JP2007202430A - Hollow yarn bundle module for cell culture - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a hollow yarn bundle module for cell culture capable of efficiently retrieving cells. <P>SOLUTION: The hollow yarn bundle module 1 for cell culture comprises a hollow yarn bundle 5 with hollow yarns made from a semipermeable membrane material and a housing 2 housed with the hollow yarn bundle 5 and equipped with a culture fluid inlet 2d and a culture fluid outlet 2e. In this module 1, cutting elements 6a and 6b for cutting at least one end of the hollow yarn bundle 5 housed in the housing 2 are formed in the housing 2, heat-shrinkable films 7 are provided to releasably seal up the cutting elements 6a and 6b, and one of the ends of the housing 2 is provided with a housing-dividing element 2f for extracting the hollow yarn bundle 5 from the housing 2. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a hollow fiber bundle module for cell culture suitable for culturing cells in the medical field and pharmaceutical manufacturing field.

  In recent years, with the advancement of cell function analysis, discovery of various stem cells, and differentiation induction techniques thereof, the possibility of treatment for various diseases that have been conventionally difficult to treat has come to be expected.

  In particular, there is an increasing expectation for regenerative medicine aiming at repairing a lost tissue and improving its function, and various efforts are being made.

  In many cases, regenerative medicine is based on a cell culture technique in which cells having a target function in a home or other home are grown in a culture apparatus and transplanted to a patient. However, there are some basic issues that have not been solved in the cell culture technology that should be the basis of the technology, which is preventing the spread of regenerative medicine.

  One of the challenges is that the technology of culture vessels and culture systems that can efficiently recover the proliferated cells while ensuring strict discriminability of cells for each patient has not been established at present. It is done.

In the prior art, the seed cells are isolated and collected from the patient's own blood or tissue, but a large amount of cells are required to grow to the number of cells required for transplantation (10 ⁷ to 10 ⁹ levels). Since blood and tissue are required, the burden on the patient is inevitable.

  As a means for culturing such a large number of cells, a culture apparatus using a module filled with a semipermeable hollow fiber membrane is widely used. This type of culture apparatus is excellent in individual identification and is suitable for performing a plurality of cultures simultaneously.

For example, in Patent Document 1, a semipermeable tubular membrane (capillary: equivalent to the “hollow fiber” of the present application) is enclosed in a glass shell to form a cell culture unit, and a nutrient medium is circulated through the capillary. A technique for growing cells attached to the capillary surface is disclosed.
JP-A-49-41579

  However, the conventional module culture is intended to collect the secretory components of the cells in the culture solution as in the case of monoclonal antibody production, and is not configured to collect the cells grown in the module. In other words, it cannot be said that consideration is given to the life and death of the cells existing inside and the maintenance of physiological activity.

  That is, in the conventional module culture, from the viewpoint of avoiding bacterial contamination and liquid leakage, it is recommended to use a module with strength and sealability that does not cause damage during the culture operation. When recovering adherent cells, it is common to detach cells by trypsin treatment, but the high sealing performance in the module structure is an obstacle, making it difficult to perform cell recovery processing, and the cell recovery rate is low, In addition, there is a problem that cells are damaged by excessive treatment, which is not practical.

  This invention is made | formed in view of the said situation, and it aims at providing the hollow fiber bundle module for cell cultures which can collect | recover efficiently, without damaging the cultured cell.

  The present invention relates to a cell having a hollow fiber bundle in which hollow fibers made of a semipermeable membrane material are bundled, and a housing in which the hollow fiber bundle and a culture solution are stored and a culture solution introduction unit and a culture solution discharge unit are provided. In the culture hollow fiber bundle module, a cutting opening for cutting at least one end of the hollow fiber bundle housed in the housing is formed in the housing, and the cutting opening is releasably sealed. A cell culture hollow provided with a sealing body for stopping, and further provided with a housing dividing part for removing a hollow fiber bundle cut at one end part to obtain a free end from the housing at the end part of the housing This is a yarn bundle module.

  The hollow fiber bundle module for cell culture in the present invention circulates a culture solution in a hollow fiber and supplies a culture solution component that has permeated through the hollow fiber membrane to cells existing outside the hollow fiber. The yarn is, for example, a single material selected from regenerated cellulose, cellulose ester, polysulfone, polyethersulfone, polyacrylonitrile, polyvinyl alcohol, polystyrene, polymethyl methacrylate or polycarbonate, or a mixture of a plurality of materials. It can consist of

  The hollow fiber bundle in the present invention means one or two or more hollow fibers arranged in the longitudinal direction in the housing. In addition, the semipermeable membrane has a selective permeation performance that allows some components of the solution to pass therethrough but does not allow other components to pass therethrough. For example, a porous membrane that can adjust the components to be passed depending on the pore diameter, etc. Is shown. Moreover, it is preferable to use the hollow fiber whose surface has been subjected to a hydrophilic treatment. This is because the cell affinity increases and the membrane permeability of the culture solution component also improves.

  In the present invention, the cutting opening may have a slit cut out in a direction orthogonal to the cylinder axis direction of the housing.

  In the present invention, a heat shrink film can be wound around the opening for cutting as the sealing body.

  In the present invention, a screw portion can be formed on the outer peripheral wall of the housing in the vicinity of the slit, and a union nut that can be screwed into the screw portion as a sealing body can be provided.

  In the present invention, a flange or an annular groove is formed in the outer peripheral wall of the housing on both sides of the slit, and a clamp that is fastened and fixed to the flange or the annular groove as a sealing body can be provided.

  In this invention, the convex part which can be fitted in the opening part for a cutting | disconnection can be formed in the inner surface of the said clamp.

  In the present invention, the sealing body can have a resin or metal cover attached to the slit.

  In the present invention, it is preferable to provide a sealing material around the slit edge.

  In the present invention, as the opening for cutting, a cylindrical portion is extended from one end of the housing in a direction orthogonal to the tube axis direction, and the lid that closes the opening at the tip of the cylindrical portion or the inside of the cylindrical portion is provided. A plug to be filled can be provided.

  In the present invention, the housing split part can be provided with a pipe joint composed of a male screw part and a female screw part.

  In the present invention, the hollow fiber bundle can supplementarily have an arbitrary number of cell-adhesive growth fiber bundles along the hollow fiber.

  According to the present invention, the hollow fiber bundle to which the cells cultured in the hollow fiber bundle module are attached can be easily taken out without damage or loss, so that the cell recovery efficiency can be improved. Have

  FIG. 1 shows the structure of a hollow fiber bundle module for cell culture according to the present invention (hereinafter abbreviated as a hollow fiber bundle module). FIG. 1 (a) is a front view, and FIG. The CC side view side view is shown. In FIG. 1 (a), the upper side from the central axis A of the housing is shown in cross section to show the internal configuration.

a Housing The hollow fiber bundle module 1 has a resin housing 2, and this housing 2 is mainly composed of a cylindrical portion 2a and caps 2b and 2c for sealing both ends of the cylindrical portion 2a.

  Each cap 2b, 2c is screwed into a threaded portion formed at the end of the cylindrical portion 2a, and a culture medium conduit (culture medium introduction) is introduced into one cap 2b. Part) 2d is provided on the central axis A, and the other cap 2c has a culture medium discharge pipe (culture liquid discharge part) 2e for discharging the culture medium introduced into the housing 2 on the central axis A. Is provided.

  Note that the culture fluid conduit 2d and the culture fluid discharge tube 2e are distinguished for convenience in order to explain the configuration of the hollow fiber bundle module 1, and the hollow fiber bundle module 1 has no directionality. When the culture solution is introduced from the above, 2e becomes the culture solution conduit and 2d becomes the culture solution discharge tube.

  Examples of the material of the housing 2 include polycarbonate, polysulfone, polypropylene, polystyrene, polyethylene, and acrylic resin. Of these, polycarbonate and polysulfone are preferably selected. This is because these materials have high heat resistance, so that high-pressure steam sterilization can be performed and efficient sterilization can be performed. In addition, since the transparency is high, there is an advantage that the cultured state of the cells can be easily confirmed with the naked eye.

  The downstream end (housing end) 2f (in the flow direction of the culture medium indicated by B in the figure) of the cylindrical portion 2a is a separate part connected to the cylindrical portion 2a via a screw portion, It has a cylindrical shape having the same inner diameter as the cylindrical portion 2a.

  The cap 2e is screwed through a screw portion 2g formed on the outer wall of the body portion of the downstream end portion 2f. Therefore, the end portion of the cylindrical portion 2a and the downstream end portion 2f connected via the screw portion 2g constitute a pipe joint and function as a housing dividing portion.

  In the figure, reference numeral 3 denotes an O-ring that seals a gap between the cap 2b and the cylinder portion 2a, a gap between the cylinder portion 2a and the downstream end 2f, and a gap between the downstream end 2f and the cap 2c.

  Further, cell insertion tubes 4a and 4b are respectively provided on the downstream side and the upstream side of the tube portion 2a, and each cell insertion tube 4a and 4b is in the direction perpendicular to the central axis A and in the opposite direction. Projecting from 2a. The arrangement of the cell insertion tubes 4a and 4b is not limited to the opposite direction, and may be formed apart in the same direction.

  Note that two cell insertion tubes 4a and 4b are provided for the purpose of effectively inserting cells without creating a dead space, and one of them may be used as a gas vent. .

  Further, slits 6a and 6b for cutting a hollow fiber bundle 5 described later are formed as cutting openings at one end portion (upstream side in the present embodiment) of the cylindrical portion 2a, and the slits 6a and 6b. Through this, a part of the hollow fiber bundle 5 housed in the housing 2 is exposed.

  FIG. 1B shows a cross section viewed from the direction of arrows CC in FIG. 1A in order to explain the shapes of the slits 6a and 6b.

  As shown in FIG. 1B, the upper slit 6a is formed in a direction orthogonal to the central axis A and by cutting out the body portion of the cylindrical portion 2a in an arc shape. The lower slit 6b is formed so as to be point-symmetric with respect to the slit 6a (centering on the central axis A). In addition, the slit 6a and the slit 6b are partitioned by connecting portions 2h and 2i.

  A heat shrink film 7 as a sealing body is attached to the outside of the slits 6a and 6b in a ring shape and is shrunk when heated to close the slits 6a and 6b. This prevents the culture fluid from leaking out of the slits 6a and 6b when the culture fluid is introduced into the housing 2 from the culture fluid conduit 2d.

  In the figure, W indicates the width of the heat shrink film 7, and 8 indicates an O-ring provided at the edge of the slits 6a and 6b.

  In the above-described embodiment, the slit 6a and the slit 6b are formed in two positions up and down, but the arrangement of each slit is not limited to the upper and lower sides, and can be arranged to face left or right or diagonally. In addition, about arrangement | positioning of a slit, it is not limited to opposing arrangement | positioning.

  Further, the number of slits can be formed at two or more locations within a range in which the strength of the connecting portion can be ensured, and these slits are arranged at equal intervals or at arbitrary intervals on the circumference of the barrel portion in the cylindrical portion 2a. can do.

  According to the housing 2 having the above-described configuration, the ICS (space in the hollow fiber) and the ECS (space formed by the outside of the hollow fiber and the housing 2) are separated from each other. Since the culture medium discharge pipe 2e communicates with the ECS and the cell insertion tubes 4a and 4b communicate with each other, the cells are introduced into the ECS of the housing 2, and the culture liquid is supplied through the culture medium conduit 2d and the culture medium discharge pipe 2e. If it is circulated, the cells do not come into direct contact with the flow of the circulating culture solution, and only the necessary components that have passed through the hollow fiber membrane holes of the hollow fiber bundle 5 having a semipermeable membrane function come into contact with the cells. Become. Thereby, the cells can be cultured without being affected by the shearing force due to the culture fluid flow.

b Hollow Fiber Bundle The hollow fiber bundle 5 is made of a bundle of a plurality of hollow fibers 5a having a semipermeable membrane function, and is contained in the culture solution for cells existing in the ECS by circulating the culture solution through the lumen. It is designed to provide essential ingredients for growth. Similarly, waste components secreted by the cells are discharged into the culture solution.

  The hollow fiber bundle 5 is selected from, for example, regenerated cellulose, cellulose ester, polysulfone, polyethersulfone, polyacrylonitrile, polyvinyl alcohol, polystyrene, polymethyl methacrylate or polycarbonate in consideration of affinity with cells. It can be composed of one type of polymer or a mixture of two or more types.

  Furthermore, by hydrophilizing the surface, a hydrophobic polymer such as polyethylene, polypropylene, PTFE (polytetrafluoroethylene), PVDF (polyvinylidene difluoride) can be used as a material. By performing the hydrophilization treatment, the cell affinity is increased and the membrane permeability of the culture solution component is also improved.

  In this illustration, the hollow fiber bundle 5 made of one polymer means that all the hollow fibers 5a are made of the polymer. Moreover, the hollow fiber bundle 5 should just have a some polymer as a result that it is comprised with a mixture. For example, each of the hollow fibers 5a may be composed of a mixture thereof, and some hollow fibers 5a are made of any one kind of polymer, and another hollow fiber 5a is made of another arbitrary polymer. In addition, a configuration in which a plurality of polymers are included when a bundle is formed as a whole is also included.

  The hollow fiber bundle 5 used in the hollow fiber bundle module 1 of the present invention is more preferably composed of regenerated cellulose, cellulose ester, polysulfone, and polyethersulfone among the above-described polymers. Because it is necessary to sterilize the hollow fiber bundle module 1 in advance in cell culture, high pressure steam sterilization is possible by using regenerated cellulose, cellulose ester, polysulfone and polyethersulfone having high heat resistance. This is because sterilization can be performed efficiently.

  Examples of the hydrophilic treatment method include treatment with polyvinyl pyrrolidone, ethylene vinyl alcohol copolymer, polyvinyl acetal diethylaminoacetate, alcohol, and the like. Examples of alcohol include ethanol and isopropanol. Among these, the polyvinylpyrrolidone treatment is preferable in that stable hydrophilicity can be imparted to the hollow fiber surface.

  The pore diameter in the semipermeable membrane of the hollow fiber 5a (hereinafter referred to as membrane pore diameter) allows the necessary components in the culture medium to pass from the ICS of the hollow fiber to the ECS, and conversely the cell secretion waste components. The ECS is not particularly limited as long as it can be passed from the ICS to the ICS and does not pass the cells to be cultured.

  However, as the membrane pore size decreases, the membrane strength increases, but the mass exchange rate decreases. On the other hand, as the membrane pore size increases, the membrane strength decreases, but the mass exchange rate increases. Therefore, in order to obtain a desired membrane strength and mass exchange rate in these characteristics, the average membrane pore diameter of the hollow fiber 5a is preferably 0.001 to 2 μm, more preferably 0.005 to 0.00. 2 μm.

  In addition, when it is necessary to add various cell growth factors and differentiation-inducing factors, the pore diameter is 0.005 considering that these factors can be enclosed in ECS and cost reduction is possible. More preferably, it is -0.01 micrometer.

  The inner diameter of the hollow fiber 5a is not particularly limited as long as the culture medium can be circulated, but a thin hollow fiber is preferable from the viewpoint of ensuring high membrane permeability, and the range is 0.1 to 3 mm. Can be used, but considering the ease of processing and the ability to suppress clogging caused by insoluble components generated in the culture solution, the one in the range of 0.2 to 1 mm is selected. More preferably.

  Further, the number and thickness of the hollow fibers 5a are not particularly limited as long as the cell culture according to the present invention can be performed. However, the number of cells required, nutrition of cultured cells, oxygen requirement, hollow fiber bundle module It is preferable to adjust appropriately in relation to the volume of the ECS in 1.

  After the hollow fiber 5a is housed in the housing 2, the hollow fiber gap is sealed with a sealing material in a state in which the openings at both ends of the hollow fiber are held, and both ends are glued and fixed. Specifically, one end portion of the bundled hollow fibers 5a is fixed to the inner wall of the barrel portion at the upstream end portion of the cylindrical portion 2a constituting the housing 2, and the other end portion is opposed to the cylindrical portion 2a. It is fixed to the body inner wall of the downstream end 2f connected by screwing. In addition, as a sealing material which fixes the hollow fiber 5a, a urethane resin, an epoxy resin, etc. can be used, for example.

  The hollow fiber bundle 5 having a semipermeable membrane function configured as described above supplies essential growth components to cells existing in the ECS by circulating the culture solution through the culture solution conduit 2d and the culture solution discharge pipe 2e. In addition, metabolic components can be removed by diffusion.

c Culture Solution The culture solution is not particularly limited as long as it contains an essential component for growing cells to be cultured, and any conventionally known culture solution can be used. The essential component for cell growth is a component indispensable for the cells to be cultured to proliferate, and is composed of various organic and inorganic substances, including dissolved gas for supplying oxygen and the like. As such a culture solution, for example, a medium prepared by adding serum or various growth factors and differentiation inducing factors to a general-purpose medium such as Dulbecco MEM, RPMI 1640, and Ham F12 is used.

  In culture, the culture solution is stored in an appropriate container and repeatedly circulated in the ICS by a pump, but this does not exclude the temporary supply of the culture solution. In the present invention, the “circulation” of the culture solution includes a transient supply.

  The volume ratio of ECS with respect to the hollow fiber bundle 5 can be appropriately set as necessary, such as cell type and cultured cell density, but is preferably 1 or more and 20 or less. By setting the number to 1 or more, it is possible to increase the amount of cells put into the module and to sufficiently supply a space in which cells grow.

  Moreover, by setting it as 20 or less, it becomes possible to avoid insufficient supply of the essential growth component to the cells in the hollow fiber bundle module 1. In order to perform cell culture most suitably, 2 or more and 6 or less are preferable. The volume of the hollow fiber bundle 5 at the above volume ratio is the volume of each hollow fiber 5a that is in contact with the ECS in the module, and the cross-sectional area formed by the outer periphery of the cross section of the hollow fiber 5a (including the hollow part). ) And the length of the hollow fiber length.

  In addition, when the fiber bundle for cell attachment growth mentioned later is used, the volume of the hollow fiber bundle 5 is a value obtained by multiplying the cross sectional area of the hollow fiber 5a and the hollow fiber length, and the cross sectional area of the fiber bundle for cell attachment growth. And a value obtained by multiplying the length of the fiber bundle for cell adhesion growth.

  It is preferable to appropriately set the number of the hollow fiber bundles 5 and the like so as to achieve such a volume ratio.

d Cell Culture Method and Cultured Cell Collection Method Next, a cell culture method using the hollow fiber bundle module 1 having the above configuration and a method for collecting cultured cells will be described.

  Since the cells are supplied with sufficient nutrients and oxygen through the hollow fiber membrane, they can grow at high density.

  The cultured cells densified in the ECS are directly attached to the surface of the hollow fiber 5a, or a cell attachment carrier, specifically, an arbitrary number of cells prepared along the hollow fiber, for example, polyolefin having cell affinity. It adheres indirectly to the surface of the hollow fiber 5a through a fiber bundle for cell adhesion growth made of a polymer material such as a resin or polyester resin, or a fiber bundle for cell adhesion growth made of an inorganic material such as ceramics or glass. . Indirect attachment also includes being present in the gap between the hollow fibers 5 a or the gap between the hollow fibers 5 a and the housing 1.

  Moreover, recovering the cultured cells means recovering cells directly attached to the surface of the hollow fiber 5a or indirectly through a cell attachment carrier, and the cells or hollow fibers 5a existing in the gap between the hollow fibers 5a and the housing. Collecting the cells present in the gap with 1.

  As a cell growth means for regenerative medicine, three-dimensional culture capable of growing while maintaining the original function of cells is required, and generally, culture using a hollow fiber bundle module is considered to have conditions for three-dimensional culture. ing. In this three-dimensional culture, it is preferable to interpose an appropriate cell adhesion carrier (the above-mentioned fiber bundle for cell adhesion growth) in the cell growth space.

  The fiber bundle for cell adhesion growth according to the present invention has a form in which fibers having cell affinity are interposed between the hollow fiber bundles 5 in the form of yarns or as strip-like fiber pieces (for example, Japanese Patent Application Laid-Open No. 2002-112763). No.) or as a sheet body wound in a spiral shape when viewed from the cross-sectional direction, and can be used in a form sandwiched between the hollow fiber bundles 5.

  In these cell culture methods, a microcarrier or gel is introduced together with cells from the cell insertion tube 4a (or 4b), and the above-mentioned culture solution is circulated to the ICS through the culture solution conduit 2d and the culture solution discharge tube 2e. The cells can be cultured. In addition, culture conditions such as culture medium circulation rate, presence / absence of medium exchange, and oxygen addition method can be appropriately set according to cell type, cell concentration, and the like.

  The constituent elements and recovery methods shown in the above a, b, c, and d are not limited to the technical means described in the respective sections, but conventionally known module structures and cell culture means, and further improvements thereof, It is possible to freely apply the improvement technique as long as it does not adversely affect the achievement of the object of the present invention.

  Although the characteristic part of the module structure of the present invention will be described based on examples, it should be understood that these descriptions are merely illustrative and not limiting.

Example About 1500 regenerated cellulose hollow fibers having an inner diameter of 200 μm and a film thickness of 10 μm (total area of the hollow fiber is about 920 cm ² ) are bundled to form a hollow fiber bundle. A fiber bundle (length: 131 mm) in which 200 PET fibers are bundled is prepared, and the hollow fiber bundle and the PET fiber bundle = 13: 2 are uniformly distributed and arranged between the hollow fibers. A fiber bundle made of fibers was placed in a polycarbonate housing (length: 177 mm, diameter: 30 mm) and sealed.

  Note that one of both ends of the hollow fiber and the cell support was fixed to the end of the cylindrical portion 2a using urethane resin, and the other was fixed to the downstream end 2f.

  The hole diameter of the hollow fiber was 0.002 to 0.005 μm, and the volume occupied by the hollow fiber and the cell support was adjusted to about 32% with respect to the volume in the housing 2.

  FIG. 2 is a culture circuit diagram of a small culture device assembled using the hollow fiber bundle module 1 of the present invention.

  Prior to cell culture, the circuit, hollow fiber bundle module 1 was EOG sterilized.

  The culture solution bottle 10 contains 100 ml of liquid medium (ThemES cell culture medium consisted of 80% (V / V) knockout (KO) DMEM, 4 mM Gultamine, 100 μM MEM-non-essential acid, 55 μM 2-mercaptoethanol [2- ME] (invitorogen) 100 U / ml penicillin and 100 μg / ml streptomycin (Sigma) were added.

  The liquid medium circulated through the small culture device CellMax (Spectrum) is discharged from the culture solution bottle 10 through the circulation pump 11, the carbon dioxide concentration is adjusted to 5% by the gas exchanger 12, and the circuit 13 is flowed at a flow rate of 50 mL / min. , 14 were circulated.

In the cell culture space 15 in the housing 2, 10 ³ U / ml LIF (ESGRO, Chemicon International Temecula. CA), 20% (v / v) of Fetal bovine serum (JRH Biosciences) was added to the liquid medium described above. .

Mouse embryonic stem cells (R1 cells) were injected at 2.9 × 10 ⁵ cells / ml into cell culture space 15 (12.2 ml) and cultured for 21 days.

  For cell recovery, the hollow fiber bundle module 1 is washed with PBS, and then the heat shrink film 7 covering the slits 6a and 6b (see FIG. 1 (a)) provided in the hollow fiber bundle module 1 is cut. The upstream end of the hollow fiber bundle 5 that can be seen through the slits 6a and 6b was cut in the diameter direction using a knife.

  Next, the hollow fiber bundle 5 can be easily extracted from the housing 2 by removing the downstream end 2f together with the cap 2c from the cylindrical portion 2a.

  When the extracted hollow fiber bundle 5 was observed, it was confirmed that the cell colonies were attached in a state of straddling several hollow fibers.

Thereafter, the cultured cells were detached from the hollow fiber using trypsin. The number of collected cells reached 1.0 × 10 ⁹ and the cell density reached 8.7 × 10 ⁷ / ml. The survival rate was 74%.

In the conventional static culture, only a cell density of about 6.0 × 10 ⁵ / ml was obtained, which means that 145 times higher density cells could be recovered.

  The collected cells were continuously cultured in a petri dish for 5 days, and it was confirmed that they could be collected aseptically.

  FIG. 3 shows a second embodiment of the hollow fiber bundle module according to the present invention.

  In addition, in each figure demonstrated below, the same code | symbol is attached | subjected about the same component as Fig.1 (a), and the description is abbreviate | omitted.

  The hollow fiber bundle module 20 shown in FIG. 3 is different from the hollow fiber bundle module 1 described above in that the cylindrical portion 2a and the downstream end portion 2f ′ are not screwed but simply connected to each other. There is to be.

  Specifically, O-rings 21 and 21 are provided around the connection side end portions of the cylindrical portion 2a and the downstream side end portion 2f ′, respectively, and straddle the both connection side end portions provided with the O-rings 21 and 21. Thus, the heat shrink film 22 is provided in a ring shape. The heat shrink film 22 is attached to both connection side ends in the same manner as the heat shrink film 7 described above, and releasably connects the tube portion 2a and the downstream end portion 2f '. The cylindrical portion 2a and the downstream end 2f 'function as a housing dividing portion.

  As described above, according to the configuration in which the downstream end 2f ′ is connected to the cylindrical portion 2a by butting, the heat shrink film 7 covering the slits 6a and 6b is cut and removed, and the upstream side of the hollow fiber bundle 5 is removed. If the heat-shrinkable film 22 is cut after the end portion is cut, there is an advantage that the hollow fiber bundle 5 can be easily pulled out in the tube axis direction A.

  FIG. 4 shows a third embodiment of the hollow fiber bundle module according to the present invention.

  The hollow fiber bundle module 30 shown in the figure is different from the hollow fiber bundle module 1 described above in that the slits 6a and 6b are not provided and the upstream end 2j of the cylindrical portion 2a can be divided. It is in.

  Specifically, the downstream end 2f ′ of the cylindrical portion 2a has the same configuration as the downstream end 2f ′ shown in FIG. 3, and the cylindrical portion 2a and the downstream end 2f ′ are connected in a state of being abutted with each other. Then, the heat shrink film 22 is attached to the abutting portion, and both are fixed.

  On the other hand, the upstream end 2j of the cylindrical portion 2a is also composed of independent parts, and the upstream end 2j is connected to the cylindrical portion 2a in a state of being abutted in the same manner as the downstream end 2f ′. Yes. O-rings 31 and 31 are attached to the connection side end portions of the cylindrical portion 2a and the upstream end portion 2j, respectively, and the heat-shrinkable film straddles both connection side end portions provided with the O-rings 31 and 31. 32 is attached and both are fixed.

  As described above, when the upstream end 2j and the downstream end 2f ′ of the cylindrical portion 2a are connected to each other by butting and can be easily separated, the cylindrical portion A gap is formed between 2a and the upstream end 2j to form a cutting opening. The upstream end of the hollow fiber bundle 5 can be cut using the opening for cutting, and further, if the heat shrink film 32 is cut, the hollow fiber bundle 5 can be pulled out in the cylinder axis direction A, Only the hollow fiber bundle 5 can be easily removed from the cylindrical portion 2a.

  Further, since the upstream end 2j and the downstream end 2f ′ have substantially the same configuration, if one of them is a cutting opening, the other becomes a housing division, and conversely, either one If the is a housing dividing portion, the other functions as a cutting opening.

  FIGS. 5A to 5E schematically show modifications of the sealing structure for sealing the slits of the hollow fiber bundle module 1 shown in FIG.

  The hollow fiber bundle module 40 shown in FIG. 5A adopts a union seal method instead of sealing with a heat shrink film.

  A flange portion 2k is provided around the upstream edge of the slits 6a and 6b, and a union screw 2k ′ is formed on the outer peripheral surface of the flange portion 2k. Further, an O-ring 41a is attached to the outer peripheral edge of the flange portion 2k. On the other hand, a flange 2m is also provided around the downstream edge of the slits 6a and 6b, and an O-ring 41b is mounted on the downstream annular side wall of the flange 2m.

  Reference numeral 42 denotes a union nut having a through-hole through which the cylindrical portion 2a passes, and having an L-shaped cross section (viewed from the front), and can be screwed to the union screw 2k 'on the inner wall of the trunk portion. A threaded portion is provided. If the union nut 42 is screwed into the union screw 2k ′ and tightened, the slits 6a and 6b can be sealed with the O-ring 41b sandwiched.

  The hollow fiber bundle module 45 shown in FIG. 5 (b) employs a ring belt seal method instead of sealing with a heat shrink film.

  The flange portions 2n and 2p are provided around the upstream edge portion and the downstream edge portion of the slits 6a and 6b, respectively, and a ring-shaped rubber belt 46 is provided so as to straddle the flange portions 2n and 2p. The slits 6a and 6b can be sealed by tightening a metal clamp band 47 from the outside of the belt 46.

  The hollow fiber bundle module 50 shown in FIG. 5 (c) employs an O-ring seal system instead of sealing with a heat shrink film.

  O-rings 51 and 51 are mounted on the upstream edge and the downstream edge of the slits 6a and 6b, respectively. By tightening a metal clamp band 52 from the outside of these O-rings 51 and 51, the slits 6a and 6b can be sealed. In the figure, 2q is an annular groove formed on the outer wall of the body portion of the cylindrical portion 2a for engaging the annular protrusions formed inward at both ends of the clamp band 52 in the tube axis direction. It is.

  A hollow fiber bundle module 55 shown in FIG. 5 (d) employs a convex clamp 56 in place of the clamp band 52 in the O-ring seal system shown in FIG. 5 (c).

  The convex clamp 56 includes convex portions 56a and 56a that can be fitted into the slits 6a and 6b. The convex clamp 56 is positioned by fitting the convex portions 56a and 56a into the slits 6a and 6b. In this state, the slits 6a and 6b can be sealed.

  The hollow fiber bundle module 60 shown in FIG. 5 (e) has one slit 6c and is configured so that the slit 6c can be sealed with one touch.

  A gasket 61 is attached along the periphery of the opening of the slit 6c, and a detachable cover 62 is attached so as to close the opening of the slit 6c. The cover 62 is formed in an arc shape, and is formed with hooks 62a and 62a protruding at both ends in the longitudinal direction. On the other hand, hook grooves 63a, 63a (only the front side is shown in the figure) are formed in the vicinity of the slit 6c corresponding to the hooks 62a, 62a on the outer wall of the barrel portion 2a.

  Therefore, if the hooks 62a and 62a of the cover 62 are fitted into the hook grooves 63a and 63a, the slit 6c can be sealed via the gasket 61.

  Next, FIGS. 6A to 6C schematically show a modification of the cutting portion of the hollow fiber bundle module 1 shown in FIG.

  In the hollow fiber bundle module 65 shown in FIG. 6A, a cylindrical part (cylindrical part) 66 as a cutting part is erected in a direction perpendicular to the central axis A in a part of the cylindrical part 2a.

  A male screw portion 66 a is formed on the outer peripheral surface of the upper end portion of the cylindrical portion 66, and a disc-shaped cap (lid body) 67 having a female screw portion that can be screwed with the male screw portion 66 a is an opening of the cylindrical portion 66. Can be attached as a lid for sealing. An O-ring 68 is attached to the opening edge of the cylindrical portion 66.

  In this configuration, the hollow fiber bundle 5 can be cut by removing the cap 67 and inserting a cutting tool such as a knife from the upper opening of the cylindrical portion 66.

  A hollow fiber bundle module 70 shown in FIG. 6 (b) includes a cylindrical portion 71 as a cutting portion in a part of the cylindrical portion 2a as in the module of FIG. 6 (a).

  However, an O-ring 72 is attached to the front end edge of the cylindrical portion 71, and a disc-shaped cap (lid body) 73 is attached from the outside of the O-ring 72. Further, hooks 73 a and 73 a that can be locked to the outer wall of the tip end portion of the cylindrical portion 71 are provided on the side surface of the cap 73. If the hooks 73 a and 73 a are locked to the cylindrical portion 71, the O-ring 72 is sandwiched. In this state, the upper opening of the cylindrical portion 71 can be sealed.

  In the hollow fiber bundle module 75 shown in FIG. 6C, a rectangular tube portion (tubular portion) 76 as a cutting portion is formed in a direction perpendicular to the central axis A on a part of the side surface of the tube portion 2a.

  A gasket 77 is attached in a rectangular shape to the front end edge of the rectangular tube portion 76, and a rectangular cap (lid body) 78 is attached from the outside of the gasket 77. The side walls of the four sides of the cap 78 are provided with hooks 78a to 78d that can be locked to the side surface of the rectangular tube portion 76. If the hooks 78a to 78d are locked to the rectangular tube portion 76, the gasket 77 is sandwiched. In this state, the opening of the rectangular tube portion 76 can be sealed.

  Instead of the lids 67, 73, 78, plugs that fit into the cylinders 66, 71, 76 and fill the cylinders can be mounted.

(a) is the front view which showed the structure of the hollow fiber bundle module based on this invention in a partial cross section, (b) is CC sectional view taken on the line of Fig.1 (a). It is a cell culture circuit diagram using a hollow fiber bundle module. It is a front view which shows 2nd embodiment of a hollow fiber bundle module. It is a front view which shows 3rd embodiment of a hollow fiber bundle module. (a)-(e) is a schematic diagram which shows the modification of the slit sealing structure which concerns on a hollow fiber bundle module. (a)-(c) is a schematic diagram which shows the modification of the cutting part which concerns on a hollow fiber bundle module.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Hollow fiber bundle module 2 Housing 2a Cylinder part 2b, 2c Cap 2d Culture solution conduit | pipe (when culture solution is circulated in B direction)
2e Culture fluid discharge tube (when culture fluid is circulated in direction B)
2f Downstream end (when culture fluid is circulated in direction B)
2g Screw part 2h, 2i Connection part 3 O-ring 4a, 4b Cell insertion tube 5 Hollow fiber bundle 6a, 6b Slit 7 Heat shrink film 8 O-ring 10 Culture solution bottle 11 Circulation pump 12 Gas exchanger 13, 14 Circuit 15 Cell culture space (ECS)

Claims (11)

A hollow fiber bundle in which hollow fibers made of a semipermeable membrane material are bundled;
In the hollow fiber bundle module for cell culture having the hollow fiber bundle and the culture medium, and the housing provided with the culture liquid introduction part and the culture liquid discharge part,
A cutting body for cutting at least one end of the hollow fiber bundle housed in the housing is formed in the housing, and a sealing body for releasably sealing the cutting opening is provided. Provided,
The cell culture hollow, further comprising a housing dividing portion for removing the hollow fiber bundle which has been cut at one end portion to obtain a free end at the end portion of the housing from the housing. Yarn bundle module.   The hollow fiber bundle module for cell culture according to claim 1, wherein the opening for cutting has a slit cut out in a direction orthogonal to the cylinder axis direction of the housing.   The hollow fiber bundle module for cell culture according to claim 1 or 2, wherein a heat shrink film is wound around the opening for cutting as the sealing body.   The hollow fiber bundle module for cell culture according to claim 2, wherein a thread portion is formed on the outer peripheral wall of the housing in the vicinity of the slit, and the unsealed nut is screwed into the thread portion as the sealing body.   3. The cell culture according to claim 2, wherein a collar or an annular groove is formed in the outer peripheral wall of the housing on both sides of the slit, and a clamp is fastened and fixed to the collar or the annular groove as the sealing body. Hollow fiber bundle module.   The hollow fiber bundle module for cell culture according to claim 5, wherein a convex portion that can be fitted into the opening for cutting is formed on an inner surface of the clamp.   The hollow fiber bundle module for cell culture according to claim 2, which has a resin or metal cover attached to the slit as the sealing body.   The hollow fiber bundle module for cell culture according to any one of claims 4 to 7, wherein a sealing material is provided around the slit edge.   The cutting opening has a cylindrical portion extending from one end portion of the housing in a direction orthogonal to the tube axis direction, and a lid that closes the opening at the tip of the cylindrical portion or the inside of the cylindrical portion is provided. The hollow fiber bundle module for cell culture according to claim 1, further comprising a plug for filling.   The hollow fiber bundle module for cell culture according to any one of claims 1 to 9, wherein the housing split part is provided with a pipe joint composed of a male screw part and a female screw part.   The hollow fiber bundle module for cell culture according to any one of claims 1 to 10, wherein the hollow fiber bundle has an arbitrary number of cell attachment growth fiber bundles along the hollow fiber. .

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