reproductive technology cycles outcome compared to the infertile population. Data about sperm analysis and assisted reproductive technology (ART) cycles outcomes for 78 men with spinal cord injury referred for sperm cryopreservation between 1998 and 2013 were retrospectively analysed and compared with a reference group consisting of every Intra Cytoplasmic Sperm Injection (ICSI) cycle performed in our in vitro fertilization unit over the 2009-2014 period. Semen was collected by penile vibratory stimulation (PVS), electroejaculation or via testis biopsies. Fresh and frozen semen parameters and ART outcomes with frozen-thawed sperm were measured. Patients were divided into three groups according to the sperm retrieval methods: 37 electroejaculations, 37 PVSs and four surgical sperm retrievals. Low ejaculate volume was observed in 33% of the patients, and oligozoospermia in 37% of the patients. Specimens from 77 of the patients contained motile sperm and were therefore frozen for future use. There was no statistical difference for any of the fresh semen parameters between all groups. Twenty of them underwent ICSI leading to five live births, while four underwent intrauterine insemination. ART with frozen-thawed spermatozoa can provide men with SCI with comparable results as in the infertile population.

Among all the strategies available in order to improve success rates in IVF cycles, a lot of work has been done on embryo culture conditions and embryo quality evaluation. Most IVF centres use conventional incubators and select embryo according to punctual morphological evaluation, but this strategy has several limitations. Recently developed commercial devices associating more stable culture conditions and time lapse observation of embryo development provide new insights into early embryo development in IVF cycles. Among them, the Embryoscope(®) appears to be the most user-friendly, performing and suited for routine daily practice. The first Embryoscope(®) for France was installed in the University Hospital of Nantes in 2011. In our experience, it takes relatively a short time to get used to this system. Moreover, its integration in routine process yielded several advantages, such as better embryo selection according to kinetic parameters and observation of abnormal cleavage events, continuing education and training, quality control and flexibility. This leaded to an overall increase in success rates in IVF cycles.

Time-lapse analysis of tripronucleated zygotes obtained in ICSI cycles showed that 75.4% cleaved into embryos. These embryos subsequently demonstrated slower developmental kinetics than normally fertilized embryos.

Objective To evaluate the impact of female smoking on early embryo development in vitro using a time-lapse system. Design Retrospective analysis of prospectively collected database. Setting Assisted reproductive technology (ART) program. Patient(s) A total of 135 infertile couples undergoing IVF-intracytoplasmic sperm injection (ICSI) cycle. Intervention(s) Embryo culture in a time-lapse incubator (Embryoscope, Unisense Fertilitech). Main Outcome Measure(s) Embryo morphokinetic parameters. Result(s) Of 135 women, 139 and 729 oocytes were cultured in 23 smokers and 112 nonsmokers, respectively. Basal characteristics and ovarian stimulation parameters were comparable in both groups. Serum antimullerian hormone level and antral follicle count were significantly lower in smokers than in nonsmokers. Conventional embryologic parameters were comparable in both groups, but time-lapse analysis showed that most cleavage events occurred significantly later in smokers than in nonsmokers, leading to poor cycle outcome in smokers. Conclusion(s) Female smoking impairs early embryo development, as reflected by time-lapse analysis. This might contribute to the dramatic decrease in pregnancy rates (PRs) in smokers undergoing IVF. Time lapse is a useful tool for the identification of deleterious components impairing early embryo development.

One week after fertilization, human embryos implant into the uterus. This event requires the embryo to form a blastocyst consisting of a sphere encircling a cavity lodging the embryo proper. Stem cells can form a blastocyst model that we called a blastoid1. Here we show that naive human pluripotent stem cells cultured in PXGL medium2 and triply inhibited for the Hippo, TGF-β and ERK pathways efficiently (with more than 70% efficiency) form blastoids generating blastocyst-stage analogues of the three founding lineages (more than 97% trophectoderm, epiblast and primitive endoderm) according to the sequence and timing of blastocyst development. Blastoids spontaneously form the first axis, and we observe that the epiblast induces the local maturation of the polar trophectoderm, thereby endowing blastoids with the capacity to directionally attach to hormonally stimulated endometrial cells, as during implantation. Thus, we propose that such a human blastoid is a faithful, scalable and ethical model for investigating human implantation and development3,4., Blastoids derived from naive PXGL-cultured human pluripotent stem cells in which Hippo, TGF-β and ERK pathways are inhibited closely recapitulate aspects of blastocyst development, form cells resembling blastocyst-stage cells and thus provide a model system for implantation and development studies.