WO2016140379A1 - Tube microcapillaire polymère pour la congélation rapide de cellules, et tube microcapillaire capable de piéger des cellules - Google Patents

Tube microcapillaire polymère pour la congélation rapide de cellules, et tube microcapillaire capable de piéger des cellules Download PDF

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Publication number
WO2016140379A1
WO2016140379A1 PCT/KR2015/002062 KR2015002062W WO2016140379A1 WO 2016140379 A1 WO2016140379 A1 WO 2016140379A1 KR 2015002062 W KR2015002062 W KR 2015002062W WO 2016140379 A1 WO2016140379 A1 WO 2016140379A1
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Prior art keywords
polymer
tube
cell
microcapillary
capillary
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PCT/KR2015/002062
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English (en)
Korean (ko)
Inventor
허윤석
김종호
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계명대학교 산학협력단
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Priority to PCT/KR2015/002062 priority Critical patent/WO2016140379A1/fr
Publication of WO2016140379A1 publication Critical patent/WO2016140379A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts

Definitions

  • the present invention relates to a polymer microcapillary tube for rapid freezing of cells, and more particularly, in a medical cell rapid freezing system consisting of cell pretreatment, cell injection, and rapid freezing and storage, A polymer microcapillary for rapid cell freezing that can be used for rapid freezing and storage after injection.
  • the present invention also relates to a microcapillary tube capable of trapping cells, and more particularly, to a cell injected for the vitrification process of rapid freezing in a medical cell rapid freezing system consisting of cell pretreatment, cell injection, and quick freezing and storage.
  • cell cryopreservation techniques can be broadly divided into two stages: slow freezing and moving rapid cooling (Vitrification, or “Ice Free” cryopreservation).
  • Slow freezing here uses a slower cooling rate than on average 2 ° C./min, resulting in ice formation during the cooling process.
  • These ice crystals act like a blade on cells, causing cell membrane damage and damage to intracellular microstructure, resulting in loss of cell survival as well as normal cell function.
  • Prior art 1 (US Pat. No. 6,500,608) is a method invention for vitrification of biological cells, and discloses a method in which cells are directly exposed to a liquid nitrogen. Because of the unknown contaminants in the refrigerant itself and recently living bacteria at very low temperatures, the US FDA does not permit the direct exposure of cells to the refrigerant for cryopreservation of eggs and embryos in artificial fertilization.
  • Prior Art 2 (Vitrification by ultra-fast cooling at a low concentration of cryoprotectants in a quartz micro-capillary: A study using murine embryonic stem cells / Xiaoming He et al / Cryobiology 56 (2008) 223-232) is a quartz capillary (Quartz). capillary) to test the rapid freezing of cells.
  • quartz Quartz
  • brittleness is very strong, there is a disadvantage that the cell breaks well during the experiment.
  • artificial fertilization In vitro fertilization
  • the broken material has a disadvantage that can never be used in clinical and commercialized.
  • the present applicant intends to propose a polymer microcapillary tube which is a key component of the medical cell rapid freezing system, which prevents direct exposure of cells, easy sealing at both ends, and prevents cell loss by breaking during cooling. do.
  • Figure 1 is a view showing the flow of the treatment process in a general medical cell rapid freezing system
  • Figure 2 is a view showing a conventional vitrification process of rapid cooling.
  • a general medical cell rapid freezing system includes a pretreatment of cells, a cell injection process, a quick freezing process, and a storage process. The cryopreserved cells are then thawed later.
  • the conventional rapid cooling vitrification process a high concentration (4-8M) cryopreservative is injected into the cell, and the step-by-step injection to recover.
  • the current vitrification process is divided into several stages to inject a high concentration of cryopreservative into the cells, and even in the thawing process after cryopreservation, the vitrification is divided into several stages to remove the cryopreservative in the cells.
  • This stepwise loading and cryopreservation removal process is very complex and time consuming and can lead to fatal damage to cells due to prolonged processing.
  • many microfluidic devices capable of trapping cells have been developed. However, the microfluidic devices are not suitable for realizing rapid freezing due to their large size and significant drop in thermal conductivity.
  • microcapillaries that are currently capable of trapping cells have not been implemented.
  • Prior art 3 discloses a microfluidic cell trap and assay device for high throughput analysis
  • prior art 4 Controlled loading of cryoprotectants (CPAs) to oocyte with linear and complex CPA profiles.
  • CPAs cryoprotectants
  • the present invention has been proposed to solve the above problems of the conventionally proposed methods, by forming a polymer hollow tube of a polymer material having excellent thermal conductivity as a polymer microcapillary tube, the cell during the treatment in the medical cell rapid freezing system It is an object of the present invention to provide a polymer microcapillary tube for rapid cell freezing, in which cells subjected to a pretreatment procedure can be injected and used for rapid freezing and storage.
  • the present invention comprises a polymer hollow tube made of a polymer having excellent thermal conductivity as a polymer microcapillary tube, but having a material and a radius and a thickness for increasing thermal conductivity, so that the cells are not directly exposed, and both ends of the polymer hollow tube It is another object of the present invention to provide a polymer microcapillary tube for quick freezing of cells, which is easy to seal, prevents breakage during the cooling process, prevents cell loss, and allows cells to be vitrified and observed.
  • the present invention enables the commercialization of the polymer microcapillary tubes required for rapid cryopreservation techniques, and provides a foundation for the development of integrated automated systems in all processes of cryopreservation. Another object is to provide a polymer microcapillary for quick freezing.
  • the present invention has been proposed to solve the above problems of the previously proposed methods, trapping the cells in the capillary tube made of a high thermal conductivity polymer material, so that the fluid flow of the cryopreservative It is an object of the present invention to provide a microcapillary tube capable of trapping cells, which can simplify the complicated steps of the vitrification process in the medical cell quick freezing system and reduce the time required.
  • the present invention provides a microcapillary built-in capillary tube, which allows cell traps, fluid control, and rapid cooling systems to provide an integrated microcapillary tube, while the position of the cell is fixed during handling of the cell during the handling. It is yet another object to provide a cell trap-capable microcapillary tube that does not have to worry about losing cell position, so that it can effectively observe cell volume changes during cryopreservant injection and be used for cell characterization studies.
  • the present invention by having a material, radius and thickness to increase the thermal conductivity of the capillary tube, it is possible to prevent breakage during rapid cooling to prevent cell loss, cell traps and fluid control and microfluidic devices and It is yet another object to provide a cell trap-capable microcapillary, which makes it possible to provide a microcapillary tube that can be integrated as a module of key components of a medical cell rapid freezing system.
  • the polymer microcapillary tube The polymer microcapillary tube,
  • a polymer hollow tube which forms a hollow in which cells which have been subjected to the cell pretreatment can be injected during the treatment in the medical cell rapid freezing system consisting of cell pretreatment, cell injection, and rapid freezing and storage;
  • the cell After the cell is injected into the hollow, it is characterized by the configuration of the polymer material having a high thermal conductivity so that cracking is prevented even during the treatment of the rapid freezing process.
  • the polymer microcapillary tube Preferably, the polymer microcapillary tube,
  • the openings at both ends of the hollow opening may be sealed by heat sealing.
  • the polymer microcapillary tube More preferably, the polymer microcapillary tube,
  • the process of rapid freezing and storage may be performed.
  • the polymer microcapillary tube Preferably, the polymer microcapillary tube,
  • the shape of the polymer hollow tube may be configured in a round shape of a cylinder.
  • the polymer hollow tube More preferably, the polymer hollow tube,
  • the outer shape may be configured in a round shape of a cylinder, and the hollow of the inner diameter may be configured in a round shape.
  • the polymer hollow tube is
  • the outer shape may be configured in a round shape of a cylinder, but one surface may be configured in a plane to facilitate observation after cell injection.
  • the polymer hollow tube is
  • the polymer microcapillary tube Preferably, the polymer microcapillary tube,
  • the shape of the polymer hollow tube may be configured in a square shape of a square pillar.
  • the polymer hollow tube More preferably, the polymer hollow tube,
  • the external shape may be configured in a square shape of a square pillar, and the hollow of the inner diameter may be configured in a square shape.
  • the polymer hollow tube is
  • the polymer microcapillary tube Preferably, the polymer microcapillary tube,
  • the cells injected into the hollow of the polymer hollow tube can be prepared using a photocurable epoxy of a transparent material.
  • the polymer microcapillary tube More preferably, the polymer microcapillary tube,
  • the polymer hollow tube can be produced by an injection molding method.
  • the polymer microcapillary tube More preferably, the polymer microcapillary tube,
  • the polymer hollow tube may be manufactured by a 3D printing process.
  • the polymer microcapillary tube More preferably, the polymer microcapillary tube,
  • the polymer hollow tube may be manufactured by one of injection molding or 3D printing processing methods, but may have a diameter of 500 microns.
  • the polymer microcapillary tube Even more preferably, the polymer microcapillary tube,
  • the polymer hollow tube may have a thickness of 500 microns.
  • the polymer microcapillary Even more preferably, the polymer microcapillary
  • the polymer hollow tube may have a diameter of 500 microns and a thickness of 500 microns. However, when the resolution is increased due to improved performance, the polymer hollow tube may be manufactured as a smaller size capillary tube.
  • the polymer microcapillary Even more preferably, the polymer microcapillary
  • the polymer microcapillary Even more preferably, the polymer microcapillary
  • Surface mirror processing may be performed by reducing the thickness and outer diameter of the polymer hollow tube to maximize thermal conductivity.
  • the polymer microcapillary Even more preferably, the polymer microcapillary
  • Polishing may be performed by reducing the thickness and outer diameter of the polymer hollow tube to maximize thermal conductivity.
  • the polymer microcapillary tube Preferably, the polymer microcapillary tube,
  • the cell is prevented from being directly exposed by injecting the cells into the hollow of the polymer hollow tube, and made of a polymer material having high thermal conductivity, and sealing the openings at both ends by heat sealing, thereby preventing cell loss by breaking during cooling. It can be done.
  • microcapillary tube capable of trapping
  • a microcapillary tube capable of trapping cells capable of trapping cells
  • Cavities in which cells and cryoprotectants can be injected for vitrification of the rapid freezing process are treated in a medical cell rapid freezing system consisting of cell pretreatment, cell injection, and rapid freezing and storage.
  • the capillary Preferably, the capillary,
  • It can be made of a polymer material having high thermal conductivity.
  • the capillary Preferably, the capillary,
  • the cells injected into the hollow may be trapped by the micro filter, and after the cryopreservant is injected, the ends of the hollow may be closed by the sealing portion through heat sealing.
  • the capillary Preferably, the capillary,
  • the cell change injected into the hollow it may be prepared using a photocurable epoxy of a transparent material.
  • the capillary is
  • Rapid prototyping (RP) using the photocurable epoxy of the transparent material can be produced.
  • the capillary is
  • It can be produced by 3D printing processing method.
  • the capillary is
  • the diameter of the hollow can be configured to 500 ⁇ m.
  • the capillary is
  • the thickness of the capillary tube can be configured to 500 ⁇ m.
  • the capillary is
  • It can be configured to a diameter of 500 ⁇ m 500 ⁇ m thickness, but can be composed of a capillary tube of a smaller size (size) when the resolution (resolution) increases due to improved performance.
  • the capillary Preferably, the capillary,
  • the thickness and outer diameter of the capillary tube may be reduced.
  • the capillary is
  • Surface mirror processing may be performed by reducing the thickness and outer diameter of the capillary tube.
  • the capillary is
  • Polishing may be performed in a manner that reduces the thickness and outer diameter of the capillary.
  • the capillary is
  • the hollow tube shape in which the hollow is formed may be configured as one of a round shape of a cylinder or a square shape of a square pillar.
  • the capillary is
  • the outer shape is configured in a round shape of a cylinder, but one surface may be configured in a plane to facilitate observation of the injected cells.
  • the capillary is
  • micro filter More preferably, the micro filter,
  • It may be composed of polystyrene beads (polystyrene beads).
  • micro filter More preferably, the micro filter,
  • the position of the cells is fixed during the cell injection process to maintain the position of the cells during handling, and to observe the volume change of the injected cells during the injection process of the cryopreservative. You can do that.
  • micro filter More preferably, the micro filter,
  • Styrene bead (Styrene bead) can be stacked in a hollow of the capillary tube and then raised to a specific transition temperature (eg, 100 °C) to allow each bead to stick to form a filter.
  • a specific transition temperature eg, 100 °C
  • micro filter Even more preferably, the micro filter,
  • the size of the micropores of the filter can be determined by the treatment time and the bead size of the particular transition temperature.
  • micro filter Even more preferably, the micro filter,
  • the diameter of the polystyrene beads can be configured to 160 ⁇ m.
  • micro filter Even more preferably, the micro filter,
  • cryopreservant may be configured to form a gap of 20-40 ⁇ m with a diameter of the hollow of the capillary so that fluid flow through the polystyrene beads can be controlled.
  • the polymer microcapillary tube for rapid freezing of cells is composed of a polymer hollow tube having excellent thermal conductivity, so that the cell pretreatment process is performed during the treatment in the medical cell rapid freezing system. Cells can be injected and used for quick freezing and storage.
  • the polymer hollow tube made of a polymer having excellent thermal conductivity is composed of a polymer microcapillary tube, and has a material, a radius, and a thickness for increasing thermal conductivity, so that the cells are not directly exposed. It is easy to seal the both ends of the, can be prevented from breaking during the cooling process to prevent cell loss, and to keep the cells in the vitrified state can be observed.
  • microcapillary tube trapping proposes a microcapillary tube trapping proposed in the present invention, by trapping the cells in the capillary tube made of a polymer material having excellent thermal conductivity, by constructing a micro-filter that allows the flow of the cryopreservative fluid in a built-in type In the medical cell rapid freezing system, it is possible to simplify the complicated steps of the vitrification process and to reduce the time required.
  • the present invention by constructing a built-in capillary tube of the micro filter, it is possible to provide a micro capillary tube integrated with the cell trap, fluid control, rapid cooling system, the position of the cell is fixed during the cell injection process, the cell during handling There is no risk of losing the location, which allows for effective observation of cell volume changes and use in cell characterization during cryopreservant injection.
  • a material, radius and thickness to increase the thermal conductivity of the capillary tube it is possible to prevent breakage during rapid cooling to prevent cell loss, and to integrate with cell traps and fluid control and microfluidic devices It may be possible to provide a microcapillary tube as a module of key components of a medical cell rapid freezing system.
  • Figure 1 shows the flow of the process in a typical medical cell rapid freezing system.
  • FIG. 2 is a view illustrating a conventional vitrification process of rapid cooling.
  • Figure 3 is a view showing the flow of processing of the refrigeration system to which the polymer microcapillary tube for rapid cell freezing according to an embodiment of the present invention.
  • Figure 4 is a view showing the configuration of the processing process of the refrigeration system to which the polymer microcapillary tube for rapid cell freezing according to an embodiment of the present invention.
  • Figure 5 is a view showing a perspective view of a round shape of the polymer microcapillary for rapid cell freezing according to an embodiment of the present invention.
  • FIG. 6 is a diagram illustrating a configuration example in which one round surface of a polymer microcapillary tube for rapid freezing of cells is processed in a plane;
  • Figure 7 is a view showing the configuration of a rectangular form of the polymer microcapillary for cell rapid freezing according to an embodiment of the present invention.
  • FIG. 8 is a view showing a configuration in which the heat sealing of the polymer microcapillary tube for rapid freezing of cells according to an embodiment of the present invention is completed.
  • Figure 9 is a view showing the configuration of a capillary microcapillary cell trap according to another embodiment of the present invention.
  • FIG. 10 is a view showing an example of the configuration processing the outer surface of the round shape in a planar microcapillary possible cell trap according to an embodiment of the present invention.
  • FIG. 11 is a view showing an example of the external configuration of the cell trap is possible microcapillary tube according to an embodiment of the present invention.
  • FIG. 12 is a view showing a state of use of the microcapillary capable of trapping cells according to another embodiment of the present invention.
  • capillary 111 hollow
  • sealing portion 120 micro filter
  • FIG 3 is a view showing the flow of the process of the refrigeration system to which the polymer microcapillary tube is applied for rapid cell freezing according to an embodiment of the present invention
  • Figure 4 is a rapid cell freezing according to an embodiment of the present invention Figure showing the configuration of the treatment process of the refrigeration system to which the polymer microcapillary tube is applied.
  • the refrigeration system to which the polymer microcapillary tube for rapid freezing of cells according to an embodiment of the present invention is applied is treated by the process of cell pretreatment, cell injection, rapid freezing, and storage Will be made.
  • the polymer microcapillary tube 100 that can be used for cell injection, rapid freezing, and storage after the cell pretreatment is performed in the medical cell rapid freezing system will be described in detail.
  • Figure 5 is a view showing a perspective view of the round shape of the polymer microcapillary for rapid cell freezing according to an embodiment of the present invention
  • Figure 6 is a polymer microcapillary for rapid cell freezing according to an embodiment of the present invention
  • FIG. 7 is a view showing an example of a configuration in which one surface of a round shape is processed into a plane
  • FIG. 7 is a diagram illustrating a rectangular configuration of a polymer microcapillary tube for rapidly freezing cells according to an embodiment of the present invention.
  • the polymer microcapillary tube 100 for rapid cell freezing according to an embodiment of the present invention may include a polymer hollow tube 102 and an opening part forming a hollow 101. 103, and may further include a heat seal (104).
  • the polymer hollow tube 102 is a hollow 101 into which cells which have been subjected to cell pretreatment can be injected during the treatment in a medical cell rapid freezing system consisting of cell pretreatment, cell injection, and rapid freezing and storage. ) Is a constitution of the microtubules.
  • the polymer hollow tube 102 is preferably made of a polymer material having high thermal conductivity so that the cell can be prevented from being broken even when the cell is injected into the hollow 101 during the rapid freezing process.
  • the polymer hollow tube 102 of the polymer microcapillary tube 100 may have a polymer shape in a cylindrical round shape.
  • the polymer hollow tube 102 constituting the outer shape in a round shape of a cylinder may be configured to form a hollow 101 of the inner diameter in a round shape.
  • the polymer hollow tube 102 having an outer shape in a cylindrical round shape may have a flat surface on one side to facilitate observation after cell injection.
  • the reason for forming a flat surface of a cylindrical shape in a cylindrical shape is to minimize the refraction of light to facilitate cell observation.
  • the polymer hollow tube 102 may have a polymer shape in a square shape of a square pillar.
  • the polymer hollow tube 102 constituting the outer shape in a square shape of the square pillar is to configure the hollow 101 of the inner diameter in a square shape.
  • the polymer hollow tube 102 may be made to easily observe the cells injected into the hollow 101 by minimizing the refraction of light through the rectangular shape of one surface.
  • the opening part 103 is a structure in which the hollows 101 at both ends are opened along the longitudinal direction of the polymer hollow tube 102. That is, the open part 103 refers to a portion in which the hollows 101 are opened at both ends along the length direction of the polymer hollow tube 102.
  • the opening 103 at both ends is sealed through a heat seal 104 which will be described later.
  • the heat seal 104 is configured to seal both ends of the polymer hollow tube 102 after cells are injected into the hollow 101 of the polymer hollow tube 102.
  • the treatment of heat sealing 104 is a treatment configuration that requires additional materials and steps for sealing at both ends in order to go from the conventional quartz capillary to the closed type. To be resolved.
  • the polymer microcapillary tube 100 is injected into the hollow 101 of the polymer hollow tube 102 to prevent the cells from being directly exposed, and is composed of a polymer material having high thermal conductivity, and the amount of heat sealing 104.
  • the opening 103 at the tip can be sealed so that it can not be broken during the cooling process so that cell loss can be prevented.
  • the polymer microcapillary tube 100 is hollow 101 through a heat sealing process after cells are injected into the hollow 101 of the polymer hollow tube 102.
  • the openings 103 at both ends of the open side can be sealed.
  • the polymer microcapillary tube 100 may be used to perform a process of rapid freezing and storage after the openings 103 at both ends of the polymer hollow tube 102 are heat sealed 104 and sealed.
  • the polymer microcapillary tube 100 may be manufactured using a photocurable epoxy of a transparent material for observation of cells injected into the hollow 101 of the polymer hollow tube 102.
  • the polymer microcapillary tube 100 may be manufactured by the injection molding method of the polymer hollow tube 102, or may be manufactured by the 3D printing processing method of the polymer hollow tube 102.
  • the polymer microcapillary tube 100 is manufactured by one of the injection or 3D printing processing method of the polymer hollow tube 102, the diameter may be configured to 500micron.
  • the polymer microcapillary tube 100 may be configured to have a thickness of 500 micron of the polymer hollow tube 102.
  • the polymer microcapillary tube 100 is composed of a diameter of 500 micron and a thickness of 500 micron of the polymer hollow tube 102, but when the resolution is increased due to the improvement in performance, the microcapillary tube of smaller size may be manufactured. Can be.
  • the polymer microcapillary tube 100 may reduce the thickness and outer diameter of the polymer hollow tube 102 to increase the thermal conductivity of the polymer hollow tube 102 having a thickness of 500 micron.
  • the polymer microcapillary tube 100 may perform surface mirror processing in such a manner as to reduce the thickness and outer diameter of the polymer hollow tube 102 to maximize thermal conductivity.
  • the polymer microcapillary tube 100 may be polished in such a manner as to reduce the thickness and outer diameter of the polymer hollow tube 102 to maximize thermal conductivity.
  • Figure 5 is a polymer microcapillary tube 100 for rapid freezing of cells according to an embodiment of the present invention, shows a perspective view of a polymer hollow tube 102 configured in a round shape of a cylinder.
  • FIG. 6A illustrates a side view of one surface of a polymer hollow tube 102 having a cylindrical round shape
  • FIG. 6B shows a polymer hollow tube 102 having a cylindrical round shape.
  • Fig. 1 a perspective view in which one surface is configured as a plane is shown.
  • 7 is a polymer microcapillary tube 100 for rapid freezing of cells according to an embodiment of the present invention.
  • 8 shows a configuration example in which the heat seal 104 of the polymer microcapillary tube 100 for rapid freezing of cells according to an embodiment of the present invention is formed.
  • the microcapillary tube capable of trapping cells according to another embodiment of the present invention may include a capillary tube 110 and a micro filter 120.
  • the capillary 110 is a cell and cryoprotectant to be injected for the vitrification process of rapid cooling in the medical cell rapid freezing system consisting of cell pretreatment, cell injection and rapid freezing and storage. It is a configuration to have a structure in which the hollow 111 at both ends in the longitudinal direction in the form of a hollow tube to form a hollow 111 that can be opened.
  • the capillary tube 110 may be made of a polymer material having high thermal conductivity so that the cell can be prevented from being broken even when the cell is injected into the hollow 111 during the rapid freezing process.
  • the capillary 110 is trapped by the micro-filter 120 which will be described later the cells injected into the hollow 111, the hollow 111 is treated by heat sealing after the cryopreservant is injected. Both ends of the open end may be finished with the seal 112.
  • the capillary 110 may be manufactured using a photocurable epoxy of a transparent material for the observation of cell changes injected into the hollow 111.
  • the capillary tube 110 may be a hollow tube made by Rapid Prototyping (RP) technology as a method of using a photocurable epoxy of a transparent material.
  • RP Rapid Prototyping
  • the advantage of RP is that we can freely make the shape we want, and by using the photocurable epoxy, the light transmittance of the material is excellent, so that the experimenter can easily observe the cells injected into the capillary 110, and also with the help of other complicated devices It is possible to implement the structure of the micro filter 120 in the fine capillary without receiving.
  • the capillary 110 may be manufactured by a 3D printing processing method, in addition to the approach of the RP technology.
  • the capillary 110 is manufactured by one of RP (Rapid Prototyping) technology or 3D printing processing technology, the diameter of the hollow 111, that is, the inner diameter can be configured to 500 ⁇ m, the thickness of the capillary 110 500 ⁇ m It can be configured as.
  • the capillary 110 may be configured to have a diameter of 500 ⁇ m ⁇ 500 ⁇ m, but may also be configured as a capillary 110 having a smaller size when resolution is increased due to performance improvement.
  • the capillary 110 may be further made to reduce the thickness and outer diameter of the capillary 110 to increase the thermal conductivity of the capillary 110 having a thickness of 500 ⁇ m.
  • the capillary tube 110 may be subjected to a surface mirror surface treatment process to reduce the thickness and outer diameter of the capillary tube 110, and another method may be a polishing process to reduce the thickness and outer diameter of the capillary tube 110. May be
  • the capillary tube 110 may be configured to have a hollow tube shape in which the hollow 111 is formed in one of a round shape of a cylinder or a square shape of a square column.
  • the capillary 110 can be implemented in various vessel models through 3D printing or RP technology.
  • the capillary 110 may be configured to have one surface in a plane to facilitate observation of the injected cells when forming the outer shape in a round shape of a cylinder.
  • the capillary 110 may be configured to form a flat surface of a cylindrical shape in a round shape, thereby minimizing the refractive index of the light to facilitate the observation of the injected cells.
  • the micro filter 120 traps cells injected into the hollow 111 formed in the capillary 110, and is formed in the hollow 111 of the capillary 110 to allow the fluid flow of the cryopreservative. . That is, the micro filter 120 is a structure capable of trapping the cells injected into the hollow 111 of the capillary 110, the position of the cells is fixed during the cell injection process to maintain the position of the cells during handling, cryopreservative During the injection process, the volume change of the injected cells can be observed.
  • the micro filter 120 may be composed of polystyrene beads.
  • the micro filter 120 is formed by stacking styrene beads in the hollow 111 of the capillary 110 and stacking the beads at a specific transition temperature (eg, 100 ° C.) to form respective filters. Do it.
  • the size of the micro holes of the filter may be determined according to the processing time and the bead size of the specific transition temperature.
  • the micro filter 120 may preferably be configured to a diameter of 160 ⁇ m polystyrene beads.
  • the micro filter 120 traps cells injected into the hollow 111 of the capillary tube 110, and the cryopreservative may control the flow of the fluid through the polystyrene beads. It is possible to form a gap (Gap) of 20-40 ⁇ m the diameter of the hollow 111 of 110.
  • FIG. 10 is a diagram illustrating a configuration example in which a rounded outer surface of a microcapillary tube according to an embodiment of the present invention is processed in a plane.
  • FIG. 10A illustrates a side view of one surface of the capillary tube 110 having a cylindrical round shape
  • FIG. 10B illustrates one surface of the capillary 110 having a cylindrical round shape.
  • the perspective view structure comprised in a plane is shown. That is, the capillary tube 110 as shown in FIG. 10 is configured to form a surface of a cylindrical round shape in a plane to minimize the refractive index of light to facilitate observation of the injected cells.
  • FIG. 11 is a view showing an example of the external configuration of a microcapillary tube capable of trapping cells according to an embodiment of the present invention.
  • FIG. 11A illustrates a capillary tube capable of trapping cells, and shows an example in which the external shape of the capillary tube 110 in which the microfilter 120 is formed in the hollow 111 in the form of a cylinder is rounded.
  • B shows an example in which the external shape of the capillary tube 110 in which the microfilter 120 is formed in the hollow 111 as a microcapillary tube capable of trapping cells is formed in a square column shape.
  • the capillary 110 may be implemented in various vessel models through 3D printing or RP technology.
  • FIG. 12 is a view showing the state of use of the microcapillary capable of trapping cells according to an embodiment of the present invention.
  • 12A illustrates an example in which a capillary tube 110 and a micro static mixing tee are connected to each other
  • FIG. 12B illustrates a cell injected into the capillary tube 110 in a visceral form.
  • the structure of the state where the cell was trapped by the micro filter 120 which becomes is shown. That is, as shown in FIG. 12, the capillary tube 110 in which the micro filter 120 is formed in a built-in shape holds cells (eg, eggs) which are connected to a micro static mixing tee and injected.
  • the cryopreservative may be passed through the polystyrene beads of the micro filter 120 to enable fluid flow.
  • the cell trap-capable microcapillary tube solves the problem that the conventional microfluidic system does not allow rapid freezing during cryopreservation after cell treatment.
  • Integration with cryopreservant injection technology enables the development of an integrated refrigeration system with an integrated microfluidic device with automatic cell pretreatment.
  • the capillary tube 110 of the built-in filter obtained by the RP technology is used for rapid cooling, as well as can be used in the study of cell characteristics by observing the change of the cell according to the cryopreservant injection profile of the cells in the prototype form.

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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne un tube microcapillaire polymère conçu pour la congélation rapide de cellules, ledit tube étant un tube polymère creux constitué d'un matériau polymère ayant une excellente conductivité thermique. En tant que tube polymère microcapillaire, des cellules ayant subi un prétraitement s'intégrant dans le cadre du traitement d'un système de congélation rapide à visée médicale, peuvent être injectées et utilisées pour être congelées rapidement et être conservées. Par ailleurs, en formant un microfiltre intégré capable de piéger les cellules dans le tube microcapillaire, qui est constitué d'un matériau polymère ayant une excellente conductivité thermique, et en permettant l'écoulement d'un fluide cryoprotecteur, le tube microcapillaire de la présente invention permet de simplifier les étapes complexes du procédé de vitrification d'un système de congélation rapide à visée médicale, et d'en réduire la durée.
PCT/KR2015/002062 2015-03-04 2015-03-04 Tube microcapillaire polymère pour la congélation rapide de cellules, et tube microcapillaire capable de piéger des cellules WO2016140379A1 (fr)

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CN114532329A (zh) * 2017-04-21 2022-05-27 富士胶片欧文科技有限公司 玻璃化装置和用于制备样品的方法

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CN114532329A (zh) * 2017-04-21 2022-05-27 富士胶片欧文科技有限公司 玻璃化装置和用于制备样品的方法
CN114532329B (zh) * 2017-04-21 2023-11-28 富士胶片欧文科技有限公司 玻璃化装置和用于制备样品的方法
CN113907065A (zh) * 2021-11-08 2022-01-11 深圳先进技术研究院 换液处理芯片和冷冻载杆
CN113907065B (zh) * 2021-11-08 2022-06-14 深圳先进技术研究院 换液处理芯片和冷冻载杆

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