WO2024045482A1 - Appareil et procédé d'activation combinée pour embryons précoces - Google Patents

Appareil et procédé d'activation combinée pour embryons précoces Download PDF

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Publication number
WO2024045482A1
WO2024045482A1 PCT/CN2023/073078 CN2023073078W WO2024045482A1 WO 2024045482 A1 WO2024045482 A1 WO 2024045482A1 CN 2023073078 W CN2023073078 W CN 2023073078W WO 2024045482 A1 WO2024045482 A1 WO 2024045482A1
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Prior art keywords
early
early embryos
liquid metal
capillary
activation
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PCT/CN2023/073078
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English (en)
Chinese (zh)
Inventor
黄海波
梅世亮
苗胜
庞焱
姜德胜
陈立国
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苏州大学
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Publication of WO2024045482A1 publication Critical patent/WO2024045482A1/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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/10—Petri dish
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/10—Hollow fibers or 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
    • 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
    • C12M35/08—Chemical, biochemical or biological means, e.g. plasma jet, co-culture

Definitions

  • the present application relates to the technical field of embryo engineering, and in particular to a joint activation device and method for early embryos.
  • early embryo activation is a common approach at various stages of the early embryo micromanipulation process.
  • cytoplasmic activation of the oocyte is accomplished by the entry of sperm and the induction of calcium oscillations.
  • PA parthenogenetic activation
  • ICSI intracytoplasmic sperm injection
  • somatic cell nuclear transfer artificial assisted activation of oocytes is essential.
  • artificial auxiliary activation methods are mainly divided into single activation and combined activation.
  • Commonly used single activations mainly include mechanical activation, electrical activation, and chemical activation (calcium ionophore A23187, ionomycin, ethanol, etc.); combined activation mainly involves simple physical activation or chemical activation first, and then protein synthesis inhibition.
  • the commonly used joint activation is to first electrically activate early embryos and then soak the early embryos in a solution containing chemicals to chemically activate early embryos.
  • the device is relatively crude and has low accuracy.
  • Electrical activation uses an activation chamber with two parallel plate electrodes to electrically activate a batch of early embryos. Due to the large distance between the macroscopic parallel plate electrodes, the electrical stimulation voltage applied to a single early embryo is also large and the distribution is not uniform. It is extremely easy to cause excessive electrical damage to the embryo, and it is impossible to achieve precise electrical stimulation of specific parts of the early embryo; it is impossible to use a set of activation devices to complete the combined activation process of electrical activation and chemical activation of the early embryo.
  • the purpose of this application is to provide a joint activation device and method for early embryos, which can be used to achieve joint stimulation including electrical stimulation and chemical stimulation of specific parts of early embryos, and to realize a set of activation devices to complete the stimulation of early embryos. joint activation, thereby increasing the activation rate of early embryos.
  • this application provides a joint activation device for early embryos, including:
  • Petri dishes used to store early-stage embryos
  • a cell suction mechanism is used to adjust the position and posture of the early embryo and fix the early embryo
  • a joint activation mechanism includes a capillary microneedle and a first needle holder, and the capillary microneedle is connected to the first needle holder; the capillary microneedle includes a liquid metal channel and a chemical substance channel; Liquid metal and wires are provided in the liquid metal channel.
  • the liquid metal is located at the tip of the capillary microneedle.
  • the wires are used to connect the excitation power supply.
  • the liquid metal and the wires constitute a liquid metal electrode for energizing the capillary microneedle.
  • the early embryo is electrically activated; the chemical substance channel is used to release chemical substances to chemically activate the early embryo.
  • the capillary microneedle includes a plurality of liquid metal channels and one chemical substance channel, and the plurality of liquid metal channels are arranged around one chemical substance channel.
  • the bottom of the culture dish is provided with an array of micropits, and the micropits are used to prevent movement of the early embryos.
  • the first needle holding instrument includes a first screw cap, a second screw cap, a first connection base, a third screw cap and a first fixed rod;
  • the first screw cap is threadedly connected to the second screw cap, the second screw cap and the third screw cap are both threadedly connected to the first connecting seat, and the first fixed rod is connected to the third screw cap.
  • the tail of the capillary microneedle penetrates the first screw cap and the second screw cap, the first connection seat is provided with a pneumatic drive inlet, and the chemical substance channel in the capillary microneedle is in contact with the pneumatic drive The entrance is connected.
  • the cell suction mechanism includes a cell suction needle and a second needle holder, the cell suction needle is connected to the second needle holder, and the second needle holder is provided with an air guide port, The air guide port is connected with the cell suction needle.
  • the diameter of the liquid metal channel is 7 ⁇ m.
  • the diameter of the chemical substance channel is 10 ⁇ m.
  • the inner diameter of the cell suction needle is 15 ⁇ m and the outer diameter is 100 ⁇ m.
  • the diameter of the micropits is 150 ⁇ m and the depth is 50 ⁇ m.
  • This application also provides a joint activation method for early embryos, which is applied to the joint activation device for early embryos, including:
  • control the joint activation mechanism After the cell suction mechanism fixes the early embryo, control the joint activation mechanism to continuously pass high-voltage direct current pulses to the target site of the early embryo to electrically activate the early embryo;
  • the joint activation mechanism is controlled to release chemical substances to chemically activate the early embryo.
  • This application provides a joint activation device for early embryos, including: a culture dish for storing early embryos; a cell suction mechanism for adjusting the position and posture of early embryos and fixing early embryos; a joint activation mechanism for joint activation
  • the mechanism includes a capillary microneedle and a first needle-holding device, and the capillary microneedle is connected to the first needle-holding device; the capillary microneedle includes a liquid metal channel and a chemical substance channel; liquid metal and wires are provided in the liquid metal channel, and the liquid metal is located in the capillary microneedle. The tip of the fine needle and the wire are used to connect the excitation power supply.
  • the liquid metal and the wire constitute a liquid metal electrode for electrical activation of early embryos; the chemical substance channel is used to release chemicals for chemical activation of early embryos.
  • the capillary microneedles of this application can achieve precise joint activation of local parts of a single early embryo, such as the nuclear substance area, and reduce excessive electrical or chemical damage to other parts of the early embryo, thereby increasing the activation rate of early embryos; and a joint activation mechanism integrates It combines electrical activation and chemical activation to improve the convenience of combined activation of early embryos; the cell holding mechanism can quickly adjust the position of cells, shorten time, and reduce factors that lead to cell death or stagnant development due to adjustment of cell position.
  • the joint activation method of early embryos provided by the embodiments of the present application corresponds to the device and has the above effects.
  • Figure 1 is a structural diagram of a joint activation device for early embryos provided by an embodiment of the present application
  • Figure 2 is a structural diagram of a culture dish provided by an embodiment of the present application.
  • Figure 3 is a structural diagram of a joint activation mechanism provided by an embodiment of the present application.
  • Figure 4 is a tip structure diagram of a capillary microneedle provided by an embodiment of the present application.
  • Figure 5 is a schematic cross-sectional view of the tip of a capillary microneedle provided by an embodiment of the present application
  • Figure 6 is an internal structural diagram of a capillary microneedle provided by an embodiment of the present application.
  • Figure 7 is a process flow chart for preparing a capillary microneedle tip provided by an embodiment of the present application.
  • Figure 8 is a structural diagram of a cell suction mechanism provided by an embodiment of the present application.
  • Figure 9 is a flow chart of a joint activation method for early embryos provided by an embodiment of the present application.
  • the reference numbers are as follows: 1 is a culture dish, 2 is a joint activation mechanism, 3 is a cell suction mechanism, 4 is a cell, 101 is a micro pit, 201 is a capillary microneedle, 202 is the first screw cap, 203 is the second screw cap.
  • 204 is the first connecting seat
  • 205 is the third screw cap
  • 206 is the first fixed rod
  • 207 is the air pressure drive inlet
  • 208 is the wire outlet
  • 209 is the first annular washer
  • 210 is the second annular washer
  • 211 is Chemical pipeline
  • 2011 is the liquid metal channel
  • 2012 is the chemical substance channel
  • 2013 is the wire
  • 301 is the cell suction needle
  • 302 is the fourth screw cap
  • 303 is the fifth screw cap
  • 304 is the second connection seat
  • 305 is The sixth screw cap
  • 306 is the second fixed rod
  • 307 is the air guide port
  • 308 is the gas pipe
  • 309 is the third annular washer
  • 310 is the fourth annular washer
  • A is the tip of the capillary microneedle
  • B is the cell suction needle. The tip of the needle.
  • the principle of electrical activation is that under the action of a short high-voltage direct current pulse in early embryos, the stability of the phosphodiester bimolecular structure of the membrane changes, and many recoverable tiny holes are formed on the cell membrane, allowing extracellular Ca+ to enter the cell and causing The intracytoplasmic Ca+ concentration increases, and the increase in intracellular Ca2+ concentration causes the disappearance of CSF activity, cyclinB is rapidly degraded, MPF activity disappears, and cells are activated to complete the second meiosis and enter the next cell cycle.
  • Chemical activation mainly includes calcium ionophore A23187, ionomycin, ethanol activation, protein synthesis inhibitors and protein phosphorylation inhibitors.
  • CaA Calcium ionophore A23187 activates early embryos mainly through the release of intracytoplasmic Ca2+.
  • Ionomycin another calcium ionophore, also functions through the release of Ca2+ in early embryos. However, it does not directly promote the release of Ca2+ in early embryos, but first uses Ca2+ channels on the cell membrane to influx from the outside of the cell into the cell, and then activates Ca2+ in the endogenous endoplasmic reticulum to enter the cytoplasm of early embryos.
  • the activation of early embryos by ethanol mainly causes the formation of IP3 in the cell membrane. Through IP3, the receptor mediates the release of endogenous Ca2+, causing the early response of early embryo activation.
  • the protein cytokines maturation-promoting factor (MPF) and cell quiescence factor (CSF) are regulators of the cell cycle, and it is precisely because of their action that oocytes will be arrested in the MII phase. These factors are very sensitive to Ca2+. When the egg-activated cytoplasmic Ca2+ rises to a certain level, MPF and CSF will be inactivated or disappear.
  • the use of protein synthesis inhibitors to activate early embryos does not work because it can cause fluctuations in Ca2+ concentration, but because it can inhibit the synthesis of these protein-based cytokines, thereby causing early embryos to leave the MII stage and resume the second meiosis.
  • Protein phosphorylation inhibitors can prevent protein phosphorylation and thereby inhibit the activities of MPF and CSF. They also inhibit the discharge of the second polar body and ensure that the chromosomes of parthenogenetically activated oocytes are diploid.
  • the core of this application is to provide a device and method for joint activation of early embryos.
  • Figure 1 is a structural diagram of a joint activation device for early embryos provided by an embodiment of the present application. As shown in Figure 1, 1 is a culture dish, 2 is a joint activation mechanism, 3 is a cell suction mechanism, and 4 is a cell cells.
  • Petri dish 1 is used to store early embryos; cell suction mechanism 3 is used to adjust the position and posture of early embryos and fix early embryos; joint activation mechanism 2, joint activation mechanism 2 includes capillary microneedles 201 and a first needle holding device , the capillary microneedle 201 is connected to the first needle holder; the capillary microneedle 201 includes a liquid metal channel 2011 and a chemical substance channel 2012; the liquid metal channel 2011 is provided with liquid metal and a wire 2013, and the liquid metal is located at the tip of the capillary microneedle 201, The wire 2013 is used to connect the excitation power supply.
  • the liquid metal and the wire 2013 constitute liquid metal for electrical activation of early embryos; the chemical channel 2012 is used for releasing chemicals to chemically activate early embryos.
  • An embodiment of the present application provides a joint activation device for early embryos, including: a culture dish for storing early embryos; a cell suction mechanism for adjusting the position and posture of early embryos and fixing early embryos; a joint activation mechanism,
  • the joint activation mechanism includes a capillary microneedle and a first needle-holding device, and the capillary microneedle is connected to the first needle-holding device;
  • the capillary microneedle includes a liquid metal channel and a chemical substance channel; the liquid metal channel is provided with liquid metal and a wire, and the liquid metal Located at the tip of the capillary microneedle, the wire is used to connect the power supply.
  • the liquid metal and the wire constitute an electrode for electrical activation of early embryos; the chemical channel is used to release chemicals to chemically activate early embryos.
  • the capillary microneedles of this application can achieve precise joint activation of local parts of a single early embryo, such as the nuclear substance area, and reduce excessive electrical or chemical damage to other parts of the early embryo, thereby increasing the activation rate of early embryos; and a joint activation mechanism integrates It combines the functions of electrical activation and chemical activation to improve the convenience of combined activation of early embryos; the cell holding mechanism can quickly adjust the position of cells, shorten time, and reduce factors that lead to cell death or stagnant development due to adjustment of cell position. .
  • FIG. 2 is a structural diagram of a culture dish provided by the embodiment of the present application. As shown in Figure 2, the bottom of the culture dish 1 is provided with an array of micro pits 101. , the micropit diameter is 150 ⁇ m and the depth is 50 ⁇ m.
  • the culture dish 1 is equivalent to the operating table of the entire device.
  • the cells 4 are placed in the culture dish 1.
  • the micro-pits 101 at the bottom prevent the early embryos from flowing easily and are convenient for grabbing.
  • FIG 3 is a structural diagram of a joint activation mechanism provided by an embodiment of the present application.
  • the joint activation mechanism 2 includes a capillary microneedle 201 and a first needle holder.
  • the first needle holding instrument includes a first screw cap 202, a second screw cap 203, a first connection base 204, a third screw cap 205, a first fixed rod 206, Pneumatic drive inlet 207, wire outlet 208, first annular gasket 209, second annular gasket 210, chemical pipeline 211.
  • A is the needle tip part of the capillary microneedle 201 .
  • the first screw cap 202 is threadedly connected to the second screw cap 203, the second screw cap 203 and the third screw cap 205 are both threadedly connected to the first connecting seat 204, the first fixed rod 206 is threadedly connected to the third screw cap 205;
  • the tail of the microneedle 201 penetrates the first screw cap 202 and the second screw cap 203.
  • the first connection seat 204 is provided with a pneumatic drive inlet 207.
  • the chemical channel 2012 in the capillary microneedle 201 is connected to the pneumatic drive inlet 207 through a chemical pipeline 211. .
  • a through hole is provided on the side wall of the capillary microneedle 201 near the tail, and the corresponding second screw cap 203 is provided with a wire outlet 208 to facilitate the extraction of the wire 2013 from the liquid metal channel 2011.
  • the embodiment of the present application does not specifically limit the number of chemical substance channels 2012 and liquid metal channels 2011 in the capillary microneedle.
  • the capillary microneedle 201 may include multiple liquid metal channels 2011 and one chemical substance channel 2012.
  • the air pressure driven inlet 207 can be connected to a microinjection pump, which increases the gas pressure in the chemical substance channel 2012 to release the chemical in the chemical substance channel 2012.
  • Figure 4 is a tip structural diagram of a capillary microneedle provided by an embodiment of the present application.
  • Figure 5 is a schematic cross-sectional view of a tip of a capillary microneedle provided by an embodiment of the present application. Combining Figures 4 and 5, it includes 6 liquids Metal channel 2011 and a chemical substance channel 2012, six liquid metal channels 2011 are arranged around a chemical substance channel 2012; the diameter of the liquid metal channel 2011 is 7 ⁇ m, and the diameter of the chemical substance channel 2012 is 10 ⁇ m.
  • Figure 6 is an internal structural diagram of a capillary microneedle provided by an embodiment of the present application. As shown in Figure 6, a liquid metal channel 2011 is provided with liquid metal and a wire 2013.
  • the liquid metal is located at the tip of the capillary microneedle 201, and the wire 2013 Leading from the tip of the needle to the through hole near the tail of the needle, the wire 2013 is used to connect to the power supply.
  • the liquid metal and the wire 2013 form an electrode that can electrically activate the cells.
  • the first fixed rod 206 of the first needle holder can be connected to a mechanical arm (not shown in the figure), and the position adjustment of the joint activation mechanism is realized by controlling the mechanical arm.
  • the first annular gasket 209 and the second annular gasket 210 in the first needle holder are used to deform and fix the tail of the capillary microneedle 201 when it is squeezed by the screw cap.
  • the capillary microneedle 201 in the embodiment of the present application is detachably connected to the first needle holder, so that the capillary microneedle 201 can be easily replaced.
  • Figure 7 is a flow chart of a process for preparing the tip of a capillary microneedle provided by an embodiment of the present application.
  • the process for preparing the tip of a capillary microneedle in the joint activation mechanism will be introduced below with reference to Figure 7 process:
  • a capillary microneedle drawing instrument is used to heat and draw the glass tube to a set shape and size. Before stretching the glass tube, the glass tube is prefabricated. The shape of the glass tube is a chemical channel with a diameter of 1mm in the middle. There are six microtubes (liquid metal channels) with a diameter of 0.7 mm surrounding the chemical channel. The glass tube is heated and drawn by a needle puller into a 10 ⁇ m chemical channel and six 7 ⁇ m liquid metal channels.
  • liquid metal from the tail of the capillary microneedle glass tube into six glass microtubes with a diameter of 7 ⁇ m.
  • a high-precision syringe is used to inject liquid metal from the tail of the capillary microneedle.
  • the liquid metal in this device is gallium-based liquid metal, but the liquid metal is not limited to gallium-based liquid metal.
  • the chemical substance channel needs to be blocked to prevent the liquid metal from flowing into the chemical substance channel.
  • the surface is quickly oxidized to form a thin solid oxide film of about 1 nm, which helps stabilize the shape of the liquid metal capillary microneedle electrode and prevents the liquid metal from flowing out from the tip of the capillary microneedle and affecting the Performance of capillary microneedle electrodes.
  • FIG 8 is a structural diagram of a cell suction mechanism provided by an embodiment of the present application.
  • the cell suction mechanism includes a cell suction needle 301 and a second needle holder.
  • the cell suction needle 301 and the second needle holder The second needle-holding device is connected with an air guide port 307, and the air guide port 307 is connected with the cell suction and holding needle 301.
  • the second needle holder includes a fourth screw cap 302, a fifth screw cap 303, a second connecting seat 304, a sixth screw cap 305, a second fixed rod 306, an air guide port 307, a gas pipe 308, a third annular gasket 309, Fourth annular washer 310.
  • B is the tip of the cell suction needle.
  • the embodiments of this application are effective for cell absorption.
  • the needle 301 is not specifically limited.
  • the tip of the cell suction needle 301 is made of a prefabricated glass hose with an inner diameter of 15 ⁇ m and an outer diameter of 100 ⁇ m.
  • the principle of negative pressure is used to suck the early embryos and keep the cells in a In a relatively stable state, the cell holding mechanism 3 can put down or hold the cells at any time. More importantly, when the cells are placed on the culture dish, the cell holding mechanism 3 can be moved to different positions around the cells, and then moved to different positions around the cells.
  • Cell suction can achieve the purpose of adjusting cell posture, which is very convenient and has high convenience and repeatability.
  • the second fixed rod 306 of the second needle holder can be connected to a mechanical arm (not shown in the figure), and the position adjustment of the cell suction mechanism 3 is realized by controlling the mechanical arm.
  • the third annular gasket 309 and the fourth annular gasket 310 in the second needle holder are used to deform and fix the tail of the cell suction needle 301 when it is squeezed by the screw cap.
  • the joint activation device for early embryos is described in detail.
  • This application also provides corresponding embodiments of the joint activation method for early embryos.
  • Figure 9 is a flow chart of a joint activation method for early embryos provided by an embodiment of the present application. As shown in Figure 9, the joint activation method for early embryos includes:
  • Target sites in early embryos include nuclear material areas.
  • the specific process is as follows:
  • Early embryo fixation stage Use mouse or pig oocytes and sperm to fuse in vitro to form fertilized eggs. Place the early embryo on the culture dish, and then fix it with the cell holding mechanism 3 to prevent the early embryo from moving or rotating.
  • (2) Activation stage of early embryos Use joint activation mechanism 2 to activate early embryos.
  • a higher voltage DC pulse is continuously supplied, with a voltage of 3.2V and a pulse width of 50 to 100 ⁇ s.
  • the permeability of the cell membrane increases.
  • the chemical solution is released through the cell membrane and enters the early embryo, quickly activating a series of metabolic reactions in the early embryo.
  • the embodiments of the present application provide a joint activation method for early embryos, which electrically activates the cell membrane of early embryos, causing the stability of the phosphodiester bimolecular structure of the membrane to change, and forming many recoverable tiny holes on the cell membrane.
  • Calcium ionophores, ionomycin, etc. released by chemical activation enter cells through tiny holes and activate a series of metabolic reactions in early embryos, thereby greatly increasing the development and survival rate of embryos. This is of great significance to the in vitro development process of early embryos.

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Abstract

La présente invention concerne un appareil d'activation combiné pour embryons précoces. L'appareil comprend : une boîte de culture pour stocker les embryons précoces ; un mécanisme de maintien de cellule pour ajuster la posture des embryons précoces et fixer les embryons précoces ; et un mécanisme d'activation combiné, comprenant une micro-aiguille capillaire et un premier instrument de maintien d'aiguille. La micro-aiguille capillaire comprend un canal métallique liquide et un canal de substance chimique ; un métal liquide et un fil sont agencés dans le canal métallique liquide pour former une électrode métallique à l'état liquide pour activer électriquement les embryons précoces ; le canal de substance chimique est utilisé pour libérer des substances chimiques pour activer chimiquement les embryons précoces. Le mécanisme d'activation combiné intègre deux fonctions d'activation électrique et d'activation chimique, de telle sorte que la commodité d'activation combinée des embryons précoces est améliorée ; la micro-aiguille capillaire peut obtenir l'activation combinée précise de parties locales telles qu'une région de substance nucléaire d'un embryon précoce unique, de telle sorte qu'une détérioration électrique excessive ou un endommagement chimique d'autres parties des embryons précoces est réduit, ce qui permet d'améliorer le taux d'activation des embryons précoces.
PCT/CN2023/073078 2022-08-30 2023-01-19 Appareil et procédé d'activation combinée pour embryons précoces WO2024045482A1 (fr)

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CN202211056138.4A CN115353956A (zh) 2022-08-30 2022-08-30 一种早期胚胎的联合激活装置和方法

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CN102119331A (zh) * 2008-06-05 2011-07-06 生命科技公司 细胞跨膜电位的激活和监测
CN104109629A (zh) * 2014-07-30 2014-10-22 苏州大学 一种压电超声显微注射器及压电超声显微注射系统
CN106754348A (zh) * 2016-12-23 2017-05-31 南京农业大学 一种电激活设备及采用其的卵母细胞或者胚胎的rna干扰新方法
CN111849768A (zh) * 2020-07-14 2020-10-30 苏州大学 一种卵细胞多效能精准电刺激装置及方法
CN115353956A (zh) * 2022-08-30 2022-11-18 苏州大学 一种早期胚胎的联合激活装置和方法

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