Mounting evidence that oocyte quality profoundly affects fertilisation an d subsequent embryo development drives the continued search for reliable predictors of oocyte developmental competence. In the present review, we provide an overall summary and analysis of potential criteria that can be used to evaluate oocyte quality. These criteria are specifically classified as morphological and cellular/molecular predictors. Traditional methods for the evaluation of oocyte quality are based on morphological classification of thefollicle, cumulus-oocytecomplex, polar body and/or meiotic spindle. Although the use of morphological characteristics as predictors of oocyte quality is controversial, such a grading system can provide valuable information for the preselection of oocytes with higher developmental competence and, therefore, may maximise embryo developmental outcome. Several intrinsic markers (such as mitochondrial status and glucose-6-phosphate dehydrogenase 1 activity) and extrinsic markers (such as apoptosis of follicular cells and levels of the transforming growth factor-beta superfamily in follicular fluid or serum) have also been reported as useful indicators of oocyte competence and embryo quality. Compared with the morphological parameters, these cellular and molecular predictors of oocyte quality may prove to be more precise and objective, although further studies and refinement of techniques are needed.

Evaluation of oocyte quality: morphological, cellular and molecular predictors.

BACKGROUND Non-invasive selection of developmentally competent human oocytes may increase the overall efficiency of human assisted reproduction and is regarded as crucial in countries where legal, social or religious factors restrict the production of supernumerary embryos. The purpose of this study was to summarize the predictive value for IVF success of morphological features of the oocyte that can be obtained by light or polarized microscopic investigations. METHODS Studies about oocyte morphology and IVF/ICSI outcomes were identified by using a systematic literature search. RESULTS Fifty relevant articles were identified: 33 analysed a single feature, 9 observed multiple features and investigated the effect of these features individually, 8 summarized the effect of individual features. Investigated structures were the following: meiotic spindle (15 papers), zona pellucida (15 papers), vacuoles or refractile bodies (14 papers), polar body shape (12 papers), oocyte shape (10 papers), dark cytoplasm or diffuse granulation (12 papers), perivitelline space (11 papers), central cytoplasmic granulation (8 papers), cumulus-oocyte complex (6 papers) and cytoplasm viscosity and membrane resistance characteristics (2 papers). None of these features were unanimously evaluated to have prognostic value for further developmental competence of oocytes. CONCLUSIONS No clear tendency in recent publications to a general increase in predictive value of morphological features was found. These contradicting data underline the importance of more intensive and coordinated research to reach a consensus and fully exploit the predictive potential of morphological examination of human oocytes.

Predictive value of oocyte morphology in human IVF: a systematic review of the literature

Visual morphology assessment is routinely used for evaluating of embryo quality and selecting human blastocysts for transfer after in vitro fertilization (IVF). However, the assessment produces different results between embryologists and as a result, the success rate of IVF remains low. To overcome uncertainties in embryo quality, multiple embryos are often implanted resulting in undesired multiple pregnancies and complications. Unlike in other imaging fields, human embryology and IVF have not yet leveraged artificial intelligence (AI) for unbiased, automated embryo assessment. We postulated that an AI approach trained on thousands of embryos can reliably predict embryo quality without human intervention. We implemented an AI approach based on deep neural networks (DNNs) to select highest quality embryos using a large collection of human embryo time-lapse images (about 50,000 images) from a high-volume fertility center in the United States. We developed a framework (STORK) based on Google’s Inception model. STORK predicts blastocyst quality with an AUC of >0.98 and generalizes well to images from other clinics outside the US and outperforms individual embryologists. Using clinical data for 2182 embryos, we created a decision tree to integrate embryo quality and patient age to identify scenarios associated with pregnancy likelihood. Our analysis shows that the chance of pregnancy based on individual embryos varies from 13.8% (age ≥41 and poor-quality) to 66.3% (age <37 and good-quality) depending on automated blastocyst quality assessment and patient age. In conclusion, our AI-driven approach provides a reproducible way to assess embryo quality and uncovers new, potentially personalized strategies to select embryos.

/pdf/deep-learning-enables-robust-assessment-and-selection-of-zky4c3ge3q.pdf

Deep learning enables robust assessment and selection of human blastocysts after in vitro fertilization.

The cryopreservation of spermatozoa was introduced in the 1960s as a route to fertility preservation. Despite the extensive progress that has been made in this field, the biological and biochemical mechanisms involved in cryopreservation have not been thoroughly elucidated to date. Various factors during the freezing process, including sudden temperature changes, ice formation and osmotic stress, have been proposed as reasons for poor sperm quality post-thaw. Little is known regarding the new aspects of sperm cryobiology, such as epigenetic and proteomic modulation of sperm and trans-generational effects of sperm freezing. This article reviews recent reports on molecular and cellular modifications of spermatozoa during cryopreservation in order to collate the existing understanding in this field. The aim is to discuss current freezing techniques and novel strategies that have been developed for sperm protection against cryo-damage, as well as evaluating the probable effects of sperm freezing on offspring health.

Sperm cryopreservation: A review on current molecular cryobiology and advanced approaches.

STUDY QUESTION
Can a deep learning model predict the probability of pregnancy with fetal heart (FH) from time-lapse videos?

/pdf/deep-learning-as-a-predictive-tool-for-fetal-heart-pregnancy-4lfpu3yke3.pdf

Deep learning as a predictive tool for fetal heart pregnancy following time-lapse incubation and blastocyst transfer

BACKGROUND To examine the relationship between the meiotic spindle, the first cleavage plane and any resulting influence on embryonic development parameters. METHODS Sibling oocytes (n = 246) were allocated to either a control [polar body (PB)-aligned] or a treatment (spindle-aligned) microinjection group by use of a random numbers table. Spindles were identified by PolScope((R)) and the early embryo development parameters, and angle of first cleavage plane in relation to a defined animal-vegetal pole were analysed. RESULTS Most oocytes (92.7%) had a visible spindle at the time of microinjection; however, 62.6% of first PBs (1PBs) were not located above the spindle (average deviation 37.3 +/- 33.2 degrees; range 0-176.6), with 6.9% of 1PBs in the opposite hemisphere to the spindle. The second PBs (2PBs) can also have an unpredictable deviation from the position of the meiotic spindle (12.5 +/- 16.7 degrees; range 0-91.8). This increased when the 1PB was above the spindle, forming a physical barrier to extrusion (average 24.7 +/- 16.1 degrees; range 7.9-91.8). Embryos developing from the spindle-aligned microinjection group had significantly more blastomeres per embryo (P = 0.044), a higher morphology score per embryo (P = 0.008) and a significantly higher average embryo score parameter (P = 0.003), with more embryos developing without any detectable fragmentation (P < 0.05) than the PB-aligned control group. Non-fragmented embryos undergo meridional cleavage, with a small angle between the spindle location and first cleavage plane (16.4 +/- 14.0 degrees ) compared with embryos with some degree of fragmentation (P = 0.002). This angle increased with the degree of fragmentation, with worst quality embryos having a spindle:cleavage angle of 45.1 +/- 17.7 degrees. CONCLUSIONS The 1PB and, to a lesser degree, the 2PB can be unreliable predictors of the exact meiotic spindle location in human oocytes. Embryos from spindle-aligned oocytes have an increase in all measured development parameters over control siblings. When the animal pole is defined as the meiotic spindle location, non-fragmented embryos tend to develop from a meridional cleavage; with the most fragmented embryos developing from a more equatorial initial cleavage plane. This study proposes that the spindle accurately marks the animal pole in human oocytes, and provides evidence linking the meiotic spindle location to the first cleavage plane and resulting early embryo development parameters in human embryos.

Meiotic spindle location and identification and its effect on embryonic cleavage plane and early development

STUDY QUESTION What is the inter-observer and intra-observer agreement between embryologists when selecting a single Day 5 embryo for transfer? SUMMARY ANSWER The inter-observer and intra-observer agreement between embryologists when selecting a single Day 5 embryo for transfer was generally good, although not optimal, even among experienced embryologists. WHAT IS KNOWN ALREADY Previous research on the morphological assessment of early stage (two pronuclei to Day 3) embryos has shown varying levels of inter-observer and intra-observer agreement. However, single blastocyst transfer is now becoming increasingly popular and there are no published data that assess inter-observer and intra-observer agreement when selecting a single embryo for Day 5 transfer. STUDY DESIGN, SIZE, DURATION This was a prospective study involving 10 embryologists working at five different IVF clinics within a single organization between July 2013 and November 2015. PARTICIPANTS/MATERIALS, SETTING, METHODS The top 10 embryologists were selected based on their yearly Quality Assurance Program scores for blastocyst grading and were asked to morphologically grade all Day 5 embryos and choose a single embryo for transfer in a survey of 100 cases using 2D images. A total of 1000 decisions were therefore assessed. For each case, Day 5 images were shown, followed by a Day 3 and Day 5 image of the same embryo. Subgroup analyses were also performed based on the following characteristics of embryologists: the level of clinical embryology experience in the laboratory; amount of research experience; number of days per week spent grading embryos. The agreement between these embryologists and the one that scored the embryos on the actual day of transfer was also evaluated. Inter-observer and intra-observer variability was assessed using the kappa coefficient to evaluate the extent of agreement. MAIN RESULTS AND THE ROLE OF CHANCE This study showed that all 10 embryologists agreed on the embryo chosen for transfer in 50 out of 100 cases. In 93 out of 100 cases, at least 6 out of the 10 embryologists agreed. The inter-observer and intra-observer agreement among embryologists when selecting a single Day 5 embryo for transfer was generally good as assessed by the kappa scores (kappa = 0.734, 95% CI: 0.665-0.791 and 0.759, 95% CI: 0.622-0.833, respectively). The subgroup analyses did not substantially alter the inter-observer and intra-observer agreement among embryologists. The agreement when Day 3 images were included alongside Day 5 images of the same embryos resulted in a change of mind at least three times by each embryologist (on average for <10% of cases) and resulted in a small decrease in inter-observer and intra-observer agreement between embryologists (kappa = 0.676, 95% CI: 0.617-0.724 and 0.752, 95% CI: 0.656-808, respectively).The assessment of the inter-observer agreement with regard to morphological grading of Day 5 embryos showed only a fair-to-moderate agreement, which was observed across all subgroup analyses. The highest overall kappa coefficient was seen for the grading of the developmental stage of an embryo (0.513; 95% CI: 0.492-0.538). The findings were similar when the individual embryologists were compared with the embryologist who made the morphological assessments of the available embryos on the actual day of transfer. LIMITATIONS, REASONS FOR CAUTION All embryologists had already completed their training and were working under one organization with similar policies between the five clinics. Therefore, the inter-observer agreement might not be as high between embryologists working in clinics with different policies or with different levels of training. WIDER IMPLICATIONS OF THE FINDINGS The generally good, although not optimal uniformity between participating embryologists when selecting a Day 5 embryo for transfer, as well as, the surprisingly low agreement when morphologically grading Day 5 embryos could be improved, potentially resulting in increased pregnancy rates. Future studies need to be directed toward technologies that can help achieve this. STUDY FUNDING/COMPETING INTERESTS None declared. TRIAL REGISTRATION NUMBER Not applicable.

/pdf/inter-observer-and-intra-observer-agreement-between-f18yn67hec.pdf

Inter-observer and intra-observer agreement between embryologists during selection of a single Day 5 embryo for transfer: a multicenter study

Purpose: To assess the ability of various facets of embryo culture (microscope stage warmers, volumes of culture media, culture vessel lids, and type of culture incubator) to maintain a constant temperature in vitro.

Methods: Ability to maintain 37.0°C in the microenvironment of gametes was recorded by digital thermocouple in the chosen facets of in vitro culture.

Results: Stage warmers are highly variable in their ability to maintain the set temperature (range 33.8°C–37.0°C after 60 s). Temperature loss in culture media is both volume and vessel dependent, and the direct heat transfer culture incubator (MINC) has superior temperature maintenance compared with a large volume air convection incubator (FORMA), where temperature regain from 35.0°C to 37.0°C took 5.5 min compared to >20 min.

Conclusions: There are large measurable differences in the ability to maintain set temperature that depend on the stage warmer used, volume of media, use of vessel lids, and the type of incubator chosen for IVF culture.

Objective Assessments of Temperature Maintenance Using In Vitro Culture Techniques

https://www.fertstert.org/article/S0015-0282(02)04343-1/pdf

Simon Cooke

Papers

Deep learning as a predictive tool for fetal heart pregnancy following time-lapse incubation and blastocyst transfer

Meiotic spindle location and identification and its effect on embryonic cleavage plane and early development

Inter-observer and intra-observer agreement between embryologists during selection of a single Day 5 embryo for transfer: a multicenter study

Objective Assessments of Temperature Maintenance Using In Vitro Culture Techniques

Improvement in early human embryo development using new formulation sequential stage-specific culture media.