阅读说明：本技术 移植用室及移植用器件 (Transplantation chamber and transplantation device ) 是由 长谷川和弘 竹上龙太 神长邦行 望月勇辅 于 2018-06-28 设计创作，主要内容包括：根据本发明,提供一种移植用室及移植用器件,所述移植用室具有在内部与外部的边界的至少一部分处包含多孔膜的免疫隔离膜,上述多孔膜包含聚合物,并且在膜内具有孔径最小的层状的致密部位,孔径在厚度方向上从上述致密部位朝向上述多孔膜的一个表面A及另一个表面B这两者连续地增加,上述表面B附近的孔隙率为65％以上,上述表面B的平均孔径大于上述表面A的平均孔径,上述表面B配置于上述内部侧,移植用室具有上述表面B彼此对置并且上述多孔膜彼此接合的接合部,所述移植用器件在上述移植用室内包有生物学结构物。本发明的移植用室不易产生物质透过性的降低,并且免疫隔离膜彼此的接合部的强度高。(According to the present invention, there are provided a chamber for transplantation having an immunoisolation membrane comprising a porous membrane at least a part of the boundary between the inside and the outside, the porous membrane comprising a polymer and having a layered dense portion having the smallest pore diameter in the membrane, the pore diameter continuously increasing in the thickness direction from the dense portion toward both of one surface a and the other surface B of the porous membrane, the porosity in the vicinity of the surface B being 65% or more, the average pore diameter of the surface B being larger than the average pore diameter of the surface a, the surface B being disposed on the inside, the chamber for transplantation having a junction where the surfaces B face each other and the porous membranes are joined to each other, and a device for transplantation having a biological structure enclosed in the chamber for transplantation. The transplantation chamber of the present invention is less likely to cause a decrease in the permeability of a substance, and has a high strength at the junction between the immunoisolatory membranes.)

1. A chamber for transplantation, wherein,

having an immunoisolation membrane at least a portion of the boundary of the inner portion and the outer portion,

the immunoisolation membrane comprises a porous membrane,

the porous membrane comprises a polymer that is selected from the group consisting of,

the porous membrane has a layered dense site with the smallest pore size within the membrane,

the pore diameter continuously increases in the thickness direction from the dense sites toward both the one surface A and the other surface B of the porous film,

the porosity in the vicinity of the surface B is 65% or more,

the average pore size of the surface B is larger than the average pore size of the surface a,

the surface B is disposed on the interior side,

the transplantation chamber has a junction where the surfaces B face each other and the porous membranes are joined to each other.

2. The transplant chamber of claim 1 wherein,

the bonding is bonding using welding performed via a thermoplastic resin having a melting point lower than that of the polymer.

3. The transplant chamber of claim 2 wherein,

the thermoplastic resin is polyethylene.

4. The chamber for transplantation according to any one of claims 1 to 3,

the average pore diameter of the compact part is 0.02-1.5 μm.

5. The chamber for transplantation according to any one of claims 1 to 4,

the average pore diameter of the surface B is 3.0-20 μm.

6. The chamber for transplantation according to any one of claims 1 to 5,

the average pore diameter of the surface A is 1.0-2.5 μm.

7. The chamber for transplantation according to any one of claims 1 to 6,

the dense portion is located at a distance of 15% or more and less than 50% of the thickness of the porous film from the surface A of the porous film.

8. The chamber for transplantation according to any one of claims 1 to 7,

the thickness of the dense part is 0.5-30 μm.

9. The chamber for transplantation according to any one of claims 1 to 8,

the thickness of the porous membrane is 10 to 50 μm.

10. The chamber for transplantation according to any one of claims 1 to 9,

the porous membrane comprises polysulfone or polyethersulfone and at least one hydrophilic polymer.

11. A transplant device comprising the chamber for transplant according to any one of claims 1 to 10 and a biological structure encapsulated therein.

12. The device for transplantation according to claim 11,

the biological structure releases a physiologically active substance.

13. The device for transplantation according to claim 12,

the physiologically active substance is insulin.

Technical Field

The present invention relates to a chamber for transplantation comprising an immunoisolation membrane and a device for transplantation comprising the chamber for transplantation.

Background

Immunoisolation is one of the methods for preventing immune response in a recipient when a biological structure such as a cell, a tissue, or an organ is transplanted, and an immunoisolation membrane is a selectively permeable membrane that allows permeation of water, oxygen, glucose, and the like and blocks permeation of immune cells and the like associated with immune rejection. For example, the purpose of transplantation can be achieved while preventing immune rejection by using a transplantation device using an immunoisolation membrane that allows permeation of a physiologically active substance when cells that secrete the physiologically active substance are transplanted.

Non-patent document 1 describes the following: transplantation was performed using a commercially available transplantation chamber (TheraCyte (registered trademark)) formed using a porous membrane, which is a laminated membrane of a cell-retaining membrane having a pore diameter of 0.45 μm and an outer membrane of Polytetrafluoroethylene (PTFE) having a pore diameter of 5 μm.

Prior art documents

Non-patent document

Non-patent document 1: transfer, 67,665(1995)

Disclosure of Invention

Technical problem to be solved by the invention

In the transplantation chamber described in non-patent document 1, the surface of the laminated membrane having a small pore size and cell-retaining property is the inner space side. In this way, the structure of the minimum pore diameter required for immunoisolation on the surface of the immunoisolation membrane is likely to cause a decrease in substance permeability due to adsorption of proteins and the like. When the immunoisolatory membranes are joined to each other to form the chamber for transplantation, the surfaces having the smaller pore size are joined to each other, but the strength of the joined portion tends to be lower than in the case where the surfaces having the larger pore size are joined to each other.

The present invention addresses the problem of providing a chamber for transplantation and a device for transplantation, which are less likely to cause a decrease in the permeability of a substance and have high strength at the junction between immunoisolatory membranes.

Means for solving the technical problem

The present inventors have conducted extensive studies to solve the above problems, and have found that a minute structure of an immunoisolation membrane, which is less likely to cause a decrease in permeability to a substance, is used to produce a chamber for transplantation, thereby completing the present invention.

Namely, the present invention provides the following <1> to <13 >.

<1> a chamber for transplantation, wherein,

having an immunoisolation membrane at least a portion of the boundary of the inner portion and the outer portion,

the above-mentioned immunoisolation membrane comprises a porous membrane,

the above-mentioned porous film comprises a polymer,

the porous membrane has a layered dense portion having the smallest pore diameter in the membrane,

the pore diameter continuously increases from the dense portion toward both the one surface A and the other surface B of the porous film in the thickness direction,

the porosity in the vicinity of the surface B is 65% or more,

the average pore diameter of the surface B is larger than that of the surface a,

the surface B is disposed on the inner side,

the transplantation chamber has a junction portion where the surfaces B face each other and the porous membranes are joined to each other.

<2> the chamber for transplantation according to <1>, wherein,

the joining is joining by welding using a thermoplastic resin having a melting point lower than that of the polymer.

<3> the chamber for transplantation according to <2>, wherein,

the thermoplastic resin is polyethylene.

<4> the chamber for transplantation according to any one of <1> to <3>, wherein,

the dense part has an average pore diameter of 0.02 to 1.5 μm.

<5> the chamber for transplantation according to any one of <1> to <4>, wherein,

the surface B has an average pore diameter of 3.0 to 20 μm.

<6> the chamber for transplantation according to any one of <1> to <5>, wherein,

the average pore diameter of the surface A is 1.0 to 2.5 μm.

<7> the chamber for transplantation according to any one of <1> to <6>, wherein,

the dense portion is located at a distance of 15% or more and less than 50% of the thickness of the porous film from the surface A of the porous film.

<8> the chamber for transplantation according to any one of <1> to <7>, wherein,

the thickness of the dense part is 0.5 to 30 μm.

<9> the chamber for transplantation according to any one of <1> to <8>, wherein,

the porous film has a thickness of 10 to 50 μm.

<10> the chamber for transplantation according to any one of <1> to <9>, wherein,

the porous membrane comprises polysulfone or polyethersulfone and at least one hydrophilic polymer.

<11> a device for transplantation, wherein the chamber for transplantation of any one of <1> to <10> contains a biological structure.

<12> the device for transplantation according to <11>, wherein the biological structure releases a physiologically active substance.

<13> the transplantation device according to <12>, wherein the physiologically active substance is insulin.

Effects of the invention

According to the present invention, it is possible to provide a chamber for transplantation in which a decrease in the permeability of a substance is unlikely to occur and the strength of the joint between the immunoisolatory membranes is high. The transplantation device having the biological structure enclosed in the transplantation chamber of the present invention has high strength and can be used for a long period of time.

Drawings

Fig. 1 is a graph showing pore size distributions of porous membranes of examples 1 and 4 and comparative example 1.

Detailed Description

The present invention will be described in detail below.

In the present specification, "to" is used in a sense including numerical values described before and after "to" as a lower limit value and an upper limit value.

Laboratory for transplantation

The transplantation chamber is a container for transplanting the biological structure into a recipient. The transplantation chamber can enclose the biological structure therein.

The transplantation chamber of the present invention has an immunoisolation membrane at the boundary between the inside and the outside (the boundary separating the inside and the outside of the transplantation chamber). The immunoisolation membrane comprises a porous membrane comprising a polymer.

< immunoisolation Membrane >

In the present specification, an immunoisolation membrane refers to a membrane for immunoisolation.

Immunoisolation is one of the methods to prevent immunological rejection of the recipient at the time of transplantation. Here, the immunological rejection is a rejection of the transplanted biological structure by the recipient. The biological structures are sequestered from immunological rejection in the recipient by immunoisolation. The immune rejection includes an immune rejection based on a cellular immune response and an immune rejection based on a humoral immune response.

The immunoisolation membrane is a selectively permeable membrane that allows the permeation of nutrients such as oxygen, water, and glucose, and prevents the permeation of immune cells and the like associated with immune rejection. Examples of the immune cells include macrophages, dendritic cells, neutrophils, eosinophils, basophils, natural killer cells, various T cells, B cells, and other lymphocytes.

Depending on the application, the immunoisolation membrane preferably prevents the permeation of high molecular weight proteins such as immunoglobulins (e.g., IgM and IgG) and complements, and preferably allows physiologically active substances having a relatively low molecular weight such as insulin to permeate.

The permselectivity of the immunoisolation membrane may be adjusted depending on the application. The immunoisolation membrane may be a membrane that blocks selective permeability of substances having a molecular weight of, for example, 500kDa or more, 100kDa or more, 80kDa or more, or 50kDa or more. For example, the immunoisolatory membranes are preferably capable of preventing the penetration of the smallest IgG (molecular weight of about 160kDa) of the antibodies. The immunoisolation membrane may be a membrane that blocks selective permeability of a substance having a sphere size of 500nm or more, 100nm or more, 50nm or more, or 10nm or more.

The transplantation chamber of the present invention includes 1 or more immunoisolatory membranes at the boundary between the inside and the outside. The immunoisolation membrane may comprise only a porous membrane, or may comprise another layer such as a hydrogel membrane. Preferably, at least one surface of the immunoisolation membrane is a porous membrane, and also preferably, the immunoisolation membrane is composed of a porous membrane.

The thickness of the immunoisolation membrane is not particularly limited, but is preferably 10 μm or more. The particle size may be 500 μm or less, preferably 300 μm or less, more preferably 200 μm or less, still more preferably 100 μm or less, and particularly preferably 50 μm or less.

[ porous film ]

(Structure of porous Membrane)

The porous film means a film having a plurality of pores. The pores can be confirmed by, for example, a Scanning Electron Microscope (SEM) image or a Transmission Electron Microscope (TEM) image of the membrane cross section.

The thickness of the porous film is not particularly limited, but is preferably 10 μm or more. Further, it may be 250 μm or less, preferably 200 μm or less, more preferably 100 μm or less, and further preferably 50 μm or less. The thickness of the porous membrane can be set to 10 μm or more, whereby the strength of the membrane in the transplantation chamber can be set to a strength that does not break in the recipient. Further, the thickness of the porous membrane can be set to 250 μm or less, whereby a rigid chamber for transplantation can be provided without giving a feeling of discomfort to the recipient.

In the transplantation chamber of the present invention, the porous membrane has a layered dense site with the smallest pore size in the membrane. The pore diameter continuously increases from the dense portion toward both the one surface a and the other surface B of the porous film in the thickness direction. In the present specification, the surface of the porous membrane on the outside of the transplantation chamber is referred to as surface a, and the surface on the inside thereof is referred to as surface B. The pore diameter is determined by the average pore diameter of a dividing line of a membrane section described later.

The surface of the film means a main surface (main surface or back surface indicating the area of the film) and does not mean a surface in the thickness direction of the film edge. The surface of the porous film may be the surface with other layers. In addition, in the immunoisolation membrane, the porous membrane preferably has a structure that is uniform in the direction within the membrane (direction parallel to the membrane surface) with respect to the pore size, the pore size distribution (difference in pore size in the thickness direction), and the like.

Since the porous membrane has a pore size distribution in the thickness direction, the transplantation chamber of the present invention can have an increased lifetime. This is because an effect such as multistage filtration using a plurality of membranes having substantially different pore diameters can be obtained, and deterioration of the membranes can be prevented.

The pore diameter may be measured from a photograph of a cross section of the membrane obtained by an electron microscope. The porous membrane is cut with a microtome or the like, and a photograph of the cross section of the porous membrane can be obtained as a slice of a thin film that can be observed in the cross section.

In the present specification, the pore diameters in the thickness direction of the membrane were compared with pore diameters in 19 dividing lines obtained by dividing a SEM photograph of a cross section of the membrane into 20 pieces in the thickness direction of the membrane. More than 50 holes intersecting or connecting with the dividing line are successively selected, the respective hole diameters are measured, and the average value is calculated as the average hole diameter. Here, the pore diameter is calculated by calculating the area of the pore from an SEM photograph of a cross section of the membrane by image processing, without using the length of the selected portion where the pore and the dividing line intersect, and calculating the obtained area as the area of a perfect circle. At this time, the holes were large and 50 or more dividing lines could not be selected, and 50 were measured by expanding the field of view of the SEM photographs of the film cross sections. The comparison of the pore diameters in the thickness direction of the membrane was performed by comparing the obtained average pore diameters for each dividing line.

The layered dense site having the smallest pore diameter means a layered site of a porous film including a dividing line having the smallest average pore diameter among the dividing lines in the cross-sectional photograph of the film. The dense sites may include 2 or more parting lines. For example, when 2 or more continuous dividing lines having an average pore diameter within 1.1 times the minimum average pore diameter are continuous, the dense site includes the 2 or more continuous dividing lines. In this specification, the thickness of the dense site is a product of the number of dividing lines included in the dense site and 20 times the thickness of the film.

In the present specification, the average pore diameter of the dense sites is defined as the minimum pore diameter of the porous membrane. The minimum pore diameter of the porous membrane is preferably 0.02 to 1.5. mu.m, more preferably 0.02 to 1.3. mu.m. This is because the minimum pore size of the porous membrane can prevent at least the permeation of normal cells. Here, the average pore diameter of the dense site is determined by ASTM F316-80.

The porous membrane has dense sites within the membrane. The phrase "having dense sites in the film" means that the dense sites are not in contact with the surface of the film, and the dense sites are not sites including the division line closest to any surface of the film. By using a porous membrane having a structure in which dense sites are present in the membrane, the permeability of a substance that is intended to be allowed to permeate through the porous membrane is less likely to decrease than in the case of using a porous membrane having dense sites that are in contact with the surface. While not being bound by any theory, it is believed that this is because it becomes difficult to cause adsorption of proteins by having dense sites within the membrane.

The dense portion is preferably located on one surface side of the porous membrane than the central portion of the thickness of the porous membrane. Specifically, the dense site may be located at a distance of less than 50% of the thickness of the porous film from the surface a of the porous film, preferably at a distance of 40% or less, and more preferably at a distance of 35% or less. The dense portion may be located at a distance of 10% or more, preferably 13% or more, more preferably 15% or more, further preferably more than 15%, and particularly preferably 20% or more of the thickness of the porous film from the surface a of the porous film. The distance may be determined from the film sectional photograph. In the present specification, the surface of the porous membrane on the side closer to the dense site is referred to as "surface X". The surface X is preferably located on the inner side in the transplantation chamber, but may be the surface (surface a) on the outer side in the transplantation chamber.

In the porous membrane of the transplantation chamber of the present invention, the pore diameter continuously increases from the dense site toward both the surface a and the surface B in the thickness direction. The phrase "the pore diameter continuously increases in the thickness direction" means that the difference in the average pore diameter between the above-mentioned dividing lines adjacent to the thickness direction is increased so as to be 50% or less, preferably 40% or less, and more preferably 30% or less of the difference between the maximum average pore diameter (maximum pore diameter) and the minimum average pore diameter (minimum pore diameter). By "continuously increasing" is meant essentially no decrease and an increase uniformly, but sites of decrease may also be produced unexpectedly. For example, when 2 dividing lines of SEM photographs of the membrane cross section used for the comparison of the pore diameter in the thickness direction of the membrane are combined from the surface, the average value of the average pore diameter of the combined dividing lines uniformly increases (uniformly decreases from the surface toward the dense site), and it can be determined that "the pore diameter continuously increases from the dense site toward the surface of the membrane in the thickness direction".

The structure of the porous membrane in which the pore diameter continuously increases in the thickness direction can be realized by, for example, a production method described later.

In the present specification, in SEM photographs of a membrane cross section used for comparison of pore diameters in the thickness direction of the membrane, the average pore diameter at a dividing line closest to the surface a of the porous membrane is referred to as the average pore diameter of the surface a, and the average pore diameter at a dividing line closest to the surface B is referred to as the average pore diameter of the surface B. In the transplantation chamber of the present invention, the average pore size of the surface B is larger than that of the surface a. Since the surface B, i.e., the surface inside the transplantation chamber, has a larger pore diameter, adsorption of the included biological structure or the like is less likely to occur. Further, since the surfaces B having a large pore diameter are bonded to each other, the porous films can be strongly bonded to each other. This is because, when surfaces having a large pore diameter are joined, an adhesive agent serving as a joining member or a thermoplastic resin provided for welding easily enters the inside of the pore, and the volume fraction of the joining member at the joint portion can be increased, thereby increasing the joining strength.

The average pore diameter of the surface B may be 0.15 to 100. mu.m, preferably 1.0 to 50 μm, and more preferably 3.0 to 20 μm. The average pore diameter of the surface A may be 0.05 to 10 μm, preferably 0.3 to 5.0. mu.m, and more preferably 1.0 to 2.5. mu.m. The average pore diameter of the surface B is preferably 1.1 times or more, more preferably 1.5 times or more, and still more preferably 2 times or more the average pore diameter of the surface a. Further, the amount is preferably 8 times or less, more preferably 5 times or less, and still more preferably 3 times or less.

In the present specification, the average pore diameter of the dividing line having the largest average pore diameter among the dividing lines is defined as the maximum pore diameter of the porous membrane. The maximum pore diameter of the porous membrane is preferably 0.15 μm or more and 100 μm or less, more preferably 1.0 μm to 50 μm, and still more preferably 2.0 μm to 21 μm. The porous membrane of the transplantation chamber of the present invention preferably has a maximum pore size on the surface B.

The ratio of the average pore diameter of the dense portion to the maximum pore diameter of the porous membrane (the ratio of the minimum pore diameter to the maximum pore diameter of the porous membrane and the value obtained by dividing the maximum pore diameter by the minimum pore diameter, which may also be referred to as "anisotropy ratio" in the present specification) is preferably 3 or more, more preferably 4 or more, and still more preferably 5 or more. In order to increase the average pore diameter except for the dense sites and to improve the substance permeability of the porous membrane. The anisotropy ratio is preferably 25 or less, and more preferably 20 or less. This is because the effect of the multi-stage filtration described above can be effectively obtained in the range of the anisotropy ratio of 25 or less.

In the transplantation chamber of the present invention, the porosity in the vicinity of the surface B is 65% or more. With such porosity, the porous films are easily bonded to each other. In the present specification, the vicinity of the surface B indicates a portion from the surface B to a thickness of 5 times of the thickness of the porous film. The porosity can be determined from the density in the vicinity of the surface B, and specifically, is a value determined by the procedure described in examples. The porosity in the vicinity of the surface B is preferably 67% or more, more preferably 70% or more. The porosity in the vicinity of the surface B is usually 90% or less. By setting the porosity to 90% or less, it is possible to prevent the graft chamber from being damaged in the recipient, or the pores from being largely deformed to induce angiogenesis, or to cause a decrease in the selective permeability, while maintaining the required strength of the porous membrane.

(element distribution of porous Membrane)

The porous film preferably satisfies the formulae (I) and (II) on at least one surface.

B/A≤0.7 (I)

A≥0.015 (II)

In the formula, a represents the ratio of N element (nitrogen atom) to C element (carbon atom) in the surface of the film, and B represents the ratio of N element to C element in a depth of 30nm from the same surface.

The formula (II) indicates that a certain amount or more of N element is present on at least one surface of the porous film, and the formula (I) indicates that the N element in the porous film is unevenly present at a distance of less than 30nm from the surface.

Since the surface satisfies the formula (I) and the formula (II), the biocompatibility of the porous membrane, particularly the biocompatibility on the surface side satisfying the formula (I) and the formula (II), is improved.

The porous film may have only one surface satisfying the formulae (I) and (II) or both surfaces satisfying the formulae (I) and (II), but preferably both surfaces satisfy the formulae (I) and (II). When only one of the surfaces satisfies the formulae (I) and (II), the surface may be medial or lateral, preferably medial in the transplantation chamber described later. When only one of the surfaces satisfies the formulae (I) and (II) and the porous film has the surface X, the surface satisfying the formulae (I) and (II) is preferably the surface X.

In the present specification, the ratio of N element to C element (a value) on the film surface and the ratio of N element to C element (B value) in a depth of 30nm from the surface are calculated using XPS measurement results XPS measurement is X-ray photoelectron spectroscopy, and is a method of analyzing the composition of elements constituting the film surface by irradiating X-rays to the film surface and measuring the kinetic energy of photoelectrons emitted from the film surface, under the condition that the monochromatized Al-K α rays described in examples are used, the a value is calculated from the result at the start of sputtering, and the B value is calculated from the result calculated as the time 30nm from the surface of the film measured by the sputtering rate.

The ratio B/A is preferably 0.03 or more, more preferably 0.05 or more, as long as it is 0.02 or more.

A is preferably 0.050 or more, and more preferably 0.080 or more. Further, a may be 0.20 or less, preferably 0.15 or less, and more preferably 0.10 or less.

B is preferably 0.002 to 0.08, more preferably 0.003 to 0.07, as long as it is 0.001 to 0.10.

The element distribution of the porous film, particularly the distribution of the N element, can be controlled by the moisture concentration contained in the temperature-controlled wet air, the time for blowing the temperature-controlled wet air, the temperature of the coagulation liquid, the immersion time, the temperature of the glycol bath for washing, the time for immersing the glycol bath for washing, the speed of the porous film production line, and the like in the method for producing the porous film described later. The distribution of the N element can be controlled by the water content in the film-forming dope.

(composition of porous Membrane)

The porous membrane contains a polymer. The porous membrane is preferably composed essentially of a polymer.

The polymer forming the porous membrane is preferably biocompatible. Herein, "biocompatible" is meant to include non-toxicity, non-allergy inducing properties, and does not include the encapsulated nature of the polymer in vivo.

The number average molecular weight (Mn) of the polymer is preferably 1,000 to 10,000,000, more preferably 5,000 to 1,000,000.

Examples of polymers include thermoplastic or thermosetting polymers, preferably thermoplastic polymers. Specific examples of the polymer include cellulose acylate such as polysulfone and cellulose acetate, nitrocellulose, sulfonated polysulfone, polyethersulfone, polyvinylidene fluoride, polyacrylonitrile, a styrene-acrylonitrile copolymer, a styrene-butadiene copolymer, a saponified ethylene-vinyl acetate copolymer, polyvinyl alcohol, polycarbonate, an organosiloxane-polycarbonate copolymer, polyester carbonate, organopolysiloxane, polyphenylene oxide, polyamide, polyimide, polyamideimide, polybenzimidazole, an ethylene-vinyl alcohol copolymer, Polytetrafluoroethylene (PTFE), and the like. These may be homopolymers, copolymers, polymer blends, or polymer alloys from the viewpoints of solubility, optical properties, electrical properties, strength, elasticity, and the like.

Among these, polysulfone, polyethersulfone and cellulose acylate are preferable, and polysulfone is more preferable.

When polysulfone or polyethersulfone is used as the polymer, the porous membrane preferably further contains a hydrophilic polymer. Examples of the hydrophilic polymer include polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxyethyl cellulose, and the like, and among these, polyvinylpyrrolidone is preferable. By combining hydrophobic polysulfone or polyethersulfone with hydrophilic polymers, biocompatibility can be improved.

The porous film may contain other components than the above components as additives.

Examples of the additive include metal salts of inorganic acids such as sodium chloride, lithium chloride, sodium nitrate, potassium nitrate, sodium sulfate, and zinc chloride, metal salts of organic acids such as sodium acetate and sodium formate, other polymers such as polyethylene glycol, polyelectrolyte such as sodium polystyrene sulfonate and polyvinyl benzyl trimethyl ammonium chloride, and ionic surfactants such as sodium dioctyl sulfosuccinate and sodium alkylmethyltaurate. The additive may act as an expanding agent for the porous structure. As additives, metal salts are preferably used. The porous membrane containing polysulfone or polyethersulfone preferably contains lithium chloride.

The porous film is preferably a film formed of one composition as a single layer, and is preferably not a multilayer laminated structure. By forming the porous membrane as a single layer from one composition, the chamber for transplantation can be manufactured inexpensively in a simple process.

(method for producing porous film)

The method for producing the porous film is not particularly limited as long as the porous film having the above structure can be formed, and any general polymer film forming method can be used. The polymer film forming method includes a stretching method, a casting method and the like, and a casting method is preferable.

For example, in the casting method, the porous film having the above-described structure can be produced by adjusting the type and amount of the solvent used for the film formation dope and the drying method after casting.

The porous film produced by the casting method can be produced by a method including the following steps (1) to (4), for example.

(1) A film-forming dope containing a polymer, an additive as needed, and a solvent as needed is cast onto a support in a dissolved state.

(2) Temperature-controlled wet wind is blown to the surface of the cast liquid film.

(3) The film obtained after blowing the temperature-controlled wet air was immersed in the solidification liquid.

(4) The support was peeled off as needed.

The temperature of the temperature-controlled wet air may be 4 to 60 ℃, preferably 10 to 40 ℃. The relative humidity of the temperature-controlled humid air may be 15% to 100%, preferably 25% to 95%. The temperature-controlled wet air may be blown at a wind speed of 0.1 m/sec to 10 m/sec for 0.1 sec to 30 sec, preferably 1 sec to 10 sec. The temperature-controlled wet air may be blown at a wind speed of 0.1 m/sec to 10 m/sec for 0.1 sec to 30 sec, preferably 1 sec to 10 sec.

The average pore diameter and the position of the dense portion can be controlled by the concentration of moisture contained in the temperature-controlled moist air and the time for blowing the temperature-controlled moist air. The average pore diameter of the dense portion can also be controlled by the water content in the film forming dope.

By blowing the temperature-controlled wet air onto the surface of the liquid film as described above, evaporation of the solvent can be controlled, and coagulation can be caused from the surface of the liquid film into the film. In this state, by immersing the polymer in a coagulating liquid containing a solvent which is low in solubility of the polymer but compatible with the solvent of the polymer, the coagulated phase can be fixed to the fine pores, and fine pores other than the fine pores can be formed.

In the process of dipping in the solidification liquid, the temperature of the solidification liquid is only required to be-10 ℃ to 80 ℃. During this period, the temperature is changed to adjust the time from the formation of the condensed phase on the support surface side of the dense portion to the solidification, and to control the size of the pore diameter until the pore diameter reaches the support surface side. When the temperature of the coagulation liquid is increased, the formation of the condensed phase becomes fast and the time for coagulation becomes long, so that the pore diameter toward the support surface side tends to be increased. On the other hand, when the temperature of the solidification liquid is lowered, the formation of the condensed phase becomes slow and the time for solidification is shortened, so that the pore diameter toward the support surface side is difficult to be increased.

As the support, a plastic film or a glass plate may be used. Examples of the material of the plastic film include polyesters such as polyethylene terephthalate (PET), polycarbonates, acrylic resins, epoxy resins, polyurethanes, polyamides, polyolefins, cellulose derivatives, and silicones. The support is preferably a glass plate or PET, and more preferably PET.

The film-forming dope may contain a solvent. The solvent may be a solvent having high solubility of the polymer to be used (hereinafter, may be referred to as "good solvent"), depending on the polymer to be used. The good solvent is preferably replaced with the solidification solution quickly when the solvent is immersed in the solidification solution. Examples of the solvent include N-methyl-2-pyrrolidone, dioxane, tetrahydrofuran, dimethylformamide, dimethylacetamide and a mixed solvent thereof when the polymer is polysulfone, etc., dioxane, N-methyl-2-pyrrolidone, dimethylformamide, dimethylacetamide and dimethylsulfoxide or a mixed solvent thereof when the polymer is polyacrylonitrile, etc., dimethylformamide, dimethylacetamide and a mixed solvent thereof when the polymer is polyamide, etc., acetone, dioxane, tetrahydrofuran, N-methyl-2-pyrrolidone and a mixed solvent thereof when the polymer is cellulose acetate, etc. Among these, N-methyl-2-pyrrolidone is preferably used.

In addition to the good solvent, the film-forming dope is preferably a solvent (hereinafter, sometimes referred to as "non-solvent") having low solubility but compatibility with the polymer solvent. Examples of the non-solvent include water, cellosolves, methanol, ethanol, propanol, acetone, tetrahydrofuran, polyethylene glycol, and glycerol. Among these, water is preferably used.

The polymer concentration of the film-forming dope may be 5 mass% or more and 35 mass% or less, preferably 10 mass% or more and 30 mass% or less. By setting the content to 35% by mass or less, sufficient permeability (for example, permeability to water) can be provided to the obtained porous membrane, and by setting the content to 5% by mass or more, a porous membrane in which a permeable substance is selectively formed can be secured. The amount of the additive to be added is not particularly limited as long as the uniformity of the film-forming dope is not impaired by the addition, and is 0.5% by volume or more and 10% by volume or less with respect to a normal solvent. When the film-forming raw solution contains the non-solvent and the good solvent, the ratio of the non-solvent to the good solvent is not particularly limited as long as the mixed solution is kept in a uniform state, but is preferably 1.0 to 50% by mass, more preferably 2.0 to 30% by mass, and still more preferably 3.0 to 10% by mass.

In the membrane forming stock solution for producing a porous membrane containing a polymer selected from the group consisting of polysulfone and polyethersulfone and polyvinylpyrrolidone, polyvinylpyrrolidone is contained in an amount of preferably 50 to 120 mass%, more preferably 80 to 110 mass%, based on the total mass of polysulfone and polyethersulfone. When the film-forming stock solution contains lithium chloride as an additive, the lithium chloride is contained in an amount of preferably 5 to 20 mass%, more preferably 10 to 15 mass%, based on the total mass of the polysulfone and the polyether sulfone.

As the coagulating liquid, a solvent having low solubility of the polymer to be used is preferably used. Examples of such solvents include alcohols such as water, methanol, ethanol, and butanol; glycols such as ethylene glycol and diethylene glycol; aliphatic hydrocarbons such as ether, n-hexane, and n-heptane; glycerol such as glycerin, etc. Examples of the preferable coagulating liquid include water, alcohols, and a mixture of 2 or more of these. Among these, water is preferably used.

It is also preferable that the coagulation liquid is immersed in the solution and then washed with a solvent different from the used coagulation liquid. The washing can be performed by immersion in a solvent. Diethylene glycol is preferred as the cleaning solvent. Diethylene glycol is used as the cleaning solvent, and the distribution of the N element in the porous membrane can be adjusted by adjusting either or both of the temperature and the immersion time of diethylene glycol for immersing the membrane. In particular, when polyvinylpyrrolidone is used in the film-forming stock solution of the porous film, the remaining amount of polyvinylpyrrolidone in the film can be controlled. A further washing with water may be carried out after the washing with diethylene glycol.

As a method for producing a porous film, reference can be made to Japanese patent application laid-open No. 4-349927, Japanese patent application laid-open No. 4-068966, Japanese patent application laid-open No. 4-351645, Japanese patent application laid-open No. 2010-235808, and the like.

[ other layers ]

The immunoisolation membrane may include layers other than a porous membrane.

As the other layer, a hydrogel film can be cited. The hydrogel membrane is preferably biocompatible, and examples thereof include alginate gel membrane, agarose gel membrane, polyisopropylacrylamide membrane, cellulose-containing membrane, cellulose derivative (for example, methylcellulose) -containing membrane, and polyvinyl alcohol membrane. As the hydrogel film, an alginate gel film is preferable. Specific examples of the alginate gel film include polyion composite films of alginic acid-poly-L-lysine-alginic acid.

< construction of transplantation Chamber, etc. >

The immunoisolation membrane is disposed at least partially on the boundary between the inside and the outside of the transplantation chamber. With this arrangement, the biological structure contained in the transplantation chamber can be protected from immune cells and the like present outside, and nutrients such as water, oxygen, and glucose can be absorbed from outside to inside of the transplantation chamber.

The immunoisolation membrane may be disposed over the entire boundary between the inside and the outside of the transplantation chamber, or may be disposed over a part of the entire boundary corresponding to an area of, for example, 1 to 99%, 5 to 90%, 10 to 80%, 20 to 70%, 30 to 60%, 40 to 50%. The immunoisolation membrane is preferably disposed substantially over the entire boundary between the inside and the outside of the transplantation chamber. The face on which the immunoisolation film is disposed may be a continuous portion or may be divided into 2 or more portions.

When the immunoisolation membrane is not disposed over the entire boundary between the inside and the outside of the transplantation chamber, the remaining part may be formed of a material such as an impermeable membrane that does not allow penetration of nutrients such as oxygen, water, and glucose, in addition to cells.

The transplantation chamber has a junction portion where the surfaces B face each other and the porous membranes are joined to each other. In the junction, the immunoisolatory membranes are joined to each other. The part of the immunoisolation membrane to be joined is not particularly limited, and is preferably an end of the immunoisolation membrane. In particular, it is preferable that the ends are joined to each other. In the present specification, the term "end portion" as used in the film means an outer peripheral portion or a portion thereof having a constant width that actually comes into contact with a side surface (edge) made of the thickness of the film. The immunoisolatory membranes are preferably joined to each other over the entire periphery except for an injection port or the like described later. For example, the chamber for transplantation is also preferably configured as follows: the structure in which 2 immunoisolatory membranes are opposed and joined at the outer peripheries thereof or the line-symmetric structure of 1 immunoisolatory membrane is folded into 2 parts and joined at the opposed outer peripheries.

The joining can be performed by bonding with an adhesive, welding, or the like.

For example, the surfaces B may be bonded to each other with a curable adhesive while facing each other. Since the surfaces of the larger holes face each other, the adhesive easily enters the inside of the holes, and the volume ratio of the joining member at the joining portion becomes high, so that the joining strength is easily improved. Examples of the adhesive include known adhesives such as epoxy adhesives, silicon adhesives, acrylic adhesives, and urethane adhesives.

The surfaces B are opposed to each other, and a thermoplastic resin is sandwiched between the porous films, and the portions are heated, whereby the both can be bonded. In this case, as the thermoplastic resin, a resin having a melting point lower than the melting point of the polymer (the most contained polymer) contained in the porous film is preferably used. This is because, at the time of bonding, the resin having a lower melting point can be melted first and intruded into the pores of the porous film. Since the surfaces of the larger hole diameters face each other, a large amount of resin can be infiltrated into the holes, and the volume ratio of the joining members in the joining portion can be increased. Specific examples of the thermoplastic resin include polyethylene, polypropylene, polyurethane, polyvinyl chloride, polytetrafluoroethylene, polyethylene terephthalate, and polycarbonate. Among them, polyethylene, polypropylene, polyurethane, polyvinyl chloride, and polytetrafluoroethylene are preferable, and polyethylene, polyurethane, and polyvinyl chloride are more preferable.

The porous membranes in the immunoisolation membranes may be welded to each other with the surfaces B in direct contact with each other without interposing another material therebetween. By such welding, a transplantation chamber free from problems caused by interlayer resin and the like can be obtained. Further, by performing welding in a state where the surfaces B having large apertures are in direct contact with each other, the depth of the joint can be further increased. When a porous membrane containing a polymer selected from the group consisting of polysulfone and polyethersulfone is used, the porous membranes can be fused and integrated with each other by heating to a temperature that is less than the melting point of the polymer and is not lower than the glass transition temperature. Specifically, the heating for welding may be 190 ℃ or more and less than 340 ℃, and preferably 230 ℃ or more and less than 340 ℃.

The form of the chamber for transplantation is not limited, and may be any form such as a bag, balloon, cartridge, microcapsule, or tympanum. For example, a drum-shaped transplantation chamber can be formed by bonding an immunoisolation film to the top and bottom of a silicone ring. When used as a transplantation device described later, the shape of the transplantation chamber is preferably a shape capable of preventing the migration of the migration chamber in the recipient. Specific examples of the shape of the transplantation chamber include a cylindrical shape, a disk shape, a rectangular shape, an oval shape, a star shape, and a circular shape. The chamber for transplantation may be in the form of a sheet, a twisted wire, a spiral, or the like. The implantation chamber may enclose the biological structure and be formed into the shape described above for the first time when used as an implantation device described later.

The chamber for transplantation may include biocompatible plastic or the like for maintaining the shape or strength as a container. For example, the boundary between the inside and the outside of the transplantation chamber may be formed of a porous membrane or a biocompatible plastic. Alternatively, from the viewpoint of strength, the transplantation chamber in which the porous membrane is actually disposed on the entire boundary between the inside and the outside may further include a biocompatible plastic having a mesh structure outside the boundary between the inside and the outside.

< injection opening >

It is also preferable that the implantation chamber is provided with an injection port or the like for injecting the biological structure or the like into the implantation chamber. The injection port may be provided with a tube leading to the inside of the transplantation chamber.

The tube may contain a thermoplastic resin, for example. The thermoplastic resin is preferably a resin having a melting point lower than that of the polymer material of the porous film.

Specific examples of the thermoplastic resin used for the pipe include polyethylene, polypropylene, polyurethane, polyvinyl chloride, polytetrafluoroethylene, polyethylene terephthalate, and polycarbonate. Among them, polyethylene, polypropylene, polyurethane, polyvinyl chloride, and polytetrafluoroethylene are preferable, and polyethylene, polyurethane, and polyvinyl chloride are particularly preferable.

The tube is sandwiched between the immunoisolation membranes so as to be in contact with a part of the porous membrane, and then joined to the part. The joining can be performed by thermal welding, adhesion using an adhesive, or the like. Among them, thermal welding is preferably performed.

When the heat welding is performed, the tube preferably contains a thermoplastic resin having a melting point lower than that of the polymer material of the porous film. This is because when the tube containing the thermoplastic resin having a lower melting point is thermally welded to the porous film, the tube material can be first melted into the pores of the porous film when heated.

In the case of bonding, as the adhesive, an epoxy-based, silicon-based, acrylic, urethane-based adhesive or the like can be used. For example, when a tube containing a resin material having a melting point lower than that of the polymer material of the porous film is used, bonding by adhesion can be performed.

< use of Chamber for transplantation >

The chamber for implantation encloses the biological structure and is used to implant the biological structure into a recipient. By using the chamber for transplantation, immunological rejection of the recipient to the transplanted biological structure can be prevented. That is, immunoisolatory membranes can be used to protect biological structures from the recipient's immune system. In the present specification, the recipient refers to a living body to be transplanted. The subject is preferably a mammal, more preferably a human.

[ biological Structure ]

The biological structure refers to a structure derived from a living body. Examples of the living body include viruses, bacteria, yeasts, fungal cells, insects, plants, and mammals. The living body is preferably a mammal in general. Examples of the mammal include cattle, pigs, sheep, cats, dogs, and humans. The biological structure is preferably a structure derived from any one of mammals.

Examples of the biological structure include organs, tissues, and cells. Among these, cells are preferable as the biological structure. The number of cells may be 1 or more, but preferably more. The plurality of cells may be separated cells or aggregates.

The biological structure may be directly obtained from a living body. In particular, when the biological structure is a cell, the biological structure may be obtained directly from a living body, or may be differentiated and induced from cells such as embryonic stem cells (ES cells), induced pluripotent stem cells (iPS cells), and mesenchymal stem cells. The cell may be a precursor cell.

As the biological structure, it is preferable to release a physiologically active substance as one mode. Examples of the physiologically active substance include various hormones, various cytokines, various enzymes, and other various factors in vivo. More specific examples include insulin, dopamine, factor VIII, and the like.

Here, insulin is a polypeptide in which an a chain of 21 amino acid residues and a B chain of 30 amino acid residues are linked via a disulfide bond (molecular weight is about 6000). in the living body of mammals, insulin is secreted from β cells on langerhans islets of pancreas. in the present invention, when insulin-secreting cells are used as a biological structure, insulin to be secreted may be human-type insulin or insulin of other mammalian types (for example, porcine-type insulin. insulin may be insulin produced by a gene recombination method. as a method for obtaining genetically recombinant insulin, for example, a variety of insulin analogs can be used as described in menguixianzhu (see 270-p, tianxiaojian, okuan and "present and future insulin preparations", Medieal reviewco., ltd., 2002): various insulin analogs (see, for example, h.c. lee, j.w.yjn, etal., 271, 488-483, 271, 488-2000).

The biological structure is preferably an insulin-secreting cell, which is a cell capable of secreting insulin in response to a change in blood glucose level, but the insulin-secreting cell is not particularly limited, and examples thereof include pancreatic β cells present in Langerhans' islets of langerhans of pancreas, pancreatic β cells, which may be human pancreatic β cells, and pancreatic β cells such as pigs and mice, and the method of extracting pancreatic β cells from pigs can be referred to the disclosure of Japanese patent laid-open publication No. 2007-195573, and the insulin-secreting cells may be cells derived from human stem cells (for example, Kawasaki, regenerative medicine, Vol.1, Vol.2, pp.57-61, and 2002) or cells derived from small intestinal epithelial cells (for example, Katsukui et al, regenerative medicine, Vol.1, Vol.2, pp.63-68, and 2002), or insulin-secreting cells incorporating a gene encoding insulin (for example, H.C.Lee., YJ.488, Vol.2002, Vol.483, Vol.2, Vol.s.408, and Langhua).

Device for transplantation

The transplantation device is a composite body including at least a transplantation chamber and a biological structure. In the transplantation device, the biological structure is enclosed in the transplantation chamber.

In the transplantation device, the transplantation chamber may contain only the biological structure or may contain the biological structure and other structures or components other than the biological structure. For example, the biological structure may be contained in the transplantation chamber together with the hydrogel, preferably in a state of being contained in the hydrogel. Alternatively, the device for transplantation may contain a protein or peptide such as a pH buffer, an inorganic salt, an organic solvent, or albumin.

In the implant device, the biological structure may include only one type, or may include 2 or more types. For example, the bioactive substance may contain only a biological structure that releases a bioactive substance for transplantation or exerts a function for other transplantation purposes, or may further contain a biological structure that assists the function of the biological structure.

The implant device may be any device that is implanted into, for example, the abdominal cavity or the subcutaneous space. Alternatively, the grafting device may be a vascular access device. For example, when insulin-secreting cells are used as biological structures, insulin can be secreted in accordance with changes in blood glucose level by transplanting blood and membranes in direct contact.

As for the transplantation device and the transplantation chamber, protein nucleases, those described in Vol.45, pp.2307-2312, (Daihuajiuzi, 2000), Japanese patent application laid-open (JP-A) 2009-522269, Japanese patent application laid-open (JP-A) No. 6-507412, and the like can be referred to.