scholarly journals Towards Improving Embryo Selection: Simultaneous Next Generation Sequencing Of DNA And RNA From A Single Trophectoderm Biopsy

2018 ◽  
Author(s):  
Noga Fuchs Weizman ◽  
Brandon A. Wyse ◽  
Ran Antes ◽  
Zenon Ibarrientos ◽  
Mugundhine Sangaralingam ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Clinical Sample ◽  
Reproductive Technologies ◽  
Embryo Selection ◽  
Human Embryos ◽  
Acid Oxidation ◽  
Time To Pregnancy ◽  
Initial Development ◽  
Transcriptomic Data ◽  
Fertility Treatments

ABSTRACTImproved embryo selection is crucial in optimizing the results from assisted reproduction. Preimplantation genetic screening reduces time to pregnancy and miscarriages. Correlating the transcriptome of an embryo, with fertility treatments and outcomes, holds promise in improving the overall results. We developed a novel method for embryo selection in fertility treatments that integrates embryonic genomic and transcriptomic data and evaluated it in this pilot study.A total of 21 embryos donated for research were included. Three were used for the initial development and optimization of sample processing and sequencing. Thereafter, 18 embryos were used to demonstrate the clinical safety and reproducibility of our method. Two trophectoderm biopsies were taken from each embryo: one was processed as a clinical sample for genomic profiling (control, n=18), while the other biopsy (n=18) was split and utilized for independent, simultaneous genomic and transcriptomic analysis, here termed Preimplantation Genetic and Transcriptomic Testing (PGT2).High quality genomic and transcriptomic data were obtained from all analyzed samples. The concordance between genomic data obtained with PGT2 and control samples was 100% with clinical grade quality metrics. Euploid embryos showed downregulation of genes involved in anaerobic metabolism, oxidative phosphorylation, and fatty-acid oxidation. This is the first study to provide full genomic and transcriptomic profiles from a single TE biopsy from human embryos in a clinical setting unleashing the potential of improving embryo selection and outcomes in infertility treatments. Clinical trials are needed to correlate transcriptomic data with outcomes.SUMMARYDespite advances in assisted reproductive technologies, the success rate has remained relatively constant. Under the age of 35, there is a 40% chance of delivering a child per embryo transfer, which decreases with increasing maternal age. Prioritizing embryos for transfer is based on morphological assessment and, in some cases, incorporates genetic testing as well. Selection of euploid embryos for transfer shortens the time to pregnancy and reduces the risk for miscarriages. Adding the mRNA analysis to the genomic assessment of an embryo has the potential of improving the outcomes of fertility treatments.

The birth of a healthy child in the assisted reproductive technologies program after autologous co-culture of embryo with cumulus cells and a new CAT transfer technology. Case report

GYNECOLOGY ◽  
2021 ◽  
Vol 23 (3) ◽  
pp. 270-274
Author(s):  
Gunai R. Asfarova ◽  
Veronika I. Smol'nikova ◽  
Natalia P. Makarova ◽  
Iuliia S. Drapkina ◽  
Anastasiia P. Sysoeva ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Healthy Child ◽  
Assisted Reproductive Technologies ◽  
Cumulus Cells ◽  
Reproductive Technologies ◽  
Biologically Active ◽  
Human Embryos ◽  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
Art Programs

Cumulus cells are essential during oocytes growth and development, as well as during their maturation and fertilization. Research results have shown that embryo co-cultivation with autologous cumulus cells increases the frequency of blastocyst formation, and also improves the effectiveness of ART programs. Embryo transfer in such programs is recommended to be carried out using the CAT technology (Cumulus-Aided embryo Transfer), which includes embryo cultivation on a layer of cumulus cells and embryo transfer with a certain amount of diluted cumulus cells. Patient G., 38 years old, came to the department with infertility for 15 years and recurrent implantation failure in history. The patient had ART program with autologous co-cultivation of embryos with cumulus cells and a new CAT transfer technology. The patient fell pregnant and gave birth to a healthy child. Autologous cumulus cells can be a source of biologically active substances and improve embryological parameters and implantation rate in ART programs. Embryo co-cultivation with cumulus cells is especially important for patients with recurrent implantation failure. This technique can become an alternative for optimizing human embryos culturing.

scholarly journals Preimplantation Genetic Testing for Chromosomal Abnormalities: Aneuploidy, Mosaicism, and Structural Rearrangements

Genes ◽  
2020 ◽  
Vol 11 (6) ◽  
pp. 602 ◽  
Author(s):  
Manuel Viotti
Keyword(s):  
Genetic Testing ◽  
Clinical Outcomes ◽  
Assisted Reproductive Technologies ◽  
Chromosomal Abnormalities ◽  
Reproductive Technologies ◽  
Chromosomal Mosaicism ◽  
Human Embryos ◽  
Chromosomal Anomalies ◽  
Structural Rearrangements ◽  
Preimplantation Genetic Testing

There is a high incidence of chromosomal abnormalities in early human embryos, whether they are generated by natural conception or by assisted reproductive technologies (ART). Cells with chromosomal copy number deviations or chromosome structural rearrangements can compromise the viability of embryos; much of the naturally low human fecundity as well as low success rates of ART can be ascribed to these cytogenetic defects. Chromosomal anomalies are also responsible for a large proportion of miscarriages and congenital disorders. There is therefore tremendous value in methods that identify embryos containing chromosomal abnormalities before intrauterine transfer to a patient being treated for infertility—the goal being the exclusion of affected embryos in order to improve clinical outcomes. This is the rationale behind preimplantation genetic testing for aneuploidy (PGT-A) and structural rearrangements (-SR). Contemporary methods are capable of much more than detecting whole chromosome abnormalities (e.g., monosomy/trisomy). Technical enhancements and increased resolution and sensitivity permit the identification of chromosomal mosaicism (embryos containing a mix of normal and abnormal cells), as well as the detection of sub-chromosomal abnormalities such as segmental deletions and duplications. Earlier approaches to screening for chromosomal abnormalities yielded a binary result of normal versus abnormal, but the new refinements in the system call for new categories, each with specific clinical outcomes and nuances for clinical management. This review intends to give an overview of PGT-A and -SR, emphasizing recent advances and areas of active development.

P–354 Analysis of pregnancy and miscarriage rates in anti-centromere antibodies (ACA)-positive patients treated with ART

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y Kida ◽  
M Tokoro ◽  
H Kitasaka ◽  
T Yoshimura ◽  
N Fukunaga ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Antinuclear Antibody ◽  
Assisted Reproductive Technologies ◽  
Formation Rate ◽  
Reproductive Technologies ◽  
Pregnancy Rates ◽  
Human Embryos ◽  
Antibody Testing ◽  
Miscarriage Rate ◽  
Significant Difference

Abstract Study question Do ACA have an effect on pregnancy and miscarriage rates of human embryos? Summary answer The present results suggest that in ACA-positive cases, the pregnancy rate per transfer was significantly lower, although the miscarriage rate was not affected. What is known already We have previously shown that patients with high levels of anti-centromere antibody (ACA), (one of the anti-nuclear antibodies (ANA)), frequently have dispersal of the female chromosomes in the cytoplasm. Additionally, we reported that the clinical outcome was characterized by a low oocyte maturation rate following ovum pick up and high multiple pronuclear formation rate after fertilization. However, the post-implantation course of embryos with ACA-positive cases has not yet been reported. Therefore, in this study, we analyzed the pregnancy and miscarriage rates in ACA-positive patients treated with Assisted Reproductive Technologies (ART). Study design, size, duration 6581 patients who underwent embryo transfer after antinuclear antibody testing between January 2014 and February 2020 were included in the analysis. Participants/materials, setting, methods The subjects were classified into three groups: ANA-negative (without ACA or any other ANA), ACA-positive (with only ACA) and ANA-positive (with ANA but not ACA). The cycle in which the gestational sac was confirmed was considered a positive pregnancy. The pregnancy and miscarriage rates were compared among the groups using “Ryan Test” for statistical analysis. Main results and the role of chance Of the 6581 eligible cases, the incidence of antinuclear antibody were 71.3% (4695/6581; ANA-negative), 0.9% (61/6581; ACA-positive) and 27.7% (1825/6581; ANA-positive). The pregnancy rates based on the total number of embryo transfer cycles for each were ANA-negative: 31.5% (5283/16792), ACA-positive: 17.6% (41/233), and ANA-positive: 32.4% (1891/5833). The pregnancy rates were significantly lower in the ACA-positive group than in the other groups. The miscarriage rate was 29.4% (1553/5283) in ANA-negative, 31.7% (13/41) in ACA-positive, and 28.0% (529/1891) in ANA-positive, with no significant difference between the three groups. Limitations, reasons for caution Retrospective analysis Wider implications of the findings: ACA-positive patients may benefit from a treatment strategy to increase the absolute number of oocytes by obtained in order to increase the chances of normal fertilization and attainment of implantation. Trial registration number none

scholarly journals Impact of coronavirus disease-2019 pandemic on assisted reproductive technologies: Potentialities of melatonin in management

2021 ◽  
Vol 2 ◽  
pp. 112-118
Author(s):  
Shruti R. Hansda ◽  
Jayita Pal Chowdhury ◽  
Rakesh Verma ◽  
Chandana Haldar
Keyword(s):  
Health Care Services ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Effective Vaccine ◽  
Human Beings ◽  
Female Reproduction ◽  
Effective Measure ◽  
Fertility Treatments ◽  
New Variant ◽  
The Masses

The world is still grappling with the threat due to the emergence of the coronavirus disease-2019 (COVID-19) caused by a new variant of coronavirus. The high transmission of the virus among humans has led to a pandemic and there is also emergence of mutant strain of virus (severe acute respiratory syndrome coronaviruses 2 Variant VOC 202012/01) within communities. With the uncertainties in the development, efficacy and reach of an effective vaccine among the masses, it has become a mammoth task for human beings to carry on with the dayto-day task and lifestyle. This has severely impacted the health-care services including the fertility treatments. The assisted reproductive technology (ART) services have accordingly molded itself to cater to patients. There are various guidelines and regulations introduced to provide fertility services on the basis of priority of the patients seeking treatments, keeping in compliance to the safety measures for the healthcare staffs and patients. On the other hand, couples wish to pursue their way to parenthood even in the testing times. Melatonin, a neurohormone, is a multipotent molecule associated with male and female reproduction and is being protectively involved in the various reproductive processes. It maintains the circadian rhythmicity of various hormones and has potentials in treating COVID-19 patients. Its use in IVF procedures can be protective and effective measure to deal with the uncertainties of time. Its immune enhancing role can be utilized in fulfilling the patients’ health and achieving fertility goals contributing to the success of ART.

scholarly journals To understand the reproductive tract microbiome associated with infertility through metagenomics analysis

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Manisha Vajpeyee ◽  
Lokendra Bahadur Yadav ◽  
Shivam Tiwari ◽  
Parikshit Tank
Keyword(s):  
Reproductive Tract ◽  
Assisted Reproductive Technologies ◽  
Live Birth Rate ◽  
Reproductive Technologies ◽  
Female Reproductive Tract ◽  
Human Reproduction ◽  
Main Body ◽  
Fertility Treatments ◽  
Vaginal Tract ◽  
Human Illness

Abstract Background Knowledge of the microbiome is in its infancy in health and human illness, especially concerning human reproduction. We will be better able to treat dysbiosis of the reproductive tract clinically if it is better explained and understood. It has been shown that altered vaginal microbiota affects parturition, and its function is uncertain in assisted reproductive technologies. However, the effects of recognized microbes such as Mycoplasma tuberculosis, Chlamydia trachomatis, and Neisseria gonorrhoeae are well established, resulting in subclinical changes which are considered to be risk factors for infertility and poor reproductive outcomes. Main body Recent studies indicate that the vaginal tract comprises several different organisms of the microbiome. Some microbiota can play an important role not only in the reproductive tract but also in overall health. The microbiome of the female reproductive tract has been identified mainly based on studies that examine vaginal samples across many reproductive technologies, using a metagenomics approach. Conclusion Alteration of reproductive tract microbiota or presence of certain microbiota irrespective of the level of pathogenicity may interfere with fertilization, implantation, and subsequent embryo development. This may lead to failed fertility treatments and reduced live birth rate (LBR).

scholarly journals Pre-Implantation Mouse Embryos Cultured In Vitro under Different Oxygen Concentrations Show Altered Ultrastructures

2020 ◽  
Vol 17 (10) ◽  
pp. 3384
Author(s):  
Manuel Belli ◽  
Paolo Rinaudo ◽  
Maria Grazia Palmerini ◽  
Elena Ruggeri ◽  
Sevastiani Antonouli ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Inner Cell Mass ◽  
Culture Media ◽  
Cell Mass ◽  
Reproductive Technologies ◽  
Mouse Embryos ◽  
Human Embryos ◽  
Fertilization In Vitro ◽  
Vitro Fertilization

Assisted Reproductive Technologies routinely utilize different culture media and oxygen (O2) concentrations to culture human embryos. Overall, embryos cultured under physiological O2 tension (5%) have improved development compared to embryos cultured under atmospheric O2 conditions (20%). The mechanisms responsible for this remain unclear. This study aimed to evaluate the effect of physiologic (5%) or atmospheric O2 (20%) tension on the microscopic ultrastructure of pre-implantation mouse embryos using Transmission Electron Microscopy (TEM). Embryos flushed out of the uterus after natural mating were used as the control. For use as the control, 2-cells, 4-cells, morulae, and blastocysts were flushed out of the uterus after natural fertilization. In vitro fertilization (IVF) was performed using potassium simplex optimized medium (KSOM) under different O2 tensions (5% and 20%) until the blastocyst stage. After collection, embryos were subjected to the standard preparative for light microscopy (LM) and TEM. We found that culture in vitro under 5% and 20% O2 results in an increase of vacuolated shaped mitochondria, cytoplasmic vacuolization and presence of multi-vesicular bodies at every embryonic stage. In addition, blastocysts generated by IVF under 5% and 20% O2 showed a lower content of heterochromatin, an interruption of the trophectodermal and inner cell mass cell membranes, an increased density of residual bodies, and high levels of glycogen granules in the cytoplasm. In conclusion, this study suggests that in vitro culture, particularly under atmospheric O2 tension, causes stage-specific changes in preimplantation embryo ultrastructure. In addition, atmospheric (20%) O2 is associated with increased alterations in embryonic ultrastructure; these changes may explain the reduced embryonic development of embryos cultured with 20% O2.

scholarly journals MODERN APPROACHES TO CULTIVATION AND AUTOANALYSIS OF HUMAN EMBRYO MORPHODYNAMICS IN VIVO

2021 ◽  
pp. 35-43
Author(s):  
O. V. Shurygina ◽  
G. B. Nemkovskiy ◽  
D. Y. Rusakov ◽  
D. S. Gromenko ◽  
M. I. Taxants ◽  
...  
Keyword(s):  
Human Embryo ◽  
Visual Information ◽  
Assisted Reproductive Technologies ◽  
Medical Center ◽  
Time Lapse ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Human Embryos ◽  
The Moment

Relevance: Currently, it is extremely important to identify predictors of the development of a competent embryo that determine its implantation potential. In this case, the predictors are predictive parameters that should be assessed together to rank and select human embryos. We introduced the concept of «human embryo morphodynamic profile» to standardize the description of the development of human embryos cultured in vitro. We identified a set of morphokinetic states that are included in the profile and located on the time scale depending on the moment of their registration. All timing cutoffs (points) are given in chronological order relative to the moment of fertilization. The purpose of the study was to implement an information system utilizing artificial intelligence technologies for an automated formation of the morphodynamic profile of a human embryo based on time-lapse photography of the process of human embryo cultivating to the blastocyst stage. Materials and methods: Visual information about the pre-implantation development of human embryos to the blastocyst stage (0 - 6 days from insemination) was collected using an «Embryovisor» incubator for IVF laboratories with a time-lapse (hyperlapse) video fixation system (LLC «WESTTRADE LTD,” Russia). The embryos were cultivated individually in special microwells of WOW dishes (Vitrolife, Sweden). Visual information about cultured human embryos was collected, marked, and prepared at the Laboratory of assisted reproductive technologies (ART) of the Clinical Hospital IDK CJSC “Medical Company IDK” (Group of Companies “Mother and Child,” Samara, Russia) and the medical center “Semya” (Ufa, Russia). The morphodynamic profile was marked using the EmbryoVisor software (customized version). Graphics and markup information was uploaded to the SberCloud cluster. A convolutional neural network for solving the multiclass classification task was implemented on the Christofari supercomputer of the SberCloud cluster. Results: Based on the available database, we have developed a system for forming the morphodynamic profile of a human embryo, taking into account the placement of markers of fixed morphokinetic states. Conclusion: The ability to record major morphodynamic events and assess them allows a more comprehensive approach to evaluating and ranking developing embryos and selecting the most promising embryo for implantation.

scholarly journals Primate models for assisted reproductive technologies

Reproduction ◽  
2004 ◽  
Vol 128 (3) ◽  
pp. 293-299 ◽  
Author(s):  
Laura Hewitson
Keyword(s):  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Human Embryos ◽  
Gene Therapies ◽  
Monkey Model ◽  
Human Fertilization ◽  
Molecular Events ◽  
Cytoskeletal Rearrangements ◽  
Nuclear Remodeling ◽  
Human Primate

Although the deliberate creation of human embryos for scientific research is complicated by ethical and practical issues, a detailed understanding of the cellular and molecular events occurring during human fertilization is essential, particularly for understanding infertility. It is clear from cytoskeletal imaging studies of mouse fertilization that this information cannot be extrapolated to humans because of unique differences in centrosomal inheritance. However, the cytoskeletal rearrangements during non-human primate fertilization are very similar to humans, providing a compelling animal model in which to examine sperm–egg interactions. In order to address this key step in primate fertilization and to avoid the complexities in working with fertilized human zygotes, studies are now exploring the molecular foundations of various assisted fertilization techniques in a monkey model. While intracytoplasmic sperm injection with ejaculated or testicular sperm is quite successful in primate models, there are some specific differences when compared with standard IVF that warrant further investigation, particularly in regards to nuclear remodeling, genomic imprinting, Y-chromosome deletions and developmental outcomes. Similarly, primate models have been useful for examining spermatid function during fertilization but these have met with limited success. One area of primate reproductive research that has yet to be mastered is reproductive cloning. Genetically identical primates would provide the ultimate approach for accelerating stem cell-based therapies for a number of neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease, as well as targeted gene therapies for various metabolic disorders.

Sign in / Sign up

Export Citation Format

Share Document