JP2018139527A - Culture medium production method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method of producing a culture medium comprising fibers aligned in one direction which has good productivity.SOLUTION: The method of producing a culture medium comprises: a preparation step of preparing a first surface, a second surface on the opposite side thereto, at least one through-hole penetrating from the first surface to the second surface, and a frame that has a first bonding part on the first surface; a deposition step of forming a fiber assembly by discharging fiber raw material liquid from a nozzle, creating fibers while depositing the fibers around the peripheral surface of a winding rotary body; a transfer step of transferring the fiber assembly to the first surface of the frame body via the first bonding part while rotating the winding rotary body; and a mounting step of mounting the frame to which the fiber assembly has been transferred to the substrate so as to face the first surface.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a method for producing a culture medium, and particularly relates to an improvement in productivity of a culture medium having fibers arranged in one direction.

  In recent years, fiber base materials have attracted attention as a medium for culturing biological tissues and microorganisms (see Patent Document 1). The fiber substrate is, for example, a woven fabric, a knitted fabric, or a non-woven fabric, and has a three-dimensional structure. Therefore, biological tissues and microorganisms can be cultured in a state close to a physiological environment in vitro.

Special table 2010-517590

  When directionality is seen in the growth of biological tissues and microorganisms, it is desirable that the fibers constituting the fiber base material are arranged in a certain direction. This is because biological tissues and microorganisms are easy to grow. However, usually, the fiber base material is held in shape by entanglement of fibers, and does not have the above arrangement.

  One aspect of the present invention includes a first surface, a second surface opposite to the first surface, and one or more through holes penetrating from the first surface to the second surface. A preparation step of preparing a frame having a first adhesive portion on one surface, and a fiber raw material liquid is ejected from a nozzle to generate the fiber, and the fiber is circulated around the peripheral surface of the winding rotary body And depositing the fiber assembly on the first surface of the frame body while rotating the winding rotary body, and depositing the first adhesive portion to form the fiber assembly. And a mounting step of mounting the frame body onto which the fiber assembly has been transferred, on a substrate so that the first surface faces the substrate.

  According to the production method of the present invention, a culture medium having fibers arranged in one direction can be produced with high productivity.

It is a perspective view which shows typically the culture medium which concerns on this invention. It is a side view which shows typically the winding rotary body in each process of the manufacturing method which concerns on this invention, a frame, and a board | substrate ((a)-(c)). It is the perspective view (a) and top view (b) which show an example of the winding rotary body which concerns on this invention. It is a side view which shows the further another example of the winding rotary body which concerns on this invention. It is a schematic top view of the one part area | region of the fiber assembly for demonstrating the arrangement | sequence of a fiber.

[Culture medium]
The medium produced by the method according to the present embodiment is suitably used for, for example, a potential measuring device for measuring these potentials in a state where biological tissue or microorganisms are retained.
An example of the medium is shown in FIG. FIG. 1 is a perspective view schematically showing the culture medium 100.
The medium 100 includes a substrate 110, a frame body 120 mounted on the substrate 110, and a fiber assembly 130 interposed between the substrate 110 and the frame body 120. However, the fiber assembly 130 is disposed not in the entire mounting surface 110X on which the frame body 120 of the substrate 110 is mounted, but in a range facing one main surface (first surface 120X) of the frame body 120. ing. The culture medium 100 may be accommodated in a holder or the like as necessary and disposed in the potential measurement device.

(Fiber assembly)
The fiber aggregate 130 is an aggregate of a plurality of fibers 131. In the fiber assembly 130, the plurality of fibers 131 are arranged in one direction. That the plurality of fibers 131 are arranged in one direction means that in the fiber assembly 130, the fibers 131 do not intersect with each other, or the average angle at which the fibers 131 intersect each other is more than 0 ° and not more than 60 °. Say something. In this way, when the plurality of fibers 131 are arranged, the fibers 131 are likely to extend along the arrangement direction of the fibers 131, so that stress on the biological tissue and microorganisms is reduced. Therefore, it becomes easy for biological tissues and microorganisms to grow along the arrangement direction of the fibers 131.

  Here, the average angle at which the fibers 131 intersect can be determined from the intersection of the fibers 131 in the average length direction. The average length direction of the fibers 131 can be determined based on, for example, an SEM photograph when the fiber assembly 130 is viewed from the normal direction. FIG. 5 is a schematic top view of the fiber assembly for explaining the arrangement of the fibers. In FIG. 5, the state of the fiber assembly 130 in the SEM photograph which image | photographed the fiber assembly 130 from the normal line direction is imitated. A square region R having a predetermined size (for example, 100 μm × 100 μm) is set when the fiber assembly 130 composed of a plurality of fibers 131 is viewed from the normal direction. At this time, the region R is determined so that twelve or more fibers 131 are included in the region R, and 50% or more of the fibers 131 located in the region R intersect two opposite sides of the region R. In this region R, the direction of a straight line (dotted line in FIG. 5) connecting a point where a certain fiber 131 intersects the two opposite sides is defined as the average length direction of the fiber 131.

  For example, in the region R, the average angle at which the fibers 131 intersect is selected from a plurality of (for example, 20) fibers 131 that are arbitrarily selected, and two fibers 131 are arbitrarily selected. An angle at which the average length direction intersects (for example, θ1 in FIG. 5) is obtained. Another two fibers 131 are selected, and an angle (for example, θ2 in FIG. 5) at which the average length direction of each fiber 131 intersects is obtained. Such an operation is performed on the remaining selected fibers 131 (for example, 16 pieces). And the average of each angle is calculated and it is set as the average angle which the fibers 131 cross.

  The ratio of the area of the fiber 131 to the unit area of the fiber assembly 130 can be selected from 10 to 90%. For example, when used for cardiomyocyte culture or a potential measurement device, the fiber assembly 130 is very thin, and the ratio of the fibers 131 per unit area is 20 to 50%, and is uniformly dispersed at 30 to 40%. It is preferable to deposit. The ratio of the area of the fibers 131 is a region of a predetermined area (for example, an ellipse having a minor axis of 3 mm and a major axis of 6 mm) on one main surface (for example, the upper surface) of the fiber assembly 130. Can be obtained by binarizing an image obtained with an optical microscope or the like, calculating the area occupied by the fibers 131, and converting the area to a ratio (%) per unit area.

  The material of the fiber 131 is not particularly limited as long as it can be used as a biological tissue or a microorganism culture medium. Among these, the fiber 131 is different from the block polymer containing a polystyrene block and a polybutadiene block in that the affinity to biological tissues and microorganisms is high, and stressing the biological tissues and microorganisms is difficult when culturing. And a styrene resin. The fiber 131 may contain various additives as required.

  The block polymer may be, for example, a diblock body in which a polybutadiene (PB) block and a polystyrene (PS) block are connected, but a polyblock body of a triblock body or more in which PB blocks and PS blocks are alternately connected. Is preferred. The block polymer preferably contains a PS block at least at the end from the viewpoint of ensuring affinity with the styrene resin. The PB block increases the flexibility and elongation of the resulting fiber 131.

  The content of the PB block in the block polymer is, for example, 10 to 30% by mass, preferably 15 to 30% by mass, and more preferably 20 to 30% by mass or 20 to 25% by mass. When the content of the PB block is in such a range, the affinity with the styrene resin is increased, and the homogeneous fibers 131 are easily generated. Further, the obtained fiber 131 has high flexibility and elongation. Further, when the fiber 131 is generated by the electrospinning method, high spinnability is ensured.

  As the styrene resin, a polymer different from the above block polymer is used. Examples of the styrene resin include polystyrene (styrene homopolymer), and a copolymer of styrene and another copolymerizable monomer. A styrene resin may be used alone or in combination of two or more.

  From the viewpoint of achieving both flexibility and easy formation of the fibers 131, the mass ratio of the block polymer to the styrene resin (= block polymer: styrene resin) is, for example, 2: 1 to 1: 5, preferably 1. : 1-1: 4. In particular, when the fiber assembly 130 is formed by an electrospinning method using a solution, when the mass ratio is in such a range, the block polymer and the styrene resin are easily dissolved in a solvent, and high spinnability is ensured. You can also.

For example, the average fiber diameter of the fibers 131 is preferably 0.5 to 10 μm, more preferably 1 to 5 μm, and particularly preferably 1.5 to 4 μm.
The average fiber diameter is an average value of the diameters of the fibers 131. The diameter of the fiber 131 is a diameter of a cross section perpendicular to the length direction of the fiber 131. If such a cross section is not circular, the maximum diameter may be considered as the diameter. Further, the width in the direction perpendicular to the length direction of the fibers 131 when viewed from the normal direction of one main surface of the fiber assembly 130 may be regarded as the fiber diameter. The average fiber diameter is, for example, an average value of the diameters of arbitrary portions of arbitrary 10 fibers included in the fiber assembly 130.

(Frame)
The frame 120 includes a first surface 120X, a second surface 120Y opposite to the first surface 120X, and one or more through holes 121 penetrating from the first surface 120X to the second surface 120Y. The fiber assembly 130 is disposed on the surface of the first surface 120X so as to cover at least a part of the through hole 121. That is, the fiber assembly 130 (fiber 131) is exposed from the opening (first opening) on the first surface 120X side of the through hole 121.

  When the frame body 120 is mounted on the substrate 110, the first opening of the through hole 121 is closed by the substrate 110 via the fiber assembly 130, and at least one recess is formed on the mounting surface 110X of the substrate 110. The A culture solution containing biological tissue or microorganisms is injected into the recess from the opening (second opening) on the second surface 120Y side of the through hole 121. The injected biological tissue or microorganism grows using the fiber assembly 130 as a scaffold. Since the fibers 131 constituting the fiber assembly 130 are arranged in a state along one direction, the biological tissue or microorganism can grow along the length direction of the fiber 131 with less stress. .

  The material of the frame 120 is not particularly limited, and may be made of glass or resin (including elastomer). The size of the frame 120 is not particularly limited as long as the entire surface of the first surface 120X can face the substrate 110 and does not interfere with the wiring of electrodes (described later) arranged on the substrate 110.

  The number of through holes 121 is not particularly limited, and may be set as appropriate according to the size and use of the frame body 120. The shape and size of the first opening and the second opening are not particularly limited, and may be set as appropriate according to the application. The shape and size of the first opening and the second opening may be the same or different. The shape of the depression formed by the through hole 121 is not particularly limited. For example, when both the first opening and the second opening are circular, the shape of the recess may be a columnar shape or a mortar shape. Especially, it is preferable that the said hollow is a mortar shape with a large 2nd opening at the point which a culture solution can inject easily.

(substrate)
The substrate 110 is insulative and includes, for example, a plurality of electrodes (first electrode) (not shown) and a plurality of micro electrodes (second electrodes) electrically connected to the first electrode. The plurality of first electrodes are insulated from each other. The plurality of second electrodes 203 are formed at predetermined intervals in a matrix manner and are insulated from each other.

  The plurality of first electrodes are disposed so as not to contact the fiber assembly 130, while the plurality of second electrodes are disposed so as to contact at least a part of the fiber assembly 130. By measuring the voltage between the first electrode and the second electrode, the potential of the fiber assembly 130 (ie, biological tissue or microorganism) can be measured. In this way, by measuring changes over time in the potential of the fiber assembly 130 or changes in conditions, it is possible to evaluate the state or function of biological tissue or microorganisms based on this change in potential. . At this time, since the biological tissue or the microorganism is in a state of less stress, highly accurate evaluation is possible. Furthermore, by applying a voltage between the first electrode and the second electrode, it is possible to stimulate biological tissue or microorganisms (electrical signals) and promote their growth.

  The substrate 110 is not particularly limited as long as it is insulative, and may be appropriately selected depending on the application. Examples of 110 include a glass plate, a quartz plate, and an acrylic plate. The first electrode is not particularly limited and may be appropriately selected depending on the application. Examples of the first electrode include an ITO (indium tin oxide) electrode and a platinum electrode.

  The second electrode only needs to be able to measure the potential of a biological tissue or a microorganism, and can be appropriately selected according to the application. The size of the second electrode, the distance between the adjacent second electrodes, and the number of second electrodes can be appropriately selected according to the type of biological tissue or microorganism, the size of the sample, and the like. The length of one side of the second electrode (diameter in the case of a disk) is, for example, 10 to 100 μm, and may be 15 to 60 μm. The distance between adjacent second electrodes (distance between the centers of the second electrodes) is, for example, 50 to 1000 μm, and may be 50 to 500 μm.

(Adhesive part)
The fiber assembly 130 and the frame body 120 are bonded via a first bonding portion 140A formed on the frame body 120. At this time, a part of the fiber assembly 130 is held so as to be embedded in the first adhesive portion 140A. Since the fiber assembly 130 is an assembly of fibers 131 arranged in one direction, the material of the first adhesive portion 140A easily enters between the fibers 131 and easily penetrates to the substrate 110 side. Therefore, the first adhesive portion 140 </ b> A can also function as an adhesive portion between the frame body 120 and the substrate 110. However, in FIG. 1, for convenience, the first bonding portion 140 </ b> A is arranged on the frame body 120 side with respect to the fiber assembly 130. Note that the first bonding portion 140A is not formed in the region corresponding to the first opening.

The material (adhesive A) of the first adhesive portion 140A is not particularly limited, and examples thereof include a pressure sensitive adhesive, a hot melt adhesive, a curable adhesive, and the like.
The pressure-sensitive adhesive is applied to the frame body 120 and adheres the frame body 120 and the fiber assembly 130 (further, the substrate 110; the same applies hereinafter) due to its adhesiveness. The material of the pressure sensitive adhesive is not particularly limited, and examples thereof include silicone resin. Examples of the silicone resin include dimethyl silicone and methylphenyl silicone.

  The hot melt adhesive is applied to the frame 120 while being heated, and is cooled, thereby bonding the frame 120 and the fiber assembly 130 together. The material of the hot melt adhesive is not particularly limited. For example, a thermoplastic resin such as a polyester such as polyurethane or polyethylene terephthalate, a copolymer polyester such as urethane-modified copolymer polyester, polyamide, or polyolefin (for example, polypropylene or polyethylene) is used. It is included as a main component (component occupying 50% by mass or more).

  The curable adhesive is applied to the frame body 120, polymerized by ultraviolet irradiation or heating, and cured to bond the frame body 120 and the fiber assembly 130. The kind of curable adhesive is not specifically limited, A thermosetting resin, an ultraviolet curable resin, etc. are mentioned. Examples of these resins include acrylic resins and epoxy resins. When using a curable adhesive, it is preferable to make a curable adhesive into a semi-hardened state before a transfer process. In this case, after the transfer process or the mounting process, a curing operation is further performed to completely cure the curable adhesive.

  Among them, as the adhesive A, a pressure-sensitive adhesive and a hot-melt adhesive are preferable because a special step for curing can be omitted, and a heating device for melting the adhesive is unnecessary. In terms, pressure-sensitive adhesives are preferred. In the case where the adhesive A includes a hot-melt adhesive and / or a curable adhesive, the two members are “bonded via the first bonding portion 140 </ b> A”. "It is bonded via a cured product of agent A". The same applies to the adhesive B described later.

  The first adhesive portion 140A is formed on the first surface 120X of the frame body 120. 140 A of 1st adhesion parts may be formed in the whole surface other than the 1st opening of the 1st surface 120X, and may be formed partially.

  An adhesive part (second adhesive part 140 </ b> B) may be further formed between the substrate 110 and the frame body 120. In this case, the second adhesive portion 140B is formed on the mounting surface 110X on which the frame body 120 of the substrate 110 is mounted. 140A of 1st adhesion parts and 140B of 2nd adhesion parts can mutually contact, enclosing the fiber 131, respectively. Thereby, the adhesiveness between the frame body 120 and the substrate 110 is further enhanced, and the fiber aggregate 130 interposed therebetween is easily fixed. The material (adhesive B) of the 2nd adhesion part 140B is not specifically limited, For example, the adhesive agent similar to the adhesive agent A is mentioned. Among these, from the same viewpoint as described above, the adhesive B is preferably a pressure-sensitive adhesive. Moreover, it is preferable that the adhesive agent A and the adhesive agent B contain the adhesive material of the same material from an adhesive viewpoint.

  The second adhesive portion 140B may be formed on the entire surface other than the portion facing the first opening of the mounting surface 110X, or may be formed partially. In the illustrated example, the second bonding portion 140B is formed on the entire surface other than the portion facing the first opening of the first surface 120X.

The mass per unit area of the bonding portions (140A, 140B) is not particularly limited. Among these, from the viewpoint of preventing the cultivation of biological tissues and microorganisms while ensuring the adhesion between the fiber assembly 130 and the frame 120, and further the adhesion between the substrate 110 and the frame 120, Is preferably 0.5 to 100 mg / cm ² .

[Method for producing medium]
In this embodiment, in order to manufacture the culture medium 100 including the fiber assembly 130 having high alignment properties, the fiber 131 is wound up by the winding rotary body while spinning. Thereby, the fiber assembly 130 formed on the peripheral surface of the winding rotary body has high alignment in one direction. Further, the fiber assembly 130 is transferred to the first surface 120X of the frame body 120 in a state where the arrangement of the fibers 131 in one direction is maintained. Thereafter, the frame body 120 including the fiber assembly 130 is mounted on the substrate 110 so that the first surface 120X faces the first body 120X. As a result, the fiber assembly 130 is disposed between the substrate 110 and the frame body 120 while maintaining high alignment when wound on the winding rotary body.

  The mounting surface 110 </ b> X of the substrate 110 has a sufficiently larger area than the frame body 120 for the convenience of electrical wiring. On the other hand, the fiber assembly 130 should just be arrange | positioned so that it may expose from the 1st opening formed of the several through-hole 121 formed in the frame 120. FIG. Therefore, by transferring the fiber assembly 130 to the frame body 120 instead of the substrate 110, the fiber assembly 130 can be arranged only in a necessary portion without performing precise positioning, etc. Will improve.

  Hereinafter, the manufacturing method of this embodiment is demonstrated, referring drawings. 2A to 2C are side views schematically showing the winding rotary body 10, the frame body 120, the substrate 110, and the like in each step of the present embodiment.

  The method for producing a culture medium according to the present embodiment includes a first surface 120X, a second surface 120Y opposite to the first surface 120X, and one or more through holes 121 penetrating from the first surface 120X to the second surface 120Y. And preparing a frame 120 having the first adhesive portion 140A on the first surface 120X, and discharging the raw material liquid of the fiber 131 from the nozzle 51 to generate the fiber 131, and the fiber 131 is deposited so as to circulate on the circumferential surface of the winding rotary body 10, and the fiber assembly 130 is formed on the frame 120 by rotating the winding rotary body 10 while rotating the winding rotary body 10. The transfer process of transferring the first surface 120X to the first surface 120X via the first adhesive portion 140A and the frame body 120 to which the fiber assembly 130 has been transferred are mounted on the substrate 110 so that the first surface 120X faces each other. A mounting process.

  In the above manufacturing method, for example, the fiber 131 is generated by discharging the adhesive portion forming portion that forms the first adhesive portion 140A on the first surface 120X of the frame body 120 and the raw material liquid of the fiber 131 from the nozzle 51. In addition, the fiber 131 is deposited so as to circulate on the circumferential surface of the winding rotary body 10, and the fiber assembly 130 is rotated while rotating the winding rotary body 10, and a deposition portion that forms the fiber aggregate 130. The transfer portion that transfers the first surface 120X of the frame body 120 through the first adhesive portion 140A and the frame body 120 to which the fiber assembly 130 has been transferred so that the first surface 120X faces each other. And a mounting unit mounted on 110.

(1) Preparatory Step A frame body including a first surface 120X, a second surface 120Y on the opposite side, and one or more through holes 121 penetrating from the first surface 120X to the second surface 120Y. 120 and a substrate 110 for mounting the frame body 120 are prepared.
A first adhesive portion 140A is formed on the first surface 120X. 140 A of 1st adhesion parts are formed in at least one part other than the 1st opening of the 1st surface 120X by printing, a dispenser, etc., for example.

(2) Deposition process (FIG. 2 (a))
The fibers 131 are generated from the raw material liquid 132, and the fibers 131 are deposited while rotating around the circumferential surface of the winding rotary body 10 one or more times. Thereby, the fiber assembly 130 in which the fibers 131 are oriented in one direction is formed on the peripheral surface of the winding rotary body 10.

  The method (spinning method) for producing the fibers 131 from the raw material liquid 132 is not particularly limited, and may be appropriately selected according to the type of the fibers 131 to be produced. Examples of the spinning method include a solution spinning method, a melt spinning method, and an electrospinning method.

  The fibers 131 may accumulate along the circumferential surface of the winding rotary body 10 and may overlap in the normal direction of the circumferential surface. This overlap in the normal direction of the fibers is often observed particularly when fibers are produced by electrospinning. The fiber 131 that is deposited first tends to move due to stimulation of the fiber 131 that is deposited later, and the orientation is easily disturbed. Further, as will be described later, when the winding rotary body 10 includes the convex portion 10P, the fibers 131 that are deposited in advance may loosen between the convex portions 10P, and the orientation may be disturbed. According to this embodiment, after transferring the fiber assembly 130 deposited on the winding rotary body 10 to the frame body 120, the frame body 120 is further mounted on the substrate 110 so that the fiber assembly 130 faces. . Therefore, in the obtained culture medium 100, the fibers 131 that are deposited later are disposed on the second surface 120Y side of the frame body 120. In other words, the fibers 131 with less disorder in the orientation are arranged so as to be more in contact with the culture solution. Therefore, stress on biological tissues and microorganisms is reduced, and their growth is further promoted.

  The solution spinning method is a method in which a solution obtained by dissolving the raw material of the fiber 131 in a solvent is used as the raw material liquid 132. The solution spinning method using a solvent includes a so-called wet spinning method and a dry spinning method. In the wet spinning method, the raw material liquid 132 is discharged into the coagulating liquid, and the fiber 131 is formed by a chemical reaction between the raw material of the fiber 131 and the coagulating liquid, or by replacing the solvent with the coagulating liquid. In the dry spinning method, the fiber 131 is formed by discharging the raw material liquid 132 into the air and then removing the solvent by heating or the like. Among these, the dry spinning method is preferable in that the fibers 131 are easily deposited in a state where they are arranged in one direction.

  The melt spinning method is a method in which a melt obtained by heating and melting the raw material of the fiber 131 is used as the raw material liquid 132. The obtained raw material liquid 132 is solidified into a fiber shape by being discharged into the air and then cooled. In this case, normally, a solvent for dissolving the raw material of the fiber 131 is not used. Therefore, the melt spinning method is preferable in that the solvent removal operation can be omitted.

  The electrospinning method is common to the solution spinning method in that a solution obtained by dissolving the raw material of the fiber 131 in a solvent is used as the raw material liquid 132. However, in the electrospinning method, the raw material liquid 132 is discharged into the air while applying a high voltage. The solvent contained in the raw material liquid 132 volatilizes in the process until it reaches the peripheral surface of the winding rotary body 10.

  In the electrospinning method, in order to apply a high voltage to the raw material liquid 132, the raw material liquid 132 is charged positively or negatively. At this time, the discharge end of the raw material liquid 132 discharged into the air is attracted to the take-up rotary body 10 by grounding the winding rotary body 10 or charging it with a polarity opposite to that of the raw material liquid 132. And adhere to the peripheral surface. Then, by rotating the winding rotary body 10 while discharging the raw material liquid 132, the fibers 131 are deposited while circling around the circumferential surface of the winding rotary body 10, as in the solution spinning method and the melt spinning method. A fiber assembly 130 including fibers 131 that cover at least a part of the circumferential surface of the winding rotary body 10 and are arranged in one direction is formed.

(Raw material liquid)
The raw material liquid 132 used in the solution spinning method or the electrospinning method includes the raw material of the fiber 131 and a solvent. The raw material liquid 132 used in the melt spinning method includes a raw material of the melted fiber 131.

  The solvent is not particularly limited as long as it dissolves the raw material of the fiber 131 and can be removed by volatilization or the like, and can be appropriately selected from water and an organic solvent according to the type of raw material and production conditions. As the solvent, an aprotic polar organic solvent is preferable. Examples of such a solvent include amides such as N, N-dimethylformamide (DMF), N, N-dimethylacetamide (DMAc), and N-methyl-2-pyrrolidone (NMP) (chain or cyclic amide). A sulfoxide such as dimethyl sulfoxide; These solvent may be used individually by 1 type, and may be used in combination of 2 or more type.

  Although the solid content concentration of the raw material liquid 132 can be adjusted according to the kind of solvent etc., it is 5-50 mass%, for example, and may be 10-30 mass%. The raw material liquid 132 may further contain an additive as necessary.

(Winding rotary body)
An example of the winding rotary body 10 is shown in FIG. FIG. 3A is a perspective view of the winding rotary body 10, and FIG. 3B is a plan view thereof. FIG. 3B also shows a part of the fiber assembly 130 deposited on the peripheral surface of the winding rotary body 10.

  The configuration of the winding rotary body 10 (or a rotary base 11 described later) is not particularly limited as long as it can rotate, and may be a drum shape or a belt stretched by a plurality of rolls. Good. In the latter case, the belt is rotated by rotating at least one roll. Examples of the material of the winding rotary body 10 include metal materials, various resins, various rubbers, ceramics, and combinations thereof. When the winding rotary body 10 is a belt, the belt may be a metal belt or a resin belt. When the fiber 131 is spun by the electrospinning method, the resin belt preferably has conductivity. The outer shape of the winding rotary body 10 may be, for example, a cylinder or a prism.

The fibers 131 are deposited on the circumferential surface of the winding rotator 10 while being arranged in a direction that circulates around the circumferential surface of the winding rotator 10 (hereinafter, array direction D _F ). The arrangement direction _DF is, for example, a direction along the rotation direction of the winding rotary body 10 (that is, the direction perpendicular to the rotation axis A of the winding rotary body 10). The angle θ _F (where θ _F ≦ 90 °) formed by the arrangement direction D _F and the rotation axis A may be, for example, 60 ° or more and 90 ° or less. The arrangement direction _DF is the longitudinal direction of the fibers 131 when the fibers 131 are viewed from the normal direction of the peripheral surface of the winding rotary body 10. The longitudinal direction of the fiber 131 may be obtained by taking an approximate straight line of the fiber 131 when viewed from the normal direction of the peripheral surface of the winding rotary body 10. Angle theta _F is the average value of the angle formed between the arrangement direction D _F of a plurality of fibers 131 and the rotation axis A. The arrangement direction _DF of the plurality of fibers 131 deposited on the winding rotary body 10 may be different from each other within the above range.

  A plurality of belt-like convex portions 10 </ b> P extending in the direction along the rotation axis A of the winding rotator 10 may be disposed on the circumferential surface of the winding rotator 10. Thereby, the aggregate (fiber aggregate 130) of the fibers 131 arranged so as to circulate around the circumferential surface of the winding rotary body 10 is easily peeled off from the winding rotary body 10. As a result, the fiber assembly 130 can be easily transferred to the frame 120 while maintaining the arrangement of the fibers 131. The ends of the plurality of convex portions 10P may be connected by ribs 10R extending in a direction intersecting with the rotation axis A.

Extending direction _{D P} of the convex portion 10P is not limited to the case is parallel to the rotation axis A, the stretching direction _{D P} and the rotation axis A an angle of theta _P (However, θ _P <90 °), for example, 0 It is not less than 30 ° and not more than 30 °. Among them, from the viewpoint of the releasability of the fiber assembly 130, the angle theta _P is 0 ° or more and 20 ° or less.

The stretching direction _{D P} is a direction intersecting the arrangement direction _{D F} of the fiber 131. The angle between the stretching direction _{D P} to the arrangement direction _{D F} theta (However, theta ≦ 90 °), for example, 60 ° or more and 90 ° or less. Incidentally, the stretching direction D _P, when viewed protrusion 10P in the normal direction of the peripheral surface of the winding rotary member 10, a longitudinal centerline L _CP of the convex portion 10P is the direction of stretching. If the center line L _CP contains a curve, the extending direction D _P is the minimum rectangular center line surrounding the center line L _CP is the direction of stretching.

The shape of the convex portion 10P is not particularly limited as long as it is a belt shape. The strip, the length of the extending direction D _P of the convex portion 10P is a shape longer than the length in the direction perpendicular to the extending direction D _P. Examples of the shape when the convex portion 10P is viewed from the normal direction of the peripheral surface of the winding rotary body 10 include a rectangle and a trapezoid.

  The number of the convex portions 10P is not particularly limited and may be two or more. Especially, from a viewpoint of the peelability of the fiber assembly 130, it is preferable that 3 or more are arranged on the peripheral surface of the winding rotary body 10, and 10 or more are preferably arranged. Further, from the same viewpoint, it is preferable that the convex portions 10P are arranged at equal intervals. As will be described later, when the fiber assembly 130 is cut in a state of being wound around the winding rotary body 10 prior to the transfer process of the fiber assembly 130 to the frame 120, the fiber assembly after cutting is cut. The fiber assembly 130 is cut between the protrusions 10P so that at least a part of the body 130 is in contact with the protrusions 10P. Thereby, the arrangement of the fibers 131 is easily maintained. In this case, it is preferable that the interval between the convex portions 10P of the planned cutting location C (see FIG. 2A) is smaller than the interval between the convex portions 10P in other portions.

The length (width) in the short direction of the convex portion 10P is not particularly limited. Among them, from the viewpoint of the peelability of the fiber assembly 130, the total area of all the convex portions 10P contacting the peripheral surface of the winding rotary body 10 is 10% or more of the surface area of the peripheral surface of the winding rotary body 10, It is preferable to determine the width of each convex portion 10P so that it is 80% or less, particularly 30% or more and 70% or less. The length of the extending direction D _P of the convex portion 10P is not particularly limited. Especially, it is preferable that the convex part 10P is extended over the area | region where the fiber 131 can accumulate on the surrounding surface of the winding rotary body 10 at least.

  The height of the convex portion 10P is not particularly limited. Especially, it is preferable that the height of the convex part 10P is not excessively high at the point which suppresses the slack of the fiber 131 and maintains the arrangement | sequence in one direction. From the viewpoint of peelability of the fiber assembly 130 and suppression of slackness of the fibers 131, the height of the convex portion 10P is preferably 100 to 5000 μm. The height of the convex portion 10 </ b> P is an average value in the normal direction of the peripheral surface of the winding rotary body 10.

  The material of the convex part 10P is not specifically limited, Various resin materials are mentioned. Especially, it is preferable that the convex part 10P is provided with a silicone rubber layer at least in a contact part with the fiber 131. This is because the peelability of the fiber assembly 130 is further improved. On the other hand, since the silicone rubber has appropriate adhesiveness, the fiber assembly 130 is prevented from peeling from the peripheral surface of the winding rotary body 10 before the transfer process.

  Silicone rubber is a thermosetting compound whose main chain is formed by silicon-oxygen bonds (siloxane bonds). Examples of the silicone rubber include methyl silicone rubber, vinyl-methyl silicone rubber, phenyl-methyl silicone rubber, dimethyl silicone rubber, and fluorosilicone rubber. The entire convex portion 10P may be formed of silicone rubber. In addition, when the fiber 131 is produced | generated by the electrospinning method, it is preferable that the convex part 10P is provided with electroconductivity.

  From the viewpoint of handleability, it is preferable that the convex portion 10 </ b> P is arranged in a state where it can be attached to and detached from the winding rotary body 10. For example, as shown in FIG. 4, a concavo-convex sheet 12 including a support sheet 12 a and a silicone rubber 12 b arranged in a strip shape on the surface of the support sheet 12 a is prepared, and the concavo-convex sheet 12 is placed around the rotating base 11. You may turn. At this time, the silicone rubber 12b corresponds to the convex portion 10P. With this configuration, the protrusions 10P can be easily arranged and can be easily replaced when the protrusions 10P deteriorate.

  The material of the support sheet 12a is not particularly limited, and examples thereof include polyester such as polyethylene terephthalate, polyimide, and the like. When the fiber 131 is produced by an electrospinning method, the support sheet 12a is also preferably provided with conductivity. The thickness of the support sheet 12a is not particularly limited, and may be set as appropriate according to the material of the support sheet 12a. Examples of the silicone rubber 12b include the compounds described above.

  A heating step of heating at least one of the fiber assembly 130 and the frame 120 may be provided after the deposition step and before the transfer step. By heating the fiber assembly 130 before the transfer step, the fiber assembly 130 is transferred to the frame 120 in a softened state. Thereby, the adhesiveness of the fiber assembly 130 and the frame 120 improves. In addition, by heating the frame body 120 before the transfer process, the heat is transmitted to the fiber assembly 130 after the transfer to be softened. Thereby, the adhesiveness of the fiber assembly 130 and the frame 120 improves. Especially, the method of heating the frame 120 is preferable at the point which can suppress deterioration of the fiber 131. FIG.

  The heating method is not particularly limited, but is preferably a non-contact type in that the arrangement of the fibers 131 can be maintained. Examples of the non-contact type heating device include known devices such as a halogen lamp. The heating temperature may be appropriately set in consideration of the softening point or melting point of the fiber 131. For example, the heating temperature is adjusted so that the fiber 131 has a temperature of 80 to 140 ° C.

(3) Transfer process (FIG. 2B)
The fiber assembly 130 is transferred to the frame body 120 while rotating the winding rotary body 10.
Prior to the transfer step, the fiber assembly 130 may be cut at the planned cutting position C in a state of being wound around the winding rotary body 10. For example, the planned cutting position C is set according to the shape and size of the frame 120 (or the first opening). The fiber assembly 130 is cut in a direction along the rotation axis A, for example. The fiber assembly 130 is transferred to the frame 120 by using the cut portion as a trigger.

  The frame body 120 is placed on a stage 53 supported by, for example, the XZ stage 52 and is transported. The XZ stage 52 conveys the stage 53, that is, the frame body 120 placed on the stage 53, in a direction perpendicular to the rotation axis A of the winding rotary body 10 (X-axis direction) and an up-down direction (Z-axis direction). be able to.

  From the viewpoint of further improving productivity, it is preferable that the transfer process is performed on the plurality of frames 120 collectively or continuously. In this case, the plurality of frame bodies 120 may be arranged on the stage 53 along the Y-axis direction (direction along the rotation axis A of the winding rotary body 10) or arranged along the X-axis direction. Also good. In addition, a transfer process may be performed on an aggregate of a plurality of integrally formed frame bodies 120. In this case, the assembly of the frame bodies 120 is separated into individual frame bodies 120 after the transfer process and before the mounting process. According to this method, the fiber assembly 130 can be transferred to a plurality of frames 120 at the same time, and most of the fibers 131 accumulated on the winding rotary member 10 are used for transferring the frame 120. , Productivity further improves.

(4) Mounting process (FIG. 2 (c))
The frame body 120 to which the fiber assembly 130 has been transferred is mounted on the substrate 110 so that the first surface 120X is opposed to the frame body 120. At this time, the first bonding portion 140A and the fiber assembly 130 may be interposed between the frame body 120 and the substrate 110, and the second bonding portion 140B may be further interposed. Even when the transfer process is performed on the plurality of frame bodies 120 at once or continuously, one frame body 120 is mounted on one substrate 110.

  According to the method of the present invention, a culture medium having a fiber assembly having fibers arranged in one direction can be produced with high productivity. Furthermore, since the medium obtained by the present invention comprises fibers arranged in one direction, it is particularly useful as a medium for culturing a biological tissue or microorganism having a direction of growth.

DESCRIPTION OF SYMBOLS 10: Winding rotary body 10P: Convex part 10R: Rib 11: Rotating base 12: Concavity and convexity sheet 12a: Support sheet 12b: Silicone rubber 51: Nozzle 52: XZ stage 53: Stage 100: Medium 110: Substrate 110X: Mounting surface 120 : Frame body 120X: First surface 120Y: Second surface 121: Through hole 130: Fiber assembly 131: Fiber 132: Raw material liquid 140A: First adhesive portion 140B: Second adhesive portion

Claims (6)

A first surface, a second surface opposite to the first surface, and one or more through holes penetrating from the first surface to the second surface, the first surface having a first surface A preparation step of preparing a frame having an adhesive portion;
A deposition step of forming a fiber assembly by discharging a fiber raw material liquid from a nozzle to generate the fiber, and depositing the fiber so as to circulate on a peripheral surface of a winding rotary body,
A transfer step of transferring the fiber assembly to the first surface of the frame body through the first adhesive portion while rotating the winding rotary body;
And a mounting step of mounting the frame body, onto which the fiber assembly has been transferred, on a substrate such that the first surface faces the medium.   The method for producing a culture medium according to claim 1, wherein in the mounting step, the first surface of the frame body and the substrate are bonded via the first bonding portion. The substrate includes a second adhesive portion;
The method for producing a culture medium according to claim 1, wherein in the mounting step, the first surface of the frame body and the substrate are bonded via the second bonding portion.   The manufacturing method of the culture medium as described in any one of Claims 1-3 in which a said 1st adhesion part contains a pressure sensitive adhesive.   The manufacturing method of the culture medium of Claim 3 with which a said 2nd adhesion part contains a pressure sensitive adhesive. In the transfer step, transfer of the fiber assembly is performed collectively or continuously to the plurality of frames.
The method for producing a culture medium according to any one of claims 1 to 5, wherein in the mounting step, one frame body is mounted on one substrate.

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