JP2019024341A - Scaffold for culture - Google Patents

Abstract

To provide scaffolds for culture that align in a single direction and comprise a fiber excellent in elasticity toward the sequence direction.SOLUTION: A scaffold for culture comprises a basal plate, a first surface, a second surface to the opposite side thereof, and a plurality of through holes penetrated from the first surface to the second surface, while comprising a frame body mounted in the basal plate so as to face the first surface, and a plurality of fiber assemblies which comprise a plurality of fibers aligned in a first direction and which intervene between the basal plate and the first surface, a plurality of first openings formed in the first surface by the plurality of through holes are arranged side by side at least along the first direction, the plurality of fiber assemblies are arranged side by side along at least the first direction, and at least a part of the two fiber assemblies adjacent in the first direction are respectively exposed from the two first openings adjacent in the first direction.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a culture scaffold, and more particularly to a culture scaffold comprising fibers arranged in one direction.

  In recent years, fiber base materials have attracted attention as a culture scaffold 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 one direction and can expand and contract. 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-described arrangement and stretchability.

  One aspect of the present invention includes a substrate, a first surface, a second surface on the opposite side, and a plurality of through holes penetrating from the first surface to the second surface, A plurality of fiber assemblies including a frame body mounted on the substrate so that the first surface faces and a plurality of fibers arranged in a first direction, and interposed between the substrate and the first surface A plurality of first openings formed in the first surface by the plurality of through-holes are arranged at least along the first direction, and the plurality of fiber assemblies are , Arranged at least along the first direction, and at least a part of the two fiber assemblies adjacent in the first direction from two adjacent first openings in the first direction, respectively. The exposed scaffold for culture.

  According to the culture scaffold according to the present invention, since the fibers arranged in one direction and excellent in stretchability in the arrangement direction are provided, the growth of biological tissues and microorganisms is promoted.

1 is a perspective view schematically showing a culture scaffold according to the present invention. It is sectional drawing which shows typically the scaffold for culture | cultivation which concerns on this invention. It is a top view when the scaffold for culture | cultivation which concerns on one Embodiment of this invention sees through a board | substrate from the board | substrate side. It is a side view which shows typically the winding rotary body, the frame, a board | substrate, etc. in each process of manufacturing the scaffold for culture | cultivation which concerns on invention. It is a perspective view which shows an 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 scaffold]
The culture scaffold 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 and microorganisms are held.
An example of the culture scaffold is shown in FIG. FIG. 1 is a perspective view schematically showing a culture scaffold 100.

The culture scaffold 100 includes a substrate 110, a frame body 120 mounted on the substrate 110, and a plurality of fiber assemblies 130 (130 </ b> A and 130 </ b> B) interposed between the substrate 110 and the frame body 120. However, each fiber assembly 130 is arranged 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. Has been. The culture scaffold 100 may be housed in a holder or the like as necessary, and placed in a potential measuring device or the like.
Hereinafter, each component will be described first.

(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 (first 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. As described above, when the plurality of fibers 131 are arranged, the fibers 131 are likely to extend along the first direction _DF , so that stress on a biological tissue or a microorganism is reduced. Therefore, it becomes easy for biological tissues and microorganisms to grow along the first direction _DF .

  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. 6 is a schematic top view of the fiber assembly for explaining the arrangement of the fibers. In FIG. 6, 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. 6) connecting a point where a certain fiber 131 intersects the two opposite sides is 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. 6) is obtained. Another two fibers 131 are selected, and an angle (for example, θ2 in FIG. 6) 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 scaffold for culturing biological tissue or microorganisms. 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 two or more through holes 121 (121A, 121B) 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 121a, see FIG. 2) 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 121a of the through hole 121 is blocked by the substrate 110 via the fiber assembly 130, and at least one depression is formed on the mounting surface 110X of the substrate 110. Is done. 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.

  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 the through holes 121 is not particularly limited as long as it is 2 or more, and may be set as appropriate according to the size and use of the frame body 120. However, the plurality of through holes 121 (first openings 121a) are arranged side by side in a direction along at least the first direction _DF . This is because, for example, the angle (provided that ≦ 90 °) between a straight line formed by connecting the centers of two adjacent first openings 121a and the first direction _DF is 0 ° or more and 30 ° or less. Say. The frame body 120 only needs to have at least two first openings 121a satisfying such a relationship. A plurality of first openings 121a may be arranged in a direction intersecting with the first direction _DF as in the illustrated example. In other words, the first opening 121a in which the angle (provided that ≦ 90 °) between the straight line formed by connecting the centers of the two adjacent first openings 121a and the first direction _DF is 60 ° or more and 90 ° or less. There may be.

  The shape and size of the first opening 121a 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 121a 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 121a 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 the substrate 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)
It is preferable that the substrate 110, the frame body 120, and the fiber assembly 130 are bonded via the bonding portion 140. Since the fiber aggregate 130 is an aggregate of fibers 131 arranged in one direction, for example, when an adhesive is applied to the frame body 120, the fiber aggregate 130 penetrates between the fibers 131 and penetrates to the substrate 110 side, An adhesive portion 140 that adheres the substrate 110 is formed. Similarly, when an adhesive is applied to the substrate 110, the adhesive portion 140 penetrates to the frame body 120 side and bonds the frame body 120 and the substrate 110. In any case, a part of the fiber assembly 130 is held so as to be embedded in the bonding portion 140.

  In the illustrated example, for the sake of convenience, the adhesive portion 140 is sandwiched between the fiber assembly 130 and the adhesive portion on the frame 120 side (first adhesive portion 140A) and the adhesive portion on the substrate 110 side (second adhesive portion). 140B). The first adhesive portion 140A and the second adhesive portion 140B may face each other or may partially face each other.

  The material (adhesive) of the adhesion part 140 is not specifically limited, For example, a pressure sensitive adhesive, a hot-melt-type adhesive, a curable adhesive, etc. are mentioned. Among these, a pressure sensitive adhesive is preferable. This is because the pressure-sensitive adhesive can be easily deformed or moved by an external load. The bonding portion 140 including the pressure-sensitive adhesive is deformed (stretched) or moved so as to follow the expansion and contraction of the fibers 131.

  The pressure-sensitive adhesive is applied to the frame body 120 or the substrate 110, and adheres the frame body 120, the fiber assembly 130, and the substrate 110 by 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 mass per unit area of the adhesion part 140 (140A, 140B) is not specifically 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 .

  Hereinafter, a configuration example of the culture scaffold 100 according to the present embodiment will be described with reference to the drawings. However, the structure of the culture scaffold 100 is not limited to this. FIG. 3 is a plan view of the culture scaffold 100 as seen through the substrate 110 from the substrate 110 side. In FIG. 3, the bonding portion 140 is hatched for convenience.

  Biological tissues or microorganisms grow while expanding and contracting in the fiber arrangement direction. At this time, if the stretchability of the fibers in the arrangement direction is not sufficient, the growth of biological tissues or microorganisms is hindered, and furthermore, the fibers cannot break the stretch of biological tissues or microorganisms, and the fibers may break. is there. In particular, when one fiber assembly covers a plurality of first openings arranged in the fiber arrangement direction in a lump, growth of biological tissue or microorganisms is likely to be hindered.

  A plurality of first openings are formed in one frame. However, each of the plurality of first openings independently forms a scaffold for biological tissue or microorganisms. Therefore, when a biological tissue or microorganism grows, the fibers exposed from the plurality of first openings (exposed fibers) try to expand and contract independently. On the other hand, the fibers constituting the fiber assembly that collectively covers the plurality of first openings arranged in the fiber arrangement direction are continuous between the plurality of first openings. For this reason, independent expansion and contraction of the exposed fiber is likely to be hindered.

Therefore, (in the illustrated example, the first fiber assembly 130A and the second fiber aggregates 130B) a plurality of fiber assemblies respectively corresponding to at least a plurality arranged in a first direction _{D F} first opening 121Aa and 121Ba Arrange to do. Thereby, in each 1st opening 121a, it becomes possible for an exposed fiber to expand-contract independently in the 1st direction _DF , and the growth of a biological tissue or microorganisms is accelerated | stimulated.

The first fiber assembly 130A and the second fiber assembly 130B are arranged side by side along the first direction _DF so that the fibers 131 forming each fiber assembly 130 are arranged in the same direction. The That is, the first direction D one end of _F and intersecting direction and one end portion in a direction intersecting the first direction D _F of the second fiber aggregates 130B of the first fiber assembly 130A, first The first openings (121Aa and 121Ba) adjacent to each other in the direction _DF are relative to each other.

The plurality of fiber assemblies 130 are arranged side by side along the first direction _DF , for example, an angle formed between each of the opposed end portions and the first direction _DF (however, ≦ 90 °) is 60 ° or more and 90 ° or less. Each fiber assembly 130 may collectively cover a plurality of first openings 121a arranged in a direction intersecting the first direction _DF , as in the illustrated example.

  Such first fiber assembly 130A and second fiber assembly 130B are, for example, a base material including an assembly of fibers 131 corresponding to a plurality of fiber assemblies 130 (hereinafter simply referred to as base material M130). Is formed on the peripheral surface of the winding rotary body 10 and then cut at a predetermined position before or after being transferred to the frame body 120. Alternatively, a plurality of fiber assemblies 130 formed as separate bodies from the beginning may be arranged at predetermined positions on the substrate 110 or the frame body 120.

Adhesive 140, between the first opening 121Aa and 121Ba which are adjacent in the first direction _{D F,} it is preferably arranged in a line in a direction (second direction) crossing the first direction _{D F.} Furthermore, it is preferable that the bonding portion 140 is arranged in a line shape in the vicinity of the end portions of the fiber assemblies 130A and 130B as in the illustrated example. Thereby, the board | substrate 110, the fiber assembly 130, and the frame 120 can be adhere | attached, without impairing the elasticity of the exposed fiber exposed from each 1st opening 121a of the fiber 131. FIG. The second direction is, for example, a direction in which an angle formed with the first direction _DF (however, ≦ 90 °) is 60 ° or more and 90 ° or less. The arrangement direction of the line-shaped adhesive portion 140 is the direction of the center line in the longitudinal direction of the adhesive portion 140. When the center line includes a curve, the arrangement direction of the line-shaped adhesive portion 140 may be obtained by taking an approximate straight line of the center line when viewed from the normal direction of the main surface of the frame 120.

[Method for producing culture scaffold]
Hereinafter, a method for producing the culture scaffold of the present embodiment will be described with reference to the drawings. Here, as an example, a scaffold for culturing is manufactured by forming the base material M130 on the peripheral surface of the winding rotary body 10 and then cutting it at a predetermined position before or after transferring it to the frame body 120. I will give you. However, the manufacturing method of the culture scaffold is not limited to this. FIGS. 4A to 4C are side views schematically showing the winding rotary body 10, the frame body 120, the substrate 110, and the like in each step of manufacturing the culture scaffold of the present embodiment.

  The culture scaffold according to the present embodiment includes, for example, a first surface 120X, a second surface 120Y opposite to the first surface 120X, and one or more through holes penetrating from the first surface 120X to the second surface 120Y. 121, and preparing the 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, The fiber 131 is deposited so as to circulate on the peripheral surface of the winding rotary body 10 to form the base material M130, and the base material M130 is attached to the first of the frame body 120 while the winding rotary body 10 is rotated. A transfer step of transferring to the surface 120X via the first adhesive portion 140A and the transferred base material M130 in a region not facing the first opening 121a formed by the through hole 121. On the axis of rotation A The fiber assembly forming step of forming a plurality of fiber assemblies 130 by cutting in the opposite direction and the frame 120 including the plurality of fiber assemblies 130 are placed on the substrate 110 so that the first surface 120X faces each other. And a mounting process for mounting.

  Alternatively, the culture scaffold according to the present embodiment includes, for example, the first surface 120X, the second surface 120Y on the opposite side, and one or more penetrating from the first surface 120X to the second surface 120Y. A preparation step of preparing a frame 120 having a first adhesive portion 140A on the first surface 120X, and a raw material liquid of the fiber 131 is discharged from the nozzle 51 to generate the fiber 131. At the same time, the fiber 131 is deposited so as to circulate on the circumferential surface of the winding rotary body 10 to form a base material M130, and the base material M130 deposited on the circumferential surface of the winding rotary body 10 Cutting at a position in a direction along the rotation axis A of the winding rotary body 10 to form a plurality of fiber assemblies 130, while rotating the winding rotary body 10, the plurality of fiber assemblies 130 in turn, On the first surface 120X of the frame 120, A transfer step of transferring via one adhesive portion 140A, and a mounting step of mounting the frame body 120 including the plurality of fiber assemblies 130 on the substrate 110 so that the first surface 120X faces each other. Manufactured by the second method.

(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 predetermined positions other than the 1st opening 121a of the 1st surface 120X by printing, a dispenser, etc., for example.

The first bonding portion 140A is, for example, between the first openings 121a adjacent to each other in the first direction _DF of the frame body 120 (for example, between the first openings 121Aa and the first openings 121Ba). It is formed in a line shape in the direction intersecting with _F. Two first adhesive portions 140A may be formed in a line shape in the direction along the rotation axis A between the first openings 121a adjacent to the first direction _DF . This aspect is particularly suitable when the culture scaffold 100 is manufactured by the first method of cutting the base material M130 after the transfer step. Since the base material M130 can be cut | disconnected by the part corresponding between the two 1st adhesion parts 140A formed in the line form, cutting | disconnection operation | work becomes easy.

(2) Deposition process (FIG. 4 (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 base material M130 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 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 fiber 131 is 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 base material M130 that covers at least a part of the peripheral surface of the winding rotary body 10 and includes fibers 131 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. 5 is a perspective view of the winding rotary body 10.
The configuration of the winding rotary body 10 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. 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 (first direction D _F ) around the circumferential surface of the winding rotator 10. The first direction _DF is, for example, a direction along the rotation direction of the winding rotator 10 (that is, the direction perpendicular to the rotation axis of the winding rotator 10). The angle θ _F (where θ _F ≦ 90 °) formed by the first direction D _F and the rotation axis may be, for example, 60 ° or more and 90 ° or less. The first direction _DF is the longitudinal direction of the fiber 131 when the fiber 131 is 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. The angle θ _F is an average value of angles formed between the first direction D _F of the plurality of fibers 131 and the rotation axis. The first directions _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 of the winding rotator 10 may be disposed on the circumferential surface of the winding rotator 10. Thereby, the aggregate (base material M130) of the fibers 131 arranged so as to circulate around the circumferential surface of the winding rotary body 10 is easily separated from the winding rotary body 10. As a result, the fibers 131 can be easily transferred to the frame body 120 while maintaining the arrangement. The ends of the plurality of convex portions 10P may be connected by ribs 10R that extend in a direction intersecting the rotation axis.

  The shape of the convex portion 10P is not particularly limited as long as it is a belt shape. The band shape is a shape in which the length of the protruding portion 10P in the extending direction is longer than the length in the direction perpendicular to the extending direction. 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. Among these, from the viewpoint of the peelability of the base material M130 (fiber assembly 130), three or more are preferably arranged on the peripheral surface of the winding rotary body 10, and ten or more are preferably arranged. Further, from the same viewpoint, it is preferable that the convex portions 10P are arranged at equal intervals.

  The length (width) in the short direction of the convex portion 10P is not particularly limited. In particular, from the viewpoint of the peelability of the base material M130 (fiber assembly 130), the total area of the protrusions 10P that contacts the circumferential surface of the winding rotator 10 is the surface area of the circumferential surface of the winding rotator 10. It is preferable to determine the width of each convex portion 10P so that it is 10% or more and 80% or less, particularly 30% or more and 70% or less. The length in the extending direction 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 the peelability of the base material M130 (fiber assembly 130) and the suppression of the 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 base material M130 (fiber assembly 130) is further improved. On the other hand, since the silicone rubber has appropriate adhesiveness, the base material M130 (fiber assembly 130) is prevented from peeling off 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.

  A heating step of heating at least one of the fibers 131 and the frame 120 may be provided after the deposition step and before the transfer step. Thereby, the adhesiveness of the base material M130 (fiber assembly 130) and the frame 120 is improved. 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) Fiber assembly formation process The base material M130 is cut | disconnected in the direction in alignment with the rotating shaft A of the winding rotary body 10, and the several fiber assembly 130 is formed.
The fiber assembly forming process may be performed after the transfer process and before the mounting process. In this case, the base material M130 is cut at the planned cutting position C in a state where it is transferred to the frame body 120. Alternatively, the fiber assembly forming process may be performed after the deposition process and before the transfer process. In this case, the base material M <b> 130 is cut at the planned cutting location C while being wound around the winding rotary body 10. In any case, the planned cutting location C is set according to the length of the first opening 121a in the first direction _DF .

These methods, a plurality of fiber assemblies 130 arranged in the first direction D _F, 1 single together are arranged to the frame body 120 a plurality of first opening 121a arranged in the first direction D _F so as to cover each , Can be arranged in one step.

(4) Transfer process (FIG. 4B)
The base material M130 or the plurality of fiber assemblies 130 are transferred to the frame body 120 while rotating the winding rotary body 10. Accordingly, the plurality of fibers 131 are transferred to 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 131 should just be arrange | positioned so that it may be exposed from the 1st opening 121a. Therefore, by transferring the base material M130 or the plurality of fiber aggregates 130 to the frame body 120 instead of the substrate 110, the fibers 131 can be arranged only in necessary portions without performing precise alignment or the like. Productivity.

  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 base material M130 or the plurality of fiber assemblies 130 can be collectively transferred to the plurality of frame bodies 120, and most of the fibers 131 accumulated on the winding rotary body 10 are formed on the frame body 120. Since it is used for transfer, productivity is further improved.

(5) Mounting process (FIG. 4C)
The frame body 120 on which the fiber assembly 130 is arranged is mounted on the substrate 110 so that the first surface 120X faces the first body 120X. 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.

  Since the culture scaffold of the present invention comprises fibers that are arranged in one direction and excellent in stretchability in the arrangement direction, in particular, as a culture scaffold for culturing a biological tissue or microorganism that has a direction of growth. Useful.

DESCRIPTION OF SYMBOLS 10: Winding rotary body 10P: Convex part 10R: Rib 51: Nozzle 52: XZ stage 53: Stage 100: Culture scaffold 110: Board | substrate 110X: Mounting surface 120: Frame 120X: 1st surface 120Y: 2nd Surface 121, 121A, 121B: Through-hole 121a, 121Aa, 121Ba: First opening 130: Fiber aggregate 130A: First fiber aggregate 130B: Second fiber aggregate M130: Base material 131: Fiber 132: Raw material liquid 140: Adhesive part 140A: First adhesive part 140B: Second adhesive part

Claims (3)

A substrate,
A first surface, a second surface opposite to the first surface, and a plurality of through holes penetrating from the first surface to the second surface, the first surface facing each other. A frame mounted on the substrate;
A plurality of fiber assemblies including a plurality of fibers arranged in a first direction and interposed between the substrate and the first surface;
The plurality of first openings formed in the first surface by the plurality of through holes are arranged side by side along at least the first direction,
The plurality of fiber assemblies are arranged side by side along at least the first direction,
The culture scaffold, wherein at least a part of the two fiber assemblies adjacent in the first direction are exposed from the two first openings adjacent in the first direction.   The culture scaffold according to claim 1, further comprising a bonding portion that includes a pressure-sensitive adhesive and bonds the substrate, the fiber assembly, and the frame. The culture scaffold according to claim 2, wherein the adhesive portion is disposed in a line shape in a second direction intersecting the first direction between the first openings adjacent in the first direction.

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