NON-INVASIVE METHODS FOR STUDYING THE POTENTIAL OF HUMAN EMBRYO FOR IMPLANTATION

Culture Media ◽

Uterine Cavity ◽

Married Couples ◽

Preimplantation Embryos ◽

Vitro Fertilization ◽

Non Invasive

В статье проводится анализ представленных на сегодня инвазивных и неинвазивных методов исследования преимплантационных эмбрионов. Показана эффективность преимплантационного генетического тестирования эмбрионов до переноса в полость матки. Также рассмотрены альтернативные менее инвазивные варианты изучения жизнеспособности эмбрионов, которые могли бы являться маркерами успешной имплантации. Проблема бесплодного брака с каждым годом становится все более и более значимой. Для части супружеских пар единственной возможностью рождения ребенка становится лечение методами вспомогательных репродуктивных технологий, эффективность которых остается на сегодняшний день не более 50 %. Особенно важным является поиск новых методик, позволяющих повысить результативность процедур экстракорпорального оплодотворения. В этом направлении крайне интересным является изучение неизвазивных методов оценки имплантационного потенциала эмбрионов. В анализе представлены работы по изучению протеома, метаболома и транскриптома эмбриона. Понимание молекулярного состава культуральных сред, в которых происходило развитие эмбриона до пятых суток культивирования, позволит глубже понять физиологию раннего развития, а также установить неивазивные критерии отбора эмбриона с лучшим имплантационным потенциалом и тем самым повысить эффективность проводимых программ вспомогательных репродуктивных технологий The article analyzes the currently presented invasive and non-invasive methods for studying preimplantation embryos. The efficiency of preimplantation genetic testing of embryos before transfer to the uterine cavity has been shown. Also considered are alternative less invasive options for studying the viability of embryos, which could be markers of successful implantation. The problem of sterile marriage is becoming more and more significant every year. For some married couples, the only possibility of having a child is treatment with methods of assisted reproductive technologies, the effectiveness of which remains at most 50% today. It is especially important to search for new techniques to improve the effectiveness of in vitro fertilization procedures. In this direction, it is extremely interesting to study non-invasive methods for assessing the implantation potential of embryos. The analysis presents works on the study of the proteome, metabolome and transcriptome of the embryo. Understanding the molecular composition of the culture media in which the development of the embryo took place until the fifth day of cultivation will allow a deeper understanding of the physiology of early development and also establish non-invasive criteria for the selection of embryos with the best implantation potential and thereby increase the efficiency of the programs of assisted reproductive technologies

Thickness of endometrium: predictor of the effectiveness of IVF/ICSI programs (literature review)

GYNECOLOGY ◽

10.26442/2079-5696_20.1.113-116 ◽

2018 ◽

Vol 20 (1) ◽

pp. 113-116

Author(s):

L A Bagdasaryan ◽

I E Korneyeva

Keyword(s):

Endometrial Thickness ◽

Molecular Genetic ◽

Uterine Cavity ◽

Genetic Characteristics ◽

Vitro Fertilization ◽

Need To Evaluate ◽

The Relationship

The aim of the study is to systematically analyze the data available in the modern literature on the relationship between endometrial thickness and the frequency of pregnancy in the program of assisted reproductive technologies (ART). Materials and methods. The review includes data from foreign and domestic articles found in PubMed on this topic. Results. The article presents data on the relationship between the thickness of the endometrium and the frequency of pregnancy in ART programs. The greatest number of studies is devoted to the evaluation of the relationship between the thickness of the endometrium and the frequency of pregnancy on the day of the ovulation trigger. Data are presented on the existence of a correlation between the thickness of the endometrium measured on the day of the ovulation trigger and the frequency of clinical pregnancy, as well as data on the need to evaluate the structure of the endometrium and the state of subendometric blood flow. The importance of multilayered (three-layered) endometrium as a prognostic marker of success in in vitro fertilization/intracytoplasmic sperm injection programs in the ovum is emphasized. The conclusion. The thickness of the endometrium can not be used as an argument for canceling the cycle or abolishing embryo transfer to the uterine cavity. Further studies in this direction are needed with a study of the morphological and molecular genetic characteristics of the endometrium, which in the future will allow us to evaluate the relationship between the thickness of the endometrium and the probability of pregnancy.

In vivo measurement of pH and CO2 levels in the uterus of sows through the estrous cycle and after insemination

Scientific Reports ◽

10.1038/s41598-021-82620-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Octavio López-Albors ◽

Pedro José Llamas-López ◽

Joaquín Ángel Ortuño ◽

Rafael Latorre ◽

Francisco Alberto García-Vázquez

Keyword(s):

Estrous Cycle ◽

Invasive Approach ◽

Vitro Fertilization ◽

Non Invasive ◽

Physiological Values

AbstractThe pH–CO2–HCO3− system is a ubiquitous biological regulator with important functional implications for reproduction. Knowledge of the physiological values of its components is relevant for reproductive biology and the optimization of Assisted Reproductive Technologies (ARTs). However, in situ measurements of these parameters in the uterus are scarce or null. This study describes a non-invasive method for in situ time-lapse recording of pH and CO2 within the uterus of non-anesthetized sows. Animals were at three different reproductive conditions, estrous with no insemination and two hours after insemination, and diestrous. From pH and CO2 data, HCO3− concentration was estimated. The non-invasive approach to the porcine uterus with novel optical probes allowed the obtaining of in situ physiological values of pH, CO2, and HCO3−. Variable oscillatory patterns of pH, CO2 and HCO3− were found independently of the estrous condition. Insemination did not immediately change the levels of uterine pH, CO2 (%) and HCO3− concentration, but all the values were affected by the estrous cycle decreasing significantly at diestrous condition. This study contributes to a better understanding of the in vivo regulation of the pH-CO2-HCO3− system in the uterus and may help to optimize the protocols of sperm treatment for in vitro fertilization.

Children born with assisted technology: a focus on parents’ reproductive health

Consilium Medicum ◽

10.26442/20751753.2021.7.200900 ◽

2021 ◽

pp. 539-543

Author(s):

Daria A. Kinsht ◽

◽

Mariia K. Soboleva ◽

Keyword(s):

Reproductive Health ◽

Medical Center ◽

Newborn Infants ◽

Uterine Cavity ◽

Perinatal Outcomes ◽

Singleton Pregnancy ◽

Aim. To assess the main indicators of the initial reproductive health of parents, somatic health of the mother and their impact on the health of children born with singleton pregnancy ART. Materials and methods. The study included all newborn infants from singleton ART who were born at the Avicenna Medical Center (Novosibirsk) over the period 2006–2017 (n=409) and their parents (n=818). All stages of overcoming infertility (from the moment the parents go to the clinic to the birth of children) are considered in the context of one center, in a relatively homogeneous social group, with ensuring continuity at all stages. The average experience of infertility in couples was 7.2±0.2 years. The groups were formed depending on the type of infertility and the method of assisted reproductive technologies (ART) used: 205 children were born using in vitro fertilization (IVF), 204 children were born using the method of Intracytoplasm Sperm Injection (ICSI). The method of IVF and transfer of embryos into the uterine cavity is more often used in women with tuboperitoneal, endocrine types of infertility (premature ovarian failure syndrome), as well as in infertility associated with endometriosis. More serious reproductive problems (severe forms of male infertility, a combination of several types of infertility) in most cases lead to the use of more serious technological methods of ART, in particular, the addition of IVF and embryo transfer by the ICSI method. The reasons underlying infertility in most cases lead not only to the choice of the ART method, but also determine the characteristics of the course of pregnancy and the development of the intrauterine fetus. Pregnancy in the IVF group is significantly more often complicated by the threat of termination and premature birth, while ICSI pregnancy more often leads to impaired development of the intrauterine fetus. Conclusion. Features of medical support of women during the preparation for ART and, of course, during pregnancy, with a comprehensive, interdisciplinary correction of expected complications in each of the groups, will improve the perinatal outcomes of induced pregnancy and will contribute to the birth of healthy offspring.

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

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph17103384 ◽

2020 ◽

Vol 17 (10) ◽

pp. 3384

Author(s):

Manuel Belli ◽

Paolo Rinaudo ◽

Maria Grazia Palmerini ◽

Elena Ruggeri ◽

Sevastiani Antonouli ◽

...

Keyword(s):

Inner Cell Mass ◽

Culture Media ◽

Cell Mass ◽

Mouse Embryos ◽

Human Embryos ◽

Fertilization In Vitro ◽

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.

CHRONIC ENDOMETRITIS AS A FACTOR OF FAILURE ATTEMPT IN ASSISTED REPRODUCTIVE TECHNOLOGIES

Likarska sprava ◽

10.31640/jvd.7-8.2019(2) ◽

2019 ◽

pp. 13-21

Author(s):

O. V. Onysko ◽

О. О. Korchynska ◽

Stefania Andrashchikova ◽

Sylvia Zhultakova ◽

Alena Shlosserova

Keyword(s):

Plasma Cells ◽

Proteolytic Enzymes ◽

Uterine Cavity ◽

Reproductive Function ◽

Careful Study ◽

Chronic Endometritis ◽

Increase in frequency of disorders of women's reproductive function justifies a careful study of the underlying pathological processes. In recent years, increased attention in solving fertility problems is paid to endometrial pathology when chronic endometritis plays an important role. Current worksarefeeding an in-depth study of the etiology and pathogenesis of chronic endometritis. However, results of research pose new questions with an imaginary comprehensive study of this problem. Chronic endometritis is one of the causes of infertility and may play a role in certain complications of pregnancy and childbirth (30.3 % in patients with repeated failures of in vitro fertilization, 9.3 % with recurrent miscarriages, 9.8 % in infertility). Clinically, chronic endometritis is often asymptomatic or accompanied by nonspecific symptoms such as pelvic pain, dyspareunia, abnormal uterine bleeding, and discharges. Previously, scientists considered the uterine cavity as a sterile container. But a team of Spanish scientists sought to test for the presence of intrauterine microflora, which differs from the vaginal. As a result, the study of endometrial fluid and vaginal samples from the same patients revealed different bacterial compositions. Microflora in the intrauterine fluid was classified as Lactobacillus, and women with non-lactobacillary flora of the endometrium had a significantly lower frequency of implantation. At the initial stage of endometrial inflammation is an activation of neutrophils and macrophages, increased cytokines synthesis, extracellular matrix degradation by proteolytic enzymes. Disorders of microcirculation and sclerotic processes in the area of injury lead to the ischemia and tissue hypoxia, which activates the processes of sclerosis and angiogenesis. Chronization of the process increases the expression of chemokines and adhesion molecules, that is lead to migration of B-lymphocytes from the bloodstream and their differentiation into plasma cells, increases the activity of matrix metalloproteinases, changes local expression of estrogen and progesterone receptors. Lack of a pronounced specific clinical picture of chronic endometritis, its later detection causes impaired reproductive function. Untimely diagnosis and treatment of chronic endometritis has significant consequences in terms of the possibility of implantation of a fertilized egg in the programs of assisted reproductive technologies.

Blastomere removal affects homeostatic control leading to obesity in male mice

Reproduction ◽

10.1530/rep-20-0253 ◽

2021 ◽

Vol 161 (1) ◽

pp. 61-72

Author(s):

Magdalena Kotlarska ◽

Dawid Winiarczyk ◽

Wiesława Florek ◽

Marta Ziętek ◽

Jolanta Pęczkowicz-Szyszka ◽

...

Keyword(s):

Gene Expression ◽

Energy Homeostasis ◽

Receptor Gene ◽

Preimplantation Embryos ◽

Ghrelin Receptor ◽

Blastomere Biopsy

Preimplantation embryos are particularly vulnerable to environmental perturbations, including those related to assisted reproductive technologies. Invasive embryo manipulations, such as blastomere biopsy, are applied worldwide in clinical settings for preimplantation genetic testing. Mouse models have previously shown that blastomere biopsy may be associated with altered phenotypes in adult offspring. The aim of the present study was to investigate the specific contribution of blastomere removal to the physiological, behavioral, and molecular regulators of energy homeostasis, as compared to sham manipulation (re-introducing the blastomere into the embryo after its removal) and in vitro culture. Mice derived from 8-cell embryos subjected to blastomere removal displayed: (i) higher body weight and adiposity, (ii) increased food intake and sucrose preference, (iii) decreased time of immobility in the tail suspension test, and (iv) resistance to weight loss after social isolation or following 3 days of physical exercise – compared to mice derived from sham biopsy or from in vitro-cultured embryos. Mice generated after blastomere removal also had increased circulating leptin and leptin gene expression in adipose tissue, as well as increased ghrelin receptor gene expression in the hypothalamus, compared to control mice. The effects of blastomere biopsy on offspring phenotype were sexually dimorphic, with females not being affected. These results indicate that blastomere deprivation, rather than other perturbations of the blastomere biopsy procedure, programs male embryos to develop physiological, behavioral, and molecular dysregulation of energy homeostasis, leading to postnatal obesity.

Do assisted reproductive technologies and in vitro embryo culture influence the epigenetic control of imprinted genes and transposable elements in children?

Human Reproduction ◽

10.1093/humrep/deaa310 ◽

2020 ◽

Author(s):

J Barberet ◽

C Binquet ◽

M Guilleman ◽

A Doukani ◽

C Choux ◽

...

Keyword(s):

Dna Methylation ◽

Embryo Culture ◽

Culture Media ◽

Randomized Study ◽

Vitro Fertilization ◽

Epigenetic Control ◽

Culture Influence

Abstract STUDY QUESTION Do assisted reproductive technologies (ART) and in vitro embryo culture influence the epigenetic control of imprinted genes (IGs) and transposable elements (TEs) in children? SUMMARY ANSWER Significant differences in the DNA methylation of IGs or transposon families were reported between ART and naturally conceived children, but there was no difference between culture media. WHAT IS KNOWN ALREADY There is concern that ART may play a role in increasing the incidence of adverse health outcomes in children, probably through epigenetic mechanisms. It is crucial to assess epigenetic control, especially following non-optimal in vitro culture conditions and to compare epigenetic analyses from ART-conceived and naturally conceived children. STUDY DESIGN, SIZE, DURATION This follow-up study was based on an earlier randomized study comparing in vitro fertilization outcomes following the use of two distinct culture media. We compared the epigenetic profiles of children from the initial randomized study according to the mode of conception [i.e. ART singletons compared with those of a cohort of naturally conceived singleton children (CTL)], the type of embryo culture medium used [global medium (LifeGlobal) and single step medium (Irvine Scientific)] and the mode of in vitro fertilization (i.e. IVF versus ICSI). PARTICIPANTS/MATERIALS, SETTING, METHODS A total of 57 buccal smears were collected from 7- to 8-year-old children. The DNA methylation profiles of four differentially methylated regions (DMRs) of IGs (H19/IGF2: IG-DMR, KCNQ1OT1: TSS-DMR, SNURF: TSS-DMR, and PEG3: TSS-DMR) and two TEs (AluYa5 and LINE-1) were first assessed by pyrosequencing. We further explored IGs and TEs’ methylation changes through methylation array (Human MethylationEPIC BeadChip referred as EPIC array, Illumina). MAIN RESULTS AND THE ROLE OF CHANCE Changes in the IGs’ DNA methylation levels were found in ART children compared to controls. DNA methylation levels of H19/IGF2 DMR were significantly lower in ART children than in CTL children [52% versus 58%, P = 0.003, false discovery rate (FDR) P = 0.018] while a significantly higher methylation rate was observed for the PEG3 DMR (51% versus 48%, P = 0.007, FDR P = 0.021). However, no differences were found between the culture media. After observing these targeted modifications, analyses were performed at wider scale. Again, no differences were detected according to the culture media, but imprinted-related DMRs overlapping promoter region near the genes major for the development (MEG3, BLCAP, and DLX5) were detected between the ART and CTL children. LIMITATIONS, REASONS FOR CAUTION The sample size could seem relatively small, but the high consistency of our results was ensured by the homogeneity of the cohort from the initial randomized study, the standardized laboratory techniques and the robust statistical analyses accounting for multiple testing. WIDER IMPLICATIONS OF THE FINDINGS Although this study did not report DNA methylation differences depending on the culture medium, it sheds light on epigenetic changes that could be observed in some children conceived by ART as compared to CTL children. The clinical relevance of such differences remains largely unknown, and it is still unclear whether such changes are due to some specific ART procedures and/or to parental infertility. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by funding from the Agence Nationale pour la Recherche (‘CARE’-ANR JCJC 2017). The authors have no conflicts of interest. TRIAL REGISTRATION NUMBER Not concerned.

The history of assisted reproductive technologies: from prohibition to recognition

History of science and technology ◽

10.32703/2415-7422-2021-11-2-315-328 ◽

2021 ◽

Vol 11 (2) ◽

pp. 315-328

Author(s):

Maryna Petrushko ◽

Volodymyr Piniaiev ◽

Taisiia Yurchuk

Keyword(s):

In Vitro Fertilization ◽

Chromosomal Abnormalities ◽

Reproductive Potential ◽

Married Couples ◽

Infertility Treatment ◽

Hereditary Diseases ◽

The birth of children after infertility treatment of married couples with the help of assisted reproductive technologies has become a reality after many years of basic research on the physiology of reproductive system, development of oocyte’s in vitro fertilization methods and cultivation of embryos at pre-implantation stages. Given the widespread use of assisted reproductive technologies in modern medical practice and the great interest of society to this problem, the aim of the study was to trace the main stages and key events of assisted reproductive technologies in the world and in Ukraine, as well as to highlight the activities of outstanding scientists of domestic and world science who were at the origins of the development of this area. The paper used historical methods to study and interpret the texts of primary sources and present scientific historical events. In addition, the current trends in assisted reproductive technologies are covered based on the results of our own, more than 30 years of experience in the field of reproductive biology and medicine, and the achievements of world scientists. As a result of the work, it has been shown that despite certain ethical and social biases, the discovery of individual predecessor scientists became the basis for the efforts of Robert Edwards and Patrick Steptoe to ensure birth of the world's first child, whose conception occurred outside the mother's body. There are also historical facts and unique photos from our own archive, which confirm the fact of the first successful oocyte in vitro fertilization and the birth of a child after the use of assisted reproductive technologies in Ukraine. Over the last 20 years, assisted reproductive technologies have continued to grow, addressing many other issues of reproductive potential preservation and infertility treatment. State of the art methods of assisted reproductive technologies include the development of cryopreservation method of gametes and embryos by vitrification, genetic screening of embryos in order to prevent the hereditary diseases transmission and embryo transfer with chromosomal abnormalities, the birth of a child “from three parents” in severe cases of mutations in the mitochondrial genome, etc.

Does every family couple that being treated with assisted reproductive technologies need to use preimplantation genetic testing for aneuploidy (PGT-A)?

Reports of Vinnytsia National Medical University ◽

10.31393/reports-vnmedical-2019-23(2)-25 ◽

2019 ◽

Vol 23 (2) ◽

pp. 316-320

Author(s):

O. L. Lyovkina ◽

S. S. Derii ◽

Y. R. Kuzmenko

Keyword(s):

In Vitro Fertilization ◽

Reproductive Medicine ◽

Literature Search ◽

Current Status ◽

Vitro Fertilization ◽

Further Development

The increase in the frequency of using assisted reproductive technologies (ART) dictates the need for pre-implantation diagnosis of embryos to determine which of them are euploid and recommended for embryo transfer (ET), since the main cause of implantation failures when using in vitro fertilization (IVF) is ET aneuploid or mosaic ETs or mosaic imitations that use in vitro fertilization (IVF) is ET aneuploid or mosaic imitative failures when using in vitro fertilization (IVF) is ET aneuploid or mosaic imitations. . For this purpose, the PGT-A technology was created, since the appearance of which and its further development there have been discussions about the feasibility of using PGT-A for each pair, which turned to the use of ART. The purpose of this literature review is to describe the current status of PGT-A and determine the prospects for its widespread adoption in the practice of reproductive medicine. A literature search was carried out in the PubMed and Cochrane databases for the last 10 years. An analysis of the literature has shown that IVF with PGT-A technology has significant advantages over traditional IVF; The technology has a number of technical and financial limitations, which makes it difficult to massively introduce technology into the practice of reproductive medicine, therefore at this stage there should be clear indications for using IVF with PGT-A.

Revista Brasileira de Ginecologia e Obstetrícia / RBGO Gynecology and Obstetrics ◽

First Baby Born in Brazil after Simultaneous Diagnosis through Non-Invasive and Conventional PGT-A

10.1055/s-0041-1736302 ◽

2021 ◽

Vol 43 (11) ◽

pp. 878-882

Author(s):

Marcos Iuri Roos Kulmann ◽

Márcia Riboldi ◽

Carolina Martello ◽

Adriana Bos-Mikich ◽

Gerta Frantz ◽

...

Keyword(s):

Culture Media ◽

False Negative ◽

Media Culture ◽

Current Application ◽

Vitro Fertilization ◽

Non Invasive ◽

Discordant Diagnosis ◽

History Of

AbstractNon-invasive preimplantation genetic testing for aneuploidies (niPGT-A) aiming to assess cell-free embryonic DNA in spent culture media is promising, especially because it might overcome the diminished rates of implantation caused by the inadequate performance of trophectoderm (TE) biopsy. Our center is part of the largest study to date assessing the concordance between conventional PGT-A and niPGT-A, and we report here the delivery of the first baby born in Brazil using niPGT-A. The parents of the baby were admitted to our center in 2018. They did not present history of infertility, and they were interested in using in vitro fertilization (IVF) and PGT-A in order to avoid congenital anomalies in the offspring. A total of 11 (3 day-5 and 8 day-6) expanded blastocysts were biopsied, and the spent culture media (culture from day-4 to day-6) from 8 day-6 blastocysts were collected for niPGT-A. Overall, 7 embryos yielded informative results for trophectoderm (TE) and media samples. Among the embryos with informative results, 5 presented concordant diagnosis between conventional PGT-A and niPGT-A, and 2 presented discordant diagnosis (1 false-positive and one false-negative). The Blastocyst 4, diagnosed as 46, XY by both niPGT-A and conventional PGT-A, was warmed up and transferred, resulting in the birth of a healthy 3.8 kg boy in February 2020. Based on our results and the recent literature, we believe that the safest current application of niPGT-A would be as a method of embryo selection for patients without an indication for conventional PGT-A. The approximate 80% of reliability of niPGT-A in the diagnosis of ploidy is superior to predictions provided by other non-invasive approaches like morphology and morphokinetics selection.