Assessment Of Embryo Viability Research Articles

Determination of pyruvate and lactate as potential biomarkers of embryo viability in assisted reproduction by capillary electrophoresis with contactless conductivity detection.

Human-assisted reproduction is increasing in importance due to the constantly rising number of couples suffering from infertility issue. A key step in in vitro fertilization is the proper assessment of embryo viability in order to select the embryo with the highest likelihood of resulting in a pregnancy. This study proposes a method based on CE with contactless conductivity detection for the determination of pyruvate and lactate in spent culture media used in human-assisted reproduction. A fused-silica capillary of 64.0 cm total length and 50 μm inner diameter was used. The inner capillary wall was modified by the coating of successive layers of the ionic polymers polybrene and dextran sulfate to reverse EOF. The BGE was composed of 10 mM MES/lithium hydroxide, pH 6.50. The sample was injected by pressure 50 mbar for 18 s, separation voltage was set to -24 kV, and capillary temperature to 15°C. The presented method requires only 2 μL of the culture medium, with LODs for pyruvate and lactate of 0.03 and 0.02 μM, respectively. The results demonstrated the method's suitability for the analysis of spent culture media to support embryo viability assessment by light microscopy, providing information about key metabolites of the energy metabolism of a developing embryo.

Noninvasive embryo viability assessment by quantitation of human haptoglobin alpha-1 fragment in the in vitro fertilization culture medium: an additional tool to increase success rate

To find new candidate molecules to assess embryo viability in a noninvasive manner. Prospective, blinded study with randomized sample collection. University research center. Ninety embryos implanted in 53 randomly selected patients (mean ± SD age, 32.3 ± 5.1years) were analyzed. Superovulation treatment was initiated by the administration of the GnRh agonist triptorelin and individual dosages of recombinant FSH. Ovulation was induced by the injection of hCG. Oocytes were fertilized by intracytoplasmic sperm injection. Liquid chromatography coupled mass spectrometric quantification of the α-1 fragment of human haptoglobin in the culture medium. A novel polypeptide marker was found that might be helpful to differentiate between potentially viable and nonviable embryos. This molecule was identified with tandem mass spectrometry as the α-1 fragment of human haptoglobin. Significant correlation was found in the amount of the peptide fragment and the outcome of pregnancy. In the culture media of embryos that were assigned in the biochemical assay as nonviable (according to the amount of the haptoglobin fragment), there were no pregnancies detected; this assay revealed a 100% successful selection of the nonviable embryos. In the group assigned as viable, the rate of pregnancy was 54.7%. Viability of the embryo during the IVF process is assessed by microscopic inspection, resulting in a pregnancy rate of 25%-30%. Detection and quantitation of the α-1 haptoglobin fragment of the culture medium proved to be a useful additional method for identifying nonviable embryos, increasing the success rate to 50%.

Time-lapse embryo imaging technology

The purpose of the review is to summarize recent developments in time-lapse technologies and early embryo morphokinetics and to discuss their impact on current clinical outcomes. Contemporary embryo culture and selection methodologies that are based on classical morphology are clearly limited in providing the most suitable embryo for a successful pregnancy. Noninvasive observation of embryo development by capturing the images with a time-lapse device has recently been proposed to be a better method of embryo viability assessment. Such methodologies have been shown to increase the quality and the quantity of information on the viability without disturbing the culture conditions. Commercial availability of different time-lapse devices for human embryos facilitated the use of morphokinetics as an additional tool in human embryo selection. The application of such technologies has already shown positive results on clinical outcome by increasing our scope of traditional embryo selection, leading to higher implantation and clinical pregnancy rates. Additional benefit can come from the less-disturbed incubation environment that is created by all-in-one incubators. Such devices can also be very important research tools in order to observe and analyze the effect of different patient-specific or clinical conditions on embryo development parameters that are not available through classical embryo scoring.

Metabolomic Assessment of Embryo Viability

Preimplantation embryo metabolism demonstrates distinctive characteristics associated with the developmental potential of embryos. On this basis, metabolite content of culture media was hypothesized to reflect the implantation potential of individual embryos. This hypothesis was tested in consecutive studies reporting a significant association between culture media metabolites and embryo development or clinical pregnancy. The need for a noninvasive, reliable, and rapid embryo assessment strategy promoted metabolomics studies in vitro fertilization (IVF) in an effort to increase success rates of single embryo transfers. With the advance of analytical techniques and bioinformatics, commercial instruments were developed to predict embryo viability using spectroscopic analysis of surplus culture media. However, despite the initial promising results from proof-of-principal studies, recent randomized controlled trials using commercial instruments failed to show a consistent benefit in improving pregnancy rates when metabolomics is used as an adjunct to morphology. At present, the application of metabolomics technology in clinical IVF laboratory requires the elimination of factors underlying inconsistent findings, when possible, and development of reliable predictive models accounting for all possible sources of bias throughout the embryo selection process.

Morphological Assessment of Embryo Viability

Morphological assessment is discussed in the context of significant literature at all stages of in vitro development, beginning with the oocyte and culminating at the blastocyst stage. Current evidence is used to debate the inclusion of commonly observed morphological features in grading schemes. The biological rationale behind observed phenomena such as multinucleation and fragmentation are also explored. Current limitations as well as technological advancements that increase our ability to assess viability are highlighted. Particular attention is paid to the relationship between developmental timing and assessment schemes. Failure to standardize assessment timing and inclusion criteria is glaring weaknesses of the literature that currently make consensus unattainable. Mounting evidence suggests that the future of static assessment is very likely to be influenced by information gathered from preimplantation genetic screening and other invasive techniques as well as from continuous monitoring tools such as time lapse.

High-throughput analysis of ovarian granulosa cell transcriptome.

The quality of follicular oocytes depends on interactions with surrounding granulosa cells. Development of molecular techniques and methods enables better understanding of processes underlying mammalian reproduction on cellular level. The success in reproductive biology and medicine in different species depends on reliable assessment of oocyte and embryo viability which presently mainly bases on embryo morphology. Although successful pregnancies have been achieved using this approach, its precision still should be improved and completed with other, more objective, and accurate assessment strategies. Global profiling of gene expression in follicular cumulus cells using microarrays is continuously leading to the establishment of new biomarkers which can be used to select oocytes with highest developmental potential. Even more potential applications and greater precision could be achieved using next generation sequencing (NGS) of granulosa and cumulus cell RNA (RNA-seq). However, due to the high cost, this method is not used as frequently as microarrays at the moment. In any case, high-throughput technologies offer the possibilities and advantages in ovarian somatic cell analysis on scale that has not been noted so far. The aim of this work is to present current directions and examples of global molecular profiling of granulosa cells and underline its impact on reproductive biology and medicine.

Ammonium Concentration of Spent Medium Provides a Noninvasive Assessment of Embryonic Developmental Potential in IVF

Currently, morphological criteria are used to select embryos for transfer in in vitro fertilization (IVF)-embryo transfer. However, the implantation rate is only about 30%, indicating a need for a more efficient method of selecting high-quality embryos. This study investigated the relationship between medium ammonium concentration and IVF implantation rates and evaluated the possibility of using ammonium concentration to provide an objective, noninvasive, and simple means of assessment of embryo viability. On day 3 after fertilization, we sampled the spent medium bathing embryos obtained from patients undergoing IVF or intracytoplasmic sperm injection (ICSI) at the Reproductive Medical Center of Sun Yat-sen Memorial Hospital, between September 2010 and January 2012. The concentration of ammonium was determined using a dry chemistry system. The ammonium concentration increased significantly during the period of culture of all embryos, but the increase was significantly greater in the group that, subsequently, did not achieve pregnancy (P < .001). An receiver-operating characteristic (ROC) curve indicated that ammonium concentration was highly predictive of embryo implantation failure (area under ROC curve = 0.838). Failure to achieve implantation had a predictive sensitivity of 71% and specificity of 79.6%. The accuracy of prediction of successful or failed implantation was 75.4%. In both IVF and ICSI, the embryo implantation rate decreased significantly as the medium ammonium concentration increased (P < .05). Measurement of the ammonium concentration in the spent medium may provide a new research direction for exploring a simple, rapid, and low-cost method for reliable prediction of embryo implantation with high sensitivity and specificity.

Whole-genome sequencing technology for the assessment of embryo genetics and viability

Whole-genome sequencing technology for the assessment of embryo genetics and viability

Embryo implantation

Implantation is an essential step in the development of a pregnancy, but often fails in humans. In assisted reproductive technologies, implantation failure continues to impair treatment outcomes, with distressing results for patients and physicians. Morphokinetics, comprehensive chromosome screening, and the analysis of embryo-derived products detectable in spent culture media offer new means of assessing embryo viability. However, all await validation in randomized controlled trials. Genomic, transcriptomic, and secretomic technologies are similarly being exploited to define specific biomarkers of endometrial receptivity with the aim of identifying novel therapeutic interventions. However, to date no single, clinically relevant molecular marker capable of indicating endometrial receptivity has been reported. Recent work continues to describe the key signalling pathways which result in acceptance or rejection of the implanting embryo. In-vitro studies have revealed that the decidualized endometrium plays an important role in natural embryo selection, which could change our understanding of the aetiology and treatment of reproductive failure. Recent developments in analytical techniques have initiated a search for biomarkers of embryo quality and endometrial receptivity, and in-vitro studies have revealed novel roles for the decidualized endometrium as a biosensor of embryo quality.

Oocyte insemination techniques are related to alterations of embryo developmental timing in an oocyte donation model

Because of the different intrinsic characteristics of the classic IVF and intracytoplasmic sperm injection (ICSI) techniques, the timing of zygote development can be influenced by the method of fertilization. However, there is no information about the relevance of the insemination procedure on embryo-quality parameters as measured through their developmental dynamics. The aim of this work was to determine if the insemination technique, IVF or ICSI, influences embryo developmental kinetics by examining 1203 embryos from 178 couples undergoing oocyte donation with IVF or ICSI. Using time-lapse information, this work calculated several developmental kinetic variables, from pronuclear fading (PNF) to expanded blastocyst, and also the proportion of optimal embryos in a best time range with a predicted higher implantation potential. Embryo development after ICSI was slightly faster than after IVF; however, when PNF, rather than time of insemination, was established as t0, the differences between the two procedures disappeared. The percentage of optimal embryos showed a trend towards higher values in IVF-derived embryos; however, the difference was not statistically significant. With these results and through the time-lapse monitoring system, it is concluded that it is the fertilization method which determines embryo developmental kinetics if insemination time is used as the starting point.A key step in assisted reproduction is the assessment of oocyte and embryo viability to determine the embryo(s) most likely to implant. Current embryo assessment strategies in clinical settings largely rely on embryo morphology and cleavage rates, and although these systems have been successful in improving pregnancy rates, their precision is far from ideal as they are based on visual information. In contrast, automated image analysis may add objectivity to the process of embryo selection and consequently, lead to an improvement in the implantation rates. Timing of zygote development can be influenced by the method of fertilization or by in-vitro culture conditions, so it has been suggested that the fertilization procedure influences the length of time elapsed between fertilization and the first cleavage. In this report, we show the results from using a time-lapse monitoring system to determine the timing of key events during embryo development both in IVF and ICSI. Due to the different intrinsic characteristics of the classic IVF and ICSI techniques, the selection of a critical time point is essential so as to maximize the differences between the two methods. For that reason, we searched for evidence in the data obtained from the image analysis for a link connecting embryo cleavage and the fertilization technique and also to find whether the kinetic of development derived from classic IVF or ICSI is also related to a predicted higher implantation.

Improving embryo selection using a computer-automated time-lapse image analysis test plus day 3 morphology: results from a prospective multicenter trial

To assess the first computer-automated platform for time-lapse image analysis and blastocyst prediction and to determine how the screening information may assist embryologists in day 3 (D3) embryo selection. Prospective, multicenter, cohort study. Five IVF clinics in the United States. One hundred sixty women ≥ 18 years of age undergoing fresh IVF treatment with basal antral follicle count ≥ 8, basal FSH <10 IU/mL, and ≥ 8 normally fertilized oocytes. A noninvasive test combining time-lapse image analysis with the cell-tracking software, Eeva (Early Embryo Viability Assessment), was used to measure early embryo development and generate usable blastocyst predictions by D3. Improvement in the ability of experienced embryologists to select which embryos are likely to develop to usable blastocysts using D3 morphology alone, compared with morphology plus Eeva. Experienced embryologists using Eeva in combination with D3 morphology significantly improved their ability to identify embryos that would reach the usable blastocyst stage (specificity for each of three embryologists using morphology vs. morphology plus Eeva: 59.7% vs. 86.3%, 41.9% vs. 84.0%, 79.5% vs. 86.6%). Adjunctive use of morphology plus Eeva improved embryo selection by enabling embryologists to better discriminate which embryos would be unlikely to develop to blastocyst and was particularly beneficial for improving selection among good-morphology embryos. Adjunctive use of morphology plus Eeva also reduced interindividual variability in embryo selection. Previous studies have shown improved implantation rates for blastocyst transfer compared with cleavage-stage transfer. Addition of Eeva to the current embryo grading process may improve the success rates of cleavage-stage ETs.

A method for semi-automatic grading of human blastocyst microscope images

The precise assessment of embryo viability is an extremely important factor for the optimization of IVF treatments. In order to assess embryo viability, several embryo scoring systems have been developed. However, they rely mostly on a subjective visual analysis of embryo morphological features and thus are subject to inter- and intra-observer variation. In this paper, we propose a method for image segmentation (the dividing of an image into its meaningful constituent regions) and classification of human blastocyst images with the aim of automating embryo grading. The delineation of the boundaries (segmentation) of the zona pellucida, trophectoderm (TE) and inner cell mass (ICM) were performed using advanced image analysis techniques (level set, phase congruency and fitting of ellipse methods). The fractal dimension and mean thickness of TE and ICM image texture descriptors (texture spectrum and grey-level run lengths) were calculated to characterize the main morphological features of the blastocyst with the aim of automatic grading using Support Vector Machine classifiers. The fractal dimension calculated from the delineated TE boundary provided a good indication of cell number (presented a 0.81 Pearson correlation coefficient with the number of cells), a feature closely associated with blastocyst quality. The classifiers showed different accuracy levels for each grade. They presented accuracy ranges from 0.67 to 0.92 for the embryo development classification, 0.67-0.82 for the ICM classification and 0.53-0.92 for the TE classification. The value 0.92 was the highest accuracy achieved in the tests with 73 blastocysts. Semi-automatic grading of human blastocysts by a computer is feasible and may offer a more precise comparison of embryos, reducing subjectivity and allowing embryos with apparently identical morphological scores to be distinguished.

An investigation into the relationship between the metabolic profile of follicular fluid, oocyte developmental potential, and implantation outcome

To determine whether metabolomic analysis of follicular fluid could prove a useful noninvasive technique for the selection of viable oocytes and embryos. Metabolomic analysis based on proton nuclear magnetic resonance ((1)H NMR) performed on follicular fluid collected from in vitro fertilization (IVF) patients. A university research center and a private fertility clinic. Fifty-eight women undergoing IVF treatment. Follicular fluid collected at the time of oocyte retrieval. Metabolomic profile, assessment of oocyte developmental potential and embryo viability. The metabolomic profile of follicular fluid from follicles where the oocyte resulted in a fertilized egg that failed to cleave (n = 9) was distinctly different from that where oocytes developed into early cleavage-stage embryos. Discriminating metabolites included glucose, lactate, choline/phosphocholine, and lipoproteins. Comparison of follicular fluid from women who subsequently had a positive β human chorionic gonadotropin (n = 10) to those who were unsuccessful in achieving a pregnancy (n = 12) revealed metabolic differences that were correlated to cycle outcome. Differences in the metabolite composition of follicular fluid correlate with the developmental competence of the human oocyte. Therefore, metabolomic profiling of follicular fluid may prove to be an important technique in gamete/embryo selection.

Development of a novel analytical approach combining the quantification of amino acids, organic acids and glucose using HPLC-UV-Vis and HPLC-MS with screening viaNMR

A simple, rapid, sensitive and selective procedure based on the combination of HPLC-UV-Vis and HPLC-MS has been developed and single laboratory partially validated for the determination of a set of 13 analytes present in a commercially available IVF medium utilising small sample volumes (20–30 mL). The composition fingerprint of the complex sample obtained by NMR spectroscopy in 11 minutes provided identification based on a screening of the metabolomic profile. HPLC-MS allowed the glucose–sodium adduct to be measured accurately and the working and linear ranges achieved were 0.028–0.389 mmol L � 1 with a detection limit of 13 mM. HPLC-UV-Vis allowed accurate concentrations of pyruvic and lactic acids with linear ranges over 0.005–0.1 mmol L � 1 with a limit of detection of 28 mM for pyruvic acid to be determined in 8 minutes, while lactic acid presented a linear range over 0.1–2 mmol L � 1 with a limit of detection of 1.2 mM possible. The use of HPLC-UV-Vis allowed the chromatographic separation of 8 amino acids (aspartate, glutamate, serine, glycine, asparagine, glutamine, alanine, and proline), the dipeptide alanyl-glutamine and taurine previous to a chemical derivatization, providing a total run time of 40 minutes. The method was partially validated to show a linear range of 0.028–0.280 mmol L � 1 with detection limits ranging between 1 and 30 mM. Development of the analytical approach provided determination and quantification of a set of 13 analytes from a very complex sample. Although well established analytical techniques were used here, combinatory methodologies were partially validated for the first time to this purpose. The novelty of the combination of techniques relies on a screening tool and a strategy to the future evaluation and an improved assessment of human embryo viability.

Non-Invasive Assessment of Spent Bovine Culture Media Using Proton Nuclear Magnetic Resonance.

Early embryonic mortality is a great determinant of reproductive efficiency in mammals, including cattle. In in vitro production, approximately 90% of oocytes undergo fertilization following in vitro maturation; however, of these fertilized oocytes approximately 30-40% will reach the blastocyst stage. Factors involved in early embryonic mortality include oocyte and sperm quality, genetic factors, environmental and oxidative stresses, and largely in in vitro production, sub-optimal culture conditions. Current methods of embryo viability evaluation depend on morphological assessment, which in spite of its importance, is a poor predictor of embryo viability. Development of refined biological techniques like metabolomics has enabled us to explore the health of a cell based on its secreted metabolite constituents. Advanced analytical techniques, such as proton nuclear magnetic resonance (H1 NMR) has little chemical bias, gives detailed structural information of isolated metabolites and allow metabolites to be measured simultaneously with minimal sample preparation. We hypothesize that, embryos developing at different rates differ in their metabolomic signatures. The specific objective of study was to determine the metabolomic signatures of slow growing and fast growing embryos at timed stages of development, thereby determining possible biomarkers of embryo competency. Oocytes were collected from ovaries obtained from a local abattoir and then matured and fertilized in vitro using standard protocols. Presumptive zygotes were either placed for individual culture in 25 μl media drops. Media samples from fast growing embryos were collected at 2-cell (31 hours post fertilization), 4-cell (42 hours post fertilization), 8-cell (49 hours post fertilization) or 16-cell (72 hours post fertilization) and from slow growing embryos, which required additional 8 to 12 hours to reach equivalent embryo stage. The 18 μl media collection was diluted in 600 μl of 100% deuterium oxide (D2O) which included the internal reference standard TSP (sodium 3-(trimethyl-2,2,3,3)-1-propionic acid-d4). H1 NMR analysis was performed using a 600MHz Bruker NMR spectrometer. Preliminary data indicates distinct differences between metabolomic fingerprints of plain media, 2-cell, 4-cell, 8-cell, and 16-cell slow growing and fast growing groups. Specifically, slow growing embryos had lower lactate uptake and increased pyruvate production, while fast growing embryos demonstrated higher levels of pyruvate uptake. Moreover, in the presence of hydrogen peroxide, a reactive oxygen species, pyruvate can be decarboxylated to produce acetate, a metabolite which is higher in slow growing embryos compared to fast growing embryos. The data suggests that slow growing embryos have a slower metabolism and lower utilization of media substrates, as well as an increased response to oxidative stress. The results provide evidence towards the use of metabolomics for the development of a non-invasive tool for assessing embryo viability and the discovery of potential embryo viability biomarkers. (poster)

A review of the promises and pitfalls of oocyte and embryo metabolomics

Embryo viability assessment is one of the most important and challenging tasks in IVF. Evaluation of embryo quality is critical when selecting the best embryo(s) to transfer or cryopreserve. Until recently, the only instrument used for embryo evaluation was the inverted light microscope, which provided information based on morphological characteristics. Developmental and morphological information gained from microscopic assessment have been positively associated with IVF outcomes, including pregnancy and implantation rates. However, based on general statistics, it is clear that IVF currently still results in relatively low pregnancy rates, while simultaneously being associated with relatively high multiple implantation rates. Only with novel embryo assessment and selection procedures would it be possible to improve these outcomes. Accordingly, it has been proposed that it is possible to test the culture environment of a developing embryo to gain valuable information regarding its viability. Different approaches have been used. These include the measurement of oxygen consumption by the embryo and testing of the soluble HLA-G in the environment, as it was proposed that secretion of HLA-G is associated with higher implantation rates. Amino acid turnover, which appears to be correlated to blastocyst development, can be measured as an indication of embryo viability. Other approaches, such as time-lapse video observation or cumulus cell gene expression analysis, may be used in the future to gain a broader understanding of embryo viability. Proteomics and metabolomics are also useful tools for assessment of embryo developmental potential. Results from recent studies on predicting embryo viability by analyzing the metabolome of different stage embryos are promising, as increases in pregnancy and implantation rates were obtained using the metabolomic profile for embryo selection. Several novel approaches are currently being developed to aid in viability assessment. These need to be evaluated in prospective clinical trials, while considering their practicality in the clinical laboratory.

Receiver operating characteristic (ROC) analysis of day 5 morphology grading and metabolomic Viability Score on predicting implantation outcome.

Assessment of embryo viability is a key component of in vitro fertilization (IVF) and currently relies largely on embryo morphology and cleavage rate. In this study, we used receiver operating characteristic (ROC) analysis to compare the Viability Score (generated by metabolomic profiling of spent embryo culture media using near infrared (NIR) spectroscopy) to morphologic grading for predicting pregnancy in women undergoing single embryo transfer (SET) on day 5. A total of 198 spent embryo culture media samples were collected in four IVF centers located in the USA, Europe and Australia. First, 137 samples (training set) were analyzed by NIR to develop an algorithm that generates a Viability Score predictive of pregnancy for each sample. Next, 61 samples (validation set) were analyzed by observers blinded to embryo morphology and IVF outcome, using the Day 5 algorithm generated with the training set. Pregnancy was defined as fetal cardiac activity (FCA) at 12 weeks of gestation. The Area Under the Curve (AUC) was greater for the metabolomic Viability Score compared to Morphology [Training set: 0.75 versus 0.55, p = 0.0011; Validation set: 0.68 versus 0.50, P = 0.021], and for a Composite score (obtained using a model combining Viability Score with morphologic grading), compared to morphology alone [0.74 versus 0.50, p = 0.004]. Our findings suggest that Viability Score alone or in combination with morphologic grading has the potential to be a better classifier for pregnancy outcome than morphology alone in women undergoing SET on day 5.

A review on automatic analysis of human embryo microscope images.

Over the last 30 years the process of in vitro fertilisation (IVF) has evolved considerably, yet the efficiency of this treatment remains relatively poor. The principal challenge faced by doctors and embryologists is the identification of the embryo with the greatest potential for producing a child. Current methods of embryo viability assessment provide only a rough guide to potential. In order to improve the odds of a successful pregnancy it is typical to transfer more than one embryo to the uterus. However, this often results in multiple pregnancies (twins, triplets, etc), which are associated with significantly elevated risks of serious complications. If embryo viability could be assessed more accurately, it would be possible to transfer fewer embryos without negatively impacting IVF pregnancy rates. In order to assist with the identification of viable embryos, several scoring systems based on morphological criteria have been developed. However, these mostly rely on a subjective visual analysis. Automated assessment of morphological features offers the possibility of more accurate quantification of key embryo characteristics and elimination of inter- and intra-observer variation. In this paper, we describe the main embryo scoring systems currently in use and review related works on embryo image analysis that could lead to an automatic and precise grading of embryo quality. We summarise achievements, discuss challenges ahead, and point to some possible future directions in this research field.

Analysis of day 2 and 5 embryo culture media samples using an in-clinic near infrared (NIR) spectroscopy procedure

OBJECTIVE: Non-invasive profiling of embryo culture media using NIR spectroscopy assesses changes of the chemical composition of the embryo's surrounding medium to provide information about the potential viability of embryos. The objective of this study was to assess the media from Day 2 and 5 embryos and examine whether a NIR spectroscopy procedure [ViaMetrics E™] could assist in deciding which embryo to transfer on the same day. DESIGN: In clinic testing of embryo culture media. MATERIALS AND METHODS: From May to October 2009 a ViaMetrics-E Test Program was conducted in 13 IVF clinics in 4 countries. Embryos were cultured individually [20-40μl] and selected for transfer using morphology. After transfer, up to 6 embryo media samples were assessed per patient using the embryo viability assessment procedure. Over 2400 spectra were collected and analyzed, including 539 transferred embryos. Analysis was performed to examine: (i) the quality of the spectra and (ii) the clinical outcomes associated with the spectra and their subsequent Viability Scores. Clinical outcome was measured when the embryo(s) transfer number equaled Fetal Heart Beats (ie. 100% implantation rate). RESULTS: Due to the introduction of air bubbles prior to assessment of media, 12.5% (range 4-25% per clinic) of embryo transfer samples could not be analyzed. Of the remaining spectra, 175 Day 2 and 189 Day 5 transfers led to a 100% implantation rate, ie. the Viability Score could be correlated to implantation potential of individual embryos. For both transfer days a positive correlation was observed between an increasing Viability Score and increasing implantation rate [Day 2: R=0.77 and Day 5: R=0.98]. CONCLUSION: As a result of this study, procedural changes have been made to improve spectral quality and eliminate air bubbles. Importantly, NIR assessment of the culture media in the clinic appears to generate Viability Scores that constitute a parameter, in addition to morphology, for a rapid non-invasive assessment of embryo viability.

Assessment of day 5 morphology grading and metabolomic viability score as predictors of implantation outcome in women undergoing in vitro fertilization (IVF)

OBJECTIVE: Embryo morphology and cleavage rate are widely used for embryo viability assessment in the IVF laboratory. Recently, metabolomic profiling of embryo culture media was reported to correlate with the reproductive potential of embryos, allowing the generation a numerical “Viability Score”. In this study, we used Receiver Operator Curve (ROC) analysis to compare (i) morphologic grading (ii) the Viability Score and (iii) a combination of morphology and the Viability Score as predictors of implantation potential of Day 5 embryos.