WO2022112460A1 - Système et dispositif pour caractériser des changements dans le processus de maturation d'un ovocyte, et leur utilisation - Google Patents

Système et dispositif pour caractériser des changements dans le processus de maturation d'un ovocyte, et leur utilisation Download PDF

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Publication number
WO2022112460A1
WO2022112460A1 PCT/EP2021/083078 EP2021083078W WO2022112460A1 WO 2022112460 A1 WO2022112460 A1 WO 2022112460A1 EP 2021083078 W EP2021083078 W EP 2021083078W WO 2022112460 A1 WO2022112460 A1 WO 2022112460A1
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WIPO (PCT)
Prior art keywords
electrodes
depression
egg cell
complex resistance
oocyte
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PCT/EP2021/083078
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German (de)
English (en)
Inventor
Uwe Schnakenberg
Wilhelm Jahnen-Dechent
Carlo SCHMITZ
Julia FLOEHR
Akram EL HASNI
Tom KREMERS
Yuan Cao
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Rheinisch-Westfälische Technische Hochschule (Rwth) Aachen
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Publication of WO2022112460A1 publication Critical patent/WO2022112460A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/16—Microfluidic devices; Capillary tubes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion

Definitions

  • the invention relates to a device for characterizing changes in the maturation process of an egg cell.
  • the invention also relates to a system with the inventive device and its use.
  • the artificial insemination of egg cells is an established method in reproductive medicine.
  • ZP zona pellucida
  • the oocyte and embryo quality is typically assessed through a microscopic control.
  • the microscopic control is strongly dependent on the experience and the ability and knowledge of the assessing person, ie the assessment is highly subjective.
  • the morphological assessment of the oocyte immediately before fertilization is highly subjective and can only be performed by experienced embryologists. This is described, for example, in the article "The Istanbul consensus workshop on embryo assessment", in Human Reproduction, Vol.26, No.6 pp. 1270-1283, 2011.
  • the quality is generally only assessed again 18 hours after fertilization.
  • a microfluidic device for receiving cells is known from the prior art according to WO2012/094956 A1, with the aid of which it should be possible to determine the cell size.
  • time-lapse procedure assesses the embryo, i.e. the egg cell, only after fertilization and can therefore only be used to a limited extent for ethical reasons under German legislation. In addition, this is a very expensive process, so it is rarely used.
  • PES pre-implantation genetic screening
  • embryo protection laws apply. According to a conservative interpretation, for ethical reasons only the egg cells in the pronuclear stage (18 hours after fertilization) can be evaluated and selected. This means that at this point in time a decision must already be made as to which egg cells (usually 2 - 3) are to be cultivated further up to day 3 - 5 and finally transferred back into the uterus (usually 1 - 2 embryos). This is to avoid storing embryos.
  • the other oocytes (on average 5-10 oocytes are obtained per woman after hormonal stimulation) are frozen in the pronuclear stage.
  • the process of vitrification involves risks that reduce the quality of the oocytes (not all oocytes that were of good quality before vitrification still have them afterwards). Reliable quality control of the fertilized egg cell would therefore be particularly important.
  • the aim here is to transfer only one embryo, namely the one with the best chance of a successful pregnancy and to avoid the risks of multiple pregnancies.
  • the remaining embryos are frozen or discarded. According to the Embryo Protection Act, this is not permitted in Germany.
  • the great divergence of the legal framework conditions shows the enormous importance of reliable egg cell and embryo control.
  • the invention is explained in more detail below with reference to a drawing and exemplary embodiments.
  • the drawing is a schematic representation and not to scale. The drawing does not limit the invention in any way.
  • Fig. 1 shows the relationship between zona pellucida hardness and oocyte or embryo development over time
  • Figures 2a and 2b show a schematic view of a device according to embodiments of the invention
  • FIG. 3a, 3b & 3c each a schematic view of a device according to embodiments of the invention with an oocyte
  • FIG. 4 a further schematic view with further aspects of a device according to embodiments of the invention
  • FIG. 5 is a further schematic view showing further aspects of a device according to embodiments of the invention with an oocyte.
  • a device according to embodiments of the invention with an oocyte.
  • FIG. 5 is a further schematic view showing further aspects of a device according to embodiments of the invention with an oocyte.
  • Detailed presentation of the invention In the following, the invention is presented in more detail with reference to the figures. It should be noted that different aspects are described, which can be used individually or in combination. That is, each aspect can be used with different embodiments of the invention, unless explicitly presented as a pure alternative.
  • Figure 1 shows the relationship between zona pellucida hardness and oocyte or embryo development over time.
  • time axis reference is made to the state before fertilization as well as after fertilization and the cell division taking place up to the so-called blastocyst.
  • ovum is understood to mean not only the ovum per se, but also the maturation of an embryo up to the blastocyst stage, but in particular up to the morula, and further in particular up to the multinucleate stage.
  • the invention describes a device for evaluating egg cells Ez used for artificial insemination.
  • the device for characterizing changes in the maturation process of an egg cell Ez has a substrate S. At least one recess V is provided on the substrate S for receiving an egg cell Ez. Such devices are shown schematically in Figures 2a, 3a and 4.
  • At least two electrodes are provided in the edge regions of the recess V.
  • the edge area is also to be understood as lying in the depression V.
  • Electrodes El, E2 are provided in Figures 2a, 2b, 3a and 3b, and four electrodes Ela, Elb, E2a, E2b are provided in Figures 4 and 5.
  • the exact number/arrangement and connection of the electrodes can be suitably selected by a person skilled in the art.
  • the electrodes are preferably arranged in pairs symmetrically to one another.
  • the electrodes preferably have a similar design in the edge region of the depression V, at least with respect to the depression V.
  • the electrodes can be suitably contacted for electrical control and measurement. In the illustrations, this is provided via the contact surface that is led outwards. Even if the feed to the contact surfaces and the contact surfaces themselves are shown as being of the same type, this is not a necessity for the functioning of the invention.
  • a complex resistance Z of the egg cell can be measured during operation, with a first complex resistance Zi being able to be measured at a first point in time ti and with a second complex resistance Z 2 being measured at a second point in time t 2 , which follows at the first time ti can be measured.
  • the degree of maturity or the progress of the cell division of the egg cell Ez can be inferred from the deviation of the real and/or imaginary part and/or amount of the measured first complex resistance Zi and second complex resistance Z 2 .
  • the real part can change differently from the imaginary part, so that the degree of maturity or the progress of cell division of the egg cell Ez can be inferred from the qualitative and/or quantitative change in the real part and/or imaginary part and/or the amount for the respective type of egg cell .
  • Measurements of the complex resistance can be made periodically, for example, based on a previously measured complex resistance or the change in previous complex resistances. This can be dependent on the use of the device, for example.
  • the electrodes can be connected to an impedance spectrometer, an LCR measuring bridge or a vector network analyzer, for example, in order to measure the complex electrical resistance as a function of the applied frequency - also referred to as electrical impedance.
  • EIS electrical impedance spectroscopy
  • DS dielectric spectroscopy
  • Measurements over a large frequency range can be used to study impedance changes caused by changes in the ZP glycoprotein matrix as well as electrophysiological changes in the cytoplasm.
  • the device combines ease of use, miniaturization, integration and portability.
  • EIS also allows continuous and non-destructive live observation and characterization of maturing cells and developing embryos.
  • the artificial insemination of egg cells is an established method of assisted reproduction (ART) in humans and animals.
  • Artificial insemination means in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). It is routinely used when natural mating fails, or when livestock and research mammals are to be cloned and reproduced.
  • Artificial insemination is critically dependent on the quality of the oocytes, particularly the condition of the ZP, a gelatinous outer layer of extracellular matrix that spontaneously hardens and becomes impenetrable to sperm during oocyte isolation and culture.
  • ZP2 zona pellucida protein 2
  • oocyte electrophysiology is induced by sperm PLCzeta.
  • cytoplasmic Ca 2+ oscillations are evoked during the activation of the egg cell.
  • this physiological activation can be triggered artificially, for example by lonomycin or PLCzeta treatment.
  • the Ca 2+ oscillations induced by the PLCzeta can be regarded as a quality benchmark for the activation of the egg cells.
  • the characteristic Ca 2+ waves can now also be detected by the invention using electrical impedance spectroscopy with the devices of the invention.
  • Zn 2+ secretions that occur in the oocytes after fertilization have been detected by Zn 2+ imaging.
  • Zn 2+ releases can also be detected using electrical impedance spectroscopy with the devices of the invention.
  • the current is split between displacement current in the dielectric boundary and conduction current in the extracellular shunt path.
  • the current translocated across the membrane includes, for example, capacitance contributions from the passive permittivity of the lipid membrane, mobile charges associated with membrane proteins, and excitable contributions associated with the membrane's voltage-gated ion channel kinetics. Charges and dipoles are detected in the ZP. Measurements in this frequency range can therefore be used to estimate the effective impedance (dielectric) of the cell membrane and the ZP.
  • Impedance measurements generated from measuring the cell with even higher frequency signals can be used to study intracellular effects. In this frequency range, the cell membrane and ZP become transparent. Intracellular charge distributions such as intracellular Ca 2+ ion oscillations after the fertilization of egg cells can be measured from the impedance spectra. The exact frequencies at which maximum impedance signal changes and thus signal sensitivities can be determined depend heavily on the cell type used, the structure and the configuration of the structure, the electrode surface and its material.
  • EIS has so far been a measurement method for the short-term characterization of dielectric properties and sizes of individual cells in high-throughput applications, such as in cytometry or cell sorting/flow cytometers.
  • the present invention is designed for long-term observation periods.
  • the electrodes e.g. four microelectrodes, can be located in the depression V or at its edge.
  • the electrodes can be used to measure the dielectric properties of cells.
  • the size of electrodes depends on the design of the cavity V, the size of the oocyte Ez and the fill factor to be achieved.
  • the fill factor i.e. the quotient of the volume of the cell and the measurement volume, can be adjusted with the design of the electrodes and the cavity.
  • the device has, for example, a kind of holding device made of a planar, preferably transparent (carrier) substrate S.
  • the substrate S can be glass or a transparent polymer, for example.
  • At least one insulating layer ZS1 or ZS2 is applied to the substrate S. Suitable materials can also be plastics such as polymers or paints.
  • the intermediate layer ZS1 or ZS2 can also be made from the same material as the substrate S.
  • the intermediate layer ZS1 can also be made from a photoresist such as SU-8. At least one depression V for receiving a single egg cell Ez can be arranged in this intermediate layer ZS1.
  • the device or parts thereof can also be manufactured using other techniques.
  • parts can also be produced by injection molding or with RAD-based processes such as 3D printers.
  • substrates S and/or intermediate layers ZS1, ZS2 can be produced in this way.
  • substrates S and/or intermediate layers ZS1, ZS2 can be produced in this way.
  • metallic Elements such as electrodes E1, E2; Ela, E2a, Elb, E2b, imprint or apply by other methods.
  • the choice of manufacturing process can be determined by the number used and/or by the sizes required.
  • the size of the depression can also be larger than the egg cell. Then the egg cell lies in the depression (top view according to Figure 3a / 3b).
  • the shape of the indentation V (in top view according to Figure 2b / 3b) is not necessarily round, but can also be (poly)angular. The only important thing is that the egg cell can lie in the recess V at least at the edge, so that there is spatial fixation in relation to the electrodes. It is not absolutely necessary that the egg cell Ez is smaller than the diameter of the indentation V, but the egg cell Ez can also rest on the edge of the indentation V - as shown in Figure 3c - and thus only with a partial area in the indentation V protrude.
  • the diameter of the depression V for fixing the egg cells is preferably about 30 - 70 ⁇ m, e.g. for mouse egg cells.
  • a non-round indentation V exposes areas at the top edge of the indentation V where the field lines are conducted through the conductive medium surrounding the cell without cell contact. This reduces the measuring signal level and thus the sensitivity.
  • the diameter of the indentation V is preferably smaller than the diameter of the oocyte.
  • the oocyte then lies on the upper edge of the indentation V and covers (mostly or completely) the indentations V. For a high to maximum impedance signal level, a close contact of the oocyte Ez to the edge of the indentations V is preferred, so that all electric field lines are forced through the oocyte will.
  • an outlet opening to a fluid channel can be located in the bottom area of the depression.
  • the diameter of the opening in the recess is preferably smaller than the diameter of the recess V and the egg cell Ez.
  • This can be used, for example, as a control for the introduction of an egg cell Ez, in order to control a fluid flow, for example by means of a pressure difference, in an associated fluid channel FK .
  • This can be advantageous, for example, when the egg cell Ez to be observed does not sink into the depression V of its own accord, for example it is floating in a cell medium.
  • By generating negative pressure behind the opening nutrient medium can be drawn off into the depression V and through the fluid channel.
  • the egg cell can be hydrodynamically positioned and fixed in the depression after it has been placed in a nutrient solution (hydrodynamic trap).
  • a nutrient solution hydrodynamic trap
  • an inflow can be generated by reversing the fluid flow and thus (specifically) the egg cell can be released from the cavity.
  • An exemplary nutrient solution is, for example, EmbryoMax® Advanced KSOM Embryo Medium, as is available, for example, from Sigma-Aldrich Chemie GmbH, Taufkirchen. However, this does not explicitly exclude other nutrient solutions.
  • the fluid channel can allow fluid to flow out of the depression (to fix the egg cell in the depression) and/or fluid can flow in (e.g. release the egg cell from depression V). to allow.
  • At least one electrode E1 or E1b, E2b is attached in the shown embodiments, which is preferably ring-shaped in the case of a round depression V, see Figures 2b, 3b and 5.
  • At least one further electrode E2 is applied on the bottom of the depression V, in the embodiments in FIGS. 4 and 5 in the area of the opening.
  • An AC voltage at different frequencies can be applied between (pairs of) electrodes, the current flow can be measured and the impedance can be calculated from this.
  • Figures 2b, 3b and 4 show advantageous exemplary embodiments of the electrodes. The exact layout and placement of the electrodes depends on the application.
  • the individual electrodes are contacted with one or preferably with two leads in order to enable 2-point or preferably 4-point measurements.
  • Figures 2b, 3b and 5 each show an example of a lead to an electrode.
  • the leads can be electrically isolated with an insulating material. Preferably the insulating material is transparent.
  • measurements with 3 electrodes can also be carried out, in which the third electrode is designed as a reference electrode, e.g. made of silver/silver chloride. Other suitable materials can also be used.
  • Photoresists are preferably used for the insulating layers, preferably the SU-8 photoresist is suitable, which is also applied to the carrier substrate in thick layers in the normal process (in liquid form by spin-on and then drying) or as a dry film resist (by lamination) and with lithographic processes can be exposed and structured.
  • the electrodes and the Leads preferably consist of precious metals such as gold or platinum or oxidic semiconductors such as iridium oxide and are deposited using PVD methods (evaporation, cathode sputtering) and structured using lithographic methods in combination with lift-off techniques or etching techniques. It is also possible to use transparent electrodes, for example made from an oxidic semiconductor, such as indium tin oxide (ITO) electrodes, in order to enable optical access to the egg cell from the underside in combination with a transparent carrier substrate.
  • the leads are preferably made of conductive material with a low ohmic resistance. Other suitable materials and fabrication processes, such as precision engineering or the fabrication of leads and electrodes by modified inkjet printing on substrates, can also be used, adapted to the size of the cells to be examined.
  • the electrodes can be modified to minimize their capacitive influence on the electrical double layer in the low frequency range. This capacitive behavior (and thus the limit frequency) can be shifted to the lower frequency range by increasing the effective electrode area.
  • the electrodes can advantageously be achieved by potentiostatic deposition of conductive polymers, e.g. polypyrrole polystyrene sulfonate (PPy:PSS), poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS), polyaniline or mixtures thereof (the process is called electropolymerization). These layers are porous and thus increase the effective electrode area.
  • electrodes are porous, they have a higher effective electrode area compared to the footprint.
  • Transparent layers are advantageous for the embodiments in which optical access to the egg cell is provided from the bottom of the depression (transmitted light microscopy).
  • the one surrounding barrier B has at least one depression. Since the barrier B can be optically opaque, it can also be made from a different material than the substrate S or the intermediate layer ZS1, ZS2.
  • the degree of maturity can also be assessed by means of optical systems (in reflected light or transmitted light operation) as an alternative or in addition to an impedance measurement.
  • the device according to the invention can also have control electronics for selecting specific pairs of electrodes for targeted measurements on a number of egg cells have a specific egg cell.
  • This can, for example, connect a measuring device to the respective electrodes by means of a switch.
  • Exemplary switches include, but are not limited to, electronic switches (eg, transistors) or electrical switches (DIP switches).
  • the control can advantageously be designed as a multiplexer.
  • the control electronics can be provided on a printed circuit board, for example.
  • An impedance spectrometer or an LCR measuring bridge or a vector network analyzer as well as a PC can be connected via suitable connections and cables as an example for recording, storing and processing measured values.
  • an impedance spectrometer or an LCR measuring bridge or a vector network analyzer is implemented on the control electronics, so that an integrated and mobile system can be made available.
  • the device combines ease of use, miniaturization, integration and portability. EIS also allows continuous monitoring of maturing cells and developing embryos.
  • the device is preferably placed in an incubator or in a temperature cabinet in order to keep the medium and the egg cells at a constant temperature of preferably 37° C. for the measurements.
  • a cover plate can be provided which can be placed on the barrier.
  • the device can also have a temperature control unit.
  • the barrier B can be made of a metallic material, such as aluminum, and one or more controllable heating elements, which keep the medium, including the oocytes, regulated at a predetermined temperature, preferably 37°C for human oocytes.
  • a temperature sensor can measure the temperature in the nutrient solution/medium and thus enables the predetermined temperature to be controlled using a suitable control device.
  • a heating element can be immersed in the nutrient solution or thin-film resistors can be arranged on the bottom of the carrier substrate in the area of the depression(s) V.
  • the device can have a unit for atmospheric control.
  • the device can also be flown over with a gas, such as gas mixtures containing CCk (eg, preferably 5% CO), or it can be located in a closed atmosphere.
  • a gas such as gas mixtures containing CCk (eg, preferably 5% CO)
  • the interior of the barrier B can be filled with nutrient solution, brought to the desired operating temperature of preferably 37° C., and egg cells Ez placed in the wells V.
  • the electrical impedance spectrum (real and imaginary part, phase) of a well V filled with an egg cell Ez is measured over the entire frequency range, for example 1 Hz to 1 MHz.
  • the absolute values of the impedance can also be determined over the frequency.
  • This spectrum is subtracted from a spectrum, called the reference spectrum, which is recorded simultaneously from another well V without an oocyte.
  • parasitic effects in particular polarization effects of the electrodes and influences of the leads, are filtered out.
  • Cell-specific changes can be deduced from the difference in the spectra and the frequency of the highest sensitivity can be determined, which is advantageously used for further measurements.
  • a reference spectrum of an empty well V is recorded, then the egg cell Ez is placed in this well V and the impedance is recorded again.
  • the frequency with the highest sensitivity is determined.
  • the magnitudes of the impedance signals of both spectra can be subtracted from one another.
  • cell-specific changes can be deduced from the difference in the spectra.
  • the device In a typical measurement routine, the device is used, oocytes Ez are placed in the (respective) well V of the holding device, temperature-controlled and the electrical impedance is recorded as a function of time at the frequency of the highest sensitivity. According to the invention, changes in the maturation process of an egg cell are analyzed with the impedance measurements.
  • a local minimum can be expected to occur, for example, when a (pre-)determined slope is reached, so that the time window does not necessarily begin with the local minimum, but can also include it.
  • Sperm can be added directly to the cell medium for fertilization.
  • a supply to an entire medium for a large number of egg cells Ez for example in an arrangement for several egg cells Ez
  • a supply to individual compartments can be provided.
  • fertilize an oocyte in a targeted manner by means of an intracytoplasmic sperm injection.
  • the impedance changes, e.g. the impedance increases again.
  • the measurement period can range from a few minutes, e.g. 20 minutes, to hours and even days.
  • Physiological phenomena such as the release of Zn 2+ and Ca 2+ ions occur immediately or with a time delay after fertilization.
  • the impedance can preferably be measured at the frequency with the highest sensitivity and high sampling rate, typically one measurement per second, over a period of 60 minutes after insemination in the device described here.
  • Oscillations in the ElS signal can be resolved that can be associated with characteristic voltage-gated ion channel kinetics in the membrane evoked by characteristic sperm PLCzeta-induced Ca 2+ oscillations in the cytoplasm and the release of Zn +2 ions.
  • the device is redesigned for high-frequency measurements.
  • a vector network analyzer with an intermediate 50 W matching network is preferably connected here.
  • the device characterizes individual oocytes by electrical impedance spectroscopy to assess oocyte quality and potential for fertilization and embryo development.
  • the device measures non-destructively and enables label-free long-term measurements over the entire period of artificial insemination.
  • the oocytes can be placed in the device, removed and exchanged easily and quickly. By resting on the edge of the depression V, changes in the size of the cells can be compensated for.
  • Time-resolved EIS measurements make it possible to measure changes in ZP as well as the electrophysiological processes of the oocyte. Inspection by (inverse) microscopy allows measurement of oocyte diameter as well as ZP layer thickness during ZP maturation and hardening.
  • the parallel and thus time-saving characterization of several oocytes is an added value.
  • egg cells of different qualities can be assessed in the screening process. Only the egg cells with good development, i.e. maturation potential, are selected for fertilization. This significantly increases the chances of success.
  • the device can be used for all types of oocyte evaluation and breeding programs with limited oocyte availability and will thus improve results in assisted reproduction.
  • the oocytes/embryos of the best quality are preferably selected.
  • the fertilization success can be increased by determining the optimal fertilization times, and thereby the development potential of the embryo.
  • the chance of a successful pregnancy increases and the risk of further necessary interventions decreases.
  • determining the best egg cell/embryo quality can lead to a reduction in the number of animals to be used. By increasing the fertilization rate of an individual animal, fewer female animals need to be used for egg cell retrieval.
  • the fertilization of the egg cell must be optimized. Oocytes and sperm are often only available in reduced quantities. With the help of the device, the egg cells/embryos of the best quality can be determined and thus the success of a successful pregnancy can be increased.
  • the device according to the invention can also be used in a system for characterizing contraceptives.
  • Contraceptives can, for example, act in such a way that the ZP is changed by the influence of substances in such a way that fertilization becomes impossible for predetermined periods of time.
  • both an egg cell in the device can be exposed to such a substance and its effectiveness can be tested directly by subsequently adding sperm cells to the medium.

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Abstract

L'invention concerne un dispositif de caractérisation de changements du processus de maturation d'un ovocyte (Ez), comprenant les éléments suivants : un substrat (S) ; au moins une dépression (V) pour recevoir un ovocyte (Ez) étant prévue dans le substrat (S), comprenant : au moins deux électrodes (E1, E2 ; E1a, E1b, E2a, E2b) étant prévues dans les zones de bord de la dépression (V) ; une résistance complexe (Z) de l'ovocyte (Ez) pouvant être mesurée pendant le fonctionnement au moyen des électrodes (E1, E2 ; E1a, E1b, E2a, E2b), une première résistance complexe (Z1) pouvant être mesurée à un premier moment (t1) et une deuxième résistance complexe (Z2) pouvant être mesurée à un deuxième moment (t2) postérieur au premier moment ; un degré de maturation ou la progression de la division cellulaire de l'ovocyte (Ez) pouvant être dérivé d'un écart de la partie réelle et/ou imaginaire de la première résistance complexe mesurée (Z1) et de la seconde résistance complexe (Z2). L'invention concerne également un système comprenant le dispositif selon l'invention et leur utilisation telle que représentée dans les illustrations.
PCT/EP2021/083078 2020-11-26 2021-11-26 Système et dispositif pour caractériser des changements dans le processus de maturation d'un ovocyte, et leur utilisation WO2022112460A1 (fr)

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DE102020214862.7A DE102020214862B4 (de) 2020-11-26 2020-11-26 System und Vorrichtung zur Charakterisierung von Veränderungen des Reifungsvorgangs einer Eizelle, sowie deren Verwendung
DE102020214862.7 2020-11-26

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Citations (5)

* Cited by examiner, † Cited by third party
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