WO2023210691A1 - Active ingredient-containing solution treatment device and extracorporeal active ingredient-containing solution circulation system - Google Patents
Active ingredient-containing solution treatment device and extracorporeal active ingredient-containing solution circulation system Download PDFInfo
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- WO2023210691A1 WO2023210691A1 PCT/JP2023/016454 JP2023016454W WO2023210691A1 WO 2023210691 A1 WO2023210691 A1 WO 2023210691A1 JP 2023016454 W JP2023016454 W JP 2023016454W WO 2023210691 A1 WO2023210691 A1 WO 2023210691A1
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- active ingredient
- containing solution
- cells
- blood
- processing device
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- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 235000002639 sodium chloride Nutrition 0.000 description 1
- 229910001415 sodium ion Inorganic materials 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000000101 thioether group Chemical group 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 210000005167 vascular cell Anatomy 0.000 description 1
- NLVXSWCKKBEXTG-UHFFFAOYSA-N vinylsulfonic acid Chemical compound OS(=O)(=O)C=C NLVXSWCKKBEXTG-UHFFFAOYSA-N 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
- C12M3/06—Tissue, human, animal or plant cell, or virus culture apparatus with filtration, ultrafiltration, inverse osmosis or dialysis means
Definitions
- Patent Document 2 discloses a configuration that performs a process of imparting nutritional substances to blood via hollow fibers as an extracorporeal active ingredient-containing solution circulation system.
- the active ingredient-containing solution processing device and the extracorporeal active ingredient-containing solution circulation system of Patent Document 1 use a blood component regulator in addition to a blood purification device, a problem arises in that the device becomes larger.
- the active ingredient-containing solution processing device and the extracorporeal active ingredient-containing solution circulation system of Patent Document 2 are provided with a nutritional substance supply channel and a body fluid supply channel in addition to the blood circulation channel, so they are different from Patent Document 1. Similarly, a problem arises in that the device becomes larger. Furthermore, as the device becomes larger, there is also the problem that the amount of blood to be circulated outside the body increases.
- the present invention has been made in consideration of the above points, and provides an active ingredient-containing solution processing device and an extracorporeal active ingredient-containing solution circulation system that can reduce the size of the device and suppress the amount of active ingredient-containing solution to be circulated outside the body.
- the purpose is to provide
- the active ingredient-containing solution processing device of the first aspect and a pump that circulates at least the component in the active ingredient-containing solution from the introduction part to the discharge part of the chamber are provided.
- An extracorporeal active ingredient-containing solution circulation system is provided.
- FIG. 1 is a diagram showing a schematic configuration of an extracorporeal blood circulation system 100.
- the extracorporeal blood circulation system 100 circulates at least some components of the blood outside the body of the living body LB, and performs predetermined processing on some of the blood components in the blood processing device 1.
- the extracorporeal blood circulation system 100 includes a blood processing device 1, an infusion reservoir 10, a first introduction piping system 11, a first discharge piping system 12, a second introduction piping system 13, and a second discharge piping system 14. , a first pump 21, a second pump 22, a third pump 23, and a fourth pump 24.
- the infusion fluid reservoir 10 stores infusion fluid.
- the second introduction piping system 13 includes piping that introduces the infusion discharged from the infusion reservoir 10 into the blood processing apparatus 1 .
- the second discharge piping system 14 includes piping that discharges the infusion fluid discharged from the blood processing device 1 toward the infusion fluid reservoir 10 .
- the third pump 23 is provided in the second introduction piping system 13 and sends the transfusion toward the blood processing apparatus 1 .
- the fourth pump 24 is provided in the second discharge piping system 14 and sends the infusion from the blood processing device 1 toward the infusion reservoir 10 .
- the blood processing device 1 includes a chamber 30 and a linear member F.
- the chamber 30 includes a chamber body 31, a lid 40, a fiber support (support) 50, and filters 61 and 62.
- the chamber body 31 has a cylindrical shape extending in the vertical direction.
- the chamber body 31 is open at the top.
- the chamber body 31 includes a bottom wall 32 , a peripheral wall 33 , a top wall 34 , and a processing space 35 .
- the pedestal portion 51 is located at the lower end of the fiber support 50.
- the pedestal portion 51 has a flange shape extending in the radial direction.
- the pedestal portion 51 has an annular magnet 51A facing downward.
- the support shaft 52 has a cylindrical shape extending upward from the pedestal portion 51.
- the diameter of the support shaft 52 is smaller than the diameter of the pedestal portion 51.
- a linear member F (details will be described later) is wound around the outer periphery of the support shaft 52. As shown in FIG.
- the fitting convex portion 53 has a cylindrical shape extending upward from the upper end of the support shaft 52.
- the diameter of the fitting convex portion 53 is smaller than the diameter of the support shaft 52.
- the fitting protrusion 53 is inserted into and fitted into the fitting recess 40a of the lid 40 from below.
- the fitting protrusion 53 is removable from the fitting recess 40a.
- the fitting recess 54 is recessed upward from the lower surface of the pedestal 51 .
- the fitting protrusion 32B of the bottom wall portion 32 is inserted into the fitting recess 54 from below and fitted therein.
- the through channel 55 passes through the fiber support 50 in the vertical direction.
- the lower end of the through passage 55 opens into the fitting recess 54 .
- the fitting recess 54 is removable from the fitting protrusion 32B.
- the fiber support 50 When the fitting protrusion 53 fits into the fitting recess 40a of the lid 40, the fiber support 50 is positioned on the lid 40, and the through passage 55 communicates with the through passage 40b.
- the fitting protrusion 32B fits into the fitting recess 54 the fiber support 50 is positioned in the chamber body 31, and the through passage 55 communicates with the introduction port 32D. That is, the fiber support 50 is detachably attached to the chamber 30.
- the introduction port 32D, the through channel 55, and the through channel 40b communicate with each other.
- the blood processing apparatus 1 in which the fiber support 50 is arranged in the processing space 35 of the chamber body 31 has a maximum diameter of about 2 cm and a maximum length of about 10 cm.
- the filter 61 is arranged at the boundary 60A between the through-flow path 55 and the through-flow path 40b.
- Boundary 60A is a discharge part from which blood is discharged in chamber 30.
- the filter 62 is arranged at a boundary 60B between the introduction port 32D and the through flow path 55.
- Boundary 60B is an introduction part into which blood is introduced into chamber 30.
- the filters 61 and 62 stop the linear member F inside the chamber 30.
- the mesh size of the filters 61 and 62 is smaller than the size of the linear member F.
- the filter 62 also at the boundary 60B, which is the introduction part, in consideration of causing the liquid to flow back when performing air removal or the like.
- the core part 210 has a hydrogel filled with a plurality of cells 115.
- Shell portion 220 covers the outer periphery of core portion 210 .
- the shell portion 220 is a hydrogel that is dissociable from the hydrogel forming the core portion 210 .
- FIG. 4 is a schematic diagram illustrating the manufacturing process of cell fiber F.
- the microfluidic device 140 includes an introduction port 110 for introducing a first liquid material to form a core section 210, an introduction port 120 for introducing a second liquid material for forming a shell section 220, and a shell section 220. It has an introduction port 130 for introducing the third liquid material to be gelled.
- extracellular matrix components include, but are not limited to, collagen (type I, type II, type III, type V, type XI, etc.), mouse EHS tumor extract (type IV collagen, laminin, heparan sulfate proteoglycan, etc.). (including) reconstituted basement membrane components, gelatin, agar, agarose, fibrin, glycosaminoglycans, hyaluronic acid, proteoglycans, thrombin, aprotinin, alginic acid, and the like.
- pH-responsive gelling solution examples include an alginate solution, a chitosan solution, a carboxymethyl cellulose solution, and an acrylic acid-based synthetic solution.
- the injection speed of the solution at the introduction ports 110 and 120 is not particularly limited, but if the diameter of the microfluidic device 140 is about 50 ⁇ m to 2 mm, it may be about 10 to 500 ⁇ L/min. By adjusting the injection speed of the solution at the inlets 110 and 120, the diameter of the core portion and the coating thickness of the shell portion can be adjusted as appropriate.
- the injection speed of the solution at the introduction port 130 is not particularly limited, but may be, for example, about 1 to 10 mL/min.
- the core portion 210 contains various growth factors suitable for maintaining, proliferating, or expressing functions of the cells 115, such as epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factor (TGF), insulin-like Growth factors (IGF), fibroblast growth factors (FGF), nerve growth factors (NGF), etc. may be included.
- EGF epidermal growth factor
- PDGF platelet-derived growth factor
- TGF transforming growth factor
- IGF insulin-like Growth factors
- FGF fibroblast growth factors
- NGF nerve growth factors
- an appropriate concentration can be selected depending on the type of growth factor.
- the amount of blood to be used can be adjusted using a small device. becomes possible to suppress.
- the polysaccharide is at least one selected from the group consisting of alginic acid, starch, glycogen, cellulose, xanthan gum, hyaluronic acid, carrageenan, pectin, pullulan, and salts thereof.
- the hydrophilic polymers include polyethylene glycol, polyethyleneimine, polyvinyl alcohol, polyacrylic acid, polyacrylamide, polyvinyl alcohol, polyvinylpyrrolidone, polyvinylacetamide, polyamine, poly(4-styrene sulfonic acid), poly(allylamine hydrochloride), At least one selected from the group consisting of poly(vinylsulfonic acid, sodium salt), poly(diallyldimethylammonium chloride), and poly(2-methacryloyloxyethylphosphorylcholine).
- a cell fiber F is obtained in which the core portion 210 is occupied by cells 115 and the shell portion 220 is gelled.
- the shell part 220 contains the block copolymer, so by adjusting the temperature to a temperature below LCST, the block copolymer becomes water-soluble, and by stirring the cell fiber F in water, the block copolymer becomes water-soluble. , block copolymers can be removed.
- a cellular fiber F in which the core portion 210 is filled with cells and the shell portion 220 is macroporous (porous) is obtained.
- FIG. 5 is a fluorescence image at 37° C. of a cell fiber F having a shell portion 220 with a macroporous structure in which the core portion is filled with fluorescent nanoparticles with a diameter of 100 nm.
- FIG. 6 is a fluorescence image of the cell fiber F in FIG. 5 when the temperature reached 4° C. after 1 hour. As shown in FIG. 5, it was confirmed that when the shell portion was at 37° C., the fluorescent nanoparticles with a diameter of 100 nm could not pass through the shell portion and remained in the core portion. As shown in FIG. 6, when the shell portion was at 4° C., no fluorescent nanoparticles with a diameter of 100 nm were observed in the core portion, and it was confirmed that the fluorescent nanoparticles passed through the shell portion.
- S protein spike protein
- S protein beads that can mimic the cell infection mechanism of SARS-CoV-2 will be produced.
- S protein beads bound by the affinity of Ni-NTA and His tag were prepared.
- the binding between the red fluorescent beads and the S protein was confirmed by measuring the fluorescence resonance energy transfer from the S protein to the red fluorescent beads.
- a rat was used as a living LB in the extracorporeal blood circulation system 100, the first introduction piping system 11 and the first discharge piping system 12 were connected to the rat's vein, and a solution containing 4 ⁇ g/mL of S protein beads was injected into the vein. .
- the amount of S protein beads was measured in the first introduction piping system 11 for 45 minutes every 5 minutes after the solution containing S protein beads was injected. Injection of a solution containing S protein beads and measurement of S protein beads every 5 minutes were performed for a sample in which cell fiber F was filled with VeroE6 cells and a sample in which cell fiber F was not filled with cells. .
- Figure 8 shows the relationship between the time after injection of a solution containing S protein beads and the measured amount of S protein beads for a sample filled with VeroE6 cells and a sample in which cell fiber F is not filled with cells.
- FIG. 8 it was confirmed that in the sample using VeroE6 cells, S protein beads decreased more over time than in the sample not using cells. This is because membrane proteins in VeroE6 cells come into contact with S protein beads and are instantly bound (infected) and captured, reducing the amount of S protein beads contained in the blood in the biological LB and purifying the blood. This is thought to be the result of
- FIG. 9 is a fluorescence image of a sample in which cell fiber F is not filled with cells.
- FIG. 10 is a fluorescence image of a sample in which cell fiber F was filled with VeroE6 cells. As shown in FIG. 9, in the sample in which cell fiber F was not filled with cells, capture of S protein beads was not confirmed. On the other hand, as shown in FIG. 10, in the sample in which the cell fiber F was filled with VeroE6 cells, blood purification due to the capture of S protein beads was confirmed.
- the cell fiber F which is filled with affinity cells that have an affinity for viruses and has a length of several tens of meters, is used, so that the virus can be captured without reducing the removal ability. can do.
- Physiologically active substances released into circulating blood include albumin, proteins such as enzymes, hormones such as insulin, exosomes, MicroRNA, sugars, lipids, and the like.
- Exocrine cells that release albumin, a physiologically active substance, into circulating blood as an exocrine secretion include hepatocytes.
- Exocrine cells that release enzymes, which are physiologically active substances, into circulating blood as exocrine secretions include enzyme-producing cells.
- Exocrine cells that release hormones such as insulin, which are physiologically active substances, into circulating blood as exocrine secretions include hormone-producing cells such as ⁇ cells.
- Exocrine cells that release exosomes, which are physiologically active substances, into the circulating blood include airway epithelial cells.
- physiologically active substances can be replenished into the circulating blood.
- blood exhibiting hypoalbuminemia etc. can be supplemented with albumin
- blood exhibiting lysosomal diseases etc. can be supplemented with enzymes. That is, the extracorporeal blood circulation system 100 of this embodiment functions as a physiologically active substance. According to this embodiment, there is no need to separately provide a physiologically active substance supply channel in addition to a blood circulation channel, and it is possible to downsize the device and suppress the amount of blood to be circulated outside the body.
- human albumin was detected in all four samples. Since human albumin is normally not detected in rats, it was confirmed that human albumin released from exocrine cells in cell fiber F was replenished into circulating blood via cell fiber F.
- the cell fibers F can be used to connect affinity cells that express a membrane protein that has an affinity for a specific substance contained in blood, and exocrine cells that release physiologically active substances as exocrine substances into the blood.
- the cell fiber F may include both the above-mentioned affinity cells and exocrine cells. In this case, by using the extracorporeal blood circulation system 100, both blood purification processing and physiologically active substance replenishment processing can be performed.
- the extracorporeal blood circulation system 100 has a configuration in which the infusion circulation path is provided, but a configuration in which the infusion circulation path is not provided may also be used.
- a configuration may be adopted in which the first discharge piping system 12 provided with the second pump 22 is connected to the discharge section 33B.
- blood in the processing space 35 is sent from the discharge section 33B toward the living body LB via the first discharge piping system 12 by driving the second pump 22.
- the linear member F may have a structure including a holding portion 220 that holds the cells 115 and allows at least some components of blood to pass therethrough.
- the holding portion 220 in this configuration is formed into a linear shape using, for example, alginic acid having the above-mentioned macroporous structure.
- the active ingredient-containing solution may be, for example, an infusion solution containing water, electrolytes, nutrients, etc., or an antibody production solution.
- the chamber has at least an introduction part into which the component in the active ingredient-containing solution is introduced, and a discharge part from which at least the component in the active ingredient-containing solution is discharged, Of the introduction part and the discharge part, at least the discharge part is provided with a filter for keeping the linear member inside the chamber.
- the active ingredient-containing solution processing device according to Supplementary Note 1. (Additional note 3)
- the holding part is a mixture containing a polysaccharide and a block copolymer of a polymer having a lower critical solution temperature and a hydrophilic polymer.
- the specific substance is a virus
- the affinity cells are angiotensin converting enzyme 2 (ACE2) expressing cells
- ACE2 angiotensin converting enzyme 2
- the physiologically active substance is albumin
- the exocrine cell is a hepatocyte
- the physiologically active substance is an enzyme
- the exocrine cell is an enzyme-producing cell
- the linear member is a core portion filled with a plurality of the cells; A shell part that covers the outer periphery of the core part as the holding part; is a cell fiber having The active ingredient-containing solution processing device according to any one of Supplementary Notes 1 to 7.
- An active ingredient-containing solution processing device according to any one of Supplementary Notes 1 to 8;
- An extracorporeal active ingredient-containing solution circulation system comprising: a pump that circulates at least the components of the active ingredient-containing solution from an inlet to an outlet of the chamber.
- the present invention can be applied to an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system.
- Blood processing device active ingredient-containing solution processing device
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Abstract
The purpose of the present invention is to provide an active ingredient-containing solution treatment device that can reduce the amount of an active ingredient-containing solution to be circulated in an extracorporeal manner, by reducing the size of the device. The present invention is provided with: a chamber into which at least a portion of components in the blood is introduced; and a cellular fiber that is disposed inside the chamber and that has a core part filled with multiple cells and a shell part that covers the outer circumference of the core part. The cells include: affinitive cells in which a membrane protein having affinity with a specific substance contained in a component in the blood is expressed; and/or external secretory cells that release a bioactive substance into the blood as an external secretion.
Description
本発明は、活性成分含有溶液処理装置および体外活性成分含有溶液循環システムに関する。
本願は、２０２２年４月２７日に、日本に出願された特願２０２２－０７３５０２号に基づき優先権を主張し、その内容をここに援用する。 The present invention relates to an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system.
This application claims priority based on Japanese Patent Application No. 2022-073502 filed in Japan on April 27, 2022, the contents of which are incorporated herein.
本願は、２０２２年４月２７日に、日本に出願された特願２０２２－０７３５０２号に基づき優先権を主張し、その内容をここに援用する。 The present invention relates to an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system.
This application claims priority based on Japanese Patent Application No. 2022-073502 filed in Japan on April 27, 2022, the contents of which are incorporated herein.
体外活性成分含有溶液循環システムの一例としては、例えば採血した活性成分含有溶液である血液から老廃物や余分な水分を除去する透析を行い、血液を浄化処理する血液透析装置を有する構成が知られている（例えば、特許文献１）。
As an example of an extracorporeal active ingredient-containing solution circulation system, a configuration is known that includes a hemodialysis device that performs dialysis to remove waste products and excess water from collected blood, which is an active ingredient-containing solution, and purifies the blood. (For example, Patent Document 1).
また、特許文献２には、体外活性成分含有溶液循環システムとして、中空線維を介して血液に栄養物質を付与する処理を行う構成が開示されている。
Additionally, Patent Document 2 discloses a configuration that performs a process of imparting nutritional substances to blood via hollow fibers as an extracorporeal active ingredient-containing solution circulation system.
特許文献１の活性成分含有溶液処理装置および体外活性成分含有溶液循環システムは、血液浄化デバイスに加えて血液成分調整器を用いているため、装置が大型化するという問題が生じる。特許文献２の活性成分含有溶液処理装置および体外活性成分含有溶液循環システムは、血液の循環路に加えて栄養物質の補給流路と体液の補給流路をそれぞれ設けているため、特許文献１と同様に装置が大型化するという問題が生じる。
また、装置が大型化することで、体外で循環させる血液の量が増えるという問題も生じる。 Since the active ingredient-containing solution processing device and the extracorporeal active ingredient-containing solution circulation system ofPatent Document 1 use a blood component regulator in addition to a blood purification device, a problem arises in that the device becomes larger. The active ingredient-containing solution processing device and the extracorporeal active ingredient-containing solution circulation system of Patent Document 2 are provided with a nutritional substance supply channel and a body fluid supply channel in addition to the blood circulation channel, so they are different from Patent Document 1. Similarly, a problem arises in that the device becomes larger.
Furthermore, as the device becomes larger, there is also the problem that the amount of blood to be circulated outside the body increases.
また、装置が大型化することで、体外で循環させる血液の量が増えるという問題も生じる。 Since the active ingredient-containing solution processing device and the extracorporeal active ingredient-containing solution circulation system of
Furthermore, as the device becomes larger, there is also the problem that the amount of blood to be circulated outside the body increases.
本発明は、以上のような点を考慮してなされたもので、装置を小型して体外で循環させる活性成分含有溶液の量を抑制できる活性成分含有溶液処理装置および体外活性成分含有溶液循環システムを提供することを目的とする。
The present invention has been made in consideration of the above points, and provides an active ingredient-containing solution processing device and an extracorporeal active ingredient-containing solution circulation system that can reduce the size of the device and suppress the amount of active ingredient-containing solution to be circulated outside the body. The purpose is to provide
本発明の第１の態様に従えば、少なくとも活性成分含有溶液における一部の成分が導入されるチャンバーと、前記チャンバーの内部に配置され、複数の細胞と、前記細胞を保持し前記成分を透過させる保持部とを有する線状部材と、を備え、前記細胞は、前記活性成分含有溶液における前記成分に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞と、前記活性成分含有溶液に外分泌物として生理活性物質を放出する外分泌性細胞と、の少なくとも一方を含む、活性成分含有溶液処理装置が提供される。
According to a first aspect of the present invention, there is provided a chamber into which at least some components of an active ingredient-containing solution are introduced, a plurality of cells disposed inside the chamber, and a plurality of cells that hold the cells and permeate the components. and a linear member having a holding portion, wherein the cell is an affinity cell expressing a membrane protein having an affinity for a specific substance contained in the component in the active ingredient-containing solution, and An active ingredient-containing solution processing device is provided that includes at least one of exocrine cells that release a physiologically active substance as an exocrine secretion into a solution.
本発明の第２の態様に従えば、第１の態様の活性成分含有溶液処理装置と、少なくとも前記活性成分含有溶液における前記成分を前記チャンバーの導入部から排出部に至って循環させるポンプと、を有する、体外活性成分含有溶液循環システムが提供される。
According to a second aspect of the present invention, the active ingredient-containing solution processing device of the first aspect and a pump that circulates at least the component in the active ingredient-containing solution from the introduction part to the discharge part of the chamber are provided. An extracorporeal active ingredient-containing solution circulation system is provided.
本発明では、装置を小型して体外で循環させる活性成分含有溶液の量を抑制できる活性成分含有溶液処理装置および体外活性成分含有溶液循環システムを提供することができる。
The present invention can provide an active ingredient-containing solution processing device and an extracorporeal active ingredient-containing solution circulation system that can reduce the size of the device and suppress the amount of active ingredient-containing solution that is circulated outside the body.
以下、本発明の活性成分含有溶液処理装置および体外活性成分含有溶液循環システムの実施の形態を、図１から図１７を参照して説明する。
なお、以下の実施形態は、本発明の一態様を示すものであり、この発明を限定するものではなく、本発明の技術的思想の範囲内で任意に変更可能である。また、以下の図面においては、各構成をわかりやすくするために、実際の構造と各構造における縮尺や数等を異ならせている。 DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system of the present invention will be described below with reference to FIGS. 1 to 17.
Note that the following embodiments illustrate one aspect of the present invention, and do not limit the present invention, and can be arbitrarily modified within the scope of the technical idea of the present invention. In addition, in the following drawings, in order to make each structure easier to understand, the scale, number, etc. of each structure are different from the actual structure.
なお、以下の実施形態は、本発明の一態様を示すものであり、この発明を限定するものではなく、本発明の技術的思想の範囲内で任意に変更可能である。また、以下の図面においては、各構成をわかりやすくするために、実際の構造と各構造における縮尺や数等を異ならせている。 DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system of the present invention will be described below with reference to FIGS. 1 to 17.
Note that the following embodiments illustrate one aspect of the present invention, and do not limit the present invention, and can be arbitrarily modified within the scope of the technical idea of the present invention. In addition, in the following drawings, in order to make each structure easier to understand, the scale, number, etc. of each structure are different from the actual structure.
本実施形態では、活性成分含有溶液の少なくとも一部として血液を処理する血液処理装置を活性成分含有溶液処理装置として説明し、体外において少なくとも活性成分含有溶液として血液における一部の成分を循環させる体外血液循環システムを体外活性成分含有溶液循環システムとして説明する。
In this embodiment, a blood processing device that processes blood as at least a part of an active ingredient-containing solution will be described as an active ingredient-containing solution processing device, and an extracorporeal device that circulates some components of blood as at least a part of an active ingredient-containing solution outside the body. The blood circulation system will be explained as an extracorporeal active ingredient-containing solution circulation system.
図１は、体外血液循環システム１００の概略構成を示す図である。
図１に示すように、体外血液循環システム１００は、生体ＬＢの体外において少なくとも血液における一部の成分を循環させ、血液処理装置１において血液における一部の成分に対して所定の処理を行う。 FIG. 1 is a diagram showing a schematic configuration of an extracorporealblood circulation system 100.
As shown in FIG. 1, the extracorporealblood circulation system 100 circulates at least some components of the blood outside the body of the living body LB, and performs predetermined processing on some of the blood components in the blood processing device 1.
図１に示すように、体外血液循環システム１００は、生体ＬＢの体外において少なくとも血液における一部の成分を循環させ、血液処理装置１において血液における一部の成分に対して所定の処理を行う。 FIG. 1 is a diagram showing a schematic configuration of an extracorporeal
As shown in FIG. 1, the extracorporeal
体外血液循環システム１００は、血液処理装置１と、輸液貯溜部１０と、第１導入配管系１１と、第１排出配管系１２と、第２導入配管系１３と、第２排出配管系１４と、第１ポンプ２１と、第２ポンプ２２と、第３ポンプ２３と、第４ポンプ２４とを備えている。
The extracorporeal blood circulation system 100 includes a blood processing device 1, an infusion reservoir 10, a first introduction piping system 11, a first discharge piping system 12, a second introduction piping system 13, and a second discharge piping system 14. , a first pump 21, a second pump 22, a third pump 23, and a fourth pump 24.
第１導入配管系１１は、生体ＬＢから少なくとも血液における一部の成分を血液処理装置１に導入する配管を含む。第１排出配管系１２は、血液処理装置１から排出される少なくとも血液における一部の成分を生体ＬＢに向けて排出する配管を含む。第１ポンプ２１と第２ポンプ２２は、少なくとも血液における一部の成分を第１導入配管系１１から第１排出配管系１２に至って循環させるポンプである。第１ポンプ２１は、第１導入配管系１１に設けられ少なくとも血液における一部の成分を血液処理装置１に向けて送液する。第２ポンプ２２は、第１排出配管系１２に設けられ少なくとも血液における一部の成分を生体ＬＢに向けて送液する。
The first introduction piping system 11 includes piping that introduces at least some components of blood from the living body LB into the blood processing apparatus 1. The first discharge piping system 12 includes piping that discharges at least some components of the blood discharged from the blood processing apparatus 1 toward the living body LB. The first pump 21 and the second pump 22 are pumps that circulate at least some components of blood from the first introduction piping system 11 to the first discharge piping system 12. The first pump 21 is provided in the first introduction piping system 11 and delivers at least some components of blood toward the blood processing apparatus 1 . The second pump 22 is provided in the first discharge piping system 12 and sends at least some components of blood toward the living body LB.
なお、少なくとも血液における一部の成分とは、生体ＬＢから採血される血液、または生体ＬＢから採血される血液のうち、血液から分離された特定の成分（例えば、血漿等）である。血液から分離された特定の成分を循環させる場合、第１導入配管系１１には、生体ＬＢから採血される血液のうち、血液から特定の成分を分離する分離器と、分離器から血液処理装置１に至る配管が含まれる。以下の説明では、血液から分離された特定の成分についても単に血液と呼ぶ場合がある。
Note that at least some components of the blood are blood collected from the living body LB, or specific components (for example, plasma, etc.) separated from the blood of the blood collected from the living body LB. When circulating specific components separated from blood, the first introduction piping system 11 includes a separator that separates specific components from the blood collected from the living body LB, and a blood processing device from the separator. 1 is included. In the following description, specific components separated from blood may also be simply referred to as blood.
輸液貯溜部１０は、輸液を貯留する。第２導入配管系１３は、輸液貯溜部１０から排出される輸液を血液処理装置１に導入する配管を含む。第２排出配管系１４は、血液処理装置１から排出される輸液を輸液貯溜部１０に向けて排出する配管を含む。第３ポンプ２３は、第２導入配管系１３に設けられ輸液を血液処理装置１に向けて送液する。第４ポンプ２４は、第２排出配管系１４に設けられ血液処理装置１からの輸液を輸液貯溜部１０に向けて送液する。
The infusion fluid reservoir 10 stores infusion fluid. The second introduction piping system 13 includes piping that introduces the infusion discharged from the infusion reservoir 10 into the blood processing apparatus 1 . The second discharge piping system 14 includes piping that discharges the infusion fluid discharged from the blood processing device 1 toward the infusion fluid reservoir 10 . The third pump 23 is provided in the second introduction piping system 13 and sends the transfusion toward the blood processing apparatus 1 . The fourth pump 24 is provided in the second discharge piping system 14 and sends the infusion from the blood processing device 1 toward the infusion reservoir 10 .
血液処理装置１は、導入された血液に対して所定の処理を行う。血液に対する所定の処理は、導入された血液に含まれる特定物質を捕捉して血液を浄化する処理と、導入された血液に対して生理活性物質を放出する処理の少なくとも一方である。
The blood processing device 1 performs predetermined processing on the introduced blood. The predetermined treatment for blood is at least one of a process for purifying the blood by capturing a specific substance contained in the introduced blood, and a process for releasing a physiologically active substance from the introduced blood.
血液処理装置１は、チャンバー３０と、線状部材Ｆと、を有する。チャンバー３０は、チャンバ本体３１と、蓋４０と、ファイバ支持体（支持体）５０と、フィルター６１、６２と、を有する。
チャンバ本体３１は、上下方向に延びる筒状である。チャンバ本体３１は、上側が開口している。チャンバ本体３１は、底壁部３２と、周壁部３３と、天壁部３４と、処理空間３５と、を有する。 Theblood processing device 1 includes a chamber 30 and a linear member F. The chamber 30 includes a chamber body 31, a lid 40, a fiber support (support) 50, and filters 61 and 62.
Thechamber body 31 has a cylindrical shape extending in the vertical direction. The chamber body 31 is open at the top. The chamber body 31 includes a bottom wall 32 , a peripheral wall 33 , a top wall 34 , and a processing space 35 .
チャンバ本体３１は、上下方向に延びる筒状である。チャンバ本体３１は、上側が開口している。チャンバ本体３１は、底壁部３２と、周壁部３３と、天壁部３４と、処理空間３５と、を有する。 The
The
底壁部３２は、チャンバ本体３１の下端に位置する。底壁部３２は、段部３２Ａと、嵌合突部３２Ｂと、血液導入部３２Ｃと、導入口３２Ｄと、を有する。段部３２Ａは、処理空間３５の下端から上側に突出する円柱状である。嵌合突部３２Ｂは、段部３２Ａから上側に突出する軸状である。嵌合突部３２Ｂの外径は、段部３２Ａの外径よりも小さい。血液導入部３２Ｃは、底壁部３２の下側に突出する。血液導入部３２Ｃには、第１導入配管系１１が接続される。導入口３２Ｄは、段部３２Ａ、嵌合突部３２Ｂおよび血液導入部３２Ｃを上下方向に貫通する。生体ＬＢからの血液は、第１ポンプ２１の駆動により第１導入配管系１１および血液導入部３２Ｃを介して導入口３２Ｄからチャンバー３０の内部に導入される。
The bottom wall portion 32 is located at the lower end of the chamber body 31. The bottom wall portion 32 includes a step portion 32A, a fitting protrusion 32B, a blood introduction portion 32C, and an introduction port 32D. The step portion 32A has a cylindrical shape that projects upward from the lower end of the processing space 35. The fitting projection 32B has a shaft shape that projects upward from the stepped portion 32A. The outer diameter of the fitting protrusion 32B is smaller than the outer diameter of the stepped portion 32A. The blood introduction part 32C protrudes below the bottom wall part 32. The first introduction piping system 11 is connected to the blood introduction section 32C. The introduction port 32D vertically passes through the stepped portion 32A, the fitting protrusion 32B, and the blood introduction portion 32C. Blood from the living body LB is introduced into the chamber 30 from the introduction port 32D via the first introduction piping system 11 and the blood introduction part 32C by driving the first pump 21.
周壁部３３は、底壁部３２の外縁から上側に延びる円筒状である。周壁部３３は、輸液導入部３３Ａと、輸液排出部３３Ｂと、を有する。輸液導入部３３Ａと輸液排出部３３Ｂとは、上下方向に互いに離れて配置されている。輸液導入部３３Ａは、周壁部３３における処理空間３５の下端近傍に位置する。輸液導入部３３Ａには、第２導入配管系１３が接続される。輸液貯溜部１０からの輸液は、第３ポンプ２３の駆動により第２導入配管系１３および輸液導入部３３Ａを介して処理空間３５に導入される。輸液排出部３３Ｂは、周壁部３３における輸液導入部３３Ａよりも上側の処理空間３５の上端近傍に位置する。処理空間３５の液体は、第４ポンプ２４の駆動により輸液排出部３３Ｂおよび第２排出配管系１４を介して輸液貯溜部１０に向けて送られる。
The peripheral wall portion 33 has a cylindrical shape extending upward from the outer edge of the bottom wall portion 32. The peripheral wall portion 33 has an infusion solution introduction section 33A and an infusion solution discharge section 33B. The infusion solution introduction section 33A and the infusion solution discharge section 33B are arranged vertically apart from each other. The infusion introduction part 33A is located near the lower end of the processing space 35 in the peripheral wall part 33. The second introduction piping system 13 is connected to the infusion solution introduction section 33A. The infusion from the infusion reservoir 10 is introduced into the processing space 35 via the second introduction piping system 13 and the infusion introduction part 33A by driving the third pump 23. The infusion discharge part 33B is located near the upper end of the processing space 35 above the infusion introduction part 33A in the peripheral wall part 33. The liquid in the processing space 35 is sent toward the infusion reservoir 10 via the infusion discharge part 33B and the second discharge piping system 14 by driving the fourth pump 24.
天壁部３４は、周壁部３３の上端から径方向外側に延びる円環状である。天壁部３４の上面には、処理空間３５における開口部の周囲に円環状の溝部３４ａが形成されている。溝部３４ａには、シール材３４ｂが配置されている。シール材３４ｂは、一例として、Ｏリングである。処理空間３５は、底壁部３２と周壁部３３に囲まれた空間であり、上下方向に延びる。
The top wall portion 34 has an annular shape extending radially outward from the upper end of the peripheral wall portion 33. An annular groove 34 a is formed on the upper surface of the ceiling wall 34 around the opening in the processing space 35 . A sealing material 34b is arranged in the groove 34a. The sealing material 34b is an O-ring, for example. The processing space 35 is a space surrounded by the bottom wall part 32 and the peripheral wall part 33, and extends in the vertical direction.
蓋４０は、チャンバ本体３１の上側に設けられる。蓋４０は、上側から天壁部３４にシール材３４ｂを介して接合することで、処理空間３５を液密に閉塞する。蓋４０が処理空間３５を液密に閉塞することで、処理空間３５に導入された液体が漏れることを抑制できる。蓋４０は、下側から上側に延びる嵌合凹部４０ａと、上下方向に貫通する貫通流路４０ｂと、を有している。
The lid 40 is provided above the chamber body 31. The lid 40 liquid-tightly closes the processing space 35 by joining the top wall portion 34 from above through the sealing material 34b. By liquid-tightly closing the processing space 35 with the lid 40, leakage of the liquid introduced into the processing space 35 can be suppressed. The lid 40 has a fitting recess 40a extending from the bottom to the top, and a through passage 40b penetrating in the vertical direction.
ファイバ支持体５０は、処理空間３５に配置される。図２に示すように、ファイバ支持体５０は、台座部５１と、支持軸５２と、嵌合凸部５３と、嵌合凹部５４と、貫通流路５５と、溝部５６と、を有する。
The fiber support 50 is placed in the processing space 35. As shown in FIG. 2, the fiber support 50 includes a pedestal 51, a support shaft 52, a fitting protrusion 53, a fitting recess 54, a through channel 55, and a groove 56.
台座部５１は、ファイバ支持体５０の下端に位置する。台座部５１は、径方向に延びるフランジ状である。台座部５１は、下側に臨んで円環状の磁石５１Ａを有する。支持軸５２は、台座部５１から上側に延びる円筒状である。支持軸５２の直径は、台座部５１の直径よりも小さい。支持軸５２の外周には、図１に示すように、線状部材Ｆ（詳細は後述）が巻き付けられる。
The pedestal portion 51 is located at the lower end of the fiber support 50. The pedestal portion 51 has a flange shape extending in the radial direction. The pedestal portion 51 has an annular magnet 51A facing downward. The support shaft 52 has a cylindrical shape extending upward from the pedestal portion 51. The diameter of the support shaft 52 is smaller than the diameter of the pedestal portion 51. As shown in FIG. 1, a linear member F (details will be described later) is wound around the outer periphery of the support shaft 52. As shown in FIG.
嵌合凸部５３は、支持軸５２の上端から上側に延びる円筒状である。嵌合凸部５３の直径は、支持軸５２の直径よりも小さい。嵌合凸部５３は、蓋４０の嵌合凹部４０ａに下側から挿入されて嵌合する。嵌合凸部５３は、嵌合凹部４０ａに対して離脱可能である。嵌合凹部５４は、台座部５１の下面から上側に窪んで形成される。嵌合凹部５４には、底壁部３２の嵌合突部３２Ｂが下側から挿入されて嵌合する。貫通流路５５は、ファイバ支持体５０を上下方向に貫通する。貫通流路５５の下端は、嵌合凹部５４に開口する。嵌合凹部５４は、嵌合突部３２Ｂに対して離脱可能である。
The fitting convex portion 53 has a cylindrical shape extending upward from the upper end of the support shaft 52. The diameter of the fitting convex portion 53 is smaller than the diameter of the support shaft 52. The fitting protrusion 53 is inserted into and fitted into the fitting recess 40a of the lid 40 from below. The fitting protrusion 53 is removable from the fitting recess 40a. The fitting recess 54 is recessed upward from the lower surface of the pedestal 51 . The fitting protrusion 32B of the bottom wall portion 32 is inserted into the fitting recess 54 from below and fitted therein. The through channel 55 passes through the fiber support 50 in the vertical direction. The lower end of the through passage 55 opens into the fitting recess 54 . The fitting recess 54 is removable from the fitting protrusion 32B.
嵌合凸部５３が蓋４０の嵌合凹部４０ａに嵌合したときに、ファイバ支持体５０は蓋４０に位置決めされ、貫通流路５５は貫通流路４０ｂと連通する。嵌合凹部５４に嵌合突部３２Ｂが嵌合したときに、ファイバ支持体５０はチャンバ本体３１に位置決めされ、貫通流路５５は導入口３２Ｄと連通する。つまり、ファイバ支持体５０は、チャンバー３０に対して着脱自在に取り付けられる。ファイバ支持体５０がチャンバー３０に取り付けられたときに、導入口３２Ｄ、貫通流路５５および貫通流路４０ｂは連通する。
When the fitting protrusion 53 fits into the fitting recess 40a of the lid 40, the fiber support 50 is positioned on the lid 40, and the through passage 55 communicates with the through passage 40b. When the fitting protrusion 32B fits into the fitting recess 54, the fiber support 50 is positioned in the chamber body 31, and the through passage 55 communicates with the introduction port 32D. That is, the fiber support 50 is detachably attached to the chamber 30. When the fiber support 50 is attached to the chamber 30, the introduction port 32D, the through channel 55, and the through channel 40b communicate with each other.
溝部５６は、支持軸５２において貫通流路５５から径方向外側に延びて支持軸５２の外周面に開口する。溝部５６は、貫通流路５５と処理空間３５とを径方向に繋ぐ。チャンバー３０の内部に導入された血液は、溝部５６を介して貫通流路５５と処理空間３５との間を流動可能である。溝部５６は、周方向に間隔をあけて複数（本実施形態では４つ）配置されている。周方向に複数配置された溝部５６の組みは、上下方向に間隔をあけて２組設けられている。
The groove portion 56 extends radially outward from the through passage 55 in the support shaft 52 and opens on the outer circumferential surface of the support shaft 52 . The groove portion 56 connects the through flow path 55 and the processing space 35 in the radial direction. Blood introduced into the chamber 30 can flow between the through channel 55 and the processing space 35 via the groove 56 . A plurality of grooves 56 (four in this embodiment) are arranged at intervals in the circumferential direction. Two sets of grooves 56 arranged in the circumferential direction are provided at intervals in the vertical direction.
ファイバ支持体５０がチャンバ本体３１の処理空間３５に配置された血液処理装置１は、最大直径が約２ｃｍであり、最大長さが約１０ｃｍである。
The blood processing apparatus 1 in which the fiber support 50 is arranged in the processing space 35 of the chamber body 31 has a maximum diameter of about 2 cm and a maximum length of about 10 cm.
フィルター６１は、貫通流路５５と貫通流路４０ｂとの境界６０Ａに配置されている。境界６０Ａは、チャンバー３０において血液が排出される排出部である。フィルター６２は、導入口３２Ｄと貫通流路５５との境界６０Ｂに配置されている。境界６０Ｂは、チャンバー３０において血液が導入される導入部である。フィルター６１、６２は、線状部材Ｆをチャンバー３０の内部に止める。フィルター６１、６２のメッシュサイズは、線状部材Ｆの大きさよりも小さい。線状部材Ｆが血液とともに貫通流路４０ｂに排出されないために、少なくともフィルター６１を排出部である境界６０Ａに配置することが好ましい。血液処理の準備として、空気抜き等を実施する際に液体を逆流させることを考慮して、フィルター６２を導入部である境界６０Ｂにも配置することが好ましい。
The filter 61 is arranged at the boundary 60A between the through-flow path 55 and the through-flow path 40b. Boundary 60A is a discharge part from which blood is discharged in chamber 30. The filter 62 is arranged at a boundary 60B between the introduction port 32D and the through flow path 55. Boundary 60B is an introduction part into which blood is introduced into chamber 30. The filters 61 and 62 stop the linear member F inside the chamber 30. The mesh size of the filters 61 and 62 is smaller than the size of the linear member F. In order to prevent the linear member F from being discharged into the through-flow path 40b together with the blood, it is preferable to arrange at least the filter 61 at the boundary 60A, which is the discharge portion. In preparation for blood treatment, it is preferable to arrange the filter 62 also at the boundary 60B, which is the introduction part, in consideration of causing the liquid to flow back when performing air removal or the like.
線状部材Ｆは、図３に示すように、複数の細胞１１５と、細胞１１５を保持し少なくとも血液における一部の成分を透過させる保持部２２０とを有する。本実施形態の線状部材Ｆは、コア部とシェル部とを有するコア／シェル構造である。以下の説明では、線状部材については、コア部２１０と、シェル部２２０と、を有する細胞ファイバＦと呼ぶ。
As shown in FIG. 3, the linear member F includes a plurality of cells 115 and a holding section 220 that holds the cells 115 and allows at least some components of blood to pass therethrough. The linear member F of this embodiment has a core/shell structure having a core part and a shell part. In the following description, the linear member will be referred to as a cellular fiber F having a core portion 210 and a shell portion 220.
コア部２１０は、複数の細胞１１５が充填されたハイドロゲルを有する。シェル部２２０は、コア部２１０の外周を被覆する。シェル部２２０は、コア部２１０を形成するハイドロゲルとは解離性を有するハイドロゲルである。
The core part 210 has a hydrogel filled with a plurality of cells 115. Shell portion 220 covers the outer periphery of core portion 210 . The shell portion 220 is a hydrogel that is dissociable from the hydrogel forming the core portion 210 .
細胞ファイバＦの作製方法は特に限定されないが、例えば、図４に示すような二重の同軸マイクロ流体装置（ｃｏａｘｉａｌ ｍｉｃｒｏｆｌｕｉｄｉｃ ｄｅｖｉｃｅ）１４０を用いることにより簡便に作製することができる。２つの流体を同軸となるようにコア部及びシェル部に分けて射出することができるマイクロ流体装置１４０は、例えば、Wonje Jeong, et al., Hydrodynamic microfabrication via "on the fly" photopolymerization of microscale fibers and tubes, Lab Chip, 2004, 4, 576-580 のＦｉｇ．１にも具体的に説明されている。
Although the method for producing the cell fiber F is not particularly limited, it can be easily produced, for example, by using a double coaxial microfluidic device 140 as shown in FIG. The microfluidic device 140 that can coaxially separate and inject two fluids into a core part and a shell part is described, for example, by Wonje Jeong, et al., Hydrodynamic microfabrication via "on the fly" photopolymerization of microscale fibers and Fig. tubes, Lab Chip, 2004, 4, 576-580. 1 is also explained in detail.
図４は、細胞ファイバＦの製造過程を説明する模式図である。
マイクロ流体装置１４０は、コア部２１０を形成するための第１液状体を導入する導入口１１０と、シェル部２２０を形成するための第２液状体を導入する導入口１２０と、シェル部２２０をゲル化させる第３液状体を導入する導入口１３０と、を有する。 FIG. 4 is a schematic diagram illustrating the manufacturing process of cell fiber F.
Themicrofluidic device 140 includes an introduction port 110 for introducing a first liquid material to form a core section 210, an introduction port 120 for introducing a second liquid material for forming a shell section 220, and a shell section 220. It has an introduction port 130 for introducing the third liquid material to be gelled.
マイクロ流体装置１４０は、コア部２１０を形成するための第１液状体を導入する導入口１１０と、シェル部２２０を形成するための第２液状体を導入する導入口１２０と、シェル部２２０をゲル化させる第３液状体を導入する導入口１３０と、を有する。 FIG. 4 is a schematic diagram illustrating the manufacturing process of cell fiber F.
The
コア部２１０を形成するための第１液状体としては、細胞１１５および細胞外マトリックス成分を含む溶液を例示することができる。第１液状体に含まれる細胞としては、特に限定されないが、内分泌細胞、外分泌細胞、神経系細胞、骨格筋系細胞、血液細胞、間質細胞等が挙げられる。ここで、内分泌細胞としては、例えば、神経分泌細胞、下垂体細胞、甲状腺細胞、副甲状腺細胞、膵島細胞、胃腸内分泌細胞、心筋細胞、肝細胞、腎臓細胞、脂肪細胞、副腎細胞、性腺細胞等が挙げられる。また、外分泌細胞としては、例えば、消化管上皮細胞等が挙げられる他に、生理活性物質を放出する細胞が挙げられる。生理活性物質を放出する細胞としては、アルブミンを放出する肝細胞、酵素を放出する酵素産生細胞等が挙げられる。また、神経系細胞としては、神経幹細胞、中枢神経細胞、末梢神経細胞、グリア細胞等が挙げられる。また、骨格筋系細胞としては、骨細胞、軟骨細胞等が挙げられる。また、血液細胞としては、造血幹細胞、白血球、赤血球、血小板等が挙げられる。また、間質細胞としては、繊維芽細胞、血管細胞等が挙げられる。また、血液浄化を目的として、血液に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞を用いることができる。血液に含まれる特定物質としては、ウイルスやサイトカイン等が挙げられる。これらの細胞はｉＰＳ細胞やＥＳ細胞等から分化誘導された細胞であってもよい。
An example of the first liquid for forming the core portion 210 is a solution containing the cells 115 and an extracellular matrix component. The cells contained in the first liquid body include, but are not particularly limited to, endocrine cells, exocrine cells, nervous system cells, skeletal muscle cells, blood cells, interstitial cells, and the like. Here, examples of endocrine cells include neurosecretory cells, pituitary cells, thyroid cells, parathyroid cells, pancreatic islet cells, gastrointestinal endocrine cells, cardiomyocytes, hepatocytes, kidney cells, adipocytes, adrenal cells, gonadal cells, etc. can be mentioned. Examples of exocrine cells include gastrointestinal epithelial cells and cells that release physiologically active substances. Examples of cells that release physiologically active substances include hepatocytes that release albumin, enzyme-producing cells that release enzymes, and the like. Furthermore, examples of nervous system cells include neural stem cells, central nerve cells, peripheral nerve cells, and glial cells. Further, examples of skeletal muscle cells include bone cells, cartilage cells, and the like. Further, examples of blood cells include hematopoietic stem cells, white blood cells, red blood cells, platelets, and the like. Furthermore, examples of interstitial cells include fibroblasts, vascular cells, and the like. Furthermore, for the purpose of blood purification, affinity cells expressing membrane proteins that have an affinity for specific substances contained in blood can be used. Specific substances contained in blood include viruses, cytokines, and the like. These cells may be cells induced to differentiate from iPS cells, ES cells, or the like.
細胞外マトリックス成分としては、特に限定されないが、例えば、コラーゲン（Ｉ型、ＩＩ型、ＩＩＩ型、Ｖ型、ＸＩ型など）、マウスＥＨＳ腫瘍抽出物（ＩＶ型コラーゲン、ラミニン、ヘパラン硫酸プロテオグリカンなどを含む）より再構成された基底膜成分、ゼラチン、寒天、アガロース、フィブリン、グリコサミノグリカン、ヒアルロン酸、プロテオグリカン、スロンビン、アプロチニン、アルギン酸などを例示することができる。
Examples of extracellular matrix components include, but are not limited to, collagen (type I, type II, type III, type V, type XI, etc.), mouse EHS tumor extract (type IV collagen, laminin, heparan sulfate proteoglycan, etc.). (including) reconstituted basement membrane components, gelatin, agar, agarose, fibrin, glycosaminoglycans, hyaluronic acid, proteoglycans, thrombin, aprotinin, alginic acid, and the like.
シェル部２２０を形成するための第２液状体としては、金属イオンの存在下でゲル化する溶液、温度に応答してゲル化する溶液、ｐＨに応答してゲル化する溶液、ゲル化する溶液、磁場に応答してゲル化する溶液等を用いることができる。
The second liquid for forming the shell portion 220 may include a solution that gels in the presence of metal ions, a solution that gels in response to temperature, a solution that gels in response to pH, and a solution that gels. , a solution that gels in response to a magnetic field, etc. can be used.
金属イオンの存在下でゲル化する溶液としては、二価又は三価の金属イオンの存在下でゲル化するアルギン酸溶液を、カルシウムイオンやカリウムイオンの存在下でゲル化するカラギーナン溶液、ナトリウムイオンの存在下でゲル化するアクリル酸系合成溶液等が挙げられる。
Solutions that gel in the presence of metal ions include alginic acid solutions that gel in the presence of divalent or trivalent metal ions, carrageenan solutions that gel in the presence of calcium and potassium ions, and solutions that gel in the presence of sodium ions. Examples include acrylic acid-based synthetic solutions that gel in the presence of the solvent.
温度応答性ゲル化溶液としては、ポリ（Ｎ－イソプロピルアクリルアミド）をポリエチレングリコールで架橋した温度応答性ゲル化溶液、メチルセルロース、ヒドロキシプロピルセルロース、ポリ乳酸とポリエチレングリコールの共重合体、ポリエチレングリコールとポリプロピレンオキシドのトリブロック共重合体（市販名：プルロニック（登録商標）、ポロキサマー）、アガロース、ポリビニルアルコール等が挙げられる。
Temperature-responsive gelling solutions include temperature-responsive gelling solutions in which poly(N-isopropylacrylamide) is crosslinked with polyethylene glycol, methylcellulose, hydroxypropylcellulose, copolymers of polylactic acid and polyethylene glycol, and polyethylene glycol and polypropylene oxide. (commercial name: Pluronic (registered trademark), poloxamer), agarose, polyvinyl alcohol, etc.
ｐＨ応答性ゲル化溶液としては、アルギン酸塩溶液、キトサン溶液、カルボキシメチルセルロース溶液、アクリル酸系合成溶液等が挙げられる。
Examples of the pH-responsive gelling solution include an alginate solution, a chitosan solution, a carboxymethyl cellulose solution, and an acrylic acid-based synthetic solution.
光応答性ゲル化溶液としては、骨格にアゾベンゼンとシクロデキストリンを組み合わせた合成ゲル化溶液、フマル酸アミドをスペーサーとした超分子からなるゲル化溶液、ニトロベンジル基を介して架橋されたないしは結合されているゲル化溶液等が挙げられる。
Examples of photoresponsive gelling solutions include synthetic gelling solutions that combine azobenzene and cyclodextrin in the skeleton, gelling solutions that consist of supramolecular molecules with fumaric acid amide spacers, and gelling solutions that are cross-linked or bonded via nitrobenzyl groups. Examples include gelling solutions.
磁場応答性ハイドロゲルとしては、磁性粒子を含有させた架橋ポリ（Ｎ－イソプロピルアクリルアミド）からなるゲル化溶液等が挙げられる。
Examples of magnetic field-responsive hydrogels include gel solutions made of crosslinked poly(N-isopropylacrylamide) containing magnetic particles.
一例として、コア部２１０の材料にコラーゲン溶液を用い、シェル部２２０の材料に架橋前のアルギン酸ナトリウム溶液を用いて細胞ファイバＦを製造する際には、まず、マイクロ流体装置１４０の導入口１１０から、細胞１１５を含むコラーゲン溶液を導入して射出する。また、マイクロ流体装置１４０の導入口１２０から、架橋前のアルギン酸ナトリウム溶液を導入して射出する。また、マイクロ流体装置１４０の導入口１３０から、塩化カルシウム溶液を導入して射出する。すると、シェル部のアルギン酸ナトリウム溶液がゲル化し、シェル部２２０がアルギン酸ゲルである細胞ファイバＦを製造することができる。また、細胞ファイバ２００を３７℃程度で数分から１時間程度加熱することにより、コア部２１０の細胞１１５を含むコラーゲン溶液をゲル化させることができる。
As an example, when manufacturing the cell fiber F using a collagen solution as the material for the core part 210 and a sodium alginate solution before crosslinking as the material for the shell part 220, first, from the inlet 110 of the microfluidic device 140, , a collagen solution containing cells 115 is introduced and injected. Further, a sodium alginate solution before crosslinking is introduced and injected from the inlet 120 of the microfluidic device 140. Further, a calcium chloride solution is introduced and injected from the inlet 130 of the microfluidic device 140. Then, the sodium alginate solution in the shell portion is gelled, and a cell fiber F in which the shell portion 220 is made of alginate gel can be manufactured. Further, by heating the cell fiber 200 at about 37° C. for several minutes to about an hour, the collagen solution containing the cells 115 in the core portion 210 can be gelled.
導入口１１０及び１２０における溶液の射出速度は特に限定されないが、マイクロ流体装置１４０の口径が５０μｍ～２ｍｍ程度である場合には、１０～５００μＬ／分程度であってもよい。導入口１１０及び１２０における溶液の射出速度を調節することにより、コア部の直径及びシェル部の被覆厚みを適宜調節できる。導入口１３０における溶液の射出速度は特に限定されないが、例えば１～１０ｍＬ／分程度であってもよい。
The injection speed of the solution at the introduction ports 110 and 120 is not particularly limited, but if the diameter of the microfluidic device 140 is about 50 μm to 2 mm, it may be about 10 to 500 μL/min. By adjusting the injection speed of the solution at the inlets 110 and 120, the diameter of the core portion and the coating thickness of the shell portion can be adjusted as appropriate. The injection speed of the solution at the introduction port 130 is not particularly limited, but may be, for example, about 1 to 10 mL/min.
細胞ファイバＦの外径は特に限定されず、例えば１０μｍ～２ｍｍ、例えば２００μｍ～２ｍｍ、例えば５０μｍ～１ｍｍ程度であってもよい。細胞ファイバＦの長さは特に限定されず、数ｍｍ～数ｍ程度であってもよい。細胞ファイバＦの断面形状としては、円形、楕円系、四角形や五角形等の多角形等が挙げられる。
The outer diameter of the cell fiber F is not particularly limited, and may be, for example, about 10 μm to 2 mm, for example, 200 μm to 2 mm, and, for example, about 50 μm to 1 mm. The length of the cell fiber F is not particularly limited, and may be approximately several mm to several meters. Examples of the cross-sectional shape of the cell fibers F include a circle, an ellipse, and a polygon such as a quadrangle and a pentagon.
細胞ファイバＦを培養液中で培養することにより、細胞を増殖させることができる。細胞ファイバＦは、培養液を適切に交換することにより、数か月培養することもできる。
By culturing cell fibers F in a culture solution, cells can be grown. Cell fibers F can also be cultured for several months by appropriately replacing the culture medium.
コア部２１０には、細胞１１５の維持、増殖又は機能発現等に適した各種の成長因子、例えば上皮成長因子（ＥＧＦ）、血小板由来成長因子（ＰＤＧＦ）、トランスフォーミング成長因子（ＴＧＦ）、インスリン様成長因子（ＩＧＦ）、線維芽細胞成長因子（ＦＧＦ）、神経成長因子（ＮＧＦ）等を含有させてもよい。成長因子を含有させる場合には、成長因子の種類に応じて適宜の濃度を選択することができる。
The core portion 210 contains various growth factors suitable for maintaining, proliferating, or expressing functions of the cells 115, such as epidermal growth factor (EGF), platelet-derived growth factor (PDGF), transforming growth factor (TGF), insulin-like Growth factors (IGF), fibroblast growth factors (FGF), nerve growth factors (NGF), etc. may be included. When containing a growth factor, an appropriate concentration can be selected depending on the type of growth factor.
上記のようにして得られた細胞ファイバＦは、ピンセット等を用いて容易に操作することができるが、本実施形態では、ファイバ支持体５０における支持軸５２の外周に巻き付けられる。具体的には、図２に示すように、予め培養液が貯留されたメスシリンダーＭＳにファイバ支持体５０を浸漬させておき、図示しない磁石が埋設された回転盤ＲＰをメスシリンダーＭＳの下側に設置しておく。連続的に製造される細胞ファイバＦをメスシリンダーＭＳに導入しつつ、回転盤ＲＰの回転を磁力でファイバ支持体５０に伝達してファイバ支持体５０を回転させることで、支持軸５２の外周に細胞ファイバＦを連続的に巻き付けることができる。これにより、長さ数十メートルにもおよぶ細胞ファイバＦを支持軸５２の外周に容易に巻き付けることが可能になる。
The cell fiber F obtained as described above can be easily manipulated using tweezers or the like, but in this embodiment, it is wound around the outer periphery of the support shaft 52 in the fiber support 50. Specifically, as shown in FIG. 2, the fiber support 50 is immersed in a graduated cylinder MS in which a culture solution is stored in advance, and a rotary plate RP in which a magnet (not shown) is embedded is placed under the graduated cylinder MS. Set it to . While introducing the continuously manufactured cellular fibers F into the measuring cylinder MS, the rotation of the rotary disk RP is transmitted to the fiber support 50 by magnetic force to rotate the fiber support 50, thereby causing the fibers to be attached to the outer periphery of the support shaft 52. The cell fiber F can be wound continuously. This makes it possible to easily wrap the cell fiber F, which is several tens of meters long, around the outer periphery of the support shaft 52.
支持軸５２の外周に細胞ファイバＦが巻き付けられたファイバ支持体５０は、ファイバーカートリッジＦＣを構成する。ファイバーカートリッジＦＣは、例えば、培養液が貯留された遠沈管に浸漬させ攪拌することで、細胞１１５を培養しつつ保管することができる。そして、血液処理装置１における血液処理を実施する際に、細胞ファイバＦが巻き付けられたファイバ支持体５０は、図１に示したように、チャンバー３０に装着される。
The fiber support body 50 in which the cell fibers F are wound around the outer periphery of the support shaft 52 constitutes a fiber cartridge FC. The fiber cartridge FC can be stored while culturing the cells 115, for example, by immersing it in a centrifuge tube in which a culture solution is stored and stirring it. When carrying out blood processing in the blood processing apparatus 1, the fiber support 50 around which the cell fibers F are wound is mounted in the chamber 30, as shown in FIG.
［血液浄化処理の第１実施形態］
体外血液循環システム１００を用いて、生体ＬＢから採血される血液を浄化するには、細胞ファイバＦに充填された細胞１１５として、血液に含まれる捕捉対称となる特定物質と親和性を有する膜タンパク質を発現した親和性細胞を用いることができる。ここでは、捕捉対称となる特定物質が、細胞ファイバＦにおけるシェル部２２０を透過可能であるものとして説明する。 [First embodiment of blood purification process]
In order to purify blood collected from a living body LB using the extracorporealblood circulation system 100, a membrane protein having an affinity for a specific substance contained in the blood and to be captured is used as the cells 115 filled in the cell fiber F. can be used. Here, explanation will be given assuming that the specific substance to be captured can pass through the shell portion 220 in the cell fiber F.
体外血液循環システム１００を用いて、生体ＬＢから採血される血液を浄化するには、細胞ファイバＦに充填された細胞１１５として、血液に含まれる捕捉対称となる特定物質と親和性を有する膜タンパク質を発現した親和性細胞を用いることができる。ここでは、捕捉対称となる特定物質が、細胞ファイバＦにおけるシェル部２２０を透過可能であるものとして説明する。 [First embodiment of blood purification process]
In order to purify blood collected from a living body LB using the extracorporeal
まず、第１ポンプ２１の駆動により、生体ＬＢからの血液が、第１導入配管系１１から導入口３２Ｄ、貫通流路５５および溝部５６を介して処理空間３５に流入する。また、第３ポンプ２３の駆動により輸液貯溜部１０からの輸液が、第２導入配管系１３および輸液導入部３３Ａを介して処理空間３５に導入される。処理空間３５においては、下側から輸液が導入されることで、処理空間３５に導入された血液の濃度が低くなるとともに、濃度勾配が生じる。処理空間３５において細胞ファイバＦにおけるシェル部２２０を透過した特定物質は、膜タンパク質を介して細胞１１５と結合して捕捉される。細胞ファイバＦに特定物質が捕捉されて浄化処理された血液は、第２ポンプ２２の駆動により溝部５６から貫通流路５５、貫通流路４０ｂおよび第１排出配管系１２を介して生体ＬＢに向けて送液される。
First, by driving the first pump 21, blood from the living body LB flows into the processing space 35 from the first introduction piping system 11 through the introduction port 32D, the through channel 55, and the groove 56. Further, by driving the third pump 23, the infusion from the infusion reservoir 10 is introduced into the processing space 35 via the second introduction piping system 13 and the infusion introduction part 33A. In the processing space 35, the infusion fluid is introduced from below, so that the concentration of the blood introduced into the processing space 35 becomes low and a concentration gradient occurs. The specific substance that has passed through the shell portion 220 of the cell fiber F in the processing space 35 is captured by binding to the cell 115 via the membrane protein. The purified blood with specific substances captured by the cell fibers F is directed from the groove 56 to the living body LB via the through channel 55, the through channel 40b, and the first discharge piping system 12 by driving the second pump 22. The liquid is delivered.
このように、本実施形態では、血液成分調整器等を用いることなく、細胞ファイバＦに充填された複数の細胞１１５により捕捉対称の特定物質を捕捉するため、小型の装置により、用いる血液の量を抑制することが可能になる。
In this way, in this embodiment, in order to capture the specific substance to be captured by the plurality of cells 115 filled in the cell fiber F without using a blood component regulator or the like, the amount of blood to be used can be adjusted using a small device. becomes possible to suppress.
［血液浄化処理の第２実施形態］
特定物質が、ウイルスの場合、直径が約１００ｎｍであり、シェル部２２０を形成するための第２液状体として、例えば、アルギン酸ゲルを用いた場合には、ゲルの孔径サイズを大きく変えることが困難であり、上述したウイルス、分泌物、細菌の透過が妨げられる場合がある。シェル部２２０におけるゲルの孔径サイズを制御するためには、シェル部２２０を例えば、多糖類と、下限臨界溶液温度（ＬＣＳＴ）を有する高分子と親水性高分子とのブロック共重合体とを含む混合物とすることができる。上記多糖類とブロック共重合体とを混合することにより、その混合物において相分離を起こさせ、その後ブロック共重合体を除去することにより、マクロポーラス構造を有するシェル部２２０を形成することができる。 [Second embodiment of blood purification process]
When the specific substance is a virus, the diameter is approximately 100 nm, and when, for example, alginate gel is used as the second liquid material for forming theshell portion 220, it is difficult to greatly change the pore size of the gel. Therefore, the permeation of the above-mentioned viruses, secretions, and bacteria may be hindered. In order to control the pore size of the gel in the shell portion 220, the shell portion 220 may include, for example, a block copolymer of a polysaccharide, a polymer having a lower critical solution temperature (LCST), and a hydrophilic polymer. It can be a mixture. By mixing the polysaccharide and the block copolymer, phase separation is caused in the mixture, and then the block copolymer is removed, thereby forming the shell portion 220 having a macroporous structure.
特定物質が、ウイルスの場合、直径が約１００ｎｍであり、シェル部２２０を形成するための第２液状体として、例えば、アルギン酸ゲルを用いた場合には、ゲルの孔径サイズを大きく変えることが困難であり、上述したウイルス、分泌物、細菌の透過が妨げられる場合がある。シェル部２２０におけるゲルの孔径サイズを制御するためには、シェル部２２０を例えば、多糖類と、下限臨界溶液温度（ＬＣＳＴ）を有する高分子と親水性高分子とのブロック共重合体とを含む混合物とすることができる。上記多糖類とブロック共重合体とを混合することにより、その混合物において相分離を起こさせ、その後ブロック共重合体を除去することにより、マクロポーラス構造を有するシェル部２２０を形成することができる。 [Second embodiment of blood purification process]
When the specific substance is a virus, the diameter is approximately 100 nm, and when, for example, alginate gel is used as the second liquid material for forming the
上記多糖類は、アルギン酸、デンプン、グリコーゲン、セルロース、キサンタンガム、ヒアルロン酸、カラギーナン、ペクチンおよびプルラン、並びにこれらの塩からなる群から選ばれる少なくとも１つである。
The polysaccharide is at least one selected from the group consisting of alginic acid, starch, glycogen, cellulose, xanthan gum, hyaluronic acid, carrageenan, pectin, pullulan, and salts thereof.
上記下限臨界溶液温度を有する高分子は、ポリ（Ｎ－アルキルアクリルアミド）、ポリ（Ｎ－ビニルアルキルアミド）およびポリビニルアルキルエーテルからなる群から選ばれる少なくとも１つである。
The polymer having the above lower critical solution temperature is at least one selected from the group consisting of poly(N-alkylacrylamide), poly(N-vinylalkylamide), and polyvinylalkyl ether.
上記親水性高分子は、ポリエチレングリコール、ポリエチレンイミン、ポリビニルアルコール、ポリアクリル酸、ポリアクリルアミド、ポリビニルアルコール、ポリビニルピロリドン、ポリビニルアセトアミド、ポリアミン、ポリ（４－スチレンスルホン酸）、ポリ（アリルアミン塩酸塩）、ポリ（ビニルスルホン酸、ナトリウム塩）、ポリ（ジアリルジメチルアンモニウムクロリド）、およびポリ（２－メタクリロイルオキシエチルホスホリルコリン）からなる群から選ばれる少なくとも１つである。
The hydrophilic polymers include polyethylene glycol, polyethyleneimine, polyvinyl alcohol, polyacrylic acid, polyacrylamide, polyvinyl alcohol, polyvinylpyrrolidone, polyvinylacetamide, polyamine, poly(4-styrene sulfonic acid), poly(allylamine hydrochloride), At least one selected from the group consisting of poly(vinylsulfonic acid, sodium salt), poly(diallyldimethylammonium chloride), and poly(2-methacryloyloxyethylphosphorylcholine).
上記ＬＣＳＴを有する高分子と親水性高分子とのブロック共重合体は、それぞれの高分子に官能基を付加しておいて、官能基同士を反応させることにより、合成することができる。
例えば、マレイミド基はチオール基（ＳＨ基）と反応して安定なチオエーテル基を形成するため、親水性高分子にＳＨ基、ＬＣＳＴを有する高分子にマレイミド基を付加して両者を反応させればよい。 The block copolymer of a polymer having LCST and a hydrophilic polymer can be synthesized by adding functional groups to each polymer and allowing the functional groups to react with each other.
For example, since a maleimide group reacts with a thiol group (SH group) to form a stable thioether group, if a maleimide group is added to a hydrophilic polymer having an SH group and a polymer having LCST and the two are allowed to react. good.
例えば、マレイミド基はチオール基（ＳＨ基）と反応して安定なチオエーテル基を形成するため、親水性高分子にＳＨ基、ＬＣＳＴを有する高分子にマレイミド基を付加して両者を反応させればよい。 The block copolymer of a polymer having LCST and a hydrophilic polymer can be synthesized by adding functional groups to each polymer and allowing the functional groups to react with each other.
For example, since a maleimide group reacts with a thiol group (SH group) to form a stable thioether group, if a maleimide group is added to a hydrophilic polymer having an SH group and a polymer having LCST and the two are allowed to react. good.
上記の多糖類とブロック共重合体の混合物において相分離構造を形成した後に、ブロック共重合体を除去することで、ポアサイズが１００－１０００ｎｍで多糖類により形成されたマクロポーラス構造を有するシェル部２２０が得られる。
マクロポーラス構造を有するシェル部２２０に細胞１１５等の生体関連物質を導入する場合は、４℃～３７℃の温度範囲に調節することが必要である。 After forming a phase-separated structure in the mixture of the polysaccharide and block copolymer, the block copolymer is removed, resulting in ashell portion 220 having a macroporous structure with a pore size of 100 to 1000 nm and formed from the polysaccharide. is obtained.
When introducing biological substances such ascells 115 into the shell portion 220 having a macroporous structure, it is necessary to adjust the temperature to a range of 4°C to 37°C.
マクロポーラス構造を有するシェル部２２０に細胞１１５等の生体関連物質を導入する場合は、４℃～３７℃の温度範囲に調節することが必要である。 After forming a phase-separated structure in the mixture of the polysaccharide and block copolymer, the block copolymer is removed, resulting in a
When introducing biological substances such as
マクロポーラス構造のシェル部２２０を有する細胞ファイバＦを製造するには、マイクロ流体装置１４０の導入口１１０から、細胞懸濁液が注入され導入口１２０に向けて射出される。導入口１２０から多糖類とブロック共重合体の混合物を注入し導入口１３０に向けて射出すると、コア部２１０を細胞懸濁液が占め、上記混合物が細胞周囲を覆ってシェル部２２０を形成し、同軸のコア・シェル状の流体を形成する。導入口１３０からカルシウムイオン含有水溶液を（例えば、ＣａＣｌ２含有水溶液）を注入することで、流体のシャル部がゲル化される。その結果、コア部２１０が細胞１１５で占められ、シェル部２２０がゲル化された細胞ファイバＦが得られる。この段階では、シェル部２２０には、ブロック共重合体が含まれるため、これをＬＣＳＴ以下の温度に調節することで、ブロック共重合体は水溶性となり、細胞ファイバＦを水中で攪拌することにより、ブロック共重合体を除去できる。これにより、コア部２１０に細胞が充填され、シェル部２２０がマクロポーラス状（多孔質）となった細胞ファイバＦが得られる。得られた細胞ファイバＦにおいては、コア部の直径１００～２００μｍ、細胞充填率２～１０×１０７ｃｅｌｌｓ／ｍを目標として、コア部の直径１６０μｍ、細胞充填率２×１０７ｃｅｌｌｓ／ｍを達成できた。
To manufacture the cell fiber F having the macroporous shell portion 220, a cell suspension is injected from the introduction port 110 of the microfluidic device 140 and ejected toward the introduction port 120. When a mixture of polysaccharide and block copolymer is injected from the inlet 120 and injected toward the inlet 130, the cell suspension occupies the core part 210, and the mixture covers the cells to form the shell part 220. , forming a coaxial core-shell fluid. By injecting an aqueous solution containing calcium ions (for example, an aqueous solution containing CaCl 2 ) from the inlet 130, the shell portion of the fluid is gelled. As a result, a cell fiber F is obtained in which the core portion 210 is occupied by cells 115 and the shell portion 220 is gelled. At this stage, the shell part 220 contains the block copolymer, so by adjusting the temperature to a temperature below LCST, the block copolymer becomes water-soluble, and by stirring the cell fiber F in water, the block copolymer becomes water-soluble. , block copolymers can be removed. As a result, a cellular fiber F in which the core portion 210 is filled with cells and the shell portion 220 is macroporous (porous) is obtained. The obtained cell fiber F has a core diameter of 100 to 200 μm and a cell filling rate of 2 to 10×10 7 cells/m, with a core diameter of 160 μm and a cell filling rate of 2×10 7 cells/m. I was able to achieve it.
マクロポーラス構造のシェル部２２０を有する細胞ファイバＦを用いることで、血液に含まれるウイルスはシェル部２２０を透過できる。そのため、細胞１１５として、血液に含まれる捕捉対称となるウイルスと親和性を有する膜タンパク質を発現した親和性細胞を用いることでができる。
これにより、本実施形態では、第１実施形態で説明したように、生体ＬＢからの血液に含まれるウイルスは、処理空間３５において細胞ファイバＦにおけるシェル部２２０を透過し、膜タンパク質を介して細胞１１５と結合して捕捉される。細胞ファイバＦにウイルスが捕捉されて浄化処理された血液は、生体ＬＢに向けて送液される。 By using the cellular fiber F having themacroporous shell portion 220, viruses contained in blood can permeate the shell portion 220. Therefore, as the cells 115, affinity cells expressing a membrane protein having an affinity for the virus to be captured contained in the blood can be used.
Accordingly, in this embodiment, as described in the first embodiment, the virus contained in the blood from the living body LB permeates theshell portion 220 of the cell fiber F in the processing space 35 and enters the cell via the membrane protein. 115 and is captured. The blood that has been purified with viruses trapped in the cell fibers F is sent toward the living body LB.
これにより、本実施形態では、第１実施形態で説明したように、生体ＬＢからの血液に含まれるウイルスは、処理空間３５において細胞ファイバＦにおけるシェル部２２０を透過し、膜タンパク質を介して細胞１１５と結合して捕捉される。細胞ファイバＦにウイルスが捕捉されて浄化処理された血液は、生体ＬＢに向けて送液される。 By using the cellular fiber F having the
Accordingly, in this embodiment, as described in the first embodiment, the virus contained in the blood from the living body LB permeates the
図５は、コア部に直径１００ｎｍの蛍光ナノ粒子が充填されたマクロポーラス構造のシェル部２２０を有する細胞ファイバＦの３７℃における蛍光画像である。図６は、図５の細胞ファイバＦが１時間経過後に４℃となったときの蛍光画像である。図５に示すように、シェル部が３７℃であるときには、直径１００ｎｍの蛍光ナノ粒子はシェル部を透過できずにコア部に止まっていることを確認できた。図６に示すように、シェル部が４℃であるときには、コア部に直径１００ｎｍの蛍光ナノ粒子を確認できず、蛍光ナノ粒子はシェル部を透過したことを確認できた。
FIG. 5 is a fluorescence image at 37° C. of a cell fiber F having a shell portion 220 with a macroporous structure in which the core portion is filled with fluorescent nanoparticles with a diameter of 100 nm. FIG. 6 is a fluorescence image of the cell fiber F in FIG. 5 when the temperature reached 4° C. after 1 hour. As shown in FIG. 5, it was confirmed that when the shell portion was at 37° C., the fluorescent nanoparticles with a diameter of 100 nm could not pass through the shell portion and remained in the core portion. As shown in FIG. 6, when the shell portion was at 4° C., no fluorescent nanoparticles with a diameter of 100 nm were observed in the core portion, and it was confirmed that the fluorescent nanoparticles passed through the shell portion.
［模擬ウイルス］
特定物質としてのウイルスのうち、ＳＡＲＳ－ＣｏＶ－２の細胞感染機構を模倣可能なＳタンパク質（スパイクタンパク質）ビーズを作製する。
図７に示すように、赤色蛍光ビーズにＮｉ－ＮＴＡが付加され、緑色蛍光タンパク質にＨｉｓタグが付加されたＳタンパク質を用い、Ｎｉ－ＮＴＡとＨｉｓタグのアフィニティにより結合したＳタンパク質ビーズを作製した。赤色蛍光ビーズとＳタンパク質の結合は、Ｓタンパク質から赤色蛍光ビーズへの蛍光共鳴エネルギー移動を測定することで確認できた。 [Mock virus]
Among viruses as specific substances, S protein (spike protein) beads that can mimic the cell infection mechanism of SARS-CoV-2 will be produced.
As shown in Figure 7, using S protein in which Ni-NTA was added to red fluorescent beads and His tag was added to green fluorescent protein, S protein beads bound by the affinity of Ni-NTA and His tag were prepared. . The binding between the red fluorescent beads and the S protein was confirmed by measuring the fluorescence resonance energy transfer from the S protein to the red fluorescent beads.
特定物質としてのウイルスのうち、ＳＡＲＳ－ＣｏＶ－２の細胞感染機構を模倣可能なＳタンパク質（スパイクタンパク質）ビーズを作製する。
図７に示すように、赤色蛍光ビーズにＮｉ－ＮＴＡが付加され、緑色蛍光タンパク質にＨｉｓタグが付加されたＳタンパク質を用い、Ｎｉ－ＮＴＡとＨｉｓタグのアフィニティにより結合したＳタンパク質ビーズを作製した。赤色蛍光ビーズとＳタンパク質の結合は、Ｓタンパク質から赤色蛍光ビーズへの蛍光共鳴エネルギー移動を測定することで確認できた。 [Mock virus]
Among viruses as specific substances, S protein (spike protein) beads that can mimic the cell infection mechanism of SARS-CoV-2 will be produced.
As shown in Figure 7, using S protein in which Ni-NTA was added to red fluorescent beads and His tag was added to green fluorescent protein, S protein beads bound by the affinity of Ni-NTA and His tag were prepared. . The binding between the red fluorescent beads and the S protein was confirmed by measuring the fluorescence resonance energy transfer from the S protein to the red fluorescent beads.
［親和性細胞］
上記Ｓタンパク質ビーズを捕捉可能なアンジオテンシン変換酵素２（ＡＣＥ２）発現細胞としては、レンチウイルスベクターを用いた遺伝子導入による各種ＡＣＥ２発現細胞（ＶｅｒｏＥ６細胞、ＶｅｒｏＥ６／ＴＭＰＲＳＳ細胞、Ｈｅｌａ細胞、Ｈｅｌａ／ＡＣＥ２細胞）の作製を行った。また、上記Ｓタンパク質ビーズとの反応性評価と、フローサイトメーターによる定量比較を行った。上記の細胞のうち、Ｓタンパク質ビーズの捕捉効率が５０％以上であり、最大であったＶｅｒｏＥ６細胞をＡＣＥ２発現細胞として選定した。 [Affinity cells]
Examples of angiotensin-converting enzyme 2 (ACE2)-expressing cells capable of capturing the S protein beads include various ACE2-expressing cells (VeroE6 cells, VeroE6/TMPRSS cells, Hela cells, Hela/ACE2 cells) by gene transfer using a lentiviral vector. was created. In addition, reactivity evaluation with the above-mentioned S protein beads and quantitative comparison using a flow cytometer were performed. Among the above cells, VeroE6 cells, which had the highest S protein bead capture efficiency of 50% or more, were selected as ACE2-expressing cells.
上記Ｓタンパク質ビーズを捕捉可能なアンジオテンシン変換酵素２（ＡＣＥ２）発現細胞としては、レンチウイルスベクターを用いた遺伝子導入による各種ＡＣＥ２発現細胞（ＶｅｒｏＥ６細胞、ＶｅｒｏＥ６／ＴＭＰＲＳＳ細胞、Ｈｅｌａ細胞、Ｈｅｌａ／ＡＣＥ２細胞）の作製を行った。また、上記Ｓタンパク質ビーズとの反応性評価と、フローサイトメーターによる定量比較を行った。上記の細胞のうち、Ｓタンパク質ビーズの捕捉効率が５０％以上であり、最大であったＶｅｒｏＥ６細胞をＡＣＥ２発現細胞として選定した。 [Affinity cells]
Examples of angiotensin-converting enzyme 2 (ACE2)-expressing cells capable of capturing the S protein beads include various ACE2-expressing cells (VeroE6 cells, VeroE6/TMPRSS cells, Hela cells, Hela/ACE2 cells) by gene transfer using a lentiviral vector. was created. In addition, reactivity evaluation with the above-mentioned S protein beads and quantitative comparison using a flow cytometer were performed. Among the above cells, VeroE6 cells, which had the highest S protein bead capture efficiency of 50% or more, were selected as ACE2-expressing cells.
［血液浄化療法への実証］
体外血液循環システム１００における生体ＬＢとしてラットを用い、第１導入配管系１１および第１排出配管系１２をラットの静脈に接続し、当該静脈に４μｇ／ｍＬのＳタンパク質ビーズを含む溶液を注入した。Ｓタンパク質ビーズを含む溶液を注入してから５分毎に、第１導入配管系１１においてＳタンパク質ビーズの量を４５分間計測した。Ｓタンパク質ビーズを含む溶液の注入と、５分毎のＳタンパク質ビーズの計測は、細胞ファイバＦにＶｅｒｏＥ６細胞が充填されたサンプルと、細胞ファイバＦに細胞を充填しないサンプルとに対してそれぞれ行った。 [Demonstration for blood purification therapy]
A rat was used as a living LB in the extracorporealblood circulation system 100, the first introduction piping system 11 and the first discharge piping system 12 were connected to the rat's vein, and a solution containing 4 μg/mL of S protein beads was injected into the vein. . The amount of S protein beads was measured in the first introduction piping system 11 for 45 minutes every 5 minutes after the solution containing S protein beads was injected. Injection of a solution containing S protein beads and measurement of S protein beads every 5 minutes were performed for a sample in which cell fiber F was filled with VeroE6 cells and a sample in which cell fiber F was not filled with cells. .
体外血液循環システム１００における生体ＬＢとしてラットを用い、第１導入配管系１１および第１排出配管系１２をラットの静脈に接続し、当該静脈に４μｇ／ｍＬのＳタンパク質ビーズを含む溶液を注入した。Ｓタンパク質ビーズを含む溶液を注入してから５分毎に、第１導入配管系１１においてＳタンパク質ビーズの量を４５分間計測した。Ｓタンパク質ビーズを含む溶液の注入と、５分毎のＳタンパク質ビーズの計測は、細胞ファイバＦにＶｅｒｏＥ６細胞が充填されたサンプルと、細胞ファイバＦに細胞を充填しないサンプルとに対してそれぞれ行った。 [Demonstration for blood purification therapy]
A rat was used as a living LB in the extracorporeal
図８は、Ｓタンパク質ビーズを含む溶液を注入後の時間と、計測されたＳタンパク質ビーズ量との関係を、ＶｅｒｏＥ６細胞が充填されたサンプルと、細胞ファイバＦに細胞を充填しないサンプルとについてそれぞれ示す図である。
図８に示されるように、ＶｅｒｏＥ６細胞を用いたサンプルでは、細胞を用いないサンプルよりも、時間の経過とともにＳタンパク質ビーズが大きく減ることを確認できた。これは、ＶｅｒｏＥ６細胞における膜タンパク質がＳタンパク質ビーズと接触することで瞬時に結合（感染）して捕捉されることで、生体ＬＢ内の血液に含まれるＳタンパク質ビーズが減少して血液が浄化された結果と考えられる。 Figure 8 shows the relationship between the time after injection of a solution containing S protein beads and the measured amount of S protein beads for a sample filled with VeroE6 cells and a sample in which cell fiber F is not filled with cells. FIG.
As shown in FIG. 8, it was confirmed that in the sample using VeroE6 cells, S protein beads decreased more over time than in the sample not using cells. This is because membrane proteins in VeroE6 cells come into contact with S protein beads and are instantly bound (infected) and captured, reducing the amount of S protein beads contained in the blood in the biological LB and purifying the blood. This is thought to be the result of
図８に示されるように、ＶｅｒｏＥ６細胞を用いたサンプルでは、細胞を用いないサンプルよりも、時間の経過とともにＳタンパク質ビーズが大きく減ることを確認できた。これは、ＶｅｒｏＥ６細胞における膜タンパク質がＳタンパク質ビーズと接触することで瞬時に結合（感染）して捕捉されることで、生体ＬＢ内の血液に含まれるＳタンパク質ビーズが減少して血液が浄化された結果と考えられる。 Figure 8 shows the relationship between the time after injection of a solution containing S protein beads and the measured amount of S protein beads for a sample filled with VeroE6 cells and a sample in which cell fiber F is not filled with cells. FIG.
As shown in FIG. 8, it was confirmed that in the sample using VeroE6 cells, S protein beads decreased more over time than in the sample not using cells. This is because membrane proteins in VeroE6 cells come into contact with S protein beads and are instantly bound (infected) and captured, reducing the amount of S protein beads contained in the blood in the biological LB and purifying the blood. This is thought to be the result of
図９は、細胞ファイバＦに細胞を充填しないサンプルの蛍光画像である。図１０は、細胞ファイバＦにＶｅｒｏＥ６細胞が充填されたサンプルの蛍光画像である。
図９に示すように、細胞ファイバＦに細胞を充填しないサンプルでは、Ｓタンパク質ビーズの捕捉が確認されなかった。一方、図１０に示すように、細胞ファイバＦにＶｅｒｏＥ６細胞が充填されたサンプルでは、Ｓタンパク質ビーズの捕捉による血液浄化を確認できた。 FIG. 9 is a fluorescence image of a sample in which cell fiber F is not filled with cells. FIG. 10 is a fluorescence image of a sample in which cell fiber F was filled with VeroE6 cells.
As shown in FIG. 9, in the sample in which cell fiber F was not filled with cells, capture of S protein beads was not confirmed. On the other hand, as shown in FIG. 10, in the sample in which the cell fiber F was filled with VeroE6 cells, blood purification due to the capture of S protein beads was confirmed.
図９に示すように、細胞ファイバＦに細胞を充填しないサンプルでは、Ｓタンパク質ビーズの捕捉が確認されなかった。一方、図１０に示すように、細胞ファイバＦにＶｅｒｏＥ６細胞が充填されたサンプルでは、Ｓタンパク質ビーズの捕捉による血液浄化を確認できた。 FIG. 9 is a fluorescence image of a sample in which cell fiber F is not filled with cells. FIG. 10 is a fluorescence image of a sample in which cell fiber F was filled with VeroE6 cells.
As shown in FIG. 9, in the sample in which cell fiber F was not filled with cells, capture of S protein beads was not confirmed. On the other hand, as shown in FIG. 10, in the sample in which the cell fiber F was filled with VeroE6 cells, blood purification due to the capture of S protein beads was confirmed.
図１１は、細胞ファイバＦにＶｅｒｏＥ６細胞が１時間反応した後の蛍光画像である。図１２は、細胞ファイバＦにＶｅｒｏＥ６細胞が２時間反応した後の蛍光画像である。図１１および図１２に示されるように、反応時間が１時間よりも２時間の方がＳタンパク質ビーズの捕捉量が多いことから、反応時間に依存してＳタンパク質ビーズの捕捉量が増加することを確認できた。
FIG. 11 is a fluorescence image after VeroE6 cells reacted with cell fiber F for 1 hour. FIG. 12 is a fluorescence image after VeroE6 cells reacted with cell fiber F for 2 hours. As shown in FIGS. 11 and 12, the amount of S protein beads captured is larger when the reaction time is 2 hours than when the reaction time is 1 hour, indicating that the amount of S protein beads captured increases depending on the reaction time. I was able to confirm.
［ウイルス感染］
血液中の特定物質として、コロナウイルスのスパイクタンパク質エンベロープを持つCoV S-protein シュードタイプレンチウイルスを用いて血液を循環させた。図１３は、コロナウイルスのシュードタイプレンチウイルスを用い、細胞ファイバＦに細胞を充填しないサンプルにおいて、循環６日後の蛍光画像である。図１４は、コロナウイルスのシュードタイプレンチウイルスを用い、細胞ファイバＦにＶｅｒｏＥ６細胞を用いたサンプルにおいて、循環６日後の蛍光画像である。
図１３に示されるように、細胞ファイバＦに細胞を充填しないサンプルでは、明確なウイルス感染は確認できなかった。図１４に示されるように、細胞ファイバＦにＶｅｒｏＥ６細胞を用いたサンプルでは、ウイルスがＶｅｒｏＥ６細胞の核内に移行して蛍光が生じておりウイルス感染を確認できた。 [Virus infection]
As a specific substance in the blood, a CoV S-protein pseudotype lentivirus with the coronavirus spike protein envelope was used to circulate the blood. FIG. 13 is a fluorescence image after 6 days of circulation in a sample using coronavirus pseudotyped lentivirus and without filling cell fiber F with cells. FIG. 14 is a fluorescence image after 6 days of circulation in a sample using coronavirus pseudotype lentivirus and using VeroE6 cells as cell fiber F.
As shown in FIG. 13, clear virus infection could not be confirmed in the sample in which cell fiber F was not filled with cells. As shown in FIG. 14, in the sample in which VeroE6 cells were used as cell fiber F, the virus migrated into the nucleus of the VeroE6 cells and fluorescence was generated, confirming virus infection.
血液中の特定物質として、コロナウイルスのスパイクタンパク質エンベロープを持つCoV S-protein シュードタイプレンチウイルスを用いて血液を循環させた。図１３は、コロナウイルスのシュードタイプレンチウイルスを用い、細胞ファイバＦに細胞を充填しないサンプルにおいて、循環６日後の蛍光画像である。図１４は、コロナウイルスのシュードタイプレンチウイルスを用い、細胞ファイバＦにＶｅｒｏＥ６細胞を用いたサンプルにおいて、循環６日後の蛍光画像である。
図１３に示されるように、細胞ファイバＦに細胞を充填しないサンプルでは、明確なウイルス感染は確認できなかった。図１４に示されるように、細胞ファイバＦにＶｅｒｏＥ６細胞を用いたサンプルでは、ウイルスがＶｅｒｏＥ６細胞の核内に移行して蛍光が生じておりウイルス感染を確認できた。 [Virus infection]
As a specific substance in the blood, a CoV S-protein pseudotype lentivirus with the coronavirus spike protein envelope was used to circulate the blood. FIG. 13 is a fluorescence image after 6 days of circulation in a sample using coronavirus pseudotyped lentivirus and without filling cell fiber F with cells. FIG. 14 is a fluorescence image after 6 days of circulation in a sample using coronavirus pseudotype lentivirus and using VeroE6 cells as cell fiber F.
As shown in FIG. 13, clear virus infection could not be confirmed in the sample in which cell fiber F was not filled with cells. As shown in FIG. 14, in the sample in which VeroE6 cells were used as cell fiber F, the virus migrated into the nucleus of the VeroE6 cells and fluorescence was generated, confirming virus infection.
このように、本実施形態では、上記血液浄化処理の第１実施形態と同様の作用・効果が得られることに加えて、１００ｎｍ以上の大きな直径を有するウイルス、分泌物、細菌であっても、シェル部２２０を透過させてコア部２１０の親和精細胞と結合させて血液浄化処理を実施することができる。そのため、本実施形態では、ウイルスが血流に侵入し全身へ回ってしまったウイルス血症等に対して効果的である。また、特定物質はウイルスに限定されず、特定物質と親和性を有する膜タンパク質を発現した親和性細胞を選択することで、例えば、Ｃ型肝炎、ＨＩＶ、ラッサ熱、エボラ熱、ジカ熱等の感染症にも適用可能である。
In this way, in this embodiment, in addition to obtaining the same actions and effects as in the first embodiment of the blood purification treatment, even viruses, secretions, and bacteria having a large diameter of 100 nm or more can be treated. Blood purification can be performed by passing through the shell portion 220 and combining with the affinity sperm cells of the core portion 210. Therefore, this embodiment is effective against viremia, etc., in which the virus enters the bloodstream and spreads throughout the body. In addition, the specific substance is not limited to viruses, and by selecting affinity cells that express membrane proteins that have affinity for the specific substance, for example, hepatitis C, HIV, Lassa fever, Ebola fever, Zika fever, etc. It is also applicable to infectious diseases.
また、ウイルスを含む血液を浄化するには、ビーズに抗体を担持させることも考えられるが、この構成では、ウイルスの除去能が徐々に低下する。これに対して、本実施形態では、ウイルスと親和性を有する親和性細胞が充填され長さが数十メートルにもおよぶ細胞ファイバＦを用いているため、除去能が低下することなくウイルスを捕捉することができる。
Additionally, in order to purify blood containing viruses, it is possible to have beads carry antibodies, but with this configuration, the ability to remove viruses gradually decreases. In contrast, in this embodiment, the cell fiber F, which is filled with affinity cells that have an affinity for viruses and has a length of several tens of meters, is used, so that the virus can be captured without reducing the removal ability. can do.
［生理活性物質の放出］
上記実施形態では、循環する血液に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞を、細胞ファイバＦが有する構成について説明した。本実施形態では、血液に外分泌物として生理活性物質を放出する外分泌性細胞を細胞ファイバＦが有する構成について説明する。 [Release of physiologically active substances]
In the embodiment described above, a configuration in which the cell fiber F has affinity cells expressing a membrane protein having an affinity for a specific substance contained in circulating blood has been described. In this embodiment, a configuration in which the cell fiber F has exocrine cells that release physiologically active substances as exocrine substances into the blood will be described.
上記実施形態では、循環する血液に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞を、細胞ファイバＦが有する構成について説明した。本実施形態では、血液に外分泌物として生理活性物質を放出する外分泌性細胞を細胞ファイバＦが有する構成について説明する。 [Release of physiologically active substances]
In the embodiment described above, a configuration in which the cell fiber F has affinity cells expressing a membrane protein having an affinity for a specific substance contained in circulating blood has been described. In this embodiment, a configuration in which the cell fiber F has exocrine cells that release physiologically active substances as exocrine substances into the blood will be described.
循環する血液に放出される生理活性物質としては、アルブミン、酵素等のタンパク質、インスリン等のホルモン、エクソソーム、MicroRNA、糖類、脂質等が挙げられる。
循環する血液に生理活性物質であるアルブミンを外分泌物として放出する外分泌性細胞は、肝細胞が挙げられる。
循環する血液に生理活性物質である酵素を外分泌物として放出する外分泌性細胞は、酵素産生細胞が挙げられる。
循環する血液に生理活性物質であるインスリン等のホルモンを外分泌物として放出する外分泌性細胞は、β細胞等のホルモン産生細胞が挙げられる。
循環する血液に生理活性物質であるエクソソームを外分泌物として放出する外分泌性細胞は、気道上皮細胞が挙げられる。 Physiologically active substances released into circulating blood include albumin, proteins such as enzymes, hormones such as insulin, exosomes, MicroRNA, sugars, lipids, and the like.
Exocrine cells that release albumin, a physiologically active substance, into circulating blood as an exocrine secretion include hepatocytes.
Exocrine cells that release enzymes, which are physiologically active substances, into circulating blood as exocrine secretions include enzyme-producing cells.
Exocrine cells that release hormones such as insulin, which are physiologically active substances, into circulating blood as exocrine secretions include hormone-producing cells such as β cells.
Exocrine cells that release exosomes, which are physiologically active substances, into the circulating blood include airway epithelial cells.
循環する血液に生理活性物質であるアルブミンを外分泌物として放出する外分泌性細胞は、肝細胞が挙げられる。
循環する血液に生理活性物質である酵素を外分泌物として放出する外分泌性細胞は、酵素産生細胞が挙げられる。
循環する血液に生理活性物質であるインスリン等のホルモンを外分泌物として放出する外分泌性細胞は、β細胞等のホルモン産生細胞が挙げられる。
循環する血液に生理活性物質であるエクソソームを外分泌物として放出する外分泌性細胞は、気道上皮細胞が挙げられる。 Physiologically active substances released into circulating blood include albumin, proteins such as enzymes, hormones such as insulin, exosomes, MicroRNA, sugars, lipids, and the like.
Exocrine cells that release albumin, a physiologically active substance, into circulating blood as an exocrine secretion include hepatocytes.
Exocrine cells that release enzymes, which are physiologically active substances, into circulating blood as exocrine secretions include enzyme-producing cells.
Exocrine cells that release hormones such as insulin, which are physiologically active substances, into circulating blood as exocrine secretions include hormone-producing cells such as β cells.
Exocrine cells that release exosomes, which are physiologically active substances, into the circulating blood include airway epithelial cells.
上記の外分泌性細胞を有する細胞ファイバＦを用いて血液を循環させることにより、循環する血液に生理活性物質を補充することができる。例えば、低アルブミン血症等を呈する血液にはアルブミンを補充することができ、ライソゾーム病等を呈する血液には酵素を補充することができる。すなわち、本実施形態の体外血液循環システム１００は、生理活性物質として機能することになる。本実施形態によれば、血液の循環路に加えて生理活性物質の補給流路を別途設ける必要がなくなり、装置の小型化および体外で循環させる血液量の抑制を実現できる。
By circulating blood using the cell fiber F having exocrine cells described above, physiologically active substances can be replenished into the circulating blood. For example, blood exhibiting hypoalbuminemia etc. can be supplemented with albumin, and blood exhibiting lysosomal diseases etc. can be supplemented with enzymes. That is, the extracorporeal blood circulation system 100 of this embodiment functions as a physiologically active substance. According to this embodiment, there is no need to separately provide a physiologically active substance supply channel in addition to a blood circulation channel, and it is possible to downsize the device and suppress the amount of blood to be circulated outside the body.
図１５は、ラットに対して、外分泌性細胞を有する細胞ファイバＦを用いて血液を循環させた場合と、外分泌性細胞を有さない細胞ファイバＦを用いて血液を循環させた場合の血漿内のアルブミン濃度を示す図である。図１５においては、４つのサンプルに対して、それぞれヒトアルブミンを外分泌物として放出する外分泌性細胞を有する細胞ファイバＦを用いて血液を循環させた。
Figure 15 shows the blood plasma in rats when blood was circulated using cell fiber F with exocrine cells and when blood was circulated using cell fiber F without exocrine cells. It is a figure showing the albumin concentration of. In FIG. 15, blood was circulated for four samples each using cell fiber F having exocrine cells that release human albumin as an exocrine secretion.
図１５に示されるように、４つのサンプル全てにおいてヒトアルブミンが検出された。本来ラットからはヒトアルブミンが検出されないことから、細胞ファイバＦにおける外分泌性細胞から放出されたヒトアルブミンが、循環する血液に細胞ファイバＦ経由で補充されたと確認できた。
As shown in Figure 15, human albumin was detected in all four samples. Since human albumin is normally not detected in rats, it was confirmed that human albumin released from exocrine cells in cell fiber F was replenished into circulating blood via cell fiber F.
以上、添付図面を参照しながら本発明に係る好適な実施形態について説明したが、本発明は係る例に限定されないことは言うまでもない。上述した例において示した各構成部材の諸形状や組み合わせ等は一例であって、本発明の主旨から逸脱しない範囲において設計要求等に基づき種々変更可能である。
Although preferred embodiments of the present invention have been described above with reference to the accompanying drawings, it goes without saying that the present invention is not limited to such examples. The various shapes and combinations of the constituent members shown in the above example are merely examples, and can be variously changed based on design requirements and the like without departing from the gist of the present invention.
例えば、上記実施形態では、細胞ファイバＦが、血液に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞と、血液に外分泌物として生理活性物質を放出する外分泌性細胞とのいずれか一方を含む構成を例示したが、この構成に限定されない。例えば、細胞ファイバＦが上述した親和性細胞と外分泌性細胞の双方を含む構成であってもよい。この場合、体外血液循環システム１００を用いることで、血液浄化処理と、生理活性物質補充処理の双方を実施できる。
For example, in the above embodiment, the cell fibers F can be used to connect affinity cells that express a membrane protein that has an affinity for a specific substance contained in blood, and exocrine cells that release physiologically active substances as exocrine substances into the blood. Although a configuration including one of the two is illustrated, the configuration is not limited to this. For example, the cell fiber F may include both the above-mentioned affinity cells and exocrine cells. In this case, by using the extracorporeal blood circulation system 100, both blood purification processing and physiologically active substance replenishment processing can be performed.
また、上記実施形態では、体外血液循環システム１００が輸液の循環路を有する構成を例示したが、輸液の循環路が設けられない構成であってもよい。
例えば、図１６に示すように、第２ポンプ２２が設けられた第１排出配管系１２が排出部３３Ｂに接続される構成であってもよい。
図１６に示す体外血液循環システム１００では、処理空間３５の血液は、第２ポンプ２２の駆動により、排出部３３Ｂから第１排出配管系１２を介して生体ＬＢに向けて送液される。 Furthermore, in the embodiment described above, the extracorporealblood circulation system 100 has a configuration in which the infusion circulation path is provided, but a configuration in which the infusion circulation path is not provided may also be used.
For example, as shown in FIG. 16, a configuration may be adopted in which the firstdischarge piping system 12 provided with the second pump 22 is connected to the discharge section 33B.
In the extracorporealblood circulation system 100 shown in FIG. 16, blood in the processing space 35 is sent from the discharge section 33B toward the living body LB via the first discharge piping system 12 by driving the second pump 22.
例えば、図１６に示すように、第２ポンプ２２が設けられた第１排出配管系１２が排出部３３Ｂに接続される構成であってもよい。
図１６に示す体外血液循環システム１００では、処理空間３５の血液は、第２ポンプ２２の駆動により、排出部３３Ｂから第１排出配管系１２を介して生体ＬＢに向けて送液される。 Furthermore, in the embodiment described above, the extracorporeal
For example, as shown in FIG. 16, a configuration may be adopted in which the first
In the extracorporeal
また、上記実施形態では、線状部材がコア部とシェル部とを有するコア／シェル構造を有する構成を例示したが、この構成に限定されない。例えば、図１７に変形例として示すように、線状部材Ｆは、細胞１１５を保持し少なくとも血液における一部の成分を透過させる保持部２２０とを有する構成であってもよい。この構成における保持部２２０は、例えば、上述したマクロポーラス構造を有するアルギン酸等で線状に形成される。
Further, in the above embodiment, a configuration in which the linear member has a core/shell structure including a core portion and a shell portion is illustrated, but the present invention is not limited to this configuration. For example, as shown in FIG. 17 as a modification, the linear member F may have a structure including a holding portion 220 that holds the cells 115 and allows at least some components of blood to pass therethrough. The holding portion 220 in this configuration is formed into a linear shape using, for example, alginic acid having the above-mentioned macroporous structure.
また、上記実施形態では、活性成分含有溶液が血液である構成を例示したが、この構成に限定されない。活性成分含有溶液としては、例えば、水・電解質、栄養等を含む輸液、抗体産生溶液等であってもよい。
Furthermore, in the above embodiment, a configuration in which the active ingredient-containing solution is blood is illustrated, but the configuration is not limited to this. The active ingredient-containing solution may be, for example, an infusion solution containing water, electrolytes, nutrients, etc., or an antibody production solution.
以下、本発明の諸態様を付記としてまとめて記載する。
（付記１）
少なくとも活性成分含有溶液における一部の成分が導入されるチャンバーと、
前記チャンバーの内部に配置され、複数の細胞と、前記細胞を保持し前記成分を透過させる保持部とを有する線状部材と、
を備え、
前記細胞は、
前記活性成分含有溶液における前記成分に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞と、
前記活性成分含有溶液に外分泌物として生理活性物質を放出する外分泌性細胞と、
の少なくとも一方を含む、活性成分含有溶液処理装置。
（付記２）
前記チャンバーは、少なくとも前記活性成分含有溶液における前記成分が導入される導入部と、少なくとも前記活性成分含有溶液における前記成分が排出される排出部とを有し、
前記導入部と前記排出部のうち、少なくとも前記排出部に前記線状部材を前記チャンバーの内部に止めるフィルターが設けられる、
付記１に記載の活性成分含有溶液処理装置。
（付記３）
前記保持部は、多糖類と、下限臨界溶液温度を有する高分子と親水性高分子とのブロック共重合体とを含む混合物である、
付記１または付記２に記載の活性成分含有溶液処理装置。
（付記４）
前記特定物質は、ウイルスであり、
前記親和性細胞は、アンジオテンシン変換酵素２（ＡＣＥ２）発現細胞である、
付記１から３のいずれか一項に記載の活性成分含有溶液処理装置。
（付記５）
前記生理活性物質はアルブミンであり、
前記外分泌性細胞は肝細胞である、
付記１から４のいずれか一項に記載の活性成分含有溶液処理装置。
（付記６）
前記生理活性物質は酵素であり、
前記外分泌性細胞は酵素産生細胞である、
付記１から５のいずれか一項に記載の活性成分含有溶液処理装置。
（付記７）
前記チャンバー内に配置され、外周に巻き付けられた前記線状部材を支持する支持体を有し、
前記支持体は、前記チャンバーに対して着脱自在に取り付けられる、
付記１から６のいずれか一項に記載の活性成分含有溶液処理装置。
（付記８）
前記線状部材は、
複数の前記細胞が充填されたコア部と、
前記保持部として、前記コア部の外周を被覆するシェル部と、
を有する細胞ファイバである、
付記１から７のいずれか一項に記載の活性成分含有溶液処理装置。
（付記９）
付記１から８のいずれか一項に記載の活性成分含有溶液処理装置と、
少なくとも前記活性成分含有溶液における前記成分を前記チャンバーの導入部から排出部に至って循環させるポンプと、を有する、体外活性成分含有溶液循環システム。 Hereinafter, various aspects of the present invention will be collectively described as supplementary notes.
(Additional note 1)
a chamber into which at least some of the components in the active ingredient-containing solution are introduced;
a linear member disposed inside the chamber and having a plurality of cells and a holding part that holds the cells and allows the components to pass through;
Equipped with
The cell is
Affinity cells expressing a membrane protein that has an affinity for a specific substance contained in the component in the active ingredient-containing solution;
exocrine cells that release physiologically active substances as exocrine secretions into the active ingredient-containing solution;
An active ingredient-containing solution processing device comprising at least one of the above.
(Additional note 2)
The chamber has at least an introduction part into which the component in the active ingredient-containing solution is introduced, and a discharge part from which at least the component in the active ingredient-containing solution is discharged,
Of the introduction part and the discharge part, at least the discharge part is provided with a filter for keeping the linear member inside the chamber.
The active ingredient-containing solution processing device according toSupplementary Note 1.
(Additional note 3)
The holding part is a mixture containing a polysaccharide and a block copolymer of a polymer having a lower critical solution temperature and a hydrophilic polymer.
The active ingredient-containing solution processing device according to Supplementary Note 1 or 2.
(Additional note 4)
The specific substance is a virus,
The affinity cells are angiotensin converting enzyme 2 (ACE2) expressing cells,
The active ingredient-containing solution processing device according to any one ofSupplementary Notes 1 to 3.
(Appendix 5)
the physiologically active substance is albumin,
the exocrine cell is a hepatocyte;
The active ingredient-containing solution processing device according to any one ofSupplementary Notes 1 to 4.
(Appendix 6)
the physiologically active substance is an enzyme;
the exocrine cell is an enzyme-producing cell;
The active ingredient-containing solution processing device according to any one ofSupplementary Notes 1 to 5.
(Appendix 7)
a support body disposed in the chamber and supporting the linear member wound around the outer periphery;
The support body is detachably attached to the chamber,
The active ingredient-containing solution processing device according to any one ofSupplementary Notes 1 to 6.
(Appendix 8)
The linear member is
a core portion filled with a plurality of the cells;
A shell part that covers the outer periphery of the core part as the holding part;
is a cell fiber having
The active ingredient-containing solution processing device according to any one ofSupplementary Notes 1 to 7.
(Appendix 9)
An active ingredient-containing solution processing device according to any one ofSupplementary Notes 1 to 8;
An extracorporeal active ingredient-containing solution circulation system, comprising: a pump that circulates at least the components of the active ingredient-containing solution from an inlet to an outlet of the chamber.
（付記１）
少なくとも活性成分含有溶液における一部の成分が導入されるチャンバーと、
前記チャンバーの内部に配置され、複数の細胞と、前記細胞を保持し前記成分を透過させる保持部とを有する線状部材と、
を備え、
前記細胞は、
前記活性成分含有溶液における前記成分に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞と、
前記活性成分含有溶液に外分泌物として生理活性物質を放出する外分泌性細胞と、
の少なくとも一方を含む、活性成分含有溶液処理装置。
（付記２）
前記チャンバーは、少なくとも前記活性成分含有溶液における前記成分が導入される導入部と、少なくとも前記活性成分含有溶液における前記成分が排出される排出部とを有し、
前記導入部と前記排出部のうち、少なくとも前記排出部に前記線状部材を前記チャンバーの内部に止めるフィルターが設けられる、
付記１に記載の活性成分含有溶液処理装置。
（付記３）
前記保持部は、多糖類と、下限臨界溶液温度を有する高分子と親水性高分子とのブロック共重合体とを含む混合物である、
付記１または付記２に記載の活性成分含有溶液処理装置。
（付記４）
前記特定物質は、ウイルスであり、
前記親和性細胞は、アンジオテンシン変換酵素２（ＡＣＥ２）発現細胞である、
付記１から３のいずれか一項に記載の活性成分含有溶液処理装置。
（付記５）
前記生理活性物質はアルブミンであり、
前記外分泌性細胞は肝細胞である、
付記１から４のいずれか一項に記載の活性成分含有溶液処理装置。
（付記６）
前記生理活性物質は酵素であり、
前記外分泌性細胞は酵素産生細胞である、
付記１から５のいずれか一項に記載の活性成分含有溶液処理装置。
（付記７）
前記チャンバー内に配置され、外周に巻き付けられた前記線状部材を支持する支持体を有し、
前記支持体は、前記チャンバーに対して着脱自在に取り付けられる、
付記１から６のいずれか一項に記載の活性成分含有溶液処理装置。
（付記８）
前記線状部材は、
複数の前記細胞が充填されたコア部と、
前記保持部として、前記コア部の外周を被覆するシェル部と、
を有する細胞ファイバである、
付記１から７のいずれか一項に記載の活性成分含有溶液処理装置。
（付記９）
付記１から８のいずれか一項に記載の活性成分含有溶液処理装置と、
少なくとも前記活性成分含有溶液における前記成分を前記チャンバーの導入部から排出部に至って循環させるポンプと、を有する、体外活性成分含有溶液循環システム。 Hereinafter, various aspects of the present invention will be collectively described as supplementary notes.
(Additional note 1)
a chamber into which at least some of the components in the active ingredient-containing solution are introduced;
a linear member disposed inside the chamber and having a plurality of cells and a holding part that holds the cells and allows the components to pass through;
Equipped with
The cell is
Affinity cells expressing a membrane protein that has an affinity for a specific substance contained in the component in the active ingredient-containing solution;
exocrine cells that release physiologically active substances as exocrine secretions into the active ingredient-containing solution;
An active ingredient-containing solution processing device comprising at least one of the above.
(Additional note 2)
The chamber has at least an introduction part into which the component in the active ingredient-containing solution is introduced, and a discharge part from which at least the component in the active ingredient-containing solution is discharged,
Of the introduction part and the discharge part, at least the discharge part is provided with a filter for keeping the linear member inside the chamber.
The active ingredient-containing solution processing device according to
(Additional note 3)
The holding part is a mixture containing a polysaccharide and a block copolymer of a polymer having a lower critical solution temperature and a hydrophilic polymer.
The active ingredient-containing solution processing device according to
(Additional note 4)
The specific substance is a virus,
The affinity cells are angiotensin converting enzyme 2 (ACE2) expressing cells,
The active ingredient-containing solution processing device according to any one of
(Appendix 5)
the physiologically active substance is albumin,
the exocrine cell is a hepatocyte;
The active ingredient-containing solution processing device according to any one of
(Appendix 6)
the physiologically active substance is an enzyme;
the exocrine cell is an enzyme-producing cell;
The active ingredient-containing solution processing device according to any one of
(Appendix 7)
a support body disposed in the chamber and supporting the linear member wound around the outer periphery;
The support body is detachably attached to the chamber,
The active ingredient-containing solution processing device according to any one of
(Appendix 8)
The linear member is
a core portion filled with a plurality of the cells;
A shell part that covers the outer periphery of the core part as the holding part;
is a cell fiber having
The active ingredient-containing solution processing device according to any one of
(Appendix 9)
An active ingredient-containing solution processing device according to any one of
An extracorporeal active ingredient-containing solution circulation system, comprising: a pump that circulates at least the components of the active ingredient-containing solution from an inlet to an outlet of the chamber.
本発明は、活性成分含有溶液処理装置および体外活性成分含有溶液循環システムに適用できる。
The present invention can be applied to an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system.
１…血液処理装置（活性成分含有溶液処理装置）、 ２１…第１ポンプ（ポンプ）、 ２２…第２ポンプ（ポンプ）、 ３０…チャンバー、 ５０…ファイバ支持体（支持体）、 ６０Ａ…境界（排出部）、 ６１、６２…フィルター、 １００…体外血液循環システム（体外活性成分含有溶液循環システム）、 １１５…細胞、 ２１０…コア部、 ２２０…シェル部（保持部）、 Ｆ…細胞ファイバ（線状部材）
1... Blood processing device (active ingredient-containing solution processing device), 21... First pump (pump), 22... Second pump (pump), 30... Chamber, 50... Fiber support (support), 60A... Boundary ( discharge part), 61, 62...filter, 100...extracorporeal blood circulation system (extracorporeal active ingredient-containing solution circulation system), 115...cell, 210...core part, 220...shell part (holding part), F...cell fiber (wire) shaped member)
Claims (9)
- 少なくとも活性成分含有溶液における一部の成分が導入されるチャンバーと、
前記チャンバーの内部に配置され、複数の細胞と、前記細胞を保持し前記成分を透過させる保持部とを有する線状部材と、
を備え、
前記細胞は、
前記活性成分含有溶液における前記成分に含まれる特定物質と親和性を有する膜タンパク質を発現した親和性細胞と、
前記活性成分含有溶液に外分泌物として生理活性物質を放出する外分泌性細胞と、
の少なくとも一方を含む、活性成分含有溶液処理装置。 a chamber into which at least some of the components in the active ingredient-containing solution are introduced;
a linear member disposed inside the chamber and having a plurality of cells and a holding part that holds the cells and allows the components to pass through;
Equipped with
The cell is
Affinity cells expressing a membrane protein that has an affinity for a specific substance contained in the component in the active ingredient-containing solution;
exocrine cells that release physiologically active substances as exocrine secretions into the active ingredient-containing solution;
An active ingredient-containing solution processing device comprising at least one of the above. - 前記チャンバーは、少なくとも前記活性成分含有溶液における前記成分が導入される導入部と、少なくとも前記活性成分含有溶液における前記成分が排出される排出部とを有し、
前記導入部と前記排出部のうち、少なくとも前記排出部に前記線状部材を前記チャンバーの内部に止めるフィルターが設けられる、
請求項１に記載の活性成分含有溶液処理装置。 The chamber has at least an introduction part into which the component in the active ingredient-containing solution is introduced, and a discharge part from which at least the component in the active ingredient-containing solution is discharged,
Of the introduction part and the discharge part, at least the discharge part is provided with a filter for keeping the linear member inside the chamber.
The active ingredient-containing solution processing device according to claim 1. - 前記保持部は、多糖類と、下限臨界溶液温度を有する高分子と親水性高分子とのブロック共重合体とを含む混合物である、
請求項１に記載の活性成分含有溶液処理装置。 The holding part is a mixture containing a polysaccharide and a block copolymer of a polymer having a lower critical solution temperature and a hydrophilic polymer.
The active ingredient-containing solution processing device according to claim 1. - 前記特定物質は、ウイルスであり、
前記親和性細胞は、アンジオテンシン変換酵素２（ＡＣＥ２）発現細胞である、
請求項１から３のいずれか一項に記載の活性成分含有溶液処理装置。 The specific substance is a virus,
The affinity cells are angiotensin converting enzyme 2 (ACE2) expressing cells,
The active ingredient-containing solution processing device according to any one of claims 1 to 3. - 前記生理活性物質はアルブミンであり、
前記外分泌性細胞は肝細胞である、
請求項１から３のいずれか一項に記載の活性成分含有溶液処理装置。 the physiologically active substance is albumin,
the exocrine cell is a hepatocyte;
The active ingredient-containing solution processing device according to any one of claims 1 to 3. - 前記生理活性物質は酵素であり、
前記外分泌性細胞は酵素産生細胞である、
請求項１から３のいずれか一項に記載の活性成分含有溶液処理装置。 the physiologically active substance is an enzyme;
the exocrine cell is an enzyme-producing cell;
The active ingredient-containing solution processing device according to any one of claims 1 to 3. - 前記チャンバー内に配置され、外周に巻き付けられた前記線状部材を支持する支持体を有し、
前記支持体は、前記チャンバーに対して着脱自在に取り付けられる、
請求項１から３のいずれか一項に記載の活性成分含有溶液処理装置。 a support body disposed in the chamber and supporting the linear member wound around the outer periphery;
The support body is detachably attached to the chamber,
The active ingredient-containing solution processing device according to any one of claims 1 to 3. - 前記線状部材は、
複数の前記細胞が充填されたコア部と、
前記保持部として、前記コア部の外周を被覆するシェル部と、
を有する細胞ファイバである、
請求項１から３のいずれか一項に記載の活性成分含有溶液処理装置。 The linear member is
a core portion filled with a plurality of the cells;
A shell part that covers the outer periphery of the core part as the holding part;
is a cell fiber having
The active ingredient-containing solution processing device according to any one of claims 1 to 3. - 請求項１から３のいずれか一項に記載の活性成分含有溶液処理装置と、
少なくとも前記活性成分含有溶液における前記成分を前記チャンバーの導入部から排出部に至って循環させるポンプと、を有する、体外活性成分含有溶液循環システム。 The active ingredient-containing solution processing device according to any one of claims 1 to 3,
An extracorporeal active ingredient-containing solution circulation system, comprising: a pump that circulates at least the components of the active ingredient-containing solution from an inlet to an outlet of the chamber.
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