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 Test-tube embryos - mouse and human development in vitro to blastocyst stage and beyond

2019 ◽  
Vol 63 (3-4-5) ◽  
pp. 203-215 ◽  
Author(s):  
Niraimathi Govindasamy ◽  
Binyamin Duethorn ◽  
Hatice O. Oezgueldez ◽  
Yung S. Kim ◽  
Ivan Bedzhov
Keyword(s):  
Embryo Culture ◽  
Assisted Reproductive Technologies ◽  
Basic Research ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Culture Conditions ◽  
Test Tube ◽  
Human Embryos ◽  
Culture Methods

Mammalian embryogenesis is intrauterine and depends on support from the maternal environment. Therefore, in order to directly study and manipulate early mouse and human embryos, fine-tuned culture conditions have to be provided to maintain embryo growth in vitro. Over time, the establishment and implementation of embryo culture methods have come a long way, initially enabling the development of few pre-implantation stages, expanding later to support in vitro embryogenesis from fertilization until blastocyst and even ex utero development beyond the implantation stages. Designing culture conditions that enable near physiological development of early embryos without maternal input, especially during the peri- and post-implantation stages, requires overcoming numerous experimental challenges, and it is still far from optimal. Nevertheless, embryo culture methods are an essential cornerstone of both assisted reproductive technologies and basic research, and these methods provide a platform to understand life’s greatest miracle – the development of a new organism.

Ethical and Policy Issues in Human Embryo Twinning

1995 ◽  
Vol 4 (3) ◽  
pp. 268-284 ◽  
Author(s):  
Andrea L. Bonnicksen
Keyword(s):  
Human Embryo ◽  
Medical Center ◽  
Reproductive Technologies ◽  
Human Embryos ◽  
George Washington ◽  
Policy Issues ◽  
Washington University ◽  
George Washington University

In 1993, investigators from George Washington University (GWU) Medical Center separated the cells of 17 human embryos and produced 48 embryos, an average of three embryos for each original. The method, variously called twinning, cloning, embryo splitting, and blastomere separation, demonstrated that human embryos could be split to create genetically identical entities during conception. When publicized, however, the experiment brought to mind a different view of cloning repeated since the beginning of the new reproductive technologies. In the early 1970s, when research onin vitrofertilization (IVF) was in its infancy, commentators worried that cloning–defined as the duplication of persons–would be next, leading to a scenario of “boys genetically exactly like the father, girls like the mother, or individuals like some true or false hero of art, science, or sports, or like some demagogue or some saint.”

La fine del concetto di “pre-embrione” nella Convenzione di Oviedo / The end of the concept of “pre-embryo” in the Oviedo Convention

2018 ◽  
Vol 66 (6) ◽  
pp. 735-745
Author(s):  
Carlo Casini ◽  
Marina Casini
Keyword(s):  
Human Embryo ◽  
Legal Status ◽  
Human Embryos ◽  
Specific Reference ◽  
Constitutional Court ◽  
The European Union ◽  
Council Of Europe ◽  
European Court ◽  
The Moment

Il contributo si sofferma sulla questione riguardante la ricerca scientifica sugli embrioni generati in vitro. L’articolo 18 della Convenzione riguarda specificamente la sperimentazione sull’embrione in vitro e per questo esso è sottoposto ad una riflessione particolarmente approfondita. L’obiettivo è quello di capire se dalla Convenzione emergono linee idonee a definire lo statuto giuridico dell’embrione umano. Gli Autori concludono nel senso che nonostante il concetto di pre-embrione (formulato proprio per teorizzare l’insignificanza dell’embrione umano nei primi 14 giorni dalla fecondazione) sia stato accolto in alcune leggi e abbia implicitamente guidato l’interpretazione di alcuni aspetti relativi alla valutazione del valore dell’embrione, la Convenzione di bioetica lo ha definitivamente respinto con il massimo di autorevolezza. La conclusione è raggiunta attraverso l’esame dell’art. 18 considerandone anche la precedente formulazione contenuta in una bozza; mediante una interpretazione sistematica della Convenzione che esige il riconoscimento del concepito, fin dalla fecondazione, come un “essere umano”; esaminando i contributi preparatori elaborati dalla Assemblea Parlamentare del Consiglio d’Europa e del Parlamento Europeo; prendendo in considerazione gli sviluppi della Convenzione di Oviedo con specifico riferimento al tema del pre-embrione. L’indagine si avvale poi anche di ampi riferimenti alla giurisprudenza della Corte europea dei diritti dell’uomo del Consiglio d’Europa, alla giurisprudenza della Corte di Giustizia dell’Unione Europea, ad alcune recenti decisioni della Corte Costituzionale italiana. ---------- The paper focuses on the question concerning scientific research on human embryos generated in vitro. Article 18 of the Oviedo Convention specifically concerns the experimentation on the in vitro embryos and for this reason it is subject to a particularly in-depth reflection. The goal is to understand if the Convention shows suitable lines to define the legal status of the human embryo. The authors conclude that despite the concept of pre-embryo (formulated to theorize the insignificance of the human embryo in the first 14 days of fertilization) has been accepted in some laws and has implicitly guided the interpretation of some aspects related to the evaluation of the value of the embryo, the Bioethics Convention definitively rejected it with the utmost authority. The conclusion is reached through the examination of the art. 18 also considering the previous formulation contained in a draft; through a systematic interpretation of the Convention which requires the recognition of the conceived, from the moment of fertilization, as a “human being”; examining the preparatory contributions prepared by the Parliamentary Assembly of the Council of Europe and the European Parliament; taking into consideration the developments of the Oviedo Convention with specific reference to the theme of the pre-embryo. The investigation also makes use of extensive references to the jurisprudence of the European Court of Human Rights of the Council of Europe, to the jurisprudence of the Court of Justice of the European Union, to some recent decisions of the Italian Constitutional Court.

246 IN VITRO PRODUCTION OF SPRINGBOK (ANTIDORCAS MARSUPIALIS) EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 239 ◽  
Author(s):  
R. Krisher ◽  
A. Auer ◽  
K. Clark ◽  
K. Emsweller ◽  
S. Rogers ◽  
...  
Keyword(s):  
South Africa ◽  
Oocyte Maturation ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Genetic Management ◽  
Blastocyst Development ◽  
Antidorcas Marsupialis

The objective of this experiment was to develop in vitro embryo production (IVP) technologies in springbok (Antidorcas marsupialis), a southern African antelope. Springbok, a fairly common species on game farms in parts of South Africa, may be used as a model species for gamete rescue and IVP techniques to be applied to the conservation of other threatened antelope species. Springbok belong to the family bovidae, subfamily antilopinae, tribe antilopini, which comprises about twenty species in genera Gazella, Antilope, Procapra, Antidorcas, Litocranius, and Ammodorcas. In this tribe alone, there are 4 species or subspecies that are critically endangered, 3 that are endangered, and 10 that are considered vulnerable, demonstrating the need for antelope conservation efforts. In addition, our studies contributed to the South African biological resource bank, so that banked springbok semen and embryos might be used in the future for managed genetic contribution to isolated captive or wild populations via assisted reproductive technologies. Oocytes were recovered (3 replicates) from ovaries obtained at supervised culls for management purposes in South Africa, and cultured in defined Gmat or undefined TCM-199 with FCS maturation medium for 28-30 h (Brad et al. 2004 Reprod. Fertil. Dev. 16, 223). Oocytes were fertilized with frozen-thawed springbok epididymal spermatozoa in modified SOF fertilization medium with caffeine (Herrick et al. 2004 Biol. Reprod. 71, 948–958). Eighteen hours after insemination, a randomly selected subset of the zygotes were fixed to determine fertilization success. The remaining zygotes were cultured in G1/G2 media. On Day 7 of culture, embryos were analyzed for development to the morula or blastocyst stage. A total of 259 selected oocytes were collected from 50 females (5.2 selected oocytes/female on average). There was no difference in the percentage of oocytes normally fertilized (2 pronuclei, PN) between oocytes matured in Gmat (n= 43; 12%) and those matured in TCM-199 (n= 42; 10%). There were significantly (P < 0.05) more oocytes penetrated (e2 PN) when matured in TCM (50%) compared to Gmat (23%). There were no differences in embryonic cleavage or morula/blastocyst development (of total oocytes inseminated) between treatments (Gmat,n= 89, 54%, 9.0%; TCM-199, n= 85, 68%, 9.4%, respectively). In both treatments, the average blastocyst grade was 2.125 using the standard bovine grading system (Curtis, Cattle Embryo Transfer Procedure, 1991). In conclusion, in vitro oocyte maturation, fertilization, and embryo culture to the blastocyst stage is possible in springbok. Importantly, blastocysts can be produced in vitro under semi-defined conditions, demonstrating that oocyte maturation without serum does support developmental competence. This is important for the potential international movement of IVP embryos to be used for genetic management in the conservation of antelope species.

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

2021 ◽  
pp. 539-543
Author(s):  
Daria A. Kinsht ◽  
◽  
Mariia K. Soboleva ◽  
Keyword(s):  
Reproductive Health ◽  
Assisted Reproductive Technologies ◽  
Medical Center ◽  
Newborn Infants ◽  
Uterine Cavity ◽  
Perinatal Outcomes ◽  
Reproductive Technologies ◽  
Singleton Pregnancy ◽  
Vitro Fertilization

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.

71 The Effect of Two Different In Vitro Culture Media and Mice Embryo Groupings on Hatchability After 24 Hours of Culture

2018 ◽  
Vol 30 (1) ◽  
pp. 174
Author(s):  
N. C. Negota ◽  
M. L. Mphaphathi ◽  
L. P. Nethenzheni ◽  
T. L. Rammutla ◽  
N. R. Serota ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Assisted Reproductive Technologies ◽  
Culture Media ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Hatching Rate ◽  
Significant Difference ◽  
Autocrine Signalling ◽  
Blastocyst Hatching

Mammalian blastocysts must hatch out from the zona pellucida before implantation. In vitro embryo culture and grouping of mice blastocysts are conducive options of assisted reproductive technologies (ART) to speed up the hatching rate of mice embryos. The number of embryos per unit volume has the greatest impact on hatching rates due to autocrine signalling. The study aimed to determine the effect of two in vitro culture (IVC) media (TCM-199 and Ham’s F10) and embryo groupings (1, 2, 3, and 4 embryos per 50-µL droplet) after 24 h of culture on hatching rate. Breeds of C57BL/6 (n = 10) and BALB/c (n = 10) were raised until they reached maturity and bred naturally to produce the first filial generation. The photoperiod was 14 h of light followed by 10 h of darkness in the breeding house, and feed and water were provided ad libitum. Female mice were superovulated using eCG and hCG. The first filial generations from 2 breeds were used for the collection of 160 blastocysts and randomly allocated into 2 IVC media (80 embryos for TCM-199 and 80 embryos for Ham’s F10) and again subjected to 4 embryo groupings (1, 2, 3, and 4 embryos per droplet) treatments. Four replicates were done per treatment group. The general linear model of Minitab version 17 (Minitab Inc., State College, PA, USA) was used to analyse the data. The hatching rate of blastocyst stage was significantly higher for TCM-199 (56.9 ± 27.2) compared with Ham’s F10 (50.0 ± 35.1%). The comparison of all embryo groupings, 1 (20.0 ± 40.5), 2 (28.8 ± 29.7), 3 (59.1 ± 38.8), and 4 (43.8 ± 32.4%) per 50-µL droplet showed significant differences, irrespective of IVC medium and breed. In TCM-199, groupings of 1 (20.0 ± 41.0), 2 (30.0 ± 29.9), 3 (63.3 ± 40.3), and 4 (42.5 ± 33.5%) had a significant difference on blastocyst hatching percent. In Ham’s F10, groupings of 1 (20.0 ± 41.0), 2 (27.5 ± 30.2), 3 (55.0 ± 37.9), and 4 (45.0 ± 32.0%) were significantly different on blastocyst hatching rate. However, an increase in hatching rate was observed for the interaction of media and embryo groupings and especially when embryos were increased per droplet in all breeds. In conclusion, the use of TCM-199 and grouping of 3 embryos per 50-µL droplet during culture had the highest hatching rate compared with the use of Ham’s F10.

Implications of Assisted Reproductive Technologies for Pregnancy Outcomes in Mammals

2020 ◽  
Vol 8 (1) ◽  
pp. 395-413 ◽  
Author(s):  
Peter J. Hansen
Keyword(s):  
Assisted Reproductive Technologies ◽  
Genetic Manipulation ◽  
Adult Life ◽  
Reproductive Technologies ◽  
Fertilization Rate ◽  
Blastocyst Stage ◽  
In Vitro Production ◽  
Long Distance ◽  
Vitro Production

Development of assisted reproductive technologies has been driven by the goals of reducing the incidence of infertility, increasing the number of offspring from genetically elite animals, facilitating genetic manipulation, aiding preservation and long-distance movement of germplasm, and generating research material. Superovulation is associated with reduced fertilization rate and alterations in endometrial function. In vitro production of embryos can have a variety of consequences. Most embryos produced in vitro are capable of establishing pregnancy and developing into healthy neonatal animals. However, in vitro production is associated with reduced ability to develop to the blastocyst stage, increased incidence of failure to establish pregnancy, placental dysfunction, and altered fetal development. Changes in the developmental program mean that some consequences of being produced in vitro can extend into adult life. Reduced competence of the embryo produced in vitro to develop to the blastocyst stage is caused largely by disruption of events during oocyte maturation and fertilization. Conditions during embryo culture can affect embryo freezability and competence to establish pregnancy after transfer. Culture conditions, including actions of embryokines, can also affect the postnatal phenotype of the resultant progeny.

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 Invisible Vulnerabilities: Ethical, Practical and Methodological Dilemmas in Conducting Qualitative Research on the Interaction with IVF Embryos

Societies ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 7
Author(s):  
Catarina Delaunay ◽  
Amélia Augusto ◽  
Mário Santos
Keyword(s):  
Human Embryo ◽  
Assisted Reproductive Technologies ◽  
Social Stigma ◽  
Single Mother ◽  
Reproductive Technologies ◽  
Point Of View ◽  
Research Experience ◽  
Sensitive Topics ◽  
The Social

The burden of deciding the fate of the supernumerary human embryo created in vitro in the context of Assisted Reproductive Technologies rests on the beneficiary couples or individuals who conceived the parental project. The beneficiaries must also take on the responsibility of choosing whether to donate surplus embryos either to others or to scientific research, or to request their destruction. Vulnerable beings, weakened from the point of view of their identity (facing the social stigma still associated with some circumstances such as being infertile, lesbian or a single mother), are required to have skills such as reflexivity and autonomy in dramatic situations that concern their relationship with their own reproductive body. Given the urgency of this issue at the socio-anthropological level, we are conducting ethnographic research aimed at analysing how specialists and lay people objectivate, evaluate and circulate different conceptions of the human embryo in vitro. Based on our research experience within this ongoing project, we intend to discuss some ethical, practical and methodological concerns for the researcher in accessing the field and conducting fieldwork. We take into account the fact that this research is focused on sensitive topics and on individuals who can be considered people in vulnerable situations.

scholarly journals Quadrupling efficiency in production of genetically modified pigs through improved oocyte maturation

2017 ◽  
Vol 114 (29) ◽  
pp. E5796-E5804 ◽  
Author(s):  
Ye Yuan ◽  
Lee D. Spate ◽  
Bethany K. Redel ◽  
Yuchen Tian ◽  
Jie Zhou ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Reproductive Technologies ◽  
Oocyte Quality ◽  
Blastocyst Stage ◽  
Vitro Fertilization ◽  
Fourfold Increase

Assisted reproductive technologies in all mammals are critically dependent on the quality of the oocytes used to produce embryos. For reasons not fully clear, oocytes matured in vitro tend to be much less competent to become fertilized, advance to the blastocyst stage, and give rise to live young than their in vivo-produced counterparts, particularly if they are derived from immature females. Here we show that a chemically defined maturation medium supplemented with three cytokines (FGF2, LIF, and IGF1) in combination, so-called “FLI medium,” improves nuclear maturation of oocytes in cumulus–oocyte complexes derived from immature pig ovaries and provides a twofold increase in the efficiency of blastocyst production after in vitro fertilization. Transfer of such blastocysts to recipient females doubles mean litter size to about nine piglets per litter. Maturation of oocytes in FLI medium, therefore, effectively provides a fourfold increase in piglets born per oocyte collected. As they progress in culture, the FLI-matured cumulus–oocyte complexes display distinctly different kinetics of MAPK activation in the cumulus cells, much increased cumulus cell expansion, and an accelerated severance of cytoplasmic projections between the cumulus cells outside the zona pellucida and the oocyte within. These events likely underpin the improvement in oocyte quality achieved by using the FLI medium.

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