Decellularization reaction device

Abstract

Provided is a decellularization reaction device. A decellularization reaction device according to the present invention comprises: a first reaction container for storing a first decellularization solution for performing a decellularization reaction; and a first ex-situ analysis application device for extracting a first decellularized solution that is subjected to a decellularized reaction with the first heterogeneous organ tissue in the first reaction container, and measuring the physical properties of the first decellularized solution in an ex-situ manner.

Description

Technical Field

[0001] The present invention relates to a decellularization reaction device, and more particularly to a decellularization reaction device for a small number of multi-variety xenogeneic organs. Background Art

[0002] Recently, with the global aging trend, the number of patients with chronic diseases has increased, and the technology of manufacturing artificial organs using 3D bioprinting has attracted much attention.

[0003] Bioink refers to an ink material used to output artificial organs through a 3D bioprinter, which is manufactured based on the extracellular matrix obtained by decellularizing xenogeneic organs.

[0004] When the number of xenogeneic organs to be decellularized is large, a reactor form suitable for large-scale production can be adopted. However, when the amount of tissue of a xenogeneic organ that can be collected, such as a porcine cornea, is small, for example, at the level of 10 g, it is difficult to operate large-scale equipment.

[0005] In addition, there is an urgent need to propose a device that can decellularize a small amount of multi-variety xenogeneic organ tissues, such as specific tissues of the heart (such as the left atrium, left ventricle, right atrium, and right ventricle), and can compare the physical property differences between xenogeneic organ tissues. Summary of the Invention

[0006] Technical Problem

[0007] The present invention provides a decellularization reaction device that can efficiently decellularize multi-variety and small amounts of xenogeneic organ tissues.

[0008] Technical Solution

[0009] The decellularization reaction device according to an embodiment of the present invention may include: a first reaction container that stores a first decellularization solution for performing a decellularization reaction with the tissue of a first xenogeneic organ to be decellularized therein.

[0010] In addition, the decellularization reaction device may include: one or more first ex-situ analysis application devices for extracting the first decellularization solution that is undergoing a decellularization reaction with the tissue of the first xenogeneic organ to be decellularized in the first reaction container to the outside of the first reaction container and ex-situ measuring the physical properties of the first decellularization solution.

[0011] The first ex-situ analysis application device may include: a first grid chamber inserted into the first reaction container and accommodating the tissue of the first xenogeneic organ to be decellularized therein.

[0012] The first off-site analysis application device may include: a first solution circulation unit, which is connected between the first reaction vessel and the first cover covering the first reaction vessel and is in communication with each other, and is used to circulate the first decellularized solution between the first reaction vessel and the first cover.

[0013] In addition, the first off-site analysis application device may include: a first solution sampling unit, which is connected to the first solution circulation unit and is used to extract the first decellularized solution from the first solution circulation unit to the outside for sampling.

[0014] It may include: a first lifting unit, which is combined with the first cover and is used to lift the first cover in the vertical direction.

[0015] The first grid chamber may include: a first driving unit, which is used to rotate the first grid chamber in the first reaction vessel for stirring.

[0016] The first grid chamber may be configured in a hollow cylindrical shape inside.

[0017] On the outer peripheral surface of the first grid chamber, a first grid may be arranged in a grid pattern.

[0018] At the upper end of the first grid chamber, a first cover for covering the first grid chamber can be detachably combined.

[0019] The first reaction vessel may be made of a transparent material or have a transparent window so that the inside of the first reaction vessel can be observed from the outside.

[0020] The first reaction vessel may have a double-layer jacket shape composed of a first inner tube and a first outer tube.

[0021] On the first outer tube, a first coolant inlet and a first coolant outlet may be respectively provided. The first coolant inlet is used to inject coolant into the space between the first outer tube and the first inner tube, and the first coolant outlet is used to discharge the injected coolant to the outside of the first outer tube.

[0022] The first reaction vessel may have a first solution outlet for flowing the decellularized solution out of the first reaction vessel.

[0023] On the first cover, a first injection port for injecting the first decellularized solution into the inside of the first reaction vessel may be installed.

[0024] On the first cover, a first water level measuring instrument for measuring the water level of the first decellularized solution in the first reaction vessel may be installed.

[0025] The first driving unit may include: a first rotating shaft coupled to the first lid of the first mesh chamber; and a first driving motor mounted on the first cover of the first reaction vessel while being coupled to the first rotating shaft and configured to rotate the first rotating shaft.

[0026] The first solution circulation unit may include: a first circulation pipe connected between the first cover and the first solution outlet for circulating the first decellularized solution in the first reaction vessel; and a first transfer pump mounted on the first circulation pipe for transferring the first decellularized solution flowing out of the first solution outlet to the first circulation port.

[0027] The first circulation pipe may be composed of a flexible pipe capable of expansion and contraction.

[0028] The first solution sampling unit may include: a first solution sampling pipe connected to the first circulation pipe for extracting and sampling the first decellularized solution circulating in the first circulation pipe.

[0029] The absorbance (UV-Vis), conductivity, and turbidity of the first decellularized solution sampled by the first solution sampling pipe may be measured, and the physical properties of the first decellularized solution may be analyzed.

[0030] The first lifting unit may include: a first coupling plate coupled to the first cover and capable of lifting; and a first lifting cylinder coupled to one end of the first coupling plate and having a first lifting rod for lifting the first coupling plate.

[0031] The decellularization reaction device according to another implementation example of the present invention may include: a first reaction vessel storing therein a first decellularized solution for performing a decellularization reaction with the tissue of a first xenogeneic organ as a decellularization target.

[0032] In addition, the decellularization reaction device according to another implementation example may include: one or more first ex-situ analysis application devices for extracting the first decellularized solution that is undergoing a decellularization reaction with the tissue of the first xenogeneic organ as a decellularization target in the first reaction vessel to the outside and measuring the physical properties of the first decellularized solution ex-situ.

[0033] In addition, the decellularization reaction device according to another implementation example may include: a second reaction vessel storing therein a second decellularized solution for performing a decellularization reaction with the tissue of a second xenogeneic organ as a decellularization target.

[0034] The decellularization reaction device according to another implementation example may include: one or more second ex-situ analysis application devices arranged side by side with the first ex-situ analysis application device and configured to extract the second decellularized solution that is undergoing a decellularization reaction with the tissue of the second xenogeneic organ as a decellularization target in the second reaction vessel to the outside and measure the physical properties of the second decellularized solution ex-situ.

[0035] The second xenogeneic organ may be the same as or different from the first xenogeneic organ, and the second decellularized solution may be the same as or different from the first decellularized solution.

[0036] The first ex vivo analysis application device may include: a first grid chamber inserted inside a first reaction vessel and accommodating the tissue of the first xenogeneic organ to be decellularized therein; a first solution circulation unit connected between the first reaction vessel and a first lid covering the first reaction vessel and communicating with each other, and configured to circulate the first decellularized solution between the first reaction vessel and the first lid.

[0037] In addition, the first ex vivo analysis application device may include: a first solution sampling unit connected to the first solution circulation unit and configured to extract the first decellularized solution from the first solution circulation unit to the outside for sampling.

[0038] The first ex vivo analysis application device may include: a first lifting unit coupled to the first lid and configured to lift the first lid in the vertical direction; and a first driving unit coupled to the first grid chamber and configured to rotate the first grid chamber within the first reaction vessel for stirring.

[0039] In addition, the second ex vivo analysis application device may include: a second grid chamber inserted inside a second reaction vessel and accommodating the tissue of the second xenogeneic organ to be decellularized therein.

[0040] The second ex vivo analysis application device may include: a second solution circulation unit connected between the second reaction vessel and a second lid covering the second reaction vessel and communicating with each other, and configured to circulate the second decellularized solution between the second reaction vessel and the second lid.

[0041] In addition, the second ex vivo analysis application device may include: a second solution sampling unit connected to the second solution circulation unit and configured to extract the second decellularized solution from the second solution circulation unit to the outside for sampling.

[0042] It may include: a second lifting unit coupled to the second lid and configured to lift the second lid in the vertical direction.

[0043] The second grid chamber may include: a second driving unit configured to rotate the second grid chamber within the second reaction vessel for stirring.

[0044] The second grid chamber may be configured in a hollow cylindrical shape inside.

[0045] A second grid may be arranged in a grid pattern on the outer peripheral surface of the second grid chamber.

[0046] At the upper end of the second grid chamber, a second lid for covering the second grid chamber may be detachably coupled.

[0047] The second reaction vessel may be made of a transparent material or have a transparent window so that the interior of the second reaction vessel can be observed from the outside.

[0048] The second reaction vessel may have a double jacket configuration consisting of a second inner tube and a second outer tube.

[0049] A second coolant inlet and a second coolant outlet may be respectively provided on the second outer tube. The second coolant inlet is used to inject coolant into the space between the second outer tube and the second inner tube, and the second coolant outlet is used to discharge the injected coolant to the outside of the second outer tube.

[0050] The second reaction vessel may have a second solution outlet for flowing out the decellularized solution to the outside of the second reaction vessel.

[0051] A second injection port for injecting a second decellularized solution into the interior of the second reaction vessel may be installed on the second cover.

[0052] The second driving unit may include: a second rotating shaft coupled to the second cover of the second mesh chamber; and a second driving motor installed on the second cover of the second reaction vessel and coupled to the second rotating shaft for rotating the second rotating shaft.

[0053] The second solution circulation unit may include: a second circulation pipe connected between the second cover and the second solution outlet for circulating the second decellularized solution in the second reaction vessel; and a second delivery pump installed on the second circulation pipe for delivering the second decellularized solution flowing out from the second solution outlet to the second circulation port.

[0054] The second circulation pipe may be composed of a flexible pipe capable of expansion and contraction.

[0055] The second solution sampling unit may include: a second solution sampling pipe connected to the second circulation pipe for extracting and sampling the second decellularized solution circulating in the second circulation pipe.

[0056] The absorbance (UV-Vis), conductivity, and turbidity of the second decellularized solution sampled by the second solution sampling pipe may be measured, and the physical properties of the second decellularized solution may be analyzed.

[0057] The second lifting unit may include: a second coupling plate coupled to the second cover and capable of lifting; and a second lifting cylinder coupled to one end of the second coupling plate and having a second lifting rod for lifting the second coupling plate.

[0058] Beneficial effects

[0059] An implementation example of the present invention utilizes a first and a second reaction vessel; a first and a second grid chamber; a circulation structure for a first and a second decellularized solution; and a dual-machine system including a first ex situ analysis application device and a second ex situ analysis application device. Therefore, it can efficiently operate the decellularization process of multiple varieties of small amounts of xenogeneic organs, and the efficiency of physical property evaluation between tissues is also very high. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 FIG. is a schematic structural diagram of a decellularization reaction device according to an embodiment of the present invention, showing a state where the first and second covers are opened.

[0061] Figure 2 FIG. is a structural diagram of a first ex situ analysis application device in a decellularization reaction device according to an embodiment of the present invention, showing a state where the first cover is closed.

[0062] Figure 3 FIG. is a photograph of each part of a porcine heart after a trimming pretreatment process used in a decellularization reaction device according to an embodiment of the present invention.

[0063] Figure 4 FIG. is a graph showing the turbidity analysis results over time in the first ultrapure water process (DW(1)) and the turbidity analysis results of the first decellularized solution using three times of ultrapure water [DW(1)-DW(2)-DW(3)] in the decellularization process of each tissue of porcine myocardium using a decellularization reaction device according to an embodiment of the present invention.

[0064] Figure 5 FIG. is a graph showing the conductivity analysis results over time in the first ultrapure water process (DW(1)) and the conductivity analysis results of the first decellularized solution using three times of ultrapure water [DW(1)-DW(2)-DW(3)] in the decellularization process of each tissue of a porcine heart using a decellularization reaction device according to an embodiment of the present invention.

[0065] Figure 6 FIG. is a graph showing the absorbance analysis results over time in the first ultrapure water process (DW(1)) and the absorbance analysis results of the first decellularized solution of the left ventricle using three times of ultrapure water [DW(1)-DW(2)-DW(3)] in the decellularization process of each tissue of porcine myocardium using a decellularization reaction device according to an embodiment of the present invention.

[0066] Figure 7 FIG. is a photograph of the process (a)-(b)-(c) of decellularizing myocardial tissue using a decellularization reaction device according to an embodiment of the present invention, where ultrapure water and various organic solvents are used as the first decellularized solution. DETAILED DESCRIPTION OF THE INVENTION

[0067] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings so that those of ordinary skill in the art to which the present invention pertains can easily implement the present invention. As those of ordinary skill in the art to which the present invention pertains can understand, without departing from the concept and scope of the present invention, the following-described embodiments can be deformed into various forms. For the same or similar parts, the same reference numerals are used as much as possible in the drawings.

[0068] The technical terms used hereinafter are only for referring to specific embodiments and do not intend to limit the present invention. Unless the sentence clearly indicates the contrary meaning, the singular form used herein also includes the plural form. The meaning of "comprising" used in the specification embodies specific characteristics, regions, integers, steps, operations, elements, and / or components, but does not exclude the existence or addition of other specific characteristics, regions, integers, steps, operations, elements, components, and / or groups.

[0069] All terms, including technical terms and scientific terms, used hereinafter have the same meaning as those generally understood by those of ordinary skill in the art to which the present invention pertains. Terms defined in the dictionary are further interpreted to be consistent with the relevant technical literature and the currently disclosed content. Unless otherwise defined, they should not be interpreted as ideal or very formal meanings.

[0070] Figure 1 is a schematic structural diagram of a decellularization reaction device according to an embodiment of the present invention, and is a diagram showing a state in which the first and second covers are opened.

[0071] Figure 2 is a structural diagram of a first in-situ analysis application device in a decellularization reaction device according to an embodiment of the present invention, and is a diagram showing a state in which the first cover is closed.

[0072] Refer to Figure 1 and Figure 2 According to the first embodiment of the present invention, the decellularization reaction device may include a first in-situ analysis application device RA1 and a second in-situ analysis application device RA2.

[0073] The first in-situ analysis application device RA1 may be provided as at least one, which can extract the first decellularization solution that is undergoing a decellularization reaction with the first xenogeneic organ tissue to be decellularized to the outside and perform an ex-situ analysis on various physical properties of the first decellularization solution.

[0074] The second in-situ analysis application device RA2 may be at least one, and is arranged side by side with the first in-situ analysis application device RA1 at a set distance. It can extract the second decellularization solution that is undergoing a decellularization reaction with the second xenogeneic organ tissue to be decellularized to the outside and perform an ex-situ analysis on various physical properties of the second decellularization solution.

[0075] Here, ex-situ analysis may refer to extracting the first decellularized solution and the second decellularized solution from the first ex-situ analysis application device RA1 and the second ex-situ analysis application device RA2 to the outside, and respectively measuring and analyzing the physical properties of the first decellularized solution and the second decellularized solution.

[0076] The first xenogeneic organ may be the same as or different from the second xenogeneic organ, and the first decellularized solution may be the same as or different from the second decellularized solution.

[0077] In addition, the first ex-situ analysis application device RA1 may include a first grid chamber 100, a first reaction vessel 200, a first driving unit 300, a first solution circulation unit 400, and a first solution sampling unit 500.

[0078] The first grid chamber 100 may have a set size and shape, and the first xenogeneic organ tissue to be decellularized may be accommodated inside it.

[0079] In addition, the first grid chamber 100 may be inserted into the first reaction vessel 200, and the first reaction vessel 200 may store the first decellularized solution for decellularizing the first xenogeneic organ tissue in the first grid chamber 100.

[0080] The first driving unit 300 may be coupled to the first grid chamber 100, and may rotate the first grid chamber 100 in the first reaction vessel 200 for stirring.

[0081] The first solution circulation unit 400 may be connected between the first reaction vessel 200 and a first cover 210 covering the first reaction vessel 200 and communicate with each other, and is used to circulate the first decellularized solution between the first reaction vessel 200 and the first cover 210.

[0082] The first solution sampling unit 500 may be connected to the first solution circulation unit 400. In order to measure various physical properties of the first decellularized solution, the first solution sampling unit 500 may extract the first decellularized solution from the first solution circulation unit 400 to the outside for sampling.

[0083] A first cover 210 for covering the first reaction vessel 200 may be arranged above the first reaction vessel 200.

[0084] In addition, a first lifting unit 600 for lifting the first cover 210 relative to the first reaction vessel 200 in the vertical direction ( Figure 1 the Y direction in

[0085] The first grid chamber 100 can be easily rotated by the first driving unit 300 and is configured in a cylindrical shape with a hollow interior so that the acellular target first xenogeneic organ tissue can be placed and accommodated inside.

[0086] In addition, when the first grid chamber 100 is in a quadrilateral shape, the xenogeneic organ tissue will be clamped or retained at the corner parts during the rotation of the first grid chamber 100, resulting in uneven agitation of the xenogeneic organ tissue. Therefore, in order to agitate the xenogeneic organ tissue evenly and efficiently, the first grid chamber 100 is designed in a cylindrical shape or the like.

[0087] The first grid chamber 100 can be configured as a small chamber that can accommodate at most less than 25 g of the acellular target first xenogeneic organ.

[0088] On the outer peripheral surface of the first grid chamber 100, the first grid 101 can be arranged in a grid pattern. In order to cause the first xenogeneic organ tissue and the like accommodated in the first grid chamber 100 to undergo an acellular reaction, the first grid 101 can have a set size, for example, a size of about 1 mm.

[0089] In addition, the size of the first grid 101 can be set to be slightly smaller than the size of the first xenogeneic organ tissue to prevent the first xenogeneic organ tissue and the like accommodated in the first grid chamber 100 from leaking outside the first grid 101 during the acellular process.

[0090] It may include a first lid 110 that is detachably coupled to the upper end of the first grid chamber 100 and is used to cover the first grid chamber 100.

[0091] The first lid 110 can have a form such as a buckle 111 so that it can be easily coupled to the first grid chamber 100, but it is not limited thereto. Of course, the first lid 110 and the first grid chamber 100 can also be coupled by means of screw coupling or the like.

[0092] The first driving unit 300 can be coupled to the upper end of the first lid 110.

[0093] The first grid chamber 100 can be made of a material with excellent chemical resistance such as Teflon or coated with such a material according to the physical properties of the first acellular solution.

[0094] In addition, the first reaction vessel 200 can be made of a transparent material or have a transparent window (not shown) so that the progress of the acellular process occurring inside the first reaction vessel 200 can be visually observed from the outside of the first reaction vessel 200.

[0095] The first reaction vessel 200 can be arranged on the first base 10 at a set interval and is supported by the 1-1 support frame 20, and the 1-1 support frame 20 is arranged relative to the first base 10 in the vertical direction ( Figure 1 the Y direction in

[0096] The first reaction vessel 200 can be made of a material that is corrosion-resistant to the decellularized solution to be used.

[0097] In addition, the first reaction vessel 200 can have a form such as a double jacket composed of a first inner tube 201 and a first outer tube 203 to control the temperature of the decellularization process.

[0098] A first coolant inlet 205 and a first coolant outlet 206 can be respectively provided on the first outer tube 203. The first coolant inlet 205 is used to inject coolant into the space between the first outer tube 203 and the first inner tube 201, and the first coolant outlet 206 is used to discharge the coolant to the outside of the first outer tube 203.

[0099] The first reaction vessel 200 can have a first solution outlet 207 for flowing out the decellularized solution to the outside of the first reaction vessel 200.

[0100] The first cover 210 can be made of a rubber material and configured in the form of an O-ring so that when it descends through the first lifting part 600 and contacts the upper end of the first reaction vessel 200, it can buffer the impact and seal the first reaction vessel 200 at the same time.

[0101] A first injection port (not shown) for connecting the first solution injection tube 220 can be installed on the first cover 210, and the first solution injection tube 220 is used to inject the first decellularized solution into the first reaction vessel 200.

[0102] The first decellularized solution can be composed of, for example, ultrapure water, organic solvents, etc.

[0103] In addition, a first solution circulation part 400 is connected to the first cover 210, so that the first decellularized solution circulated in the first solution circulation part 400 can be injected into the first reaction vessel 200.

[0104] A first water level measuring instrument 213 for measuring the water level of the first decellularized solution in the first reaction vessel 200 can be installed on the first cover 210, and the first water level measuring instrument 213 can be operated by ultrasonic waves or the like.

[0105] The first driving part 300 can include a first rotating shaft 310 and a first driving motor 320.

[0106] The first rotating shaft 310 can be coupled with the first cover 110 of the first grid chamber 100.

[0107] In addition, the first driving motor 320 can be installed on the first cover 210 of the first reaction vessel 200 while being coupled with the first rotating shaft 310, so as to enable the first rotating shaft 310 to rotate.

[0108] The first driving motor 320 can be composed of a magnetically controlled motor or the like, so as to easily rotate the first rotating shaft 310.

[0109] The first rotating shaft 310 can be arranged at the center of the upper end portion of the first cover 110 along the vertical direction ( Figure 2 the Y direction in

[0110] ) of the upper end portion of the first cover 110, and can be coupled with the first cover 110 by being coupled with the first bolt 301 or the like.

[0111] The first rotating shaft 310 can be rotated in the clockwise direction or the counterclockwise direction by the first driving motor 320, so as to drive the first grid chamber 100 to rotate to stir the first xenogeneic organ tissue accommodated in the first grid chamber 100, thereby guiding the first xenogeneic organ tissue to smoothly achieve decellularization under the action of the first decellularized solution injected into the first reaction vessel 200.

[0112] The first solution circulation unit 400 can include a first circulation pipe 410 and a first delivery pump 420.

[0113] The first circulation pipe 410 can be connected between a first circulation port (not shown) arranged on the first cover 210 and the first solution outlet 207, for circulating the first decellularized solution in the first reaction vessel 200.

[0114] In addition, the first delivery pump 420 can be installed on the first circulation pipe 410, and can quantitatively deliver the first decellularized solution flowing out from the first solution outlet 207 to the first circulation port.

[0115] The first circulation pipe 410 can be composed of a flexible pipe, so as to easily expand and contract when the first cover 210 is lifted and lowered by the first lifting unit 600.

[0116] In addition, the first circulation pipe 410 may be connected to a first waste liquid discharge pipe 430 for discharging the waste liquid of the first decellularized solution, and a first waste liquid discharge pipe control valve 431 for controlling the discharge of the first decellularized solution may be installed on the first waste liquid discharge pipe 430.

[0117] The first solution sampling unit 500 may include a first solution sampling pipe 510, which is connected to the first circulation pipe 410 and is used for extracting and sampling the first decellularized solution circulating in the first circulation pipe 410.

[0118] The first decellularized solution sampled in the first solution sampling pipe 510 may be filled into a first small container 520 such as a vial, and then various physical properties (such as absorbance (UV-Vis), conductivity, turbidity, etc.) of the first decellularized solution filled in the first small container 520 are measured and analyzed.

[0119] A first solution sampling pipe control valve 511 for controlling the opening and closing of the first solution sampling pipe 510 may be installed on the first solution sampling pipe 510.

[0120] In addition, the first lifting unit 600 may include a first coupling plate 610 and a first lifting cylinder 620.

[0121] The first coupling plate 610 may be integrally liftably coupled to the first cover 210.

[0122] In addition, the first lifting cylinder 620 may be coupled to the lower end of the first coupling plate 610 and have a first lifting rod 621 for lifting the first coupling plate 610.

[0123] The first lifting cylinder 620 may be installed and supported on the upper end surface of the first base 10 relative to the first base 10 along the vertical direction ( Figure 1 the Y direction in

[0124] A first air supply pipe 623 for supplying air into the first lifting cylinder 620 may be connected to the first lifting cylinder 620, and a first regulator 625 for adjusting the air pressure supplied to the first air supply pipe 623 may be installed on the first air supply pipe 623.

[0125] In addition, a 1-2 support frame 30 may be installed on the first base 10, and the 1-2 support frame 30 is arranged parallel to the 1-1 support frame 20 and is supported on the first base 10.

[0126] A first guide rod 630 may be coupled to the 1-2 support frame 30. The first guide rod 630 is connected to the 1-2 support frame 30 through a first connection frame 31 and is used for guiding the lifting of the first lifting rod 621.

[0127] In addition, for ease of cleaning, the first reaction vessel 200 and the 1-1 support frame 20 can be configured to be movable and detachable, and a first groove 11 for aligning the positions of the first reaction vessel 200 and the first cover 210 can be arranged on the upper end face of the first base 10.

[0128] A first elastic member 21 can be coupled to the upper end portion of the 1-1 support frame 20, and the first elastic member 21 is used to buffer the impact generated when the first cover 210 descends and seals the first reaction vessel 200.

[0129] In addition, the second off-site analysis application device RA2 can include a second grid chamber 100A, a second reaction vessel 200A, a second driving unit 300A, a second solution circulation unit 400A, and a second solution sampling unit 500A.

[0130] The second grid chamber 100A can have a set size and shape, and the acellular target second xenogeneic organ tissue is accommodated therein.

[0131] In addition, the second reaction vessel 200A can be at least one, and is arranged side by side with the first reaction vessel 200A at a set distance. The second reaction vessel 200A can store a second decellularization solution for performing a decellularization reaction with the second xenogeneic organ tissue inserted into the second grid chamber 100A therein.

[0132] The second driving unit 300A can be coupled to the second grid chamber 100A, and can rotate the second grid chamber 100A in the second reaction vessel 200A for stirring.

[0133] In addition, the second solution circulation unit 400A can be connected between the second reaction vessel 200A and the second cover 210A covering the second reaction vessel 200A and communicate with each other, and can circulate the second decellularization solution between the second reaction vessel 200A and the second cover 210A.

[0134] The second solution sampling unit 500A can be connected to the second solution circulation unit 400A. In order to measure various physical properties of the second decellularization solution, the second solution sampling unit 500A can extract the second decellularization solution from the second solution circulation unit 400A to the outside for sampling.

[0135] A second cover 210A for covering the second reaction vessel 200A can be arranged above the second reaction vessel 200A.

[0136] In addition, a second elastic member 22 can be coupled to the second cover 210A for moving the second cover 210A relative to the second reaction vessel 200A in the vertical direction ( Figure 1a second lifting part 600A that can be lifted in the Y direction (in the figure) so that the second cover 210A can be combined with or separated from the second reaction vessel 200A.

[0137] The second grid chamber 100A can be easily rotated by the second driving part 300A and can have a cylindrical shape or the like so that the decellularized target second xenogeneic organ tissue or the like can be placed and accommodated inside.

[0138] In addition, when the second grid chamber 100A has a quadrilateral shape, the xenogeneic organ tissue will be clamped or retained at the corner parts when the second grid chamber 100A rotates, resulting in uneven stirring of the xenogeneic organ tissue. Therefore, in order to stir the xenogeneic organ tissue evenly and efficiently, the second grid chamber 100A is constructed in a cylindrical shape or the like.

[0139] The second grid 101A can be arranged in a grid pattern on the second grid chamber 100A. In order to perform a decellularization reaction on the second xenogeneic organ tissue or the like accommodated in the second grid chamber 100A, the second grid 101A can have a set size, for example, a size of about 1 mm.

[0140] The second grid chamber 100A can be constructed as a small chamber that can accommodate at most less than 25 g of the decellularized target second xenogeneic organ.

[0141] In addition, the size of the second grid 101A can be set to be slightly smaller than the size of the second xenogeneic organ tissue to prevent the second xenogeneic organ tissue or the like accommodated in the second grid chamber 100A from leaking outside the second grid 101A during the decellularization process.

[0142] A second cover 110A can be included, which is detachably combined with the upper end of the second grid chamber 100A and is used to cover the second grid chamber 100A.

[0143] The second cover 110A can have a shape such as a buckle 111A so that it can be easily combined with the second grid chamber 100A, but it is not limited thereto. Of course, the second cover 110A and the second grid chamber 100A can also be combined by means of screw connection or the like.

[0144] The second driving part 300A can be combined with the upper end of the second cover 110A.

[0145] The second grid chamber 100A can be made of a material with excellent chemical resistance such as Teflon or coated with such a material according to the physical properties of the second decellularization solution.

[0146] In addition, the second reaction vessel 200A may be made of a transparent material or have a transparent window (not shown) so that the progress of the decellularization process inside the second reaction vessel 200A can be visually observed from the outside of the second reaction vessel 200A.

[0147] The second reaction vessels 200A may be arranged on the second base 10A at set intervals and supported by the second support frame 20A, and the second support frame 20A is arranged along the vertical direction ( Figure 1 the Y direction) with respect to the second base 10A.

[0148] The second reaction vessel 200A may be made of a material that is corrosion-resistant to the decellularization solution to be used.

[0149] In addition, the second reaction vessel 200A may have a form such as a double jacket composed of a second inner tube 201A and a second outer tube 203A to control the temperature of the decellularization process.

[0150] A second coolant inlet 205A and a second coolant outlet 206A may be respectively provided on the second outer tube 203A. The second coolant inlet 205A is used to inject coolant into the space between the second outer tube 203A and the second inner tube 201A, and the second coolant outlet 206A is used to discharge the coolant to the outside of the second outer tube 203A.

[0151] The second reaction vessel 200A may have a second solution outlet 207A for the decellularization solution to flow out.

[0152] The second cover 210A may be made of a rubber material or the like and configured in the form of an O-ring so that when it descends through the second lifting part 600A and contacts the upper end of the second reaction vessel 200A, it can buffer the impact and seal the second reaction vessel 200A at the same time.

[0153] A second injection port (not shown) for connecting the second solution injection tube 220A may be installed on the second cover 210A, and the second solution injection tube 220A is used to inject the second decellularization solution into the interior of the second reaction vessel 200A.

[0154] The second decellularization solution may be composed of, for example, ultrapure water, an organic solvent, etc.

[0155] In addition, a second solution circulation part 400A may be connected to the second cover 210A so that the second decellularization solution circulated in the second solution circulation part 400A can be injected into the interior of the second reaction vessel 200A.

[0156] A second water level measuring instrument 213A for measuring the water level of the second decellularized solution in the second reaction vessel 200A can be installed on the second cover 210A, and the second water level measuring instrument 213A can be operated by ultrasonic waves or the like.

[0157] The second driving unit 300A may include a second rotating shaft 310A and a second driving motor 320A.

[0158] The second rotating shaft 310A may be coupled to the second cover 110A of the second mesh chamber 100.

[0159] In addition, the second driving motor 320A may be installed on the second cover 210A of the second reaction vessel 200A and coupled to the second rotating shaft 310A, so as to be able to rotate the second rotating shaft 310A.

[0160] The second driving motor 320A may be composed of a magnetically controlled motor or the like, so as to be able to easily rotate the second rotating shaft 310A.

[0161] The second rotating shaft 310A may be arranged at the center of the upper end portion of the second cover 110A along the vertical direction ( Figure 1 the Y direction in) with respect to the upper end portion of the second cover 110A, and may be coupled to the second cover 110A by being coupled to the second bolt 301A or the like.

[0162] The second rotating shaft 310A may be rotated in the clockwise direction or the counterclockwise direction by the second driving motor 320A, so as to drive the second mesh chamber 100A to rotate to stir the second xenogeneic organ tissue accommodated in the second mesh chamber 100A, thereby guiding the second xenogeneic organ tissue to smoothly achieve decellularization under the action of the second decellularized solution injected into the second reaction vessel 200A.

[0163] When the second mesh chamber 100A is rotated and stirred at a set rotational speed (for example, 1 to 300 rpm), in order to prevent overload of the second driving motor 320A and prevent entanglement between xenogeneic organs, stirring may be alternately performed from the forward (clockwise direction) to the reverse (counterclockwise direction), and a pause time may be set in the middle.

[0164] The second solution circulation unit 400A may include a second circulation pipe 410A and a second delivery pump 420A.

[0165] The second circulation pipe 410A may be connected between a second circulation port (not shown) arranged on the second cover 210A and the second solution outlet 207A, for circulating the second decellularized solution in the second reaction vessel 200A.

[0166] In addition, the second transfer pump 420A can be installed on the second circulation pipe 410A to quantitatively transfer the second decellularized solution flowing out from the second solution outlet 207A.

[0167] The second circulation pipe 410A can be composed of a flexible pipe so that it can be easily stretched and contracted when the second cover 210A is lifted and lowered by the second lifting part 600A.

[0168] In addition, the second circulation pipe 410A can be connected with a second waste liquid discharge pipe 430A for discharging the waste liquid of the second decellularized solution, and a second waste liquid discharge pipe control valve 431A for controlling the discharge of the second decellularized solution can be installed on the second waste liquid discharge pipe 430A.

[0169] The second solution sampling part 500A can include a second solution sampling pipe 510A, which is connected to the second circulation pipe 410A and is used for extracting and sampling the second decellularized solution circulating in the second circulation pipe 410A.

[0170] The second decellularized solution sampled by the second solution sampling pipe 510A can be filled into a second small container 520A such as a vial, and then various physical properties (such as absorbance (UV-Vis), conductivity, turbidity, etc.) of the second decellularized solution filled in the second small container 520A are measured.

[0171] A second solution sampling pipe control valve 511A for controlling the opening and closing of the second solution sampling pipe 510A can be installed on the second solution sampling pipe 510A.

[0172] In addition, the second lifting part 600A can include a second coupling plate 610A and a second lifting cylinder 620A.

[0173] The second coupling plate 610A can be integrally lifted and lowered with the second cover 210A.

[0174] In addition, the second lifting cylinder 620A can be coupled to the lower end of the second coupling plate 610A and has a second lifting rod 621A for lifting the second coupling plate 610A.

[0175] The second lifting cylinder 620A can be installed and supported on the upper end surface of the second base 10A along the vertical direction ( Figure 1 the Y direction in

[0176] A second air supply pipe 623A for supplying air into the second lifting cylinder 620A can be connected to the second lifting cylinder 620A, and a second regulator 625A for adjusting the air pressure supplied to the second air supply pipe 623A can be installed on the second air supply pipe 623A.

[0177] In addition, a second support frame 30A may be installed on the second base 10A. The second support frame 30A is arranged to be parallel to the second support frame 20A and is supported on the second base 10A.

[0178] A second guide rod 630A may be coupled to the second support frame 30A. The second guide rod 630A is connected to the second support frame 30A through a second connection frame 31A and is used to guide the lifting of the second lifting rod 621A.

[0179] In addition, for ease of cleaning, the second reaction vessel 200A and the second support frame 20A may be configured to be movable and detachable. A second groove 11A for aligning the positions of the second reaction vessel 200A and the second cover 210A may be arranged on the upper end surface of the second base 10A.

[0180] A second elastic member 21A may be coupled to the upper end portion of the second support frame 20A. The second elastic member 21A is used to buffer the impact generated when the second cover 210A descends and seals the second reaction vessel 200A.

[0181] Next, with reference to Figure 1 and Figure 2 , the operation of the decellularization reaction device according to an embodiment of the present invention will be described.

[0182] First, prepare a first xenogeneic organ tissue to be decellularized and a first decellularization solution for performing a decellularization reaction with the first xenogeneic organ tissue, and prepare a second xenogeneic organ tissue to be decellularized and a second decellularization solution for performing a decellularization reaction with the second xenogeneic organ tissue.

[0183] Here, a case where the first xenogeneic organ is different from the second xenogeneic organ and the first decellularization solution is different from the second decellularization solution is taken as an example for description. However, it is also applicable in the case where the first xenogeneic organ is the same as the second xenogeneic organ or the first decellularization solution is the same as the second decellularization solution.

[0184] Next, a case where the first xenogeneic organ tissue and the first decellularization solution are injected into the first grid chamber 100 and the first reaction vessel 200 by using the first in-situ analysis application device RA1 is taken as an example for description. However, it is also applicable in the case where the second xenogeneic organ tissue and the second decellularization solution are injected into the second grid chamber 100A and the second reaction vessel 200A.

[0185] In order to inject the first xenogeneic organ tissue to be decellularized and the first decellularization solution into the first grid chamber 100 and the first reaction vessel 200 respectively, as Figure 1As shown, the first cover 110 is set to a state of being opened from the first grid chamber 100 , and the first cover 210 is set to a state of being opened from the first reaction container 200 .

[0186] In this state, after the first decellularization object first heterogeneous organ tissue is placed inside the first grid chamber 100 and the first decellularization solution is injected into the first reaction container 200, the upper end of the first grid chamber 100 is covered by the first cover 110, and then the first cover 110 is combined with the first grid chamber 100 using the buckle 111.

[0187] In addition, the first lifting rod 621 of the first lifting cylinder 620 of the first lifting unit 600 is retracted. Figure 2 As shown, the first lifting rod 621 descends a set length, and then the first cover 210 covers the upper part of the first reaction container 200, thereby sealing the first reaction container 200.

[0188] Thus, as the first cover 210 seals the first reaction container 200, the first grid chamber 100 is Figure 2 Inserted into the interior of the first reaction container 200 as shown.

[0189] In addition, although the above description describes the case where the first decellularized solution is directly injected into the first reaction container 200, it is not limited to this. As another method of injecting the first decellularized solution, the first decellularized solution can also be injected into the first reaction container 200 through a first injection port (not shown) connected to the first solution injection tube 220.

[0190] In addition, after the first decellularization solution is injected into the first reaction container 200 , the first driving motor 320 of the first driving unit 300 is started to rotate the first rotating shaft 310 clockwise or counterclockwise at a set speed, thereby rotating the first grid chamber 100 .

[0191] Thus, as the first grid chamber 100 rotates and stirs, the first heterogeneous organ tissue in the first grid chamber 100 is stirred and, at the same time, undergoes a decellularization reaction with the first decellularization solution in the first reaction container 200 .

[0192] At this time, since the first xenogeneic organ tissue is stirred under the rotation of the first grid chamber 100, and the first decellularized solution moves along the vertical direction from the first cover 210 to the inside of the first reaction container 200 through the first circulation pipe 410, the first xenogeneic organ tissue and the first decellularized solution are fully stirred, so that the first decellularized solution of uniform concentration can be obtained in real time.

[0193] In this state, the first transfer pump 420 is started to circulate the first decellularized solution undergoing the decellularization reaction in the first reaction vessel 200 through the first circulation pipe 410 at a set speed.

[0194] For the first decellularized solution that circulates from the first solution outlet 207 of the first reaction vessel 200 to the first cover 210 through the first circulation pipe 410, sampling is performed by controlling the first solution sampling pipe control valve 511 to the open state and extracting it using the first solution sampling pipe 510.

[0195] The first decellularized solution sampled by the first solution sampling pipe 510 is loaded into a first small container 520 such as a vial, and then various physical properties (e.g., absorbance (UV-Vis), conductivity, turbidity, etc.) of the first decellularized solution loaded into the first small container 520 are measured and analyzed.

[0196] In addition, by sampling and measuring the first decellularized solution with a uniform concentration, it is easy to analyze various physical properties (e.g., absorbance (UV-Vis), conductivity, and turbidity, etc.) of the first decellularized solution, so the progress of the decellularization process can be accurately grasped.

[0197] (Example)

[0198] Next, decellularization experiments on each part (left atrium / left ventricle / right atrium / right ventricle) of a porcine heart, which is an example of the first xenogeneic organ to be decellularized, will be described.

[0199] 1) As Figure 3 shown, the porcine heart tissue is cut by part (left atrium, left ventricle, right atrium, and right ventricle), and a pretreatment process such as removing blood vessels and impurities is performed.

[0200] 2) As Figure 3 shown, after putting each part of the tissue (up to 25 g) into the first grid chamber in the first reaction vessel, the first lid of the first grid chamber is closed. The tissue of each part of the porcine heart is in a state of being locally cut with scissors, etc. If the tissue of each part of the porcine heart is cut into a size smaller than the size of the first mesh hole of the first grid chamber, it may leak out and be lost outside the first grid during the decellularization process, so attention needs to be paid.

[0201] 3) In the case of the myocardium, the decellularized solutions injected into the first reaction vessel are ultrapure water for the first time, SDS aqueous solution, Triton-X aqueous solution, EtOH, PBS aqueous solution, etc., and their types are diverse. When considering the volume of the first reaction vessel and the immersion height of the first grid chamber, the injection volume of each solution is, for example, 600 ml.

[0202] First, in order to remove impurities from the myocardium and decellularize it, ultrapure water is injected into the first reaction vessel once.

[0203] 4) Then, pressurized gas (air) is applied to the first lifting cylinder to move the first cover at the upper end of the first reaction vessel downward along the vertical direction, thereby sealing the first cover and the first reaction vessel. At this time, for safety, the first groove is used to confirm whether the first reaction vessel and the first cover are in the correct positions.

[0204] 5) The rotation speed (stirring speed) of the first rotating shaft under the action of the first drive motor in the first drive unit can be set from 0 to 500 rpm. In the case of a porcine heart, it is set to 350 rpm. The first mesh chamber is rotated and stirred when set to rotate clockwise for 10 seconds, with a pause time of 8 seconds, and then counterclockwise for 10 seconds.

[0205] 6) Meanwhile, the decellularized solution inside the first reaction vessel is circulated along the vertical direction (i.e., from the lower part to the upper part of the first reaction vessel) by using a metering first delivery pump. At this time, the circulation speed of the first decellularized solution is set to, for example, 200 ml / min.

[0206] As a result, although local decellularization of the first xenogeneic organ tissue inside the first mesh chamber is carried out, the first decellularized solution in the first reaction vessel is fully stirred because it is circulated inside along the vertical direction by the first delivery pump and is stirred by the rotation of the first mesh chamber. Therefore, a first decellularized solution with a uniform concentration can be obtained in real time.

[0207] Thus, when sampling the first decellularized solution, it becomes easy to analyze the conductivity, turbidity, absorbance (UV-Vis), etc. of the first decellularized solution. Therefore, the progress of the decellularization process of the first decellularized solution can be accurately grasped in real time.

[0208] 7) Figure 4 It is a chart of the turbidity analysis results of the first decellularized solution using ultrapure water (DW) in the decellularization process of each tissue (left atrium, right atrium, left ventricle, and right ventricle) of the porcine heart, and a chart of the turbidity of each tissue analyzed over time in each decellularization process after performing a total of three ultrapure water processes [DW(1)-DW(2)-DW(3)].

[0209] As Figure 4 shown in (a) of , the increase in turbidity, that is, the color of the first decellularized solution becoming turbid over time means that the blood and impurities contained in the tissue are discharged from the tissue, and significant differences are shown in each tissue. For example, the turbidity in the case of the left atrium is much greater than that in the case of the right atrium.

[0210] In addition, although the turbidity increases over time, it shows a tendency that the rate of increase gradually flattens out.

[0211] After the DW(1) process is completed, all the solution in the reactor is drained as waste liquid, and as Figure 4 shown in (b) of [], the ultra-pure water process is further carried out twice. After the DW(3) process, the turbidity of all tissues for the final analysis is less than 30 NTU, and it is thus judged that almost most of the impurities have been removed.

[0212] 8) Figure 5 It is a graph showing the results of conductivity analysis of the first decellularization solution of ultra-pure water (DW) in the decellularization process of each tissue (left atrium, right atrium, left ventricle, and right ventricle) of the porcine heart.

[0213] This is a graph analyzing the conductivity of each tissue over time in each decellularization process after a total of three ultra-pure water processes [DW(1)-DW(2)-DW(3)] are carried out.

[0214] In Figure 5 (a) of [], in the case of 3 minutes which is the earliest data, a very high conductivity was observed because the first decellularization solution in the first reaction vessel was not evenly mixed, but it became a stable state over time.

[0215] As Figure 5 shown in (b) of [], in the case of carrying out the DW(2)-DW(3) processes, the conductivity shown is greatly reduced as well as the turbidity, and it can thus be judged that almost most of the impurities have been removed.

[0216] 9) One revelation that can be obtained from the analysis results of the turbidity and conductivity of each tissue of the myocardium is that a high turbidity does not necessarily mean a high conductivity.

[0217] For example, although the left atrium has the highest turbidity, the conductivity of the left atrium is 280 uS / cm which is the lowest, while the conductivity of the left ventricle is about 670 uS / cm, far higher than the conductivity of the left atrium.

[0218] The correlation between turbidity and conductivity is related to the first decellularization solution (substance) of the first xenogeneic organ tissue, so subsequent related research is needed.

[0219] 10) Figure 6It is the analysis result chart of the absorbance (UV-Vis) of the first decellularized solution of the left ventricle using ultrapure water (DW) in the decellularization process of various tissues (left atrium, right atrium, left ventricle, and right ventricle) of porcine myocardium, and the chart of the absorbance (UV-Vis) of each tissue analyzed by time in each decellularization process after performing a total of three ultrapure water processes [DW(1)-DW(2)-DW(3)].

[0220] Figure 6 (a1) is the analysis result of the absorbance (UV-Vis) by time, Figure 6 (a2) is the graph representing the area (integral value) of these peaks using a chart.

[0221] Similar to the analysis results of turbidity and conductivity, although the peak area (PeakArea) increases over time, it shows a tendency that the increase rate gradually flattens out.

[0222] Figure 6 (b1) is the graph obtained by plotting (Plotting) the final data in each of the three ultrapure water decellularization processes, Figure 6 (b2) is the graph representing the area of these peaks (Area of Peak) using a chart.

[0223] As Figure 6 shown in (b1), the absorbance (UV-Vis) results decrease significantly as the process progresses, which means that it gradually approximates the intensity of the reference ultrapure aqueous solution set using the baseline (Baseline), and proves that the decellularization using ultrapure water is basically completed.

[0224] Similar results to the above are also obtained in the case of tissues other than the left ventricle (left atrium, right atrium, and right ventricle).

[0225] 11) Figure 7 is the graph taken during the decellularization process of myocardial tissue using ultrapure water and various organic solvents. It can be confirmed that during the process carried out in the order of (a)-(b)-(c), the volume of the tissue becomes smaller and its color also turns white.

[0226] (Test results of performance, etc.)

[0227] When evaluating the quality of the bioink of the first xenogeneic organ tissue after decellularization, the most basic item is biochemical assay. This test quantitatively represents the content of DNA, collagen, and GAGs in the tissue.

[0228] Table 1 shows the results of analyzing the DNA content of various tissues of a porcine heart after decellularization.

[0229] Since the DNA content of each tissue is different, the general criterion for determining that decellularization has proceeded well is that the DNA content in the tissue is less than 50 ng / mg.

[0230] As shown in Table 1, the DNA content of all tissues (left atrium, right atrium, left ventricle, and right ventricle) meets the above criteria, so it can be judged that decellularization has proceeded well.

[0231] [Table 1]

[0232] Left atrium Right atrium Left ventricle Right ventricle 8.2 ng / mg 6.7 ng / mg 7.5 ng / mg 6.2 ng / mg

[0233] If collagen and GAGs (glycosaminoglycans) are removed while discharging DNA, the physical properties of the bioink are poor. Therefore, it is necessary to analyze GAGs and collagen to confirm whether these two substances are well-preserved in the tissue. As shown in Tables 2 and 3, it can be confirmed that a large amount of GAGs and collagen are well-preserved.

[0234] [Table 2]

[0235] Left atrium Right atrium Left ventricle Right ventricle 1.7 μg / mg 2.1 μg / mg 1.6 μg / mg 1.5 μg / mg

[0236] [Table 3]

[0237] Left atrium Right atrium Left ventricle Right ventricle 53.2 μg / mg 63.1 μg / mg 30.3 μg / mg 30.1 μg / mg

[0238] From the above results, it can be seen that after decellularization of multiple varieties and small quantities of xenogeneic organs using the dual-machine system of the first and second dislodgment analysis application devices, the DNA content of the left atrium, right atrium, left ventricle, and right ventricle tissues has been reduced to an extremely low level, and collagen and GAGs in the tissue have also been very well-preserved. Thus, it can be known that high-quality bioink materials are efficiently produced.

[0239] Although the present disclosure has been illustrated by the preferred embodiments as described above, the present invention is not limited thereto, and those skilled in the art to which the present invention pertains should easily understand that various modifications and variations can be made without departing from the scope of the appended claims.

[0240] Description of Reference Numerals

[0241] 100: First reaction vessel

[0242] 200: Second reaction vessel

[0243] RA1: First dislodgment analysis application device

[0244] RA2: Second dislocated analysis application device

Claims

1. A decellularization reaction device, comprising: A first reaction vessel for storing therein a first decellularization solution for performing a decellularization reaction with the tissue of a first xenogeneic organ as the decellularization target; and One or more first ex-situ analysis application devices for extracting the first decellularization solution that is undergoing a decellularization reaction with the tissue of a first xenogeneic organ as the decellularization target from inside the first reaction vessel to the outside of the first reaction vessel, and for ex-situ measuring the physical properties of the first decellularization solution.

2. The decellularization reaction device according to claim 1, wherein The first ex-situ analysis application device comprises: A first grid chamber inserted inside the first reaction vessel and accommodating therein the tissue of a first xenogeneic organ as the decellularization target.

3. The decellularization reaction device according to claim 2, wherein The first ex-situ analysis application device comprises: A first solution circulation section connected and communicating with each other between the first reaction vessel and a first cover covering the first reaction vessel, and for circulating the first decellularization solution between the first reaction vessel and the first cover; and A first solution sampling section connected to the first solution circulation section for extracting the first decellularization solution from the first solution circulation section to the outside for sampling.

4. The decellularization reaction device according to claim 3, comprising: A first lifting section coupled to the first cover for lifting the first cover in the vertical direction.

5. The decellularization reaction device according to claim 4, wherein The first grid chamber comprises: a first driving section for rotating the first grid chamber inside the first reaction vessel for stirring.

6. The decellularization reaction device according to claim 5, wherein The first grid chamber is configured in a hollow cylindrical shape inside.

7. The decellularization reaction device according to claim 6, wherein First grids are arranged in a lattice pattern on the outer peripheral surface of the first grid chamber.

8. The decellularization reaction device according to claim 6, wherein A first cover for covering the first grid chamber can be detachably coupled to the upper end portion of the first grid chamber.

9. The decellularization reaction device according to claim 1, wherein The first reaction vessel is made of a transparent material or has a transparent window so that the inside of the first reaction vessel can be observed from the outside.

10. The decellularization reaction device according to claim 9, wherein The first reaction vessel has a double jacket shape composed of a first inner tube and a first outer tube.

11. The decellularization reaction device according to claim 10, wherein A first coolant injection port and a first coolant discharge port are respectively provided on the first outer tube. The first coolant injection port is for injecting a coolant into the space between the first outer tube and the first inner tube, and the first coolant discharge port is for discharging the injected coolant to the outside of the first outer tube.

12. The decellularization reaction device according to claim 8, wherein The first reaction vessel has a first solution outlet for flowing the decellularized solution out of the first reaction vessel to the outside.

13. The decellularization reaction device according to claim 11, wherein A first injection port for injecting a first decellularized solution into the interior of the first reaction vessel is installed on the first cover.

14. The decellularization reaction device according to claim 13, wherein A first water level detector for measuring the water level of the first decellularized solution in the first reaction vessel is installed on the first cover.

15. The decellularization reaction device according to claim 5, wherein The first driving part includes: A first rotating shaft, which is combined with the first cover of the grid chamber; and A first driving motor, which is installed on the first cover of the first reaction vessel and combined with the first rotating shaft, and is used to rotate the first rotating shaft.

16. The decellularization reaction device according to claim 12, wherein The first solution circulation part includes: A first circulation pipe, which is connected between the first cover and the first solution outlet, and is used to circulate the first decellularized solution in the first reaction vessel; and A first transfer pump, which is installed on the first circulation pipe and is used to transfer the first decellularized solution flowing out of the first solution outlet to the first circulation port.

17. The decellularization reaction device according to claim 16, wherein The first circulation pipe is composed of a flexible pipe that can be telescoped.

18. The decellularization reaction device according to claim 17, wherein The first solution sampling part includes: a first solution sampling pipe, which is connected to the first circulation pipe and is used to extract and sample the first decellularized solution circulating in the first circulation pipe.

19. The decellularization reaction device according to claim 18, wherein The absorbance (UV-Vis), conductivity and turbidity of the first decellularized solution sampled by the first solution sampling pipe are measured, and the physical properties of the first decellularized solution are analyzed.

20. The decellularization reaction device according to claim 4, wherein The first lifting part includes: A first connecting plate, which is combined with the first cover and can be lifted and lowered; and A first lifting cylinder, which is combined with one end of the first connecting plate and has a first lifting rod for lifting and lowering the first connecting plate.

21. A decellularization reaction device, comprising: A first reaction vessel, which stores a first decellularized solution for performing a decellularization reaction with the tissue of a first xenogeneic organ to be decellularized inside; One or more first ex-situ analysis application devices, which are used to extract the first decellularized solution that is performing a decellularization reaction with the tissue of a first xenogeneic organ to be decellularized in the first reaction vessel to the outside, and ex-situ measure the physical properties of the first decellularized solution; A second reaction vessel, which stores a second decellularized solution for performing a decellularization reaction with the tissue of a second xenogeneic organ to be decellularized inside; and One or more second ex-situ analysis application devices, arranged side by side with the first ex-situ analysis application device, and used to extract the second decellularization solution that is undergoing a decellularization reaction with the tissue of the decellularized target second xenogeneic organ in the second reaction vessel to the outside, and to ex-situ measure the physical properties of the second decellularization solution.

22. The decellularization reaction device according to claim 21, wherein, the second xenogeneic organ is the same as or different from the first xenogeneic organ, the second decellularization solution is the same as or different from the first decellularization solution.

23. The decellularization reaction device according to claim 21, wherein, the first ex-situ analysis application device includes: a first grid chamber, inserted inside the first reaction vessel, and accommodating the tissue of the decellularized target first xenogeneic organ inside; a first solution circulation part, connected between the first reaction vessel and the first cover covering the first reaction vessel and communicating with each other, and used to circulate the first decellularization solution between the first reaction vessel and the first cover; and a first solution sampling part, connected to the first solution circulation part, and used to extract the first decellularization solution from the first solution circulation part to the outside for sampling.

24. The decellularization reaction device according to claim 23, wherein, the first ex-situ analysis application device includes: a first lifting part, combined with the first cover, and used to lift the first cover along the vertical direction; and a first driving part, combined with the first grid chamber, and used to rotate the first grid chamber inside the first reaction vessel for stirring.

25. The decellularization reaction device according to any one of claims 21 to 24, wherein, the second ex-situ analysis application device includes: a second grid chamber, inserted inside the second reaction vessel, and accommodating the tissue of the decellularized target second xenogeneic organ inside.

26. The decellularization reaction device according to claim 25, wherein, the second ex-situ analysis application device includes: a second solution circulation part, connected between the second reaction vessel and the second cover covering the second reaction vessel and communicating with each other, and used to circulate the second decellularization solution between the second reaction vessel and the second cover; and a second solution sampling part, connected to the second solution circulation part, and used to extract the second decellularization solution from the second solution circulation part to the outside for sampling.

27. The decellularization reaction device according to claim 26, including: a second lifting part, combined with the second cover, and used to lift the second cover along the vertical direction.

28. The decellularization reaction device according to claim 27, wherein, the second grid chamber includes: a second driving part, used to rotate the second grid chamber inside the second reaction vessel for stirring.

29. The decellularization reaction device according to claim 28, wherein, the second grid chamber is configured in a hollow cylindrical shape inside.

30. The decellularization reaction device according to claim 29, wherein, A second grid is arranged in a lattice pattern on the outer peripheral surface of the second grid chamber.

31. The decellularization reaction device according to claim 30, wherein A second lid for covering the second grid chamber is detachably coupled to the upper end of the second grid chamber.

32. The decellularization reaction device according to claim 25, wherein The second reaction vessel is made of a transparent material or has a transparent window so that the interior of the second reaction vessel can be observed from the outside.

33. The decellularization reaction device according to claim 32, wherein The second reaction vessel has a double jacket configuration composed of a second inner tube and a second outer tube.

34. The decellularization reaction device according to claim 33, wherein A second coolant inlet and a second coolant outlet are respectively provided on the second outer tube. The second coolant inlet is used to inject coolant into the space between the second outer tube and the second inner tube, and the second coolant outlet is used to discharge the injected coolant to the outside of the second outer tube.

35. The decellularization reaction device according to claim 31, wherein The second reaction vessel has a second solution outlet for flowing out the decellularization solution to the outside of the second reaction vessel.

36. The decellularization reaction device according to claim 35, wherein A second injection port for injecting a second decellularization solution into the interior of the second reaction vessel is installed on the second lid.

37. The decellularization reaction device according to claim 28, wherein The second driving unit includes: A second rotating shaft coupled to the second lid of the second grid chamber; and A second driving motor installed on the second lid of the second reaction vessel and coupled to the second rotating shaft, and configured to rotate the second rotating shaft.

38. The decellularization reaction device according to claim 35, wherein The second solution circulation unit includes: A second circulation pipe connected between the second lid and the second solution outlet, configured to circulate the second decellularization solution in the second reaction vessel; and A second delivery pump installed on the second circulation pipe, configured to deliver the second decellularization solution flowing out from the second solution outlet to a second circulation port.

39. The decellularization reaction device according to claim 38, wherein The second circulation pipe is composed of a flexible pipe that can be stretched and contracted.

40. The decellularization reaction device according to claim 39, wherein The second solution sampling unit includes: a second solution sampling pipe connected to the second circulation pipe, configured to extract and sample the second decellularization solution circulating in the second circulation pipe.

41. The decellularization reaction device according to claim 40, wherein The absorbance (UV-Vis), conductivity, and turbidity of the second decellularization solution sampled by the second solution sampling pipe are measured, and the physical properties of the second decellularization solution are analyzed.

42. The decellularization reaction device according to claim 27, wherein The second lifting unit includes: A second coupling plate coupled to the second lid and capable of lifting; and A second lifting cylinder, which is coupled to one end of the second coupling plate and has a second lifting rod for lifting the second coupling plate.

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