Active ingredient-containing solution treatment device and extracorporeal active ingredient-containing solution circulation system

Abstract

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.SOLUTION: The present invention includes: a chamber into which at least a portion of components in blood is introduced; and a cellular fiber that is disposed inside the chamber and that has a core part filled with a plurality of cells and a shell part that covers the outer circumference of the core part. The cells include at least one of: affinitive cells in which a membrane protein having affinity with a specific substance contained in a component in blood is expressed; and external secretory cells that release a bioactive substance into blood as an external secretion.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to an apparatus for treating a solution containing an active ingredient and an extracorporeal system for circulating a solution containing an active ingredient.

Background Art

[0002] As an example of an extracorporeal system for circulating a solution containing an active ingredient, for example, a configuration having a hemodialysis device that performs dialysis to remove waste products and excess water from blood, which is a solution containing an active ingredient collected, and purifies the blood, is known (for example, Patent Document 1).

[0003] Further, Patent Document 2 discloses a configuration as an extracorporeal system for circulating a solution containing an active ingredient, which performs a process of imparting a nutrient substance to blood through a hollow fiber.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0005] The apparatus for treating a solution containing an active ingredient and the extracorporeal system for circulating a solution containing an active ingredient in Patent Document 1 use a blood component regulator in addition to a blood purification device, so there is a problem that the apparatus becomes large-sized. The apparatus for treating a solution containing an active ingredient and the extracorporeal system for circulating a solution containing an active ingredient in Patent Document 2 respectively provide a supply channel for nutrient substances and a supply channel for body fluid in addition to the blood circulation path, so there is a problem that the apparatus becomes large-sized as in Patent Document 1. In addition, when the apparatus becomes large-sized, there is also a problem that the amount of blood circulated extracorporeally increases.

[0006] The present invention has been made in consideration of the above points, and an object thereof is to provide an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system that can reduce the size of the device and suppress the amount of the active ingredient-containing solution circulated outside the body.

Means for Solving the Problems

[0007] According to a first aspect of the present invention, there is provided an active ingredient-containing solution treatment device including: a chamber into which at least some components in the active ingredient-containing solution are introduced; a linear member disposed inside the chamber, the linear member having a plurality of cells and a holding portion that holds the cells and allows the components to permeate therethrough, wherein the cells include at least one of an affinity cell that expresses a membrane protein having an affinity for a specific substance contained in the components in the active ingredient-containing solution and an exocrine cell that releases a physiologically active substance as an external secretion into the active ingredient-containing solution.

[0008] According to a second aspect of the present invention, there is provided an extracorporeal active ingredient-containing solution circulation system including the active ingredient-containing solution treatment device according to the first aspect and a pump that circulates at least the components in the active ingredient-containing solution from an introduction portion to a discharge portion of the chamber.

Effects of the Invention

[0009] In the present invention, it is possible to provide an active ingredient-containing solution treatment device and an extracorporeal active ingredient-containing solution circulation system that can reduce the size of the device and suppress the amount of the active ingredient-containing solution circulated outside the body.

Brief Description of the Drawings

[0010]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Figure 14

Figure 15

Figure 16

Figure 17

Embodiments for Carrying Out the Invention

[0011] Hereinafter, embodiments of the active ingredient-containing solution treatment device and the extracorporeal active ingredient-containing solution circulation system of the present invention will be described with reference to FIGS. 1 to 17. Note that the following embodiments show one aspect of the present invention, do not limit the present invention, and can be arbitrarily changed within the scope of the technical idea of the present invention. In the following drawings, in order to make each configuration easier to understand, the actual structure, scale, number, etc. in each structure are made different.

[0012] In the present embodiment, a blood treatment device that treats at least a part of the active ingredient-containing solution as blood is described as an active ingredient-containing solution treatment device, and an extracorporeal blood circulation system that circulates at least a part of the components in blood as an active ingredient-containing solution outside the body is described as an extracorporeal active ingredient-containing solution circulation system.

[0013] FIG. 1 is a diagram showing a schematic configuration of an extracorporeal blood circulation system 100. As shown in FIG. 1, the extracorporeal blood circulation system 100 circulates at least a part of the components in blood outside the living body LB, and performs a predetermined treatment on at least a part of the components in blood in the blood treatment device 1.

[0014] The extracorporeal blood circulation system 100 includes a blood treatment device 1, an infusion storage unit 10, a first introduction piping system 11, a first discharge piping system 12, a second introduction piping system 13, a second discharge piping system 14, a first pump 21, a second pump 22, a third pump 23, and a fourth pump 24.

[0015] The first introduction piping system 11 includes a pipe for introducing at least a part of the components in blood from the living body LB into the blood treatment device 1. The first discharge piping system 12 includes a pipe for discharging at least a part of the components in blood discharged from the blood treatment device 1 toward the living body LB. The first pump 21 and the second pump 22 are pumps for circulating at least a part of the components in 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 sends at least a part of the components in blood toward the blood treatment device 1. The second pump 22 is provided in the first discharge piping system 12 and sends at least a part of the components in blood toward the living body LB.

[0016] Note that at least some components in the blood are blood collected from the living body LB or specific components (e.g., plasma, etc.) separated from the blood among the blood collected from the living body LB. When circulating a specific component separated from the blood, the first introduction piping system 11 includes a separator that separates a specific component from the blood collected from the living body LB and a pipe leading from the separator to the blood treatment device 1. In the following description, a specific component separated from the blood may also be simply referred to as blood.

[0017] The infusion storage unit 10 stores the infusion. The second introduction piping system 13 includes a pipe that introduces the infusion discharged from the infusion storage unit 10 into the blood treatment device 1. The second discharge piping system 14 includes a pipe that discharges the infusion discharged from the blood treatment device 1 toward the infusion storage unit 10. The third pump 23 is provided in the second introduction piping system 13 and sends the infusion toward the blood treatment device 1. The fourth pump 24 is provided in the second discharge piping system 14 and sends the infusion from the blood treatment device 1 toward the infusion storage unit 10.

[0018] The blood treatment device 1 performs a predetermined treatment on the introduced blood. The predetermined treatment on the blood is at least one of a treatment that captures a specific substance contained in the introduced blood and purifies the blood and a treatment that releases a physiologically active substance to the introduced blood.

[0019] The blood treatment 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 is cylindrical and extends in the vertical direction. The upper side of the chamber body 31 is open. The chamber body 31 includes a bottom wall portion 32, a peripheral wall portion 33, a top wall portion 34, and a treatment space 35.

[0020] The bottom wall portion 32 is located at the lower end of the chamber body 31. The bottom wall portion 32 has a stepped portion 32A, a fitting projection 32B, a blood introduction portion 32C, and an introduction port 32D. The stepped portion 32A is cylindrical and protrudes upward from the lower end of the processing space 35. The fitting projection 32B is shaft-shaped and protrudes upward from the stepped portion 32A. The outer diameter of the fitting projection 32B is smaller than the outer diameter of the stepped portion 32A. The blood introduction portion 32C protrudes downward from the bottom wall portion 32. The first introduction piping system 11 is connected to the blood introduction portion 32C. The introduction port 32D penetrates the stepped portion 32A, the fitting projection 32B, and the blood introduction portion 32C in the vertical direction. Blood from the living body LB is introduced into the chamber 30 through the first introduction piping system 11 and the blood introduction portion 32C and through the introduction port 32D by driving the first pump 21.

[0021] The peripheral wall portion 33 is cylindrical and extends upward from the outer edge of the bottom wall portion 32. The peripheral wall portion 33 has an infusion introduction portion 33A and an infusion discharge portion 33B. The infusion introduction portion 33A and the infusion discharge portion 33B are arranged apart from each other in the vertical direction. The infusion introduction portion 33A is located near the lower end of the processing space 35 in the peripheral wall portion 33. The second introduction piping system 13 is connected to the infusion introduction portion 33A. Infusion from the infusion storage portion 10 is introduced into the processing space 35 through the second introduction piping system 13 and the infusion introduction portion 33A by driving the third pump 23. The infusion discharge portion 33B is located near the upper end of the processing space 35 above the infusion introduction portion 33A in the peripheral wall portion 33. The liquid in the processing space 35 is sent toward the infusion storage portion 10 through the infusion discharge portion 33B and the second discharge piping system 14 by driving the fourth pump 24.

[0022] The top wall portion 34 is annular and extends radially outward from the upper end of the peripheral wall portion 33. An annular groove portion 34a is formed around the opening in the processing space 35 on the upper surface of the top wall portion 34. A sealing material 34b is disposed in the groove portion 34a. The sealing material 34b is, for example, an O-ring. The processing space 35 is a space surrounded by the bottom wall portion 32 and the peripheral wall portion 33 and extends in the vertical direction.

[0023] The lid 40 is provided above the chamber body 31. The lid 40 is joined to the ceiling wall portion 34 from above via a sealing material 34b, thereby hermetically closing the processing space 35. By hermetically 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 lower side to the upper side and a through-flow path 40b penetrating in the vertical direction.

[0024] The fiber support 50 is disposed in the processing space 35. As shown in FIG. 2, the fiber support 50 has a pedestal portion 51, a support shaft 52, a fitting convex portion 53, a fitting recess 54, a through-flow path 55, and a groove portion 56.

[0025] 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 is cylindrical and extends 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.

[0026] The fitting convex portion 53 is cylindrical and extends 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 convex portion 53 is inserted and fitted into the fitting recess 40a of the lid 40 from the lower side. The fitting convex portion 53 is detachable from the fitting recess 40a. The fitting recess 54 is formed by being recessed upward from the lower surface of the pedestal portion 51. The fitting protrusion 32B of the bottom wall portion 32 is inserted and fitted into the fitting recess 54 from the lower side. The through-flow path 55 penetrates the fiber support 50 in the vertical direction. The lower end of the through-flow path 55 opens into the fitting recess 54. The fitting recess 54 is detachable from the fitting protrusion 32B.

[0027] When the fitting projection 53 fits into the fitting recess 40a of the lid 40, the fiber support 50 is positioned on the lid 40, and the through-flow path 55 communicates with the through-flow path 40b. When the fitting projection 32B fits into the fitting recess 54, the fiber support 50 is positioned on the chamber body 31, and the through-flow path 55 communicates with the inlet 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 inlet 32D, the through-flow path 55, and the through-flow path 40b communicate with each other.

[0028] The groove portion 56 extends radially outward from the through-flow path 55 on the support shaft 52 and opens to the outer peripheral surface of the support shaft 52. The groove portion 56 connects the through-flow path 55 and the treatment space 35 in the radial direction. The blood introduced into the chamber 30 can flow between the through-flow path 55 and the treatment space 35 through the groove portion 56. A plurality (four in this embodiment) of groove portions 56 are arranged at intervals in the circumferential direction. Two sets of the plurality of groove portions 56 arranged in the circumferential direction are provided at intervals in the vertical direction.

[0029] The blood treatment device 1 in which the fiber support 50 is disposed in the treatment space 35 of the chamber body 31 has a maximum diameter of about 2 cm and a maximum length of about 10 cm.

[0030] The filter 61 is disposed at the boundary 60A between the through-flow path 55 and the through-flow path 40b. The boundary 60A is a discharge portion where blood is discharged in the chamber 30. The filter 62 is disposed at the boundary 60B between the inlet 32D and the through-flow path 55. The boundary 60B is an introduction portion where blood is introduced into the chamber 30. The filters 61 and 62 hold 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 dispose at least the filter 61 at the boundary 60A which is the discharge portion. Considering the backflow of the liquid when performing air bleeding or the like as a preparation for blood treatment, it is also preferable to dispose the filter 62 at the boundary 60B which is the introduction portion.

[0031] As shown in FIG. 3, the linear member F has a plurality of cells 115 and a holding part 220 that holds the cells 115 and allows at least some components in blood to permeate therethrough. The linear member F of the present embodiment has a core / shell structure having a core part and a shell part. In the following description, the linear member is referred to as a cell fiber F having a core part 210 and a shell part 220.

[0032] The core part 210 has a hydrogel filled with a plurality of cells 115. The shell part 220 covers the outer periphery of the core part 210. The shell part 220 is a hydrogel having dissociability from the hydrogel forming the core part 210.

[0033] The method for producing the cell fiber F is not particularly limited. For example, it can be easily produced by using a double coaxial microfluidic device 140 as shown in FIG. 4. The microfluidic device 140 capable of injecting two fluids separately into the core part and the shell part so as to be coaxial is specifically described, for example, in Fig. 1 of Wonje Jeong, et al., Hydrodynamic microfabrication via "on the fly" photopolymerization of microscale fibers and tubes, Lab Chip, 2004, 4, 576-580.

[0034] FIG. 4 is a schematic diagram for explaining the manufacturing process of the cell fiber F. The microfluidic device 140 has an inlet 110 for introducing a first liquid for forming the core part 210, an inlet 120 for introducing a second liquid for forming the shell part 220, and an inlet 130 for introducing a third liquid for gelling the shell part 220.

[0035] As the first liquid for forming the core part 210, a solution containing cells 115 and extracellular matrix components can be exemplified. The cells contained in the first liquid are not particularly limited, and examples include endocrine cells, exocrine cells, nervous system cells, skeletal muscle cells, blood cells, stromal cells, etc. 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. In addition, examples of exocrine cells include gastrointestinal epithelial cells and other cells that release bioactive substances. Examples of cells that release bioactive substances include hepatocytes that release albumin and enzyme-producing cells that release enzymes. In addition, examples of nervous system cells include neural stem cells, central nervous system cells, peripheral nervous system cells, glial cells, etc. In addition, examples of skeletal muscle cells include osteocytes, chondrocytes, etc. In addition, examples of blood cells include hematopoietic stem cells, white blood cells, red blood cells, platelets, etc. In addition, examples of stromal cells include fibroblasts, vascular cells, etc. Also, for the purpose of blood purification, affinity cells expressing a membrane protein having an affinity for a specific substance contained in the blood can be used. Examples of the specific substance contained in the blood include viruses and cytokines. These cells may be cells induced to differentiate from iPS cells, ES cells, etc.

[0036] The extracellular matrix components are not particularly limited, and examples include collagen (type I, type II, type III, type V, type XI, etc.), basement membrane components reconstituted from mouse EHS tumor extract (including type IV collagen, laminin, heparan sulfate proteoglycan, etc.), gelatin, agar, agarose, fibrin, glycosaminoglycan, hyaluronic acid, proteoglycan, thrombin, aprotinin, alginic acid, etc.

[0037] As the second liquid for forming the shell part 220, 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, a solution that gels, a solution that gels in response to a magnetic field, etc. can be used.

[0038] Examples of 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 ions or potassium ions, acrylic acid-based synthetic solutions that gel in the presence of sodium ions, and the like.

[0039] Examples of temperature-responsive gelling solutions include temperature-responsive gelling solutions obtained by crosslinking poly(N-isopropylacrylamide) with polyethylene glycol, methylcellulose, hydroxypropylcellulose, copolymers of polylactic acid and polyethylene glycol, triblock copolymers of polyethylene glycol and polypropylene oxide (trade name: Pluronic (registered trademark), poloxamer), agarose, polyvinyl alcohol, and the like.

[0040] Examples of pH-responsive gelling solutions include alginate solutions, chitosan solutions, carboxymethylcellulose solutions, acrylic acid-based synthetic solutions, and the like.

[0041] Examples of photo-responsive gelling solutions include synthetic gelling solutions in which azobenzene and cyclodextrin are combined in the backbone, gelling solutions composed of supramolecules with fumaric acid amide as a spacer, gelling solutions crosslinked or bonded via a nitrobenzyl group, and the like.

[0042] Examples of magnetic field-responsive hydrogels include gelling solutions composed of crosslinked poly(N-isopropylacrylamide) containing magnetic particles, and the like.

[0043] 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, a collagen solution containing cells 115 is introduced and injected from the inlet 110 of the microfluidic device 140. Also, 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 part gels, and the cell fiber F with the shell part 220 being an alginate gel can be manufactured. Also, by heating the cell fiber 200 at about 37°C for several minutes to about 1 hour, the collagen solution containing the cells 115 in the core part 210 can be gelled.

[0044] The injection rates of the solutions at the inlets 110 and 120 are not particularly limited, but when 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 rates of the solutions at the inlets 110 and 120, the diameter of the core part and the coating thickness of the shell part can be appropriately adjusted. The injection rate of the solution at the inlet 130 is not particularly limited, and it may be about 1 to 10 mL / min, for example.

[0045] 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, for example, 50 μm to 1 mm. The length of the cell fiber F is not particularly limited and may be about several mm to several m. Examples of the cross-sectional shape of the cell fiber F include a circle, an ellipse, a polygon such as a quadrilateral or a pentagon.

[0046] By culturing the cell fiber F in a culture solution, the cells can be proliferated. The cell fiber F can also be cultured for several months by appropriately replacing the culture solution.

[0047] The core part 210 may contain 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 factor (IGF), fibroblast growth factor (FGF), nerve growth factor (NGF), etc. When containing growth factors, an appropriate concentration can be selected according to the type of growth factor.

[0048] The cell fiber F obtained as described above can be easily manipulated using forceps or the like. 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 female cylinder MS in which a culture solution has been previously stored, and a rotating disk RP embedded with a magnet (not shown) is installed below the female cylinder MS. While introducing the continuously produced cell fiber F into the female cylinder MS, the rotation of the rotating disk RP is transmitted to the fiber support 50 by magnetic force to rotate the fiber support 50, so that the cell fiber F can be continuously wound around the outer periphery of the support shaft 52. Thereby, it becomes possible to easily wind the cell fiber F having a length of several tens of meters around the outer periphery of the support shaft 52.

[0049] The fiber support 50 with the cell fiber F wound around the outer periphery of the support shaft 52 constitutes the 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. Then, when performing blood treatment in the blood treatment device 1, the fiber support 50 with the cell fiber F wound around it is mounted in the chamber 30 as shown in FIG. 1.

[0050] [First Embodiment of Blood Purification Treatment] To purify the blood collected from the living body LB using the extracorporeal blood circulation system 100, as the cells 115 filled in the cell fiber F, affinity cells expressing a membrane protein having an affinity for a specific substance that is a capture target contained in the blood can be used. Here, it is described that the specific substance that is a capture target can permeate through the shell portion 220 in the cell fiber F.

[0051] First, by driving the first pump 21, the blood from the living body LB flows into the treatment space 35 through the first introduction piping system 11, the inlet 32D, the through-flow path 55, and the groove portion 56. Also, by driving the third pump 23, the infusion from the infusion storage section 10 is introduced into the treatment space 35 through the second introduction piping system 13 and the infusion introduction section 33A. In the treatment space 35, since the infusion is introduced from below, the concentration of the blood introduced into the treatment space 35 decreases and a concentration gradient is generated. The specific substance that has permeated through the shell portion 220 in the cell fiber F in the treatment space 35 binds to the cells 115 via the membrane protein and is captured. The blood that has been purified by capturing the specific substance in the cell fiber F is sent toward the living body LB through the groove portion 56, the through-flow path 55, the through-flow path 40b, and the first discharge piping system 12 by driving the second pump 22.

[0052] Thus, in this embodiment, without using a blood component adjuster or the like, a plurality of cells 115 filled in the cell fiber F capture a specific substance that is a capture target, so that it is possible to suppress the amount of blood used with a small-sized device.

[0053] [Second Embodiment of Blood Purification Treatment] When the specific substance is a virus, its diameter is about 100 nm. When using, for example, an alginate gel as the second liquid for forming the shell part 220, it is difficult to greatly change the pore size of the gel, and the permeation of the above-mentioned virus, secretion, and bacteria may be hindered. In order to control the pore size of the gel in the shell part 220, the shell part 220 can be made of a mixture containing, for example, a polysaccharide, a block copolymer of a polymer having a lower critical solution temperature (LCST) and a hydrophilic polymer. By mixing the above polysaccharide and block copolymer, phase separation occurs in the mixture, and then by removing the block copolymer, a shell part 220 having a macroporous structure can be formed.

[0054] The above polysaccharide is at least one selected from the group consisting of alginic acid, starch, glycogen, cellulose, xanthan gum, hyaluronic acid, carrageenan, pectin, and pullulan, and salts thereof.

[0055] 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.

[0056] The above hydrophilic polymer is at least one selected from the group consisting of polyethylene glycol, polyethyleneimine, polyvinyl alcohol, polyacrylic acid, polyacrylamide, polyvinyl alcohol, polyvinylpyrrolidone, polyvinylacetamide, polyamine, poly(4-styrenesulfonic acid), poly(allylamine hydrochloride), poly(vinylsulfonic acid, sodium salt), poly(diallyldimethylammonium chloride), and poly(2-methacryloyloxyethyl phosphorylcholine).

[0057] The block copolymer of the polymer having the above LCST and the hydrophilic polymer can be synthesized by adding functional groups to the respective polymers and reacting the functional groups with each other. For example, since a maleimide group reacts with a thiol group (SH group) to form a stable thioether group, an SH group may be added to a hydrophilic polymer and a maleimide group may be added to a polymer having an LCST, and the two may be reacted with each other.

[0058] After forming a phase separation structure in the mixture of the polysaccharide and the block copolymer, by removing the block copolymer, a shell portion 220 having a macroporous structure formed of the polysaccharide with a pore size of 100 - 1000 nm can be obtained. When introducing a biological substance such as a cell 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.

[0059] To produce a cell fiber F having a shell portion 220 with a macroporous structure, a cell suspension is injected from an inlet 110 of a microfluidic device 140 and ejected toward an inlet 120. When a mixture of a polysaccharide and a block copolymer is injected from the inlet 120 and ejected toward the inlet 130, the core portion 210 is occupied by the cell suspension, and the above mixture covers the periphery of the cells to form a shell portion 220, forming a coaxial core - shell - shaped fluid. By injecting an aqueous solution containing calcium ions (for example, an aqueous solution containing CaCl2) from the inlet 130, the shell portion of the fluid is gelled. As a result, a cell fiber F in which the core portion 210 is occupied by cells 115 and the shell portion 220 is gelled is obtained. At this stage, since the shell portion 220 contains a block copolymer, by adjusting this to a temperature below the LCST, the block copolymer becomes water - soluble, and by stirring the cell fiber F in water, the block copolymer can be removed. Thereby, a cell fiber F in which the core portion 210 is filled with cells and the shell portion 220 has a macroporous (porous) shape is obtained. In the obtained cell fiber F, aiming for a core portion diameter of 100 - 200 μm and a cell filling rate of 2 - 10×10 7 cells / m, a core portion diameter of 160 μm and a cell filling rate of 2×10 7 cells / m were achieved.

[0060] By using the cell fiber F having the shell part 220 with a macroporous structure, the virus contained in the blood can permeate through the shell part 220. Therefore, as the cell 115, an affinity cell expressing a membrane protein having an affinity for the virus to be captured contained in the blood can be used. Thus, in the present embodiment, as described in the first embodiment, the virus contained in the blood from the living body LB permeates through the shell part 220 in the cell fiber F in the treatment space 35, binds to the cell 115 via the membrane protein, and is captured. The blood in which the virus is captured by the cell fiber F and purified is sent toward the living body LB.

[0061] FIG. 5 is a fluorescence image at 37° C. of the cell fiber F having the shell part 220 with a macroporous structure filled with fluorescent nanoparticles having a diameter of 100 nm in the core part. FIG. 6 is a fluorescence image when the cell fiber F in FIG. 5 reaches 4° C. after 1 hour has passed. As shown in FIG. 5, when the shell part is at 37° C., it was confirmed that the fluorescent nanoparticles having a diameter of 100 nm could not permeate through the shell part and remained in the core part. As shown in FIG. 6, when the shell part is at 4° C., the fluorescent nanoparticles having a diameter of 100 nm could not be confirmed in the core part, and it was confirmed that the fluorescent nanoparticles permeated through the shell part.

[0062] [Simulated virus] Among the viruses as specific substances, S protein (spike protein) beads capable of mimicking the cell infection mechanism of SARS-CoV-2 are produced. As shown in FIG. 7, Ni-NTA was added to the red fluorescent beads, and S protein with His tag added to the green fluorescent protein was used to produce S protein beads bound by the affinity of Ni-NTA and His tag. The binding between the red fluorescent beads and the S protein could be confirmed by measuring the fluorescence resonance energy transfer from the S protein to the red fluorescent beads.

[0063] [Affinity cell] As angiotensin-converting enzyme 2 (ACE2)-expressing cells capable of capturing the above S-protein beads, various ACE2-expressing cells (VeroE6 cells, VeroE6 / TMPRSS cells, Hela cells, Hela / ACE2 cells) were prepared by gene introduction using a lentiviral vector. In addition, reactivity evaluation with the above S-protein beads and quantitative comparison by a flow cytometer were performed. Among the above cells, VeroE6 cells with a capture efficiency of the S-protein beads of 50% or more and the highest were selected as ACE2-expressing cells.

[0064] [Demonstration for blood purification therapy] Rats were used as the living body 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 vein of the rat, and a solution containing 4 μg / mL of S-protein beads was injected into the vein. Every 5 minutes after injecting the solution containing S-protein beads, the amount of S-protein beads was measured in the first introduction piping system 11 for 45 minutes. The injection of the solution containing S-protein beads and the measurement of S-protein beads every 5 minutes were performed for a sample in which the cell fiber F was filled with VeroE6 cells and a sample in which the cell fiber F was not filled with cells, respectively.

[0065] FIG. 8 is a diagram showing the relationship between the time after injecting the solution containing S-protein beads and the measured amount of S-protein beads for a sample filled with VeroE6 cells and a sample not filled with cells in the cell fiber F, respectively. As shown in FIG. 8, it was confirmed that in the sample using VeroE6 cells, the S-protein beads decreased more significantly with the passage of time than in the sample not using cells. This is presumably the result of the membrane protein in VeroE6 cells binding (infecting) and being captured instantaneously upon contact with the S-protein beads, thereby reducing the S-protein beads contained in the blood in the living body LB and purifying the blood.

[0066] Figure 9 is a fluorescence image of a sample in which the cell fiber F is not filled with cells. Figure 10 is a fluorescence image of a sample in which the cell fiber F is filled with VeroE6 cells. As shown in Figure 9, in the sample in which the cell fiber F is not filled with cells, the capture of S protein beads was not confirmed. On the other hand, as shown in Figure 10, in the sample in which the cell fiber F is filled with VeroE6 cells, blood purification by the capture of S protein beads could be confirmed.

[0067] Figure 11 is a fluorescence image after the VeroE6 cells have reacted with the cell fiber F for 1 hour. Figure 12 is a fluorescence image after the VeroE6 cells have reacted with the cell fiber F for 2 hours. As shown in Figures 11 and 12, since the capture amount of S protein beads is larger when the reaction time is 2 hours than 1 hour, it was confirmed that the capture amount of S protein beads increases depending on the reaction time.

[0068] [Virus infection] As a specific substance in the blood, blood was circulated using a CoV S-protein pseudotyped lentivirus having the spike protein envelope of the coronavirus. Figure 13 is a fluorescence image 6 days after circulation in a sample in which the cell fiber F is not filled with cells using the pseudotyped lentivirus of the coronavirus. Figure 14 is a fluorescence image 6 days after circulation in a sample using VeroE6 cells in the cell fiber F using the pseudotyped lentivirus of the coronavirus. As shown in Figure 13, in the sample in which the cell fiber F is not filled with cells, no clear virus infection could be confirmed. As shown in Figure 14, in the sample using VeroE6 cells in the cell fiber F, the virus had migrated into the nucleus of the VeroE6 cells and fluorescence occurred, and virus infection could be confirmed.

[0069] Thus, in this embodiment, in addition to obtaining the same functions and effects as those of the first embodiment of the above blood purification treatment, even for viruses, secretions, and bacteria having a diameter of 100 nm or more, the shell portion 220 can be permeated to bind to the affinity cells of the core portion 210 to perform blood purification treatment. Therefore, in this embodiment, it is effective against viremia and the like in which the virus has invaded the bloodstream and spread throughout the body. Further, the specific substance is not limited to the virus, and by selecting affinity cells expressing a membrane protein having an affinity for the specific substance, it can be applied to infectious diseases such as hepatitis C, HIV, Lassa fever, Ebola fever, and Zika fever, for example.

[0070] In addition, in order to purify blood containing viruses, it is also conceivable to carry an antibody on beads. However, in this configuration, the virus removal ability gradually decreases. On the other hand, in this embodiment, since the cell fiber F filled with affinity cells having an affinity for the virus and having a length of several tens of meters is used, the virus can be captured without a decrease in the removal ability.

[0071] [Release of bioactive substances] In the above embodiment, the configuration in which the cell fiber F has affinity cells expressing a membrane protein having an affinity for a specific substance contained in the circulating blood has been described. In this embodiment, a configuration in which the cell fiber F has exocrine cells that release a bioactive substance as an external secretion into the blood will be described.

[0072] Examples of the bioactive substance released into the circulating blood include proteins such as albumin and enzymes, hormones such as insulin, exosomes, MicroRNA, saccharides, and lipids. Examples of the exocrine cells that release albumin, which is a bioactive substance, into the circulating blood as an external secretion include hepatocytes. Examples of the exocrine cells that release an enzyme, which is a bioactive substance, into the circulating blood as an external secretion include enzyme-producing cells. Exocrine cells that release hormones such as insulin, which are physiologically active substances, into the 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 as exocrine secretions include airway epithelial cells.

[0073] By circulating blood using the cell fiber F having the above exocrine cells, it is possible to supplement the circulating blood with physiologically active substances. For example, albumin can be supplemented to blood presenting hypoalbuminemia or the like, and an enzyme can be supplemented to blood presenting lysosomal disease or the like. That is, the extracorporeal blood circulation system 100 of the present embodiment will function as a physiologically active substance. According to the present embodiment, it is not necessary to separately provide a supply path for the physiologically active substance in addition to the blood circulation path, and miniaturization of the device and suppression of the amount of blood circulated outside the body can be achieved.

[0074] FIG. 15 is a diagram showing the albumin concentration in plasma when blood is circulated using the cell fiber F having exocrine cells for rats and when blood is circulated using the cell fiber F not having exocrine cells. In FIG. 15, for four samples, blood was circulated using the cell fiber F having exocrine cells that release human albumin as an exocrine secretion, respectively.

[0075] As shown in FIG. 15, human albumin was detected in all four samples. Since human albumin is not originally detected in rats, it was confirmed that the human albumin released from the exocrine cells in the cell fiber F was supplemented to the circulating blood via the cell fiber F.

[0076] As described above, the preferred embodiments according to the present invention have been described with reference to the accompanying drawings, but it goes without saying that the present invention is not limited to such examples. The various shapes, combinations, etc. of the respective constituent members shown in the above examples are merely examples, and various modifications can be made based on design requirements and the like without departing from the gist of the present invention.

[0077] For example, in the above embodiment, the cell fiber F was exemplified as having a configuration including either an affinity cell expressing a membrane protein having an affinity for a specific substance contained in blood or an exocrine cell that releases a physiologically active substance as an exocrine secretion into the blood. However, the configuration is not limited to this. For example, the cell fiber F may have a configuration including both the above-described affinity cell and exocrine cell. In this case, by using the extracorporeal blood circulation system 100, both blood purification treatment and physiologically active substance replenishment treatment can be performed.

[0078] Further, in the above embodiment, the extracorporeal blood circulation system 100 was exemplified as having a configuration with an infusion circulation path. However, a configuration without an infusion circulation path may also be used. For example, as shown in FIG. 16, a configuration may be adopted in which a first discharge piping system 12 provided with a second pump 22 is connected to a discharge portion 33B. In the extracorporeal blood circulation system 100 shown in FIG. 16, the blood in the treatment space 35 is fed from the discharge portion 33B through the first discharge piping system 12 toward the living body LB by driving the second pump 22.

[0079] Further, in the above embodiment, the configuration in which the linear member has a core / shell structure having a core portion and a shell portion was exemplified. However, the configuration is not limited to this. For example, as shown as a modification in FIG. 17, the linear member F may have a configuration including a holding portion 220 that holds the cells 115 and allows at least some components in the blood to permeate therethrough. The holding portion 220 in this configuration is formed linearly, for example, with alginic acid or the like having the above-described macroporous structure.

[0080] Further, in the above embodiment, the configuration in which the active ingredient-containing solution is blood was exemplified. However, the configuration is not limited to this. The active ingredient-containing solution may be, for example, an infusion containing water, electrolytes, nutrients, etc., an antibody production solution, or the like.

[0081] Hereinafter, various aspects of the present invention will be collectively described as appended notes. (Appended Note 1) A chamber into which at least some components in the active ingredient-containing solution are introduced, and A linear member disposed inside the chamber, having a plurality of cells and a holding portion that holds the cells and permits permeation of the component, comprising, wherein the cells are affinity cells that express a membrane protein having an affinity for a specific substance contained in the component in the active ingredient-containing solution, and exocrine cells that release a physiologically active substance as an exocrine secretion into the active ingredient-containing solution, an apparatus for treating an active ingredient-containing solution, comprising at least one of the above. (Appendix 2) The chamber has at least an introduction portion into which the component in the active ingredient-containing solution is introduced and a discharge portion from which the component in the active ingredient-containing solution is discharged, and among the introduction portion and the discharge portion, at least the discharge portion is provided with a filter for fixing the linear member inside the chamber. The apparatus for treating an active ingredient-containing solution according to Appendix 1. (Appendix 3) The holding portion is a mixture containing a polysaccharide, a block copolymer of a polymer having a lower critical solution temperature and a hydrophilic polymer. The apparatus for treating an active ingredient-containing solution according to Appendix 1 or Appendix 2. (Appendix 4) The specific substance is a virus, and the affinity cells are angiotensin-converting enzyme 2 (ACE2)-expressing cells. The apparatus for treating an active ingredient-containing solution according to any one of Appendices 1 to 3. (Appendix 5) The physiologically active substance is albumin, and the exocrine cells are hepatocytes. The apparatus for treating an active ingredient-containing solution according to any one of Appendices 1 to 4. (Appendix 6) The physiologically active substance is an enzyme, and the exocrine cells are enzyme-producing cells. The apparatus for treating an active ingredient-containing solution according to any one of Appendices 1 to 5. (Appendix 7) It has 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 treatment apparatus according to any one of Appendices 1 to 6. (Appendix 8) The linear member, A core part filled with a plurality of the cells, As the holding part, a shell part covering the outer periphery of the core part, It is a cell fiber having, The active ingredient-containing solution treatment apparatus according to any one of Appendices 1 to 7. (Appendix 9) The active ingredient-containing solution treatment apparatus according to any one of Appendices 1 to 8, And a pump for circulating at least the component in the active ingredient-containing solution from the introduction part to the discharge part of the chamber. An extracorporeal active ingredient-containing solution circulation system.

Explanation of symbols

[0082] 1... Blood treatment apparatus (active ingredient-containing solution treatment apparatus), 21... First pump (pump), 22... Second pump (pump), 30... Chamber, 50... Fiber support (support), 60A... Boundary (discharge part), 61, 62... Filters, 100... Extracorporeal blood circulation system (extracorporeal active ingredient-containing solution circulation system), 115... Cells, 210... Core part, 220... Shell part (holding part), F... Cell fiber (linear member)

Claims

1. A chamber into which some components in at least an active ingredient-containing solution are introduced, A linear member disposed inside the chamber, having a plurality of cells and a holding portion that holds the cells and allows the components to permeate therethrough, provided with, The cells are, Affinity cells that express a membrane protein having an affinity for a specific substance contained in the components in the active ingredient-containing solution, Exocrine cells that release a physiologically active substance as an exocrine secretion into the active ingredient-containing solution, An active ingredient-containing solution treatment device including at least one of the above.

2. The chamber has at least an introduction portion into which the components in at least the active ingredient-containing solution are introduced and a discharge portion from which the components in at least the active ingredient-containing solution are discharged, Among the introduction portion and the discharge portion, at least the discharge portion is provided with a filter for fixing the linear member inside the chamber, The active ingredient-containing solution treatment device according to claim 1.

3. The holding portion is a mixture including a polysaccharide and a block copolymer of a polymer having a lower critical solution temperature and a hydrophilic polymer, The active ingredient-containing solution treatment device according to claim 1.

4. The specific substance is a virus, The affinity cells are angiotensin-converting enzyme 2 (ACE2) expressing cells, The active ingredient-containing solution treatment device according to any one of claims 1 to 3.

5. The physiologically active substance is albumin, The exocrine cells are hepatocytes, The active ingredient-containing solution treatment device according to any one of claims 1 to 3.

6. The physiologically active substance is an enzyme, The exocrine cells are enzyme-producing cells. The active ingredient-containing solution treatment device according to any one of claims 1 to 3.

7. It has a support that is arranged in the chamber and supports the linear member wound around the outer periphery. The support is detachably attached to the chamber. The active ingredient-containing solution treatment device according to any one of claims 1 to 3.

8. The linear member has a core part filled with a plurality of the cells, and as the holding part, a shell part covering the outer periphery of the core part, and is a cell fiber having the above. The active ingredient-containing solution treatment device according to any one of claims 1 to 3.

9. The active ingredient-containing solution treatment device according to any one of claims 1 to 3, and a pump that circulates at least the component in the active ingredient-containing solution from the introduction part of the chamber to the discharge part, and has an extracorporeal active ingredient-containing solution circulation system.

Images