WO2017180021A1 - Méthode de détermination in vitro d'embryons prometteurs pour leur implantation ultérieure dans l'utérus dans le cadre d'une procédure de fécondation in vitro (fiv) - Google Patents

Méthode de détermination in vitro d'embryons prometteurs pour leur implantation ultérieure dans l'utérus dans le cadre d'une procédure de fécondation in vitro (fiv) Download PDF

Info

Publication number
WO2017180021A1
WO2017180021A1 PCT/RU2017/000015 RU2017000015W WO2017180021A1 WO 2017180021 A1 WO2017180021 A1 WO 2017180021A1 RU 2017000015 W RU2017000015 W RU 2017000015W WO 2017180021 A1 WO2017180021 A1 WO 2017180021A1
Authority
WO
WIPO (PCT)
Prior art keywords
embryos
meridional
equatorial
ivf
cleavage
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/RU2017/000015
Other languages
English (en)
Russian (ru)
Inventor
Илья Викторович СЕНЕЧКИН
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Publication of WO2017180021A1 publication Critical patent/WO2017180021A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Instruments for taking body samples for diagnostic purposes; Other methods or instruments for diagnosis, e.g. for vaccination diagnosis, sex determination or ovulation-period determination; Throat striking implements
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods
    • A61B17/42—Gynaecological or obstetrical instruments or methods
    • A61B17/425—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation
    • A61B17/435—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation for embryo or ova transplantation

Definitions

  • One of the ways to further increase the effectiveness of IVF programs may be to develop methods of objective control over the selection of the most complete and promising embryos for further implantation in the uterus. Early detection of the most promising embryos for implantation is crucial, since the transfer of early (crushing) embryos to the mother's body increases the frequency of implantation and pregnancy.
  • blastomeres As is known, after 24-36 hours after fertilization, the first division of the zygote occurs and from that moment the fertilized oocyte becomes a 2-cell embryo. The cells of the embryo at this stage are called blastomeres.
  • the human embryo On the 2nd day of development, the human embryo consists of 2, 3 or 4 blastomeres.
  • the embryo After another day, the embryo normally already consists of 6-8 blastomeres, but 4 blastomeres are allowed, if on the 2nd day the embryo was 2-cell. Up to 8-20 cell stages, all cells of a human embryo are totipotent, i.e. each of them can give rise to a whole organism.
  • the human embryo On the 4th day of development, the human embryo already consists, as a rule, of 8-16 cells, the morula stage begins. It is at this stage in vivo that the embryo enters from the fallopian tube into the uterine cavity. Morules also differ among themselves 25 in the degree of compaction of the blastomeres (C1 - C2 - C3 - C4).
  • blastocyst From the moment the cavity inside the morula reaches 50% of its volume, the embryo is called a blastocyst. The larger the blastocyst cavity and the better the internal cell mass and trophoblast are developed, the greater its potential for implantation. At this stage, one can also assess the quality of the embryo by the degree of deformation, vacuolization, fragmentation. In addition, the shape and relative sizes of the blastomeres are evaluated.
  • a two-letter designation is used - one for trophoblast, the other for embryoblast.
  • the embryo begins to increase in size.
  • blastomeres continue to divide.
  • the glossy shell thinned by stretching breaks and the process of hatching (hatching) the embryo from the glossy shell begins.
  • Known methods are those in which promising embryos are selected on the basis of division time [2], expression of one or more miRNAs by the embryo [3], based on measurement of the expression level of 45 genes, as a result of which the rate of subsequent implantation is predicted [7], based on the number and size of blastomeres on the second or third day, as well as by the percentage of fragmentation and the presence of multinucleation at the stage of 4 or 8 cells of zo [8].
  • s The closest (prototype) to the proposed method we consider a method for determining in vitro promising embryos for subsequent implantation in the uterus during the in vitro fertilization (IVF) procedure based on determining the spatial distribution of blastomeres (J Cell Sci.
  • FIG. 1 The formation of a universal tetrahedral form of 4-cell embryos as a result of successive equatorial (A), meridional (B), and meridional and equatorial (C) divisions in the second fragmentation cycle is shown.
  • the equatorial and meridional division variant (not shown in the figure) differs from the ME variant only in the division sequence.
  • the time after intracellular sperm injection (h: min) is indicated in the image field.
  • An asterisk indicates the second polar body.
  • Figure 2 shows the development paths of individual nuclei (gray lines), grouped in accordance with the order and orientation of the fission planes in the second crushing cycle: successive equatorial and meridional (A and B), meridional and equatorial (B) and equatorial and meridional (G) division of the blastomeres of 2-cell embryos.
  • Solid contrast lines with markers are averaged development paths.
  • the ellipses in fig. B and D indicate the grouping of the trajectories of individual embryos in accordance with the development rate: 1 - high, 2 - low, 3 - intermediate. 5
  • the horizontal axis is the number of blastomeres in the embryo (the order of appearance of the blastomeres); vertical axis - post-ICSI time., hours
  • embryos with the EE variant of the second division of fragmentation should be considered the least promising for transfer and, at a minimum, should be the subject of special attention of the embryologist.
  • EE embryos are characterized by a smoothed “wavy” temporal development trajectory.
  • b “Step-like” trajectory (Fig. 2).
  • the averaged time trajectories differ among themselves (the Wilcoxon paired criterion; the significance of the differences (P) between the EE and ⁇ -, EE- and ⁇ -, MM- and ⁇ -, MM- and EM-options are 0.001 ; the significance of the differences between EE and EM is 0.002; between ME and EM 5 options is 0.023).
  • embryos with different variants of the second division of crushing can be ranked as follows: MM, EM, ME, EE.
  • ME- and EM-embryos can be divided into three groups in accordance with the similarity of the trajectories (Fig. 2, C, D).
  • Embryos of the second group are characterized by longer cycle times.
  • the crushing cycles of the embryos of the third groups can be comparable or exceed the duration of the corresponding cycles of the embryos from the first groups, and in some cases the cycles of embryos from the second groups (see details in Table 2).
  • the first groups unite the embryos that formed developed blastocysts (grades 4 and 5).
  • the second groups include slowly developing embryos that have reached the morula stage or begin to cavitate the blastocyst.
  • the third groups are heterogeneous, since by the end of the observations, the embryos that make them up show the entire spectrum of blastocyst gradations, although the majority of blastocysts of the 2nd and 3rd gradations (Table 2).
  • the technical result of the invention consists in the earliest and most accurate determination of the highest quality, and, accordingly, the most promising for the formation of implantation-competent blastocysts for subsequent implantation into the uterus under the IVF protocol due to the fact that the embryos with the second crushing divisions in the planes: the equatorial then the meridional (EM) or sequential meridional (MM) are least prone to organic disturbances, and, therefore, will allow you to get your desired pregnancy and subsequently lead to the birth of a healthy baby.
  • EM meridional
  • MM sequential meridional
  • Example 1 Patient Mr., was admitted to the Center for Human Reproduction in order to conduct her IVF and PE program. The patient complains of infertility for 11 years. From the anamnesis it is known that the patient in 1989 underwent surgery for PCOS (no effect), and in 1998, laparoscopy for adhesions of the pelvic organs 3-4 degrees, bilateral salpingitis. Despite the scope of the operation - the separation of adhesions of the pelvic organs and the abdominal cavity, salpingo-ovariolysis, it was not possible to restore the patency of the fallopian tubes.
  • hypothalamic syndrome An additional examination by an endocrinologist revealed: hypothalamic syndrome; hypofunction of the ovaries; galactorrhea. Correction of endocrine disorders.
  • the time of administration of CG was determined depending on the time the lead follicle reached a diameter of at least 18 mm or 2-3 follicles not less than the ovulatory dose of Pretil was 10 thousand units.
  • Aspiration of oocytes and follicular fluid is performed 36 hours after the administration of an ovulatory dose of CG separately from the right and left ovary.
  • transvaginal puncture is performed using a vaginal puncture probe with a frequency of 5 MHz ultrasound apparatus Aloka SSD-500.
  • the patient underwent follicular puncture and aspiration of preovulatory oocytes and follicular fluid. With follicular puncture from the left ovary, 3 oocytes were obtained, from the right ovary 1.
  • the obtained preovulatory oocytes from the right and left ovaries 15 are cultured in special thermostats at a temperature of 37 degrees Celsius with a mixture of 5% CO2 in air in a culture medium Menezo B2.
  • proteolytic enzymes In follicular fluid aspirated at the time of puncture of the right and left ovaries, the activity of proteolytic enzymes (acid peptide hydrolases) is determined by the increase in acid-soluble folin-positive 20 compounds.
  • the substrate for determining the activity of enzymes is lyophilized hemoglobin.
  • Follicular fluid is poured into 2 test tubes of 1 ml each (control and experimental) and 0.5 ml of a 1% solution of lyophilized commercial hemoglobin prepared in acetate buffer (0.2 M SNZ) are added to them.
  • a working solution of the reagent which is a mixture of 0.25 L of a 2% solution of sodium carbonate in a 0.1 M solution of sodium hydroxide and 0.01 L of a 0.5% solution of copper sulfate in a 1% solution of sodium citrate (this mixture is prepared before analysis).
  • the measurement of the optical test sample is carried out against the control sample in a cell with a layer thickness of 1 cm at a wavelength of 750 nm on a spectrophotometer or photoelectric colorimeter.
  • the sperm is subjected to flotation and centrifugation, after which its fertility is assessed and oocytes are inseminated.
  • 2 cells determined the sequence of the second divisions of crushing.
  • the sequence of the second cleavage divisions in the planes of two embryos corresponded (EE), one embryo (EM).
  • Patient K entered the Center for Human Reproduction with the aim of conducting an IVF and PE program.
  • the patient complained of infertility for 19 years. From the anamnesis it is known that the patient in 1988 underwent a zaparoscopic operation that did not reveal pathology from the uterine pipes. Considering the data of laparoscopy and the ovarian hypofunction revealed during the examination (hyperestrogenism), a negative PCT, a patient after preliminary hormonal correction, 5 attempts of IISM were made (without effect). The lack of effect of the therapy required further in-depth examination. With repeated laparoscopy in 1998, the diagnosis was established: grade 2 endometriosis. Coagulation of endometrioid heterotopies was carried out, Danoval treatment was carried out for 1/2 year. In the absence of the effect of the therapy, the patient is recommended a program of IVF and PE.
  • MC Superovulation was stimulated by 0.1 mg Dekapeptil from the 2nd to the 10th day of the menstrual cycle, Humegon 75 U + 75 U 24 ampoules from the 2nd to 7th day of MC. Upon reaching maturity by follicles on day 10, MC was assigned Pretil 10,000 units. In an ultrasound study, at the time of follicular puncture, 4 leading follicles were registered in the left ovary and 3 in the right. The thickness of the endometrium was 11 mm.
  • the patient underwent follicular puncture and aspiration of preovulatory oocytes and follicular fluid. With follicular puncture from the left ovary, 3 oocytes were obtained, from the right ovary 1.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Surgery (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Pregnancy & Childbirth (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Urology & Nephrology (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Transplantation (AREA)
  • Gynecology & Obstetrics (AREA)
  • Food Science & Technology (AREA)
  • Reproductive Health (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hematology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Medical Informatics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne le domaine de la médecine humaine et notamment de la gynécologie et de la médecine reproductive et peut être utilisée dans des programmes de fécondation in vitro (FIV) et de transfert d'embryons (TE) à des fins de pronostic de leurs issues. A cet effet, on détermine au moyen d'une vidée ultra-accélérée la présence d'un plan de fractionnement chez les embryons au stade de fractionnement de 2-èmes cellules: horizontal (équatorial - E) ou vertical (méridional - M). On détermine la séquence des deuxièmes divisions de fractionnement dans ces plans qui peuvent être : Séquentiel équatorial (EE), séquentiel méridional (VM), méridional puis équatorial (ME), équatorial puis méridional (EM). Dans le cas où la séquence des deuxièmes divisions de fractionnement dans ces plans correspond à (EM) ou à (MM), ces embryons sont considérés comme prometteurs pour une implantation subséquente dans l'utérus dans le cadre d'une procédure FIV. L'invention permet la détermination la plus précoce et la plus précise des blastocystes aptes à l'implantation de qualité supérieure qui sont, de ce fait, les plus prometteurs en termes de formation à des fins de leurs implantation ultérieure dans l'utérus dans le cadre d'un protocole FIV grâce au fait que les embryons dans les séquences des deuxièmes divisions dans les plans suivants - équatorial puis méridional (EM) ou méridional séquentiel(MM) - présentent moins de risques de dysfonctionnements organiques et permettent, de ce fait, d'obtenir une grossesse désirée puis la naissance d'un enfant en bonne santé.
PCT/RU2017/000015 2016-04-11 2017-01-13 Méthode de détermination in vitro d'embryons prometteurs pour leur implantation ultérieure dans l'utérus dans le cadre d'une procédure de fécondation in vitro (fiv) Ceased WO2017180021A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
RU2016113577 2016-04-11
RU2016113577A RU2625777C1 (ru) 2016-04-11 2016-04-11 Способ определения in vitro перспективных эмбрионов для последующей имплантации в матку при проведении процедуры экстракорпорального оплодотворения (эко)

Publications (1)

Publication Number Publication Date
WO2017180021A1 true WO2017180021A1 (fr) 2017-10-19

Family

ID=59495261

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/RU2017/000015 Ceased WO2017180021A1 (fr) 2016-04-11 2017-01-13 Méthode de détermination in vitro d'embryons prometteurs pour leur implantation ultérieure dans l'utérus dans le cadre d'une procédure de fécondation in vitro (fiv)

Country Status (2)

Country Link
RU (1) RU2625777C1 (fr)
WO (1) WO2017180021A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2281777C1 (ru) * 2005-04-21 2006-08-20 Мария Львовна Семенова Способ управления качеством ооцитов и композиция для добавления в среду культивирования ооцитов
RU2474823C1 (ru) * 2011-09-26 2013-02-10 Федеральное государственное бюджетное учреждение "Ивановский научно-исследовательский институт материнства и детства имени В.Н. Городкова" Министерства здравоохранения и социального развития Российской Федерации Способ прогнозирования качества эмбрионов в программе экстракорпорального оплодотворения
WO2013181549A2 (fr) * 2012-05-31 2013-12-05 Auxogyn, Inc. Procédés de prédiction de blastocyste embryonnaire in vitro
US8754014B2 (en) * 2009-04-17 2014-06-17 Institut National De La Sante Et De La Recherche Medical (Inserm) Methods for selecting oocytes and competent embryos with high potential for pregnancy outcome

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2281777C1 (ru) * 2005-04-21 2006-08-20 Мария Львовна Семенова Способ управления качеством ооцитов и композиция для добавления в среду культивирования ооцитов
US8754014B2 (en) * 2009-04-17 2014-06-17 Institut National De La Sante Et De La Recherche Medical (Inserm) Methods for selecting oocytes and competent embryos with high potential for pregnancy outcome
RU2474823C1 (ru) * 2011-09-26 2013-02-10 Федеральное государственное бюджетное учреждение "Ивановский научно-исследовательский институт материнства и детства имени В.Н. Городкова" Министерства здравоохранения и социального развития Российской Федерации Способ прогнозирования качества эмбрионов в программе экстракорпорального оплодотворения
WO2013181549A2 (fr) * 2012-05-31 2013-12-05 Auxogyn, Inc. Procédés de prédiction de blastocyste embryonnaire in vitro

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PLUS A BERENIKA ET AL.: "Downregulation of Par3 and aPKC function directs cells towards the ICM in the preimplantation mouse embryo", JOURNAL OF CELL SCIENCE, vol. 8, 11 February 2005 (2005-02-11), pages 505 - 515 *

Also Published As

Publication number Publication date
RU2625777C1 (ru) 2017-07-18

Similar Documents

Publication Publication Date Title
Ebner et al. Selection based on morphological assessment of oocytes and embryos at different stages of preimplantation development: a review
Parrella et al. A treatment approach for couples with disrupted sperm DNA integrity and recurrent ART failure
Tšuiko et al. Genome stability of bovine in vivo-conceived cleavage-stage embryos is higher compared to in vitro-produced embryos
Kirkegaard et al. A randomized clinical trial comparing embryo culture in a conventional incubator with a time-lapse incubator
Debrock et al. Preimplantation genetic screening for aneuploidy of embryos after in vitro fertilization in women aged at least 35 years: a prospective randomized trial
EP2855694B1 (fr) Procédés de prédiction de blastocyste embryonnaire in vitro
Shu et al. Fertilization, embryo development, and clinical outcome of immature oocytes from stimulated intracytoplasmic sperm injection cycles
Ebner et al. Prognosis of oocytes showing aggregation of smooth endoplasmic reticulum
Magli et al. Atlas of human embryology: from oocytes to preimplantation embryos
Simopoulou et al. Making ICSI safer and more effective: a review of the human oocyte and ICSI practice
Ebner et al. Blood clots in the cumulus–oocyte complex predict poor oocyte quality and post-fertilization development
De Vincentiis et al. Use of metaphase I oocytes matured in vitro is associated with embryo multinucleation
Kim et al. Successful pregnancy and delivery after ICSI with artificial oocyte activation by calcium ionophore in in-vitro matured oocytes: a case report
Ebner et al. Time-lapse imaging of cytoplasmic strings at the blastocyst stage suggests their association with spontaneous blastocoel collapse
RU2625777C1 (ru) Способ определения in vitro перспективных эмбрионов для последующей имплантации в матку при проведении процедуры экстракорпорального оплодотворения (эко)
RU2593740C1 (ru) Способ регулирования степени зиготности беременности в программах экстракорпорального оплодотворения и переноса эмбрионов
Ahart et al. To freeze or not to freeze, what is the answer?
MacKenna et al. Embryo early cleavage, number of blastomeres and morphology at day three as factors to predict blastocyst development
Veeck et al. Human blastocysts in vitro
CN114752552B (zh) 冷冻胚胎的复苏、培养、优选和单囊胚移植方法以及用于复苏冷冻胚胎的无创检测方法
Takashima et al. Comparison of embryo development and pregnancy rates in continuous single and sequential media cultures of sibling embryos
Zarifovna et al. Impact of Frame-By-Frame Monitoring of Embryos on Obstetric And Perinaaaaatal Outcomes In Singleton Pregnancies In Assisted Reproductive Technology Programs
Hajder et al. Polar Body Biopsy Helps in Reducing the Total Pregnancy Loss Rates in Intracytoplasmic Sperm Injection Cycles
Kirienko et al. Cytoplasmic fragmentation of human preimplantation embryos
Jones et al. Embryo culture, assessment, selection and transfer

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17782731

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 17782731

Country of ref document: EP

Kind code of ref document: A1