Decellularization reactor

Abstract

The decellularization reaction apparatus according to the present invention includes a first reaction vessel for storing a first decellularization solution for carrying out a decellularization reaction, and a first ex-situ analysis application device for extracting the first decellularization solution undergoing a decellularization reaction with tissue of a first heterologous organ in the first reaction vessel and measuring the physical properties of the first decellularization solution ex-situ.

Description

[Technical Field]

[0001] The present invention relates to a decellularization reaction device, and more particularly to a decellularization reaction device for a wide variety of xenogeneic organs in small quantities. [Background technology]

[0002] In recent years, the global trend toward aging has led to an increase in the number of people with chronic diseases, and interest has been growing in artificial organ manufacturing technology using 3D bioprinting.

[0003] Bioink is a term used to describe ink materials used to print artificial organs using 3D bioprinters, and is manufactured from the extracellular matrix obtained after decellularizing xenogeneic organs.

[0004] When the amount of xenogeneic organs to be decellularized is large, a reactor type suitable for mass production can be used. However, when the amount of xenogeneic organ tissue that can be received is small, such as pig corneas (10 g), it is difficult to operate large-scale equipment.

[0005] Additionally, there is an urgent need for a device that can decellularize small amounts of tissue from various different organs, such as detailed cardiac tissue (left atrium, left ventricle, right atrium, right ventricle), and simultaneously compare the differences in physical properties between tissues from different organs. Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention provides a decellularization reaction device that can efficiently decellularize a variety of tissues and small amounts of tissues from different organs. [Means for solving the problem]

[0007] A decellularization reaction device according to one embodiment of the present invention may include a first reaction container for storing a first decellularization solution for carrying out a decellularization reaction with tissue of a first heterologous organ to be decellularized.

[0008] The decellularization reaction device may also include one or more first ex-situ analysis application devices for extracting the first decellularization solution undergoing a decellularization reaction with the tissue of the first heterologous organ to be decellularized within the first reaction container to the outside of the first reaction container and measuring the physical properties of the first decellularization solution ex-situ.

[0009] The first ex situ analytical application device can include a first mesh chamber that is inserted into the first reaction vessel and contains tissue of the first xenogeneic organ to be decellularized therein.

[0010] The first ex situ analysis application device can include a first solution circulation unit that is connected so as to be in communication with a first reaction container and a first cover that covers the first reaction container, and that circulates a first decellularization solution between the first reaction container and the first cover.

[0011] The first ex-situ analysis application device may also include a first solution sampling unit connected to the first solution circulation unit for extracting and sampling the first decellularization solution from the first solution circulation unit to the outside.

[0012] The cover may include a first lifting unit coupled to the first cover for vertically lifting the first cover.

[0013] The first mesh chamber may include a first drive unit for rotating the first mesh chamber so as to agitate the first mesh chamber within the first reaction vessel.

[0014] The first mesh chamber may be cylindrical with an open interior.

[0015] The first mesh chamber may have a first mesh arranged in a lattice pattern on the outer peripheral surface thereof.

[0016] A first cover for covering the first mesh chamber is detachably coupled to an upper end of the first mesh chamber.

[0017] 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.

[0018] The first reaction vessel may have a double jacket configuration consisting of a first inner tube and a first outer tube.

[0019] The first external pipe may be provided with a first refrigerant inlet for injecting a refrigerant into a space between the first external pipe and the first internal pipe, and a first refrigerant outlet for discharging the injected refrigerant to the outside of the first external pipe.

[0020] The first reaction vessel can have a first solution outlet for allowing the decellularization solution to flow out of the first reaction vessel.

[0021] The first cover may be provided with a first injection port for injecting a first decellularization solution into the first reaction container.

[0022] The first cover may be provided with a first water level measuring device for measuring the water level of the first decellularization solution in the first reaction container.

[0023] The first driving unit may include a first rotating shaft coupled to the first lid of the mesh chamber, and a first driving motor provided on the first cover of the first reaction vessel and coupled to the first rotating shaft to rotate the first rotating shaft.

[0024] The first solution circulation unit includes a first circulation pipe connected between the first circulation port of the first cover and the first solution outlet for circulating the first decellularization solution in the first reaction container, and a first transfer pump provided in the first circulation pipe for transferring the first decellularization solution flowing out from the first solution outlet to the first circulation port.

[0025] The first circulation pipe may be made of an expandable and contractible flexible pipe.

[0026] The first solution sampling unit may include a first solution sampling tube connected to the first circulation tube for extracting and sampling the first decellularization solution circulated in the first circulation tube.

[0027] The physical properties of the first decellularization solution can be analyzed by measuring the absorbance (UV-Vis), electrical conductivity, and turbidity of the first decellularization solution sampled in the first solution sampling tube.

[0028] The first lifting unit may include a first connecting plate connected to the first cover so as to be able to move up and down, and a first lifting cylinder connected to one end of the first connecting plate and having a first lifting rod for lifting and lowering the first connecting plate.

[0029] A decellularization reaction device according to another embodiment of the present invention may include a first reaction vessel for storing a first decellularization solution for carrying out a decellularization reaction with tissue of a first heterologous organ to be decellularized.

[0030] In addition, a decellularization reaction apparatus according to another embodiment may include one or more first ex-situ analysis application devices for extracting a first decellularization solution undergoing a decellularization reaction with tissue of a first heterologous organ to be decellularized within a first reaction vessel to the outside and measuring the physical properties of the first decellularization solution ex-situ.

[0031] In addition, the decellularization reaction device according to another embodiment may include a second reaction vessel for storing a second decellularization solution for carrying out a decellularization reaction with tissue of a second heterologous organ to be decellularized.

[0032] In another embodiment, the decellularization reaction device may include one or more second ex situ analysis application devices arranged alongside the first ex situ analysis application device, for extracting to the outside a second decellularization solution undergoing a decellularization reaction with tissue of a second heterologous organ to be decellularized within a second reaction vessel, and for measuring the physical properties of the second decellularization solution ex situ.

[0033] The second heterologous organ may be the same as or different from the first heterologous organ, and the second decellularization solution may be the same as or different from the first decellularization solution.

[0034] The first ex situ analysis application device includes a first mesh chamber that is inserted into the first reaction container and contains tissue of the first xenogeneic organ to be decellularized therein, and a first solution circulation unit that is connected so as to be in communication with the first reaction container and a first cover that covers the first reaction container, and that circulates a first decellularization solution between the first reaction container and the first cover.

[0035] The first ex-situ analysis application device may also include a first solution sampling unit connected to the first solution circulation unit for extracting and sampling the first decellularization solution from the first solution circulation unit to the outside.

[0036] The first ex situ analytical application device may include a first lifting unit coupled to the first cover for vertically raising and lowering the first cover, and a first driving unit coupled to the first mesh chamber for rotating the first mesh chamber so as to stir within the first reaction vessel.

[0037] The second ex situ analysis application device can include a second mesh chamber that is inserted into the second reaction vessel and contains tissue of the second xenogeneic organ to be decellularized.

[0038] The second ex situ analysis application device can include a second solution circulation section that is connected so as to be in communication with the second reaction container and a second cover that covers the second reaction container, and that circulates the second decellularization solution between the second reaction container and the second cover.

[0039] The second ex-situ analysis application device may also include a second solution sampling unit connected to the second solution circulation unit for extracting and sampling the second decellularization solution from the second solution circulation unit to the outside.

[0040] The device may include a second lifting unit coupled to the second cover for vertically lifting the second cover.

[0041] The second mesh chamber can include a second drive for rotating the second mesh chamber to provide agitation within the second reaction vessel.

[0042] The second mesh chamber may be cylindrical with an open interior.

[0043] A second mesh may be arranged in a grid pattern on the outer peripheral surface of the second mesh chamber.

[0044] A second lid for covering the second mesh chamber may be detachably coupled to an upper end of the second mesh chamber.

[0045] The second reaction vessel may be made of a transparent material or have a transparent window that allows the inside of the second reaction vessel to be observed from the outside.

[0046] The second reaction vessel may have a double jacket configuration consisting of a second inner tube and a second outer tube.

[0047] The second external pipe may be provided with a second refrigerant inlet for injecting a refrigerant into the space between the second external pipe and the second internal pipe, and a second refrigerant outlet for discharging the injected refrigerant to the outside of the second external pipe.

[0048] The second reaction vessel can have a second solution outlet for allowing the decellularization solution to flow out of the second reaction vessel.

[0049] The second cover may be provided with a second injection port for injecting a second decellularization solution into the second reaction vessel.

[0050] The second driving unit may include a second rotating shaft coupled to the second lid of the second mesh chamber, and a second driving motor provided on the second cover of the second reaction vessel and coupled to the second rotating shaft for rotating the second rotating shaft.

[0051] The second solution circulation unit can include a second circulation pipe connected between the second cover and the second solution outlet for circulating the second decellularization solution in the second reaction vessel, and a second transfer pump provided in the second circulation pipe for transferring the second decellularization solution flowing out from the second solution outlet to the second circulation port.

[0052] The second circulation pipe may be made of an expandable and contractible flexible pipe.

[0053] The second solution sampling unit may include a second solution sampling tube connected to the second circulation tube for extracting and sampling the second decellularization solution circulating in the second circulation tube.

[0054] The physical properties of the second decellularization solution can be analyzed by measuring the absorbance (UV-Vis), electrical conductivity, and turbidity of the second decellularization solution sampled in the second solution sampling tube.

[0055] The second lifting unit may include a second connecting plate connected to the second cover so as to be able to move up and down, and a second lifting cylinder connected to one end of the second connecting plate and having a second lifting rod for lifting and lowering the second connecting plate. [Effects of the Invention]

[0056] According to an embodiment of the present invention, the pure ring structure of the first and second reaction vessels, the first and second mesh chambers, and the first and second decellularization solutions, as well as the twin system of the first ex situ analysis application device and the second ex situ analysis application device, are very effective in operating the decellularization process for a variety of small quantities of xenogeneic organs, and the physical properties of each tissue can be evaluated very efficiently. [Brief explanation of the drawings]

[0057] [Figure 1] FIG. 1 is a schematic diagram of a decellularization reaction apparatus according to one embodiment of the present invention, showing a state in which the first and second covers are open. [Figure 2] 1 is a block diagram of a first ex-situ analysis application device of a decellularization reaction device according to an embodiment of the present invention, showing a state in which a first cover is closed. [Figure 3] 10 is a photograph showing a pretreatment process for treating each part of a pig's heart for use in a decellularization reaction device according to one embodiment of the present invention. [Figure 4] 1 is a graph showing the results of turbidity analysis over time in the first ultrapure water step (DW(1)) during the decellularization process for each tissue of porcine cardiac muscle using a decellularization reaction apparatus according to one embodiment of the present invention, and the results of turbidity analysis of the first decellularization solution using three ultrapure water steps [DW(1)-DW(2)-DW(3)]. [Figure 5] 1 is a graph showing the results of electrical conductivity analysis over time in the first ultrapure water step (DW(1)) during the decellularization process for each tissue of a pig heart using a decellularization reaction apparatus according to one embodiment of the present invention, and the results of electrical conductivity analysis of the first decellularization solution using three ultrapure water steps [DW(1)-DW(2)-DW(3)]. [Figure 6] 1 is a graph showing the results of absorbance analysis over time in the first ultrapure water step (DW(1)) during the decellularization process for each tissue of porcine cardiac muscle using a decellularization reaction apparatus according to one embodiment of the present invention, and the results of absorbance analysis of the first decellularization solution for the left ventricle using three ultrapure water steps [DW(1)-DW(2)-DW(3)]. [Figure 7] 1A-1C are photographs showing the process (a)-(b)-(c) of decellularization of cardiac muscle tissue using ultrapure water and various organic solvents as the first decellularization solution in a decellularization reaction apparatus according to one embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0058] Hereinafter, with reference to the accompanying drawings, embodiments of the present invention will be described so that those skilled in the art can easily carry out the present invention. As will be easily understood by those skilled in the art, the embodiments described below can be modified in various forms without departing from the concept and scope of the present invention. Whenever possible, the same or similar parts will be designated by the same reference numerals throughout the drawings.

[0059] The terminology used below is merely for the purpose of referring to particular embodiments and is not intended to limit the present invention. As used herein, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise. As used in the specification, the term "comprises" embodies a particular property, region, integer, step, operation, element, component, and / or group, and does not exclude the presence or addition of other particular properties, regions, integers, steps, operations, elements, components, and / or groups.

[0060] All terms, including technical and scientific terms, used below have the same meaning as commonly understood by a person of ordinary skill in the art to which this invention pertains. Dictionary-defined terms are additionally interpreted to have a meaning consistent with the relevant technical literature and the presently disclosed content, and are not interpreted in an ideal or very formal sense unless otherwise defined.

[0061] FIG. 1 is a schematic diagram of a decellularization reaction apparatus according to one embodiment of the present invention, showing a state in which the first and second covers are open.

[0062] FIG. 2 is a configuration diagram of a first ex-situ analysis application device of a decellularization reaction device according to one embodiment of the present invention, showing a state in which the first cover is closed.

[0063] Referring to FIGS. 1 and 2, a decellularization reactor according to a first embodiment of the present invention can include a first ex-situ analysis application device (RA1) and a second ex-situ analysis application device (RA2).

[0064] At least one first ex-situ analysis application device (RA1) is provided, which can extract the tissue of the first xenogeneic organ to be decellularized and the first decellularization solution undergoing a decellularization reaction to the outside and analyze various physical properties of the first decellularization solution ex-situ.

[0065] The second ex-situ analysis application device (RA2) is placed next to the first ex-situ analysis application device (RA1) at a set distance, and can extract the tissue of the second xenogeneic organ to be decellularized and the second decellularization solution undergoing a decellularization reaction to the outside, and analyze various physical properties of the second decellularization solution ex-situ.

[0066] Here, ex-situ analysis can refer to extracting the first decellularization solution and the second decellularization solution from the first ex-situ analysis application device (RA1) and the second ex-situ analysis application device (RA2) to the outside, and measuring and analyzing the physical properties of the first decellularization solution and the second decellularization solution, respectively.

[0067] The first heterologous organ may be the same as or different from the second heterologous organ, and the first decellularization solution may be the same as or different from the second decellularization solution.

[0068] The first ex-situ analytical application device (RA1) may include a first mesh 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.

[0069] The first mesh chamber 100 has a predetermined size and shape, and can accommodate tissue of the first xenogeneic organ to be decellularized inside.

[0070] In addition, the first reaction vessel 200 can have the first mesh chamber 100 inserted therein and store a first decellularization solution for carrying out a decellularization reaction with the tissue of the first heterologous organ in the first mesh chamber 100.

[0071] The first driving unit 300 is coupled to the first mesh chamber 100 and can rotate the first mesh chamber 100 so as to stir the contents in the first reaction vessel 200 .

[0072] The first solution circulation unit 400 is connected between the first reaction container 200 and the first cover 210 that covers the first reaction container 200 so that they are in communication with each other, and can circulate the first decellularization solution between the first reaction container 200 and the first cover 210.

[0073] The first solution sampling unit 500 is connected to the first solution circulation unit 400 and can extract and sample the first decellularization solution from the first solution circulation unit 400 to the outside to measure various physical properties of the first decellularization solution.

[0074] A first cover 210 for covering the first reaction vessel 200 may be placed on the top of the first reaction vessel 200 .

[0075] In addition, the first cover 210 may be coupled to a first lifting unit 600 for raising and lowering the first cover 210 in a vertical direction (Y direction in FIG. 1) relative to the first reaction vessel 200 so that the first cover 210 can be coupled to or detached from the first reaction vessel 200.

[0076] The first mesh chamber 100 may be easily rotated by the first driving unit 300 and may be cylindrical with an open interior so that tissue of the first heterologous organ to be decellularized can be placed inside and accommodated therein.

[0077] In addition, if the first mesh chamber 100 is made rectangular, the tissue of the foreign organ may get caught or become trapped in the sharp corners when the first mesh chamber 100 is rotated, and the tissue of the foreign organ may not be mixed evenly. Therefore, the first mesh chamber 100 is made cylindrical or the like in order to mix the tissue of the foreign organ evenly and efficiently.

[0078] The first mesh chamber 100 may be configured as a small type that can accommodate a maximum amount of the first xenogeneic organ to be decellularized of less than 25 g.

[0079] The first mesh 101 is arranged in a grid pattern on the outer peripheral surface of the first mesh chamber 100, and the first mesh 101 can have a set size, for example, about 1 mm, for the decellularization reaction of the tissue of the first heterologous organ contained within the first mesh chamber 100.

[0080] In addition, the size of the first mesh 101 may be set to be slightly smaller than the tissue of the first heterologous organ so that the tissue of the first heterologous organ contained in the first mesh chamber 100 does not escape outside the first mesh 101 during the decellularization process.

[0081] The first mesh chamber 100 may include a first cover 110 detachably coupled to an upper end of the first mesh chamber 100 to cover the first mesh chamber 100 .

[0082] The first cover 110 may have a buckle 111 or the like so that it can be easily attached to the first mesh chamber 100, but is not limited thereto, and the first cover 110 and the first mesh chamber 100 can be attached by screw connection or the like.

[0083] A first driving unit 300 may be coupled to an upper end of the first cover 110 .

[0084] The first mesh chamber 100 may be made of or coated with a material with good chemical resistance, such as Teflon, depending on the physical properties of the first decellularization solution.

[0085] In addition, the first reaction vessel 200 may be made of a transparent material or have a transparent window (not shown) so that the progress of the decellularization process taking place within the first reaction vessel 200 can be visually observed from the outside of the first reaction vessel 200.

[0086] The first reaction vessels 200 are arranged at set intervals on the first base 10 and can be supported by a first-first support frame 20 arranged in a vertical direction (Y direction in FIG. 1) relative to the first base 10.

[0087] The first reaction vessel 200 may be made of a material that is corrosion-resistant to the decellularization solution to be used.

[0088] In addition, the first reaction vessel 200 may have a double jacket configuration consisting of a first inner tube 201 and a first outer tube 203 in order to control the temperature of the decellularization process.

[0089] The first external pipe 203 may be provided with a first refrigerant inlet 205 for injecting a refrigerant into the space between the first external pipe 203 and the first internal pipe 201, and a first refrigerant outlet 206 for discharging the refrigerant to the outside of the first external pipe 203.

[0090] The first reaction container 200 may have a first solution outlet 207 for discharging the decellularization solution to the outside of the first reaction container 200.

[0091] The first cover 210 may be made of a rubber material or an O-ring type so that it can seal the first reaction vessel 200 while cushioning impact when it comes into contact with the upper end of the first reaction vessel 200 by descending using the first lifting unit 600.

[0092] The first cover 210 may be provided with a first injection port (not shown) to which a first solution injection tube 220 for injecting a first decellularization solution into the first reaction container 200 is connected.

[0093] The first decellularization solution may consist of, for example, ultrapure water, an organic solvent, or the like.

[0094] In addition, a first solution circulation unit 400 is connected to the first cover 210 , and the first decellularization solution circulating in the first solution circulation unit 400 can be injected into the inside of the first reaction container 200 .

[0095] The first cover 210 is provided with a first water level measuring device 213 for measuring the water level of the first decellularization solution in the first reaction container 200, and the first water level measuring device 213 can operate using an ultrasonic method or the like.

[0096] The first driving unit 300 may include a first rotating shaft 310 and a first driving motor 320 .

[0097] The first rotating shaft 310 may be coupled to the first lid 110 of the first mesh chamber 100 .

[0098] In addition, the first driving motor 320 is installed in the first cover 210 of the first reaction container 200 and is coupled to the first rotating shaft 310 to rotate the first rotating shaft 310 .

[0099] The first driving motor 320 may be a magnetic type motor or the like so that the first rotating shaft 310 can be easily rotated.

[0100] The first rotation axis 310 may be arranged in the center of the upper end of the first lid 110 in a direction perpendicular to the upper end of the first lid 110 (Y direction in Figure 2), and may be connected to the first lid 110 by fastening it with a first bolt 301, for example.

[0101] The first rotating shaft 310 is rotated clockwise or counterclockwise by the first drive motor 320, thereby rotating the first mesh chamber 100 and stirring the tissue of the first heterologous organ contained in the first mesh chamber 100, thereby facilitating decellularization by the first decellularization solution injected into the first reaction container 200.

[0102] When the first mesh chamber 100 is rotated at a set rotation speed (e.g., 1 to 500 rpm) to stir, stirring can be alternately performed from the forward direction (clockwise) to the reverse direction (counterclockwise) to prevent overloading of the first drive motor 320 and entanglement between different organs, and a stop time can be set midway.

[0103] The first solution circulation unit 400 may include a first circulation pipe 410 and a first transfer pump 420 .

[0104] The first circulation tube 410 is connected between a first circulation port (not shown) arranged on the first cover 210 and the first solution outlet 207, and is capable of circulating the first decellularization solution in the first reaction container 200.

[0105] Furthermore, the first transfer pump 420 is provided in the first circulation pipe 410, and is capable of quantitatively transferring the first decellularization solution flowing out from the first solution outlet 207 to the first circulation port.

[0106] The first circulation pipe 410 may be made of a flexible pipe so that it can be easily extended and contracted when the first cover 210 is raised and lowered by the first lifting unit 600 .

[0107] In addition, a first waste liquid discharge pipe 430 for discharging waste liquid of the first decellularization solution is connected to the first circulation pipe 410, and the first waste liquid discharge pipe 430 may be provided with a first waste liquid discharge pipe control valve 431 for controlling the discharge of the first decellularization solution.

[0108] The first solution sampling unit 500 may include a first solution sampling pipe 510 connected to the first circulation pipe 410 for extracting and sampling the first decellularization solution circulating from the first circulation pipe 410.

[0109] The first decellularization solution sampled by the first solution sampling tube 510 is placed in a first small container 520 such as a vial, and various physical properties of the first decellularization solution placed in the first small container 520, such as absorbance (UV-Vis), electrical conductivity, turbidity, etc., can then be measured and analyzed.

[0110] The first solution sampling pipe 510 may be provided with a first solution sampling pipe control valve 511 for controlling the opening and closing of the first solution sampling pipe 510 .

[0111] The first lifting unit 600 may include a first connecting plate 610 and a first lifting cylinder 620 .

[0112] The first coupling plate 610 may be coupled to the first cover 210 so as to be able to move up and down integrally.

[0113] In addition, the first lifting cylinder 620 may be coupled to the lower end of the first connecting plate 610 and may have a first lifting rod 621 for lifting the first connecting plate 610 up and down.

[0114] The first lifting cylinder 620 may be installed and supported on the upper end surface of the first base 10 in a direction perpendicular to the first base 10 (Y direction in FIG. 1).

[0115] A first air supply pipe 623 for supplying air into the first lifting cylinder 620 is connected to the first lifting cylinder 620, and the first air supply pipe 623 may be provided with a first regulator 625 for adjusting the pressure of the air supplied to the first air supply pipe 623.

[0116] The first base 10 may also be provided with a first-second support frame 30 that is arranged parallel to the first-first support frame 20 and supported by the first base.

[0117] A first guide rod 630 may be coupled to the first-second support frame 30 via a first connection frame 31 to guide the first lifting rod 621 in its vertical movement.

[0118] In addition, the first reaction vessel 200 and the first-1 support frame 20 are configured to be movable and detachable to facilitate cleaning, and a first groove 11 may be arranged on the upper surface of the first base 10 for aligning the first reaction vessel 200 and the first cover 210.

[0119] A first elastic member 21 may be attached to the upper end of the first-1 support frame 20 to absorb the shock generated when the first cover 210 descends to seal the first reaction container 200 .

[0120] The second ex situ analytical application apparatus (RA2) can include a second mesh 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.

[0121] The second mesh chamber 100A has a set size and shape, and is capable of accommodating tissue of a second xenogeneic organ to be decellularized therein.

[0122] In addition, the second reaction vessel 200A is arranged at least one next to the first reaction vessel 200A at a set distance, and can store a second decellularization solution for carrying out a decellularization reaction with the tissue of a second heterologous organ in the second mesh chamber 100A inserted therein.

[0123] The second driving unit 300A is coupled to the second mesh chamber 100A and can rotate the second mesh chamber 100A to stir the contents in the second reaction vessel 200A.

[0124] In addition, the second solution circulation unit 400A is connected between the second reaction vessel 200A and the second cover 210A covering the second reaction vessel 200A so that they are in communication with each other, and the second decellularization solution can be circulated between the second reaction vessel 200A and the second cover 210A.

[0125] The second solution sampling unit 500A is connected to the second solution circulation unit 400A and can extract and sample the second decellularization solution from the second solution circulation unit 400A to the outside for measuring various physical properties of the second decellularization solution.

[0126] A second cover 210A for covering the second reaction vessel 200A may be placed on top of the second reaction vessel 200A.

[0127] In addition, a second lifting unit 600A may be coupled to the second cover 210A for raising and lowering the second cover 210A in a direction vertical to the second reaction vessel 200A (Y direction in FIG. 1) so as to couple or detach the second cover 210A to or from the second reaction vessel 200A.

[0128] The second mesh chamber 100A can be easily rotated by the second driving unit 300A and can have a cylindrical shape so that tissue of the second xenogeneic organ to be decellularized can be introduced and accommodated inside.

[0129] Furthermore, if second mesh chamber 100A is made rectangular, the tissue of the foreign organ may become pinched or trapped in the sharp corners when second mesh chamber 100A is rotated, preventing the tissue of the foreign organ from being stirred evenly. This is why second mesh chamber 100A is made cylindrical or the like in order to stir the tissue of the foreign organ evenly and efficiently.

[0130] The second mesh chamber 100A has a second mesh 101A arranged in a grid pattern, and the second mesh 101A can have a set size, for example, about 1 mm, for decellularization of tissue of a second heterologous organ contained within the second mesh chamber 100A.

[0131] The second mesh chamber 100A may be configured as a small type that can accommodate a maximum amount of less than 25 g of the second xenogeneic organ to be decellularized.

[0132] In addition, the size of the second mesh 101A may be set to be slightly smaller than the tissue of the second heterologous organ so that the tissue of the second heterologous organ contained in the second mesh chamber 100A does not escape to the outside of the second mesh 101A during the decellularization process.

[0133] The second mesh chamber 100A may include a second lid 110A detachably coupled to the upper end of the second mesh chamber 100A for covering the second mesh chamber 100A.

[0134] The second cover 110A may have a buckle 111A shape so that it can be easily attached to the second mesh chamber 100A, but is not limited thereto, and the second cover 110A and the second mesh chamber 100A can be attached by screw connection or the like.

[0135] A second driving unit 300A may be coupled to the upper end of the second cover 110A.

[0136] The second mesh chamber 100A may be made of or coated with a material with good chemical resistance, such as Teflon, depending on the physical properties of the second decellularization solution.

[0137] 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 being carried out in the second reaction vessel 200A can be visually observed from the outside of the second reaction vessel 200A.

[0138] The second reaction vessels 200A are arranged at set intervals on the second base 10A and can be supported by a 2-1 support frame 20A arranged in a direction perpendicular to the second base 10A (Y direction in FIG. 1).

[0139] The second reaction vessel 200A may be made of a material that is corrosion-resistant to the decellularization solution to be used.

[0140] In addition, the second reaction vessel 200A may have a double jacket configuration consisting of a second inner tube 201A and a second outer tube 203A in order to control the temperature of the decellularization process.

[0141] The second external pipe 203A may be provided with a second refrigerant inlet 205A for injecting a refrigerant into the space between the second external pipe 203A and the second internal pipe 201A, and a second refrigerant outlet 206A for discharging the refrigerant to the outside of the second external pipe 203A.

[0142] The second reaction vessel 200A can have a second solution outlet 207A for discharging the decellularization solution.

[0143] The second cover 210A may be configured in the form of an O-ring made of a rubber material so that it can be lowered by the second lifting unit 600A and seal the second reaction vessel 200A while cushioning impact when it comes into contact with the upper end of the second reaction vessel 200A.

[0144] Second cover 210A may be provided with a second injection port (not shown) to which second solution injection tube 220A for injecting a second decellularization solution into second reaction container 200A is connected. The second decellularization solution may consist of, for example, ultrapure water, an organic solvent, or the like.

[0145] Furthermore, a second solution circulation unit 400A is connected to the second cover 210A, and the second decellularization solution circulated in the second solution circulation unit 400A can be injected into the inside of the second reaction vessel 200A.

[0146] The second cover 210A is provided with a second water level measuring device 213A for measuring the water level of the second decellularization solution in the second reaction container 200A, and the second water level measuring device 213A can operate using an ultrasonic method or the like.

[0147] The second driving unit 300A may include a second rotating shaft 310A and a second driving motor 320A.

[0148] The second rotating shaft 310A may be coupled to the second lid 110A of the second mesh chamber 100.

[0149] The second drive motor 320A is installed in the second cover 210A of the second reaction container 200A and is coupled to the second rotation shaft 310A to rotate the second rotation shaft 310A.

[0150] The second drive motor 320A may be a magnetic type motor or the like so as to easily rotate the second rotating shaft 310A.

[0151] The second rotation shaft 310A may be arranged in the center of the upper end of the second lid 110A in a direction perpendicular to the upper end of the second lid 110A (Y direction in Figure 1), and may be connected to the second lid 110A by fastening it with a second bolt 301A, for example.

[0152] The second rotating shaft 310A can be rotated clockwise or counterclockwise by the second drive motor 320A to rotate the second mesh chamber 100A and agitate the tissue of the second heterologous organ in the second mesh chamber 100A, thereby facilitating decellularization using the second decellularization solution injected into the second reaction vessel 200A.

[0153] When stirring by rotating the second mesh chamber 100A at a set rotation speed (e.g., 1 to 300 rpm), stirring can be alternately performed from the forward direction (clockwise) to the reverse direction (counterclockwise) to prevent overloading of the second drive motor 320A and to prevent entanglement between different organs, and a stop time can be set midway.

[0154] The second solution circulation unit 400A can include a second circulation pipe 410A and a second transfer pump 420A.

[0155] The second circulation tube 410A is connected between a second circulation port (not shown) arranged in the second cover 210A and the second solution outlet 207A, and is capable of circulating the second decellularization solution in the second reaction container 200.

[0156] Furthermore, second transfer pump 420A is provided in second circulation pipe 410A, and is capable of quantitatively transferring the second decellularization solution circulating in second circulation pipe 410A.

[0157] The second circulation pipe 410A may be made of a flexible pipe so that it can easily expand and contract when the second cover 210A is raised and lowered by the second lifting unit 600A.

[0158] In addition, a second waste liquid discharge pipe 430A for discharging waste liquid of the second decellularization solution is connected to the second circulation pipe 410A, and a second waste liquid discharge pipe control valve 431A for controlling the discharge of the second decellularization solution may be provided on the second waste liquid discharge pipe 430A.

[0159] The second solution sampling unit 500A is connected to the second circulation pipe 410A and can include a second solution sampling pipe 510A for extracting and sampling the second decellularization solution circulating in the second circulation pipe 410A.

[0160] The second decellularization solution sampled by the second solution sampling tube 510A is placed in a second small container 520A such as a vial, and various physical properties of the second decellularization solution placed in this second small container 520A, such as absorbance (UV-Vis), electrical conductivity, turbidity, etc., can then be measured.

[0161] The second solution sampling pipe 510A may be provided with a second solution sampling pipe control valve 511A for controlling the opening and closing of the second solution sampling pipe 510A.

[0162] The second lifting unit 600A may include a second connecting plate 610A and a second lifting cylinder 620A.

[0163] The second coupling plate 610A may be coupled to the second cover 210A so as to be able to move up and down integrally.

[0164] The second lifting cylinder 620A is coupled to the lower end of the second connecting plate 610A and may have a second lifting rod 621A for lifting the second connecting plate 610A.

[0165] The second lifting cylinder 620A may be installed and supported on the upper end surface of the second base 10A in a direction perpendicular to the second base 10A (Y direction in FIG. 1).

[0166] A second air supply pipe 623A for supplying air into the second lifting cylinder 620A is connected to the second lifting cylinder 620A, and the second air supply pipe 623A may be provided with a second regulator 625A for adjusting the pressure of the air supplied to the second air supply pipe 623A.

[0167] The second base 10A may also be provided with a 2-2 support frame 30A that is disposed parallel to the 2-1 support frame 20A and supported by the second base 10A.

[0168] A second guide rod 630A may be coupled to the 2-2 support frame 30A via a second connecting frame 31A to guide the second lifting rod 621A in its vertical movement.

[0169] In addition, the second reaction vessel 200A and the 2-1 support frame 20A are configured to be movable and detachable to facilitate cleaning, and a second groove 11A for aligning the second reaction vessel 200A and the second cover 210A may be arranged on the upper end surface of the first base 10A.

[0170] A second elastic member 21A may be coupled to the upper end of the 2-1 support frame 20A to absorb the impact generated when the second cover 210A descends to seal the second reaction container 200A.

[0171] Hereinafter, the operation of a decellularization reaction device according to one embodiment of the present invention will be described with reference to FIGS.

[0172] First, prepare tissue from a first xenogeneic organ to be decellularized and a first decellularization solution for the decellularization reaction with the tissue from the first xenogeneic organ, and then prepare tissue from a second xenogeneic organ to be decellularized and a second decellularization solution for the decellularization reaction with the tissue from the second xenogeneic organ.

[0173] Here, we will explain an example in which the first heterologous organ and the second heterologous organ are different and the first decellularization solution and the second decellularization solution are different, but the same can be applied to cases in which the first heterologous organ and the second heterologous organ are the same or the first decellularization solution and the second decellularization solution are the same.

[0174] The following describes an example in which tissue from a first heterologous organ and a first decellularization solution are injected into a first mesh chamber 100 and a first reaction vessel 200 using a first ex situ analysis application device (RA1), but the same can be applied to the case in which tissue from a second heterologous organ and a second decellularization solution are injected into a second mesh chamber 100A and a second reaction vessel 200A.

[0175] In order to inject the tissue of the first heterologous organ to be decellularized and the first decellularization solution into the first mesh chamber 100 and the first reaction vessel 200, respectively, the first lid 110 is opened from the first mesh chamber 100 and the first cover 210 is opened from the first reaction vessel 200, as shown in FIG. 1.

[0176] In this state, tissue of the first heterologous organ to be decellularized is placed inside the first mesh chamber 100, and the first decellularization solution is injected into the first reaction container 200.Then, the upper end of the first mesh chamber 100 is covered with the first lid 110, and the first lid 110 is attached to the first mesh chamber 100 using the buckle 111.

[0177] Furthermore, when the first lifting rod 621 of the first lifting cylinder 620 of the first lifting unit 600 is contracted, the first lifting rod 621 descends a set length as shown in FIG. 2, causing the first cover 210 to cover the top of the first reaction vessel 200 and seal the first reaction vessel 200.

[0178] As described above, the first cover 210 seals the first reaction vessel 200, and the first mesh chamber 100 is inserted into the first reaction vessel 200 as shown in FIG.

[0179] Furthermore, although the above describes the case where the first decellularization solution is directly injected into the first reaction vessel 200, this is not limited thereto, and other methods of injecting the first decellularization solution may be used, such as injecting the first decellularization solution into the first reaction vessel 200 through a first injection port (not shown) to which the first solution injection tube 220 is connected.

[0180] Furthermore, once the injection of the first decellularization solution into the first reaction container 200 is completed, the first drive motor 320 of the first drive unit 300 is driven to rotate the first rotation shaft 310 clockwise or counterclockwise at a set speed, thereby rotating the first mesh chamber 100.

[0181] In this way, by rotating and stirring the first mesh chamber 100, the tissue of the first heterologous organ in the first mesh chamber is stirred and undergoes a decellularization reaction with the first decellularization solution in the first reaction container 200.

[0182] At this time, the rotation of the first mesh chamber 100 stirs the tissue of the first heterologous organ, and the first decellularization solution moves vertically from the first cover 210 to the inside of the first reaction vessel 200 via the first circulation tube 410, resulting in thorough mixing of the tissue of the first heterologous organ and the first decellularization solution, thereby enabling a first decellularization solution of a homogeneous concentration to be obtained in real time.

[0183] In this state, the first transfer pump 420 is operated to circulate the first decellularization solution undergoing a decellularization reaction in the first reaction vessel 200 through the first circulation pipe 410 at a set speed.

[0184] The first decellularization solution circulating from the first solution outlet 207 of the first reaction vessel 200 to the first cover 210 via the first circulation pipe 410 is extracted and sampled from the first solution sampling pipe 510 by controlling the first solution sampling pipe control valve 511 to be in an open state.

[0185] In this way, the first decellularization solution sampled by the first solution sampling tube 510 is placed in a first small container 520 such as a vial, and various physical properties of the first decellularization solution placed in this first small container 520, such as absorbance (UV-Vis), electrical conductivity, turbidity, etc., can be measured and analyzed.

[0186] Furthermore, by sampling and measuring the first decellularization solution of a homogeneous concentration, it becomes easier to analyze various physical properties of the first decellularization solution, such as absorbance (UV-Vis), electrical conductivity, and turbidity, allowing the progress of the decellularization process to be accurately grasped. [Example]

[0187] Below, an experiment on decellularization of each part of a pig heart (left atrium / left ventricle / right atrium / right ventricle) will be described as an example of a first xenogeneic organ to be decellularized.

[0188] 1) As shown in Figure 3, the pig heart is cut into tissues for each region (left atrium, left ventricle, right atrium, right ventricle), and pre-processing steps such as removing blood vessels and foreign bodies are performed.

[0189] 2) As shown in Figure 3, place each tissue region (up to 25 g) into the first mesh chamber in the first reaction vessel, and then close the first lid of the first mesh chamber. The porcine heart tissue regions are partially cut with scissors, etc., and care must be taken when cutting the porcine heart tissue regions to a size smaller than the first mesh size of the first mesh chamber, as they may escape outside the first mesh and be lost during the decellularization process.

[0190] 3) In the case of cardiac muscle, the decellularization solution injected into the first reaction vessel varies, such as primary ultrapure water, SDS aqueous solution, Triton-X aqueous solution, EtOH, PBS aqueous solution, etc., and considering the volume of the first reaction vessel and the immersion height of the first mesh chamber, the injection volume of each solution is, for example, 600 ml.

[0191] First, primary ultrapure water is injected into the first reaction vessel for removing foreign matter and decellularizing the cardiac muscle.

[0192] 4) Then, pressurize the first lifting cylinder with gas (air) to move the first cover on the top of the first reaction vessel vertically downward so that the first cover and the first reaction vessel are sealed. At this time, for safety, use the first groove to check whether the first reaction vessel and the first cover are maintained in the correct position.

[0193] 5) The rotation speed (agitation speed) of the first rotating shaft driven by the first drive motor of the first drive unit is set to 0 to 500 rpm. In the case of pig hearts, it is set to 350 rpm, and the first mesh chamber is rotated and agitated with the settings of 10 seconds clockwise, 8 seconds pause, and 10 seconds counterclockwise.

[0194] 6) At the same time, the decellularization solution inside the first reaction vessel is circulated vertically, i.e., from the bottom to the top of the first reaction vessel, using a fixed-volume first transfer pump, and the circulation rate of the first decellularization solution is set to, for example, 200 ml / min.

[0195] Ultimately, although local decellularization progresses from the first xenogeneic organ tissue inside the first mesh chamber, the first decellularization solution in the first reaction vessel becomes well mixed due to the vertical circulation of the internal solution by the first transfer pump and the stirring caused by the rotation of the first mesh chamber, and a first decellularization solution of a homogeneous concentration can be obtained in real time.

[0196] This makes it easy to analyze the electrical conductivity, turbidity, absorbance (UV-Vis), etc. of the first decellularization solution when sampling it, allowing the progress of the decellularization process of the first decellularization solution to be accurately grasped in real time.

[0197] 7) Figure 4 shows a graph of the turbidity analysis results of the first decellularization solution using ultrapure water (DW) during the decellularization process for each tissue of a pig heart (left atrium, right atrium, left ventricle, right ventricle), and a graph of the turbidity analysis for each tissue over time during each decellularization process after a total of three ultrapure water processes [DW(1)-DW(2)-DW(3)].

[0198] As shown in Figure 4(a), an increase in turbidity, i.e., the color of the first decellularization solution becoming cloudy over time, means that the blood and impurities contained within the tissue have been released from the tissue, and significant differences were observed between tissues. For example, the turbidity of the left atrium was much higher than that of the right atrium.

[0199] Furthermore, although turbidity increased over time, the rate of increase gradually slowed.

[0200] After the DW(1) process is completed, the solution in the reactor is completely discharged as waste liquid, and two more ultrapure water processes are carried out as shown in Figure 4(b). After the DW(3) process, the final turbidity analysis showed that all tissues were less than 30 NTU, which indicates that most impurities have been removed.

[0201] 8) Figure 5 is a graph showing the results of electrical conductivity analysis of the first decellularization solution using ultrapure water (DW) during the decellularization process for each tissue of a pig heart (left atrium, right atrium, left ventricle, right ventricle).

[0202] This is a graph showing the analysis of the electrical conductivity of each tissue over time during each decellularization process after a total of three ultrapure water processes [DW(1)-DW(2)-DW(3)] were performed.

[0203] At 3 minutes, which is the earliest data point in Figure 5(a), the first decellularization solution in the first reaction vessel was not homogeneously mixed and very high electrical conductivity was observed, but this stabilized over time.

[0204] As shown in Figure 5(b), as the DW(2)-DW(3) process progressed, the electrical conductivity, as well as the turbidity, decreased significantly, indicating that most of the impurities had been removed.

[0205] 9) One implication that can be derived from the results of the analysis of tissue-specific turbidity and electrical conductivity of cardiac muscle is that simply high turbidity does not necessarily mean high electrical conductivity.

[0206] For example, the turbidity was highest in the left atrium, but the electrical conductivity was lowest at 280 uS / cm in the left atrium, while the electrical conductivity in the left ventricle was very high at approximately 670 uS / cm.

[0207] The correlation between turbidity and electrical conductivity is related to the first decellularization solution (substance) of the first xenogeneic organ tissue, so further related research is needed.

[0208] 10) Figure 6 shows a graph of the absorbance (UV-Vis) analysis results of the first decellularization solution for the left ventricle using ultrapure water (DW) during the decellularization process for each tissue (left atrium, right atrium, left ventricle, right ventricle) of porcine cardiac muscle, and a graph of the absorbance (UV-Vis) of each tissue over time during each decellularization process after a total of three ultrapure water processes [DW(1)-DW(2)-DW(3)].

[0209] Figure 6(a1) shows the results of the absorbance (UV-Vis) analysis as a function of time, and Figure 6(a2) is a graph showing the area (integral value) of such peaks.

[0210] As with the turbidity and electrical conductivity analysis results, the peak area increases over time, but the rate of increase tends to become gradually slower.

[0211] Figure 6(b1) shows plotting of the final data from each of the three ultrapure water decellularization processes, and Figure 6(b2) shows a graph of these peak areas.

[0212] As shown in Figure 6(b1), the absorbance (UV-Vis) results decreased significantly as the process progressed, which means that the intensity gradually became similar to that of the reference ultrapure water solution set as the baseline, proving that decellularization using ultrapure water was almost complete.

[0213] Similar results to those described above were also obtained for tissues other than the left ventricle (left atrium, right atrium, right ventricle).

[0214] 11) Figure 7 shows the process of decellularization of cardiac muscle tissue using ultrapure water and various organic solvents. As the process progresses from (a) to (b) to (c), the tissue volume decreases and its color turns white.

[0215] (Test results for performance, etc.) The most basic item for evaluating the quality of decellularized primary xenogeneic organ tissue bioink is biochemical assay, which quantitatively indicates the content of DNA, collagen, and GAGs in the tissue.

[0216] Table 1 shows the results of analyzing the DNA content of each tissue in decellularized pig hearts.

[0217] Because the DNA content of different tissues varies, a common standard for determining successful decellularization is when the DNA content in the tissue is less than 50 ng / mg.

[0218] As shown in Table 1, the DNA content of all tissues (left atrium, right atrium, left ventricle, right ventricle) met the above criteria, so it can be determined that decellularization was successful. [Table 1]

[0219] If collagen and GAGs (glycosaminoglycans) were removed at the same time as DNA was removed, the physical properties of the bioink would be poor. Therefore, it was necessary to analyze GAGs and collagen to confirm whether the two substances were well preserved within the tissue. As shown in Tables 2 and 3, we were able to confirm that a very large amount of GAGs and collagen was well preserved. [Table 2] [Table 3]

[0220] The results show that after decellularization of a variety of small quantities of xenogeneic organs using the twin system of the first and second ex situ analytical application devices, the DNA content of the left atrium, right atrium, left ventricle, and right ventricle tissues was reduced to an extremely low level, and the collagen and GAGs in the tissues were very well preserved, demonstrating that high-quality bioink materials were effectively produced.

[0221] Although the present disclosure has been described through the preferred embodiments as described above, it will be readily understood by those skilled in the art to which the present invention pertains that the present invention is not limited thereto, and various modifications and variations are possible without departing from the scope of the claims set forth below. [Explanation of symbols]

[0222] 100 First reaction vessel 200 Second Reaction Vessel RA1 First ex-situ analytical application device RA2 Second ex-situ analytical application device

Claims

1. a first reaction vessel for storing therein a first decellularization solution for carrying out a decellularization reaction with tissue of a first xenogeneic organ to be decellularized; and one or more first ex-situ analysis application devices for extracting the first decellularization solution undergoing a decellularization reaction with the tissue of the first xenogeneic organ to be decellularized in the first reaction vessel to the outside of the first reaction vessel and measuring the physical properties of the first decellularization solution ex-situ; A decellularization reactor comprising:

2. The first ex-situ analysis application device The decellularization reaction apparatus according to claim 1 , comprising a first mesh chamber that is inserted into the first reaction vessel and contains tissue of a first xenogeneic organ to be decellularized therein.

3. The first ex-situ analysis application device a first solution circulation unit connected between the first reaction container and a first cover covering the first reaction container so as to be in communication with each other, for circulating a first decellularization solution between the first reaction container and the first cover; and The decellularization reaction device according to claim 2 , further comprising a first solution sampling unit connected to the first solution circulation unit for extracting and sampling the first decellularization solution from the first solution circulation unit to the outside.

4. The decellularization reaction apparatus according to claim 3 , further comprising a first lifting unit coupled to the first cover for vertically lifting and lowering the first cover.

5. The decellularization reaction apparatus according to claim 4 , wherein the first mesh chamber includes a first drive unit for rotating the first mesh chamber so as to stir the contents in the first reaction vessel.

6. The decellularization reaction apparatus according to claim 5 , wherein the first mesh chamber is cylindrical and has an open interior.

7. The decellularization reaction apparatus according to claim 6 , wherein a first mesh is arranged in a lattice pattern on the outer peripheral surface of the first mesh chamber.

8. The decellularization reaction apparatus according to claim 6 , wherein a first lid for covering the first mesh chamber is detachably coupled to an upper end of the first mesh chamber.

9. The decellularization reaction apparatus 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 reactor according to claim 9 , wherein the first reaction vessel has a double jacket configuration consisting of a first inner tube and a first outer tube.

11. 11. The decellularization reaction device of claim 10, wherein the first external tube is provided with a first refrigerant inlet for injecting a refrigerant into a space between the first external tube and the first internal tube, and a first refrigerant outlet for discharging the injected refrigerant to the outside of the first external tube.

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

13. The decellularization reaction apparatus according to claim 11 , wherein the first cover is provided with a first injection port for injecting a first decellularization solution into the first reaction vessel.

14. The decellularization reaction apparatus according to claim 13 , wherein the first cover is provided with a first water level measuring device for measuring the water level of the first decellularization solution in the first reaction container.

15. The first driving unit is a first rotation shaft coupled to a first lid of the mesh chamber; and The decellularization reaction apparatus according to claim 5 , further comprising a first drive motor provided on a first cover of the first reaction vessel and coupled to the first rotation shaft for rotating the first rotation shaft.

16. The first solution circulation section a first circulation tube connected between the first cover and the first solution outlet for circulating the first decellularization solution in the first reaction container; and The decellularization reaction device according to claim 12, further comprising a first transfer pump provided in the first circulation pipe for transferring the first decellularization solution flowing out from the first solution outlet to the first circulation port.

17. The decellularization reaction apparatus according to claim 16 , wherein the first circulation pipe is made of an expandable and contractible flexible pipe.

18. 18. The decellularization reaction apparatus of claim 17, wherein the first solution sampling unit includes a first solution sampling tube connected to the first circulation tube for extracting and sampling the first decellularization solution circulating in the first circulation tube.

19. The decellularization reaction apparatus according to claim 18, wherein the absorbance (UV-Vis), electrical conductivity, and turbidity of the first decellularization solution sampled by the first solution sampling tube are measured to analyze the physical properties of the first decellularization solution.

20. The first lifting unit is a first coupling plate coupled to the first cover so as to be movable up and down; and The decellularization reaction apparatus according to claim 4 , further comprising a first lifting cylinder coupled to one end of the first binding plate and having a first lifting rod for lifting and lowering the first binding plate.

21. a first reaction vessel for storing therein a first decellularization solution for carrying out a decellularization reaction with tissue of a first xenogeneic organ to be decellularized; one or more first ex-situ analysis application devices for extracting the first decellularization solution undergoing a decellularization reaction with the tissue of the first heterologous organ to be decellularized in the first reaction vessel to the outside and measuring the physical properties of the first decellularization solution ex-situ; a second reaction vessel for storing therein a second decellularization solution for carrying out a decellularization reaction with tissue of a second xenogeneic organ to be decellularized; and one or more second ex-situ analysis application devices arranged alongside the first ex-situ analysis application device, for extracting a second decellularization solution undergoing a decellularization reaction with tissue of a second xenogeneic organ to be decellularized in the second reaction vessel to the outside and measuring the physical properties of the second decellularization solution ex-situ; A decellularization reactor comprising:

22. The second heterologous organ may be the same as or different from the first heterologous organ; 22. The decellularization reactor of claim 21, wherein the second decellularization solution is the same as or different from the first decellularization solution.

23. The first ex-situ analysis application device a first mesh chamber that is inserted into the first reaction vessel and contains tissue of a first xenogeneic organ to be decellularized; a first solution circulation unit connected between the first reaction container and a first cover covering the first reaction container so as to be in communication with each other, for circulating a first decellularization solution between the first reaction container and the first cover; and The decellularization reaction device according to claim 21 , comprising a first solution sampling unit connected to the first solution circulation unit for extracting and sampling the first decellularization solution from the first solution circulation unit to the outside.

24. The first ex-situ analysis application device a first lifting unit coupled to the first cover for vertically lifting and lowering the first cover; 24. The decellularization reactor of claim 23, comprising a first drive unit coupled to the first mesh chamber for rotating the first mesh chamber to cause agitation within the first reaction vessel.

25. The second ex-situ analysis application device 25. The decellularization reaction apparatus according to claim 21, comprising a second mesh chamber that is inserted into the second reaction vessel and contains tissue of a second xenogeneic organ to be decellularized therein.

26. The second ex-situ analysis application device a second solution circulation unit connected between the second reaction container and a second cover covering the second reaction container so as to be in communication with each other, for circulating a second decellularization solution between the second reaction container and the second cover; and The decellularization reaction device according to claim 25, further comprising a second solution sampling unit connected to the second solution circulation unit for extracting and sampling the second decellularization solution from the second solution circulation unit to the outside.

27. The decellularization reaction apparatus according to claim 26, further comprising a second lifting unit coupled to the second cover for vertically lifting and lowering the second cover.

28. 28. The decellularization reaction apparatus according to claim 27, wherein the second mesh chamber includes a second drive unit for rotating the second mesh chamber so as to stir the contents within the second reaction vessel.

29. The decellularization reaction apparatus according to claim 28 , wherein the second mesh chamber has a cylindrical shape with an open interior.

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

31. The decellularization reaction apparatus according to claim 30 , wherein a second lid for covering the second mesh chamber is detachably coupled to an upper end of the second mesh chamber.

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

33. The decellularization reactor of claim 32, wherein the second reaction vessel has a double jacket configuration consisting of a second inner tube and a second outer tube.

34. 34. The decellularization reaction apparatus of claim 33, wherein the second external tube is provided with a second refrigerant inlet for injecting a refrigerant into a space between the second external tube and the second internal tube, and a second refrigerant outlet for discharging the injected refrigerant to the outside of the second external tube.

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

36. 36. The decellularization reaction apparatus of claim 35, wherein the second cover is provided with a second injection port for injecting a second decellularization solution into the second reaction vessel.

37. The second drive unit is a second rotation shaft coupled to a second lid of the second mesh chamber; and The decellularization reaction apparatus according to claim 28 , further comprising a second drive motor provided on a second cover of the second reaction vessel and coupled to the second rotation shaft for rotating the second rotation shaft.

38. The second solution circulation section includes: a second circulation tube connected between the second cover and the second solution outlet for circulating the second decellularization solution in the second reaction vessel; and The decellularization reaction device according to claim 35, further comprising a second transfer pump provided in the second circulation pipe for transferring the second decellularization solution flowing out from the second solution outlet to the second circulation port.

39. The decellularization reaction apparatus according to claim 38, wherein the second circulation pipe is made of an expandable and contractible flexible pipe.

40. 40. The decellularization reaction apparatus of claim 39, wherein the second solution sampling unit includes a second solution sampling tube connected to the second circulation tube for extracting and sampling the second decellularization solution circulating in the second circulation tube.

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

42. The second lifting unit is a second coupling plate coupled to the second cover so as to be movable up and down; and 28. The decellularization reactor of claim 27, further comprising a second lifting cylinder coupled to one end of the second binding plate and having a second lifting rod for raising and lowering the second binding plate.

Images