How much information about embryo implantation potential is included in morphokinetic data? A prediction model based on artificial neural networks and principal component analysis.

PURPOSE: The aim of this study was to answer the question of how much information about embryo implantation potential can be obtained from morphokinetic parameters through the creation a predictive model based on morphokinetic information and using advanced data-mining and artificial intelligence methods. MATERIALS AND METHODS: Time-lapse recordings of 610 embryos were included in the analysis. For each embryo, absolute (t2, t3, t4, t5) and relative (cc2 and s2) morphokinetic parameters were collected. Additionally, fragmentation levels assessed at t2, t3, t4 and t5 time-points, presence of multinucleation, evenness of blastomeres after the first and second division and women's age were included in the analysis. RESULTS: The created predictive model of implantation was constructed on the basis of two advanced data-mining methods: principal component analysis (PCA) and artificial neural networks (ANN). The receiver operating characteristic (ROC) curve constructed for the created model demonstrated its good quality. The area under the ROC curve was AUC=0.75 with a 95% confidence interval (0.70, 0.80). Finally, the model was verified on a validation set and the results were also good, although slightly weaker: AUC=0.71, with a 95% confidence interval (0.59, 0.84). CONCLUSIONS: The combination of two data-mining algorithms: PCA and ANN may be considered a method which can extract virtually all the available information from data. This methodology is indeed efficient, but models presented in the literature are also effective and close to the limit of the maximal information which can be extracted from morphokinetic data. The limit can be determined at the level of AUC value marginally above 0.7.

Morphokinetic parameters as a source of information concerning embryo developmental and implantation potential.

OBJECTIVES: The aim of the study was to present the results of time-lapse observation and to verify whether morphokinetic parameters are associated with embryo developmental and implantation potential. MATERIAL AND METHODS: The analysed data concern the development of 1,060 embryos, 898 of which (84.72%) achieved the blastocyst stage and 307 were transferred into the uterine cavity. As a result, 126 (41.04%) biochemical pregnancies and 109 (35.50%) clinical pregnancies were observed. Time from fertilisation to further divisions into 2-9 blastomeres, first to fourth round of cleavage, second to third synchronisation parameters and the duration of stages after the first, second and third division were analysed. RESULTS: Most of the parameters in the group of embryos developed to the blastocyst stage reached lower values than in the non-developed group. Moreover, parameters in the first group clearly had less dispersion. The differences between the groups with and without a biochemical pregnancy were smaller than the differences in the analysis of development to the blastocyst stage. However, in the case of clinical pregnancy analysis, there were again larger differences between both groups. A strong correlation was found between the majority of absolute morphokinetic parameters. A weaker, but still statistically significant correlation, was established between relative and other parameters. CONCLUSIONS: Morphokinetic parameters are associated with embryo developmental and implantation potential and can be considered as predictors of their quality. However, the development of efficient pregnancy prediction models needs further research utilising information from all available parameters and using advanced biostatistical methods.

[First in the world application of next generation sequencing in preimplantation genetic diagnostics in clinical practice - a case report]. / Pierwsze w swiecie zastosowanie sekwencjonowania nastepnej generacji w diagnostyce przedimplantacyjnej w praktyce klinicznej - opis przypadku.

Preimplantation genetic diagnosis (PGD) is a well established method for detecting genetic abnormalities during the course of infertility treatment, resulting in thousands of healthy newborns delivered worldwide. PGD with next generation sequencing (NGS) provides new possibilities for diagnosis and new parameters for evaluation. The use of next-generation DNA sequencing technique has lead to great progress in the human genome analysis. The aim of this study was molecular analysis using next generation sequencing technique of embryos from a couple suffering from recurrent pregnancy losses. As a result of in vitro fertilization procedure, seven embryos were created. Seven blastomeres, one from each embryo, were analyzed. Transfer of two blastocysts in a fresh cycle resulted in the singleton pregnancy. Healthy baby girl was delivered via caesarean section after 28 weeks of gestation (weight: 1250g, Apgar score: 8/9). The reason for the premature labor was likely caused by mother's pneumonia. This is the first case of clinical use of the NGS in PGD in fresh cycle after blastomere biopsy.

Do morphokinetic data sets inform pregnancy potential?

PURPOSE: The aim of this study was to create a model to predict the implantation of transferred embryos based on information contained in the morphokinetic parameters of time-lapse monitoring. METHODS: An analysis of time-lapse recordings of 410 embryos transferred in 343 cycles of in vitro fertilization (IVF) treatment was performed. The study was conducted between June 2012 and November 2014. For each embryo, the following data were collected: the duration of time from the intracytoplasmic sperm injection (ICSI) procedure to further division for two, three, four, and five blastomeres, time intervals between successive divisions, and the level of fragmentation assessed in successive time-points. Principal component analysis (PCA) and logistic regression were used to create a predictive model. RESULTS: Based on the results of principal component analysis and logistic regression analysis, a predictive equation was constructed. Statistically significant differences (p < 0.001) in the size of the created parameter between the implanted group (the median value: Me = -5.18 and quartiles: Q 1= -5.61; Q 3 = -4.79) and the non-implanted group (Me = -5.69, Q 1 = -6.34; Q 3 = -5.16) were found. A receiver operating characteristic (ROC) curve constructed for the considered model showed the good quality of this predictive equation. The area under the ROC curve was AUC = 0.70 with a 95% confidence interval (0.64, 0.75). The presented model has been validated on an independent data set, illustrating that the model is reliable and repeatable. CONCLUSIONS: Morphokinetic parameters contain information useful in the process of creating pregnancy prediction models. However, embryo quality is not the only factor responsible for implantation, and, thus, the power of prediction of the considered model is not as high as in models for blastocyst formation. Nevertheless, as illustrated by the results of this study, the application of advanced data-mining methods in reproductive medicine allows one to create more accurate and useful models.

Reproductive outcomes of intracytoplasmic sperm injection (ICSI) in good-prognosis patients who electively decided to limit the number of oocytes used for microinjection: a two-center comparative study.

OBJECTIVES: The aim of the study was to compare the outcomes of intracytoplasmic sperm injection/embryo transfer (ICSI/ET) between two IVF centers with similar pregnancy rates and embryo transfer policy but with two different approaches to good-prognosis patients who intentionally chose to limit the number of oocytes used for ICSI. MATERIAL AND METHODS: It was a retrospective two-center comparative study A total of 218 patients after successful retrieval of >10 mature oocytes following ovarian hyperstimulation were included in the study The number of fertilized oocytes used during ICSI/ET was limited to 6 and 10 in 108 and 110 patients of the Centre for Reproductive Medicine KRIOBANK and VitroLive Fertility Clinic, respectively RESULTS: No significant differences in the implantation rate (29.93% vs. 29.54%; p=0.94) and ongoing pregnancy rate (39.81% vs. 45.45%, p=0.40) were observed between patients who electively fertilized 6 as compared to 10 oocytes, respectively However in patients who deliberately limited the number of fertilized oocytes to 6 the following were observed: i) significantly fewer embryos available for ET (2.89 ± 1.23 vs. 3.77 ± 1.48, p<0.0 1); ii) considerably lower number of frozen embryos per cycle (1.05 ± 1.30 vs. 2.00 ± 1.67, p<0.01), and iii) lower rates of cycles with embryo cryopreservation (4 7.22% vs. 72.72%, p<0.01) as compared to patients with 10 fertilized oocytes. CONCLUSIONS: Elective fertilization of 6 vs. 10 oocytes does not adversely affect fresh ICSI/ET outcome in normal-responding patients. Restricted number of oocytes used for ICSI/ET may be a favorable alternative for couples who do not wish to cryopreserve surplus human embryos.

A predictive model for blastocyst formation based on morphokinetic parameters in time-lapse monitoring of embryo development.

PURPOSE: The aim of the study was to create a predictive model of blastocyst development based on morphokinetic parameters of time-lapse embryoscope monitoring. METHODS: Time-lapse recordings of 432 embryos (obtained from 77 patients), monitored in Embryoscope, were involved in the study. Patients underwent in vitro fertilization according to standard procedure between June 2012 and April 2013. A retrospective analysis of morphokinetic features, focused on duration of time from the Intracytoplasmic Sperm Injection (ICSI) procedure to consecutive embryo division for 2, 3, 4 and 5 blastomeres, as well as time intervals between each division, was conducted. All embryos were observed for 5 days. RESULTS: Based on the distribution of analyzed morphokinetic parameters and number of embryos developed to blastocyst, a range denoting the possibility of an embryo reaching blastocyst stage was determined. According to the obtained results, univariate and multivariate logistic regression analyses were performed. Based on the times of division for two and five blastomeres and intervals between the second and third division, a multivariate predictive model was created. The predictive equation was constructed based on the parameters of logistic regression analysis (odds ratios). Statistically significant differences (p < 0.001) in the size of the prediction parameter between the group of embryos developed to blastocyst (the median value: Me = 9.95, and quartiles: Q1 = 7.59, Q3 = 12.30) and embryos that did not develop to the blastocyst stage (Me = 4.66, Q1 = 2.33, Q3 = 8.19) were found. A Receiver Operating Characteristic (ROC) curve was created for the constructed predictive model. The Area Under the Curve was AUC = 0.806 with a 95 % confidence interval (0.747, 0.864). The predictive model constructed in this study has been validated using an independent data set, which indicates that the model is reliable and repeatable. CONCLUSIONS: Time-lapse imaging presents a new diagnostic tool for parametric evaluation of embryo development, from the oocyte stage, through fertilization, up to the blastocyst stage. The assessment of morphokinetic parameters can help us to provide more accurate information about the reproductive potential of embryos. It allows for early selection of embryos with high reproductive potential and shortens embryo incubation.

Comparison of the second-generation Beckman Coulter IVD and first-generation AnshLabs ELISA assays for anti-Müllerian hormone in patients undergoing IVF treatment.

OBJECTIVES: Ovarian reserve is the main factor influencing the efficacy of infertility treatment. Currently the anti- Müllerian hormone is the main indicator of the ovarian reserve and has a wide spectrum of clinical importance. It achieved a high clinical value right after the introduction of the first commercial AMH assays in 2005. Lack further research and development of the tests and monopoly on their production have led to a significant reduction of their quality resulting in lowered veracity and usefulness. Therefore, we searched for an alternative to the Beckman Coulter assay. The objective of the study was to draw a comparison between the commonly used second-generation assay by Beckman Coulter and the ultra-sensitive first-generation assay by AnshLabs. MATERIALS AND METHODS: Serum samples (n=520) were collected from female patients undergoing routine AMH evaluation before entering an IVF program. We chose samples of patients with the lowest correlation between the AMH serum level and response to stimulation. The AMH serum levels of the patients were examined using two AMH tests, the second-generation assay by Beckman Coulter and the first-generation assay by AnshLabs. Precision and accuracy of both methods were determined and the results of AMH serum levels of 130 patients were correlated with the number of: antral follicles (AFC), follicles after stimulation, and the obtained cumulus cells. RESULTS: Both precision and accuracy of the compared methods were highly satisfactory. The coefficients of variation obtained in the study conducted on two different levels of control material were lower than 12% and the load did not exceed 9%. The study proved that both of the methods yielded comparable results. The coefficient of variation between the first-generation and the second-generation AMH assays was 0.871. CONCLUSION: Both methods might be applied in the evaluation of the ovarian reserve. The first- and second-generation assays show comparable correlation with the clinical effects of stimulation, however it seems that first-generation assays are a better alternative to the unstable second-generation kits. The results from the first-generation assays are distributed on a wider range, which facilitates clinical interpretation.

The dynamics of endometrial growth and the triple layer appearance in three different controlled ovarian hyperstimulation protocols and their influence on IVF outcomes.

The impact of endometrial growth to the triple layer, endometrial thickness, and echogenicity on IVF outcomes was investigated in the study. A retrospective analysis of 583 ICSI patients was conducted: 385 with a long GnRH agonist protocol, 114 with a short GnRH agonist, and 84 with a GnRH antagonist protocol. The progression of endometrial growth to the appearance of the triple layer, endometrial thickness, and echogenicity was compared between protocols. At least one high quality blastocyst was transferred in a double embryo transfer. The time of the appearance of the endometrial triple layer was statistically significant for the pregnancy rate only in the GnRH antagonist protocol. The endometrial thickness on the day of the appearance of the triple layer had a statistically significant influence on the pregnancy rate in the GnRH antagonist and in the long GnRH agonist protocols. The highest pregnancy rate for the long GnRH agonist and the GnRH antagonist protocols was observed when the endometrium thickness was 12-13 mm (61.6% and 58.8%, respectively). The endometrial echogenicity had a significant influence on the pregnancy rate only in the long GnRH agonist protocol. Endometrial features could be helpful parameters in IVF outcomes in particular controlled ovarian hyperstimulation protocols.

Vitrification vs. slow cooling protocol using embryos cryopreserved in the 5th or 6th day after oocyte retrieval and IVF outcomes.

Modifying cryopreservation protocols may be seen as a way to simplify cryobanking procedure and increase satisfying outcomes. The aim of the study was to evaluate the influence of slow cooling protocol and vitrification on IVF outcomes using embryos preserved in the 5th or 6th day after oocyte retrieval. The study compared 2 groups of human embryos underwent slow cooling protocol (n=189) and vitrification (n=58). All embryos were cryopreserved in the 5th or 6th day after oocyte retrieval. Pre- and postfreezing embryo evaluation was performed in 2 or 3 steps scale, respectively. The study evaluates the effectiveness of two freezing methods and influence of the freezing day, pre- and postfreezing embryo grading on clinical pregnancy rate. Study showed higher pregnancy rate after vitrification (50.4%) than slow cooling protocol (25.9%). Significantly higher pregnancy rate was observed, when embryo preserved in the 5th day after oocyte retrieval (50.3%) than in the 6th day (22.7%). Postfreezing embryos evaluation showed that high quality blastocysts gave nearly four times better pregnancy outcomes than the ones evaluated as poor quality, and three times better than the ones evaluated as moderate. Prospective trials are needed to evaluate pregnancy and neonatal outcomes after vitrification. The number of controlled studies concerning vitrification has not been large enough, yet.