Prediction of live birth - selection of embryos using morphokinetic parameters.

BACKROUND: The goal of assisted reproduction is for a couple treated with IVF techniques to end the treatment by giving birth to a healthy baby. A neccessary presumption for success is the identification of the best embryo with high implantation and developmental potential. One option is to select an euploid embryo by invasive preimplantaion genetic testing for aneuploidy (PGT-A) or it is possible to select the best embryo by non-invasive time-lapse monitoring (TLM), specifically based on morphokinetic parameters and morphological markers that are able to identify an embryo with high developmental potential. MATERIALS AND METHODS: The study involved a total of 1060 embryos (585 euploid and 475 aneuploid embryos after PGT-A) with good morphology from 329 patients in the period 01/2016-10/2021. All embryos were cultured in a time-lapse incubator, trophectoderm (TE) cells biopsies for PGT-A examination were performed on day 5 (D5) or day 6 (D6) of culture. During the study period, 225 frozen embryo transfers (FET) of one euploid embryo were performed. Based on the treatment outcome, the embryos were divided into 2 groups - euploid embryos, which led to the birth of a healthy child, and euploid embryos that did not show fetal heartbeat (FHB) after FET. RESULTS: Based on the statistical analysis of the embryos without implantation and the embryos with live birth, it is clear that the morphokinetic parameters t5 (time of division into 5 cells) and tSB (time of start of blastulation) are significantly different. CONCLUSION: The results suggest that of the morphokinetic parameters tSB and t5 are predictive indicators for selecting an embryo with high developmental potential and with a high probability of achieving the birth of a healthy child.

Differences in Morphokinetic Parameters and Incidence of Multinucleations in Human Embryos of Genetically Normal, Abnormal and Euploid Embryos Leading to Clinical Pregnancy.

The selection of the best embryo for embryo transfer (ET) is one of the most important steps in IVF (in vitro fertilisation) treatment. Preimplantation genetic testing (PGT) is an invasive method that can greatly facilitate the decision about the best embryo. An alternative way to select the embryo with the greatest implantation potential is by cultivation in a time-lapse system, which can offer several predictive factors. Non-invasive time-lapse monitoring can be used to select quality embryos with high implantation potential under stable culture conditions. The embryo for ET can then be selected based on the determined morphokinetic parameters and morphological features, which according to our results predict a higher implantation potential. This study included a total of 1027 morphologically high-quality embryos (552 normal and 475 abnormal PGT-tested embryos) from 296 patients (01/2016-06/2021). All embryos were cultivated in a time-lapse incubator and PGT biopsy of trophectoderm cells on D5 or D6 was performed. Significant differences were found in the morphological parameters cc2, t5 and tSB and the occurrence of multinucleations in the stage of two-cell and four-cell embryos between the group of genetically normal embryos and abnormal embryos. At the same time, significant differences in the morphological parameters cc2, t5 and tSB and the occurrence of multinucleations in the two-cell and four-cell embryo stage were found between the group of genetically normal embryos that led to clinical pregnancy after ET and the group of abnormal embryos. From the morphokinetic data found in the PGT-A group of normal embryos leading to clinical pregnancy, time intervals were determined based on statistical analysis, which should predict embryos with high implantation potential. Out of a total of 218 euploid embryos, which were transferred into the uterus after thawing (single frozen embryo transfer), clinical pregnancy was confirmed in 119 embryos (54.6%). Our results show that according to the morphokinetic parameters (cc2, t5, tSB) and the occurrence of multinucleations during the first two cell divisions, the best euploid embryo for ET can be selected with high probability.

A new instrument to personalize ovarian stimulation: results of the CERES study after using a novel gonadotropin - follitropin delta.

: Objective: The aim of the study CERES (CzEch REkovelle real life Study) was to gather experience with the use of a novel gonadotrophine, to evaluate the efficacy of follitropin delta in Czech clinical settings and to compare our results with the clinical trial ESTHER-1. METHODS: Individualized follitropin delta daily dose in µg based on the patient's anti-Müllerian hormone (AMH) level and body weight (AMH < 15 pmol/ L: 12 µg/ d; AMH > 15 pmol/ L: 0.10­0.19 µg/ kg/ d; max. 2 µg/ d). RESULTS: A total of 85 women (aged 24-42 years) was included in the study. The average patients age was 32.9 years, the average body weight was 67.8 kg, and the mean level of AMH was 23.2 pmol/ L. There were initiated 85 controlled ovarian stimulations with follitropin delta and 84 egg collections. Forty patients (47%) had optimal number of retrieved eggs (8-14), 75 patients (88%) had embryotransfer, 10 patients (12%) had no embryo suitable for transfer, 65 patients had single embryo transfer and 10 patients had 2 embryos for transfer. There were reported 37 clinical pregnancies (43.5% cPR - clinical pregnancy rate), 30 live births (35.3% LBR - live birth rate), 3 (3.5%) early moderate ovarian hyperstimulation syndroms (OHSS) and no hospitalization due to the treatment. CONCLUSION: Individualized ovarian stimulation optimizes ovarian response, maintains treatment efficacy and improves safety by reducing OHSS incidence. The results of the Czech population study are fully comparable with the international, randomized, assessor-blinded trial ESTHER-1.

Solved problems of reproductive medicine in the Czech Republic 2020.

INTRODUCTION: During the 30th symposium of assisted reproduction held on November 11, 2020 in Brno, the solved problems in reproductive medicine in the Czech Republic in 2020 were presented. The selected topics have concerned not only current issues in the field of clinical embryology and genetics as well as gynecology, but also legislation and ethics. Discussed topics: 1. How much time does the doctor have in the CAR (centrum of assisted reproduction) outpatient clinic per patient and how does the embryologist communicate with clients? 2. Reproduction and PGT-M in oncology patients and patients at risk with hereditary oncogenic mutations. 3. Non-invasive genetic testing of embryos from culture medium. 4. Genome editing. 5. What is the need to monitor hormonal levels in stimulation protocols? 6. Monitoring and embryo selection for transfer/kryo. 7. Is it time to change the law on donor remuneration? METHODS: The topics were prepared in advance by authorized members of our company with the task of elaborating theses, which they presented in a separate conference block. The presentation and the discussion were broadcast directly from the broadcast studio at Hotel International via an online connection. After the conference, all discussion topics and comments were incorporated. CONCLUSION: The work presents the state of the solved problems of reproductive medicine in the Czech Republic.

Non-Invasive Human Embryo Metabolic Assessment as a Developmental Criterion.

The selection of a highly-viable single embryo in assisted reproductive technology requires an acceptable predictive method in order to reduce the multiple pregnancy rate and increase the success rate. In this study, the metabolomic profiling of growing and impaired embryos was assessed on the fifth day of fertilization using capillary electrophoresis in order to find a relationship between the profiling and embryo development, and then to provide a mechanistic insight into the appearance/depletion of the metabolites. This unique qualitative technique exhibited the appearance of most non-essential amino acids and lactate, and depleting the serine, alanyl-glutamine and pyruvate in such a manner that the embryos impaired in their development secreted a considerably higher level of lactate and consumed a significantly higher amount of alanyl-glutamine. The different significant ratios of metabolomic depletion/appearance between the embryos confirm their potential for the improvement of the prospective selection of the developed single embryos, and also suggest the fact that pyruvate and alanyl-glutamine are the most critical ATP suppliers on the fifth day of blastocyst development.

Gestational surrogacy in the Czech Republic.

BACKGROUND: Gestational surrogacy, is a treatment option for women with certain clearly defined medical problems, usually an absent uterus, to help them have their own genetic children. The aim of our study was to review, evaluate and share our experience and outcomes over the last 13 years of the largest surrogacy program in the Czech republic. METHODS: A total of 75 intended mothers and 82 surrogate mothers participated in this study. A retrospective cohort study was performed. Anonymized data were collected on 130 cycles of gestational surrogate (2004-2017) directly from the Clinic database. RESULTS: We performed 130 in vitro fertilization cycles with gestational surrogacy which involved 73 fresh embryo transfers and 57 frozen embryo transfers. We achieved 57 (43.9%) pregnancies and 42 (32.3%) live births. The rate of multiple pregnancies was only 2.3 %. The most common indication for using was an absent or damaged uterus (65%), followed by medical conditions precluding pregnancy (23%) and repeated in vitro fertilization cycles or pregnancy failure (12%). CONCLUSION: In the 14 years of our experience, we have shown that treatment of young women with specific indications for gestational surrogacy is beneficial, successful and relatively free of complications. However, it is imperative to follow the medical indications for this treatment and specialist recommendations.

The attitudes of IVF patients treated in the Czech Republic towards informing children born after gamete donation.

BACKGROUND: In recent decades gamete donation has received growing attention. Data from the Czech National Registry of Assisted Reproduction show that the number of cycles using donated oocytes has been increasing every year. According to Czech law, gamete donation is anonymous. Since 2011, some members of the Czech parliament have repeatedly made requests to revoke the anonymity but anonymity is one of the preconditions for such donation in this country. The aim of this study was to find out how the gamete recipients feel towards informing their child about the circumstances of their conception and their access to the identity of the donor. METHODS: A total of 195 recipients (122 women undergoing treatment - 43 Czechs, 79 foreigners (Western Europe and the USA) and 73 male partners - 28 Czechs, 45 foreigners) participated in this survey. The data were obtained by anonymous questionnaire. RESULTS: A significant difference between the attitude of the future Czech and foreign parents regarding disclosing the mode of conception was found (P = 0.003). The vast majority of Czechs were against disclosure. The foreign recipients were somewhat more divided. Regarding the donor's identity, there was no difference in atttitude between the groups. Recipients rarely consider that the knowledge of the donor's identity will be important for their child. The recipients overall, were convinced that the psychological aspects of parenting are far more important to the child than genetics, and see no reasons for disclosing the donor´s identity. CONCLUSION: While the the foreign recipients were less adamant about non-disclosure, the overall finding was in accord with the current Czech law on anonymity and not in agreement with the proposed abolition. The recipient's attitudes towards disclosing were also culturally determined. The fact that some countries have revised their rules towards open idendity is not a rationale for such change in the Czech Republic.

Inhibin A and B levels in serum and follicular fluids of women with various reproductive failures undergoing in vitro fertilization.

OBJECTIVE: Ovarian inhibins (INH) are hormones participating in the regulation of gametogenesis. We monitored inhibin A and B levels in serum (S) and follicular fluids (FF), depending on the type of fertility failure and treatment outcome. MATERIAL AND METHODS: We examined INHA and B levels in S and FF of 72 women undergoing ovarian stimulation for in vitro fertilization, including embryo transfer We took serum samples at the time of egg collection (S1), embryo transfer (S2), and diagnostics of early pregnancy (S3). FF samples were obtained during egg collection. INH A and B levels were measured by ELISA set kit in all media. RESULTS: Healthy women had median of INHA S1 592.02pg/ml INHA S2 593.58pg/ml, INHA S3 15.17pg/ml and INH B S1 242.46pg/ml, INH B S2 and INH B S3 zero levels. Women with ovarian disorders had significantly lower levels of INH A S1 and INH A S2 (p<0.05). Women with polycystic ovaries had significant higher INH B S2 levels (p<0. 05). No statistically significant differences were found in women with endometriosis. Presence of oocyte in the dominant follicle positively correlated with INH B FF levels (p<0.05). CONCLUSIONS: WE confirmed differences in the levels of inhibins in sera depending on type of fertility failure. Inhibin B better reflected the presence of an oocyte. The potential paracrine role of inhibins needs to be examined to improve preparation for the in vitro fertilization treatment (IVF).

Hyperstimulation syndrome: the levels of inhibin A and B in sera and follicular fluids.

Ovarial hyperstimulation syndrome (OHSS) represents a serious problem encountered during in vitro fertilization (IVF). We examined 10 patients with OHSS and 50 women also hormonally stimulated in the process of IVF who had no complications. In all women, we evaluated the number of obtained oocytes and the level of inhibins A and B in sera and follicular fluid collected at the time of ovarial puncture, the day embryo transfer and on the day of positivity for hCG. The level of inhibin B in both fluids was significantly higher (t = 0.0403) in women with high quality of oocytes. The higher level of inhibin A was detected in patients with OHSS at the time of oocyte collection and on the day of embryo transfer. Inhibin B was elevated only at the time of oocyte collection. The levels of inhibin A and B were identical in follicular fluids collected from both ovaries. We observed no statistically significant differences between the levels of inhibin A and B in follicular fluids of women in the absence of OHSS. Evaluation of serum levels of inhibin A and B at the time of oocyte collection may contribute to the prognosis and prevention of OHSS.