Artificial three-dimensional tissue manufacturing device, artificial three-dimensional tissue, artificial three-dimensional tissue model, and wound repair model

Abstract

To provide apparatus for manufacturing artificial three-dimensional tissues that can readily manufacture artificial three-dimensional tissues extending in a predetermined direction, without arranging anchor potions oppositely on both sides in the contraction direction.SOLUTION: Provided is an apparatus for manufacturing an artificial three-dimensional tissue in which at least a part of the tissue protrudes in a predetermined direction, the apparatus having: a protruding portion having at least a protrusion protruding to one side in a predetermined direction; and a plurality of anchors located on both sides of the protrusion in the radial direction orthogonal to the predetermined direction and facing the other side in the predetermined direction, while located on the other side of the protrusion in a predetermined direction.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to an artificial three-dimensional tissue manufacturing apparatus, an artificial three-dimensional tissue, an artificial three-dimensional tissue model, and a wound repair model. This application claims priority to U.S. Provisional Patent Application No. 62 / 935,690, filed November 15, 2019, the contents of which are incorporated herein by reference. [Background technology]

[0002] There is active research being conducted on creating small, flat artificial skin models from cells and reproducing various skin components such as blood vessels and the stratum corneum in order to use them for wound covering treatments and as screening models for drug discovery.

[0003] Patent Document 1 discloses a method for producing tubular artificial skin tissue in which a dermal tissue layer and an epidermal tissue layer are laminated in the radial direction by using a flow path forming member suspended in the culture space of a culture tank and anchor parts provided opposite each other on a side wall coaxially facing the flow path forming member, and engaging a part of the artificial three-dimensional tissue with the anchor parts to suppress contraction in the luminal direction. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2016 / 047230 Summary of the Invention [Problem to be solved by the invention]

[0005] In the above-mentioned Patent Document 1, the anchor parts are provided opposite to the contraction direction of the artificial three-dimensional tissue, so that it is suitable for manufacturing tubular artificial three-dimensional tissue in which the anchor parts extend in opposite directions and both ends are open. However, for example, in a skin model, there is currently no effective means for manufacturing an artificial three-dimensional tissue that covers a three-dimensional object with one end closed, such as a finger, the entire hand, a prosthetic hand, or a robot, while suppressing contraction. In addition, for example, a method of wrapping a planar artificial three-dimensional tissue around a mold and then sewing it together is also possible, but this is not suitable for covering a complex three-dimensional object.

[0006] The present invention has been made in consideration of the above points, and aims to provide an artificial three-dimensional tissue manufacturing device, an artificial three-dimensional tissue, an artificial three-dimensional tissue model, and a wound repair model that can easily manufacture artificial three-dimensional tissue extending in a predetermined direction without arranging anchor portions opposite each other in the contraction direction. [Means for solving the problem]

[0007] According to a first aspect of the present invention, there is provided an apparatus for manufacturing artificial three-dimensional tissue, at least a portion of which protrudes in a predetermined direction, the apparatus having a protrusion portion having at least a protrusion protruding on one side of the predetermined direction, and a plurality of anchor portions located on the other side of the protrusion in the predetermined direction, located on both sides of the protrusion in a radial direction perpendicular to the predetermined direction, and facing the other side of the predetermined direction.

[0008] According to a second aspect of the present invention, there is provided an artificial three-dimensional tissue having a bulge portion bulging out to one side in a predetermined direction, and a side portion extending from the bulge portion to the other side in the predetermined direction and formed all around an axis extending in the predetermined direction, and having an anchor mark on a part of the side portion.

[0009] According to a third aspect of the present invention, there is provided an artificial three-dimensional tissue model having artificial three-dimensional tissue, at least a portion of which protrudes in a predetermined direction, the artificial three-dimensional tissue model comprising: a protrusion portion having at least a protrusion protruding on one side of the predetermined direction; a plurality of anchor portions located on the other side of the protrusion in the predetermined direction and located on both sides of the protrusion in a radial direction perpendicular to the predetermined direction, facing the other side of the predetermined direction; and an artificial three-dimensional tissue whose end portion is folded back by the protrusion to cover the protrusion portion and engages with the anchor portions.

[0010] According to a fourth aspect of the present invention, there is provided a wound repair model having an artificial three-dimensional tissue, at least a portion of which protrudes in a predetermined direction, comprising: a protruding portion having at least a protrusion protruding on one side of the predetermined direction; a plurality of anchor portions located on the other side of the protrusion in the predetermined direction and located on both sides of the protrusion in a radial direction perpendicular to the predetermined direction, facing the other side of the predetermined direction; an artificial three-dimensional tissue having a wound portion, an end portion folded back by the protrusion to cover the protruding portion and engaged with the anchor portion; and an extracellular matrix attached to the wound portion to repair the wound portion. Effect of the Invention

[0011] In the present invention, in the artificial three-dimensional tissue manufacturing apparatus and wound repair model, an artificial three-dimensional tissue extending in a predetermined direction can be easily manufactured without arranging anchor parts opposite each other on both sides in the contraction direction. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 is a diagram showing an embodiment of the present invention, and is a cross-sectional view of a manufacturing apparatus 1 for artificial three-dimensional tissue according to a first embodiment. [Diagram 2] FIG. 2 is an external perspective view of the mold 10 and the annular member 20. [Diagram 3] 1 is a cross-sectional view of the manufacturing apparatus 1 in which a solution of extracellular matrix M containing cells C is filled into a culture space 33. FIG. [Figure 4]FIG. 2 is a cross-sectional view of a manufacturing apparatus 1 in which an artificial three-dimensional tissue T is formed after culturing a solution of extracellular matrix M containing cells C for a predetermined period of time. [Diagram 5] FIG. 2 is a photograph of dermal tissue produced by the production apparatus 1. [Figure 6] FIG. 13 is a photograph of dermal tissue produced without using the anchor structure 22 in the production apparatus 1. [Figure 7] FIG. 13 is a photograph of dermal tissue produced by the production apparatus 1 without using the anchor structure 22 and the cap portion 40. [Figure 8] FIG. 1 is a diagram showing an embodiment of the present invention, being a cross-sectional view of a manufacturing apparatus 1 for artificial three-dimensional tissue according to a second embodiment. [Figure 9] FIG. 11 is an external perspective view of a mold 10 according to a second embodiment. [Figure 10] 1 is a cross-sectional view of the manufacturing apparatus 1 in which a solution of extracellular matrix M containing cells C is filled into a culture space 33. FIG. [Figure 11] FIG. 13 is a cross-sectional view of a bag-shaped artificial dermis tissue T that covers a model of three-jointed finger-shaped divided bodies 14A, 14B, and 14C. [Figure 12] 11 is a cross-sectional view showing the movement of the dermis tissue T swinging between an upright position and an inclined position. FIG. [Figure 13] FIG. 2 is a photograph showing dermal tissue T in an upright position. [Figure 14] FIG. 13 is a photograph showing dermal tissue T in a tilted position. [Figure 15] FIG. 11 is a cross-sectional view of an apparatus 1 for producing an artificial three-dimensional tissue according to a third embodiment. [Figure 16] FIG. 11 is a cross-sectional view of an apparatus 1 for producing an artificial three-dimensional tissue according to a third embodiment. [Figure 17] FIG. 11 is a cross-sectional view of an apparatus 1 for producing an artificial three-dimensional tissue according to a fourth embodiment. [Figure 18] 18 is a cross-sectional view taken at a center position of the flow passage 18 in FIG. 17 and perpendicular to the direction of the axis J. [Figure 19] 1 is a cross-sectional view taken along a plane including the axis J of the recess 50. FIG. [Figure 20]FIG. 11 is a cross-sectional view of an apparatus 1 for producing an artificial three-dimensional tissue according to a fourth embodiment. [Figure 21] FIG. 11 is a cross-sectional view of an apparatus 1 for producing an artificial three-dimensional tissue according to a fifth embodiment. [Figure 22] FIG. 11 is a cross-sectional view of an apparatus 1 for producing an artificial three-dimensional tissue according to a sixth embodiment. [Diagram 23] FIG. 13 is a stained photograph of the collagen sheet on day 3 after transplantation. [Figure 24] FIG. 13 is a stained photograph of the collagen sheet on day 7 after transplantation. [Diagram 25] FIG. 1 is a diagram showing the relationship between the tensile length and the tensile force of Samples A to C in a tensile test. [Figure 26] FIG. 2 is a perspective view of the appearance of dermal tissue produced using a three-joint finger model. [Figure 27] FIG. 1 is a photograph of a three-jointed finger model in an upright position. [Figure 28] FIG. 1 is a photograph of a three-jointed finger model in a reclined position. [Figure 29] FIG. 2 is an external perspective view of an auxiliary mold 10A. [Diagram 30] 13 is a cross-sectional view perpendicular to the axis J of the holder tilted so that the opening of the recess 82 faces upward. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Hereinafter, an embodiment of the manufacturing apparatus for artificial three-dimensional tissue and the wound repair model of the present invention will be described with reference to Figs. In this embodiment, an example of producing artificial dermal tissue as an artificial three-dimensional tissue will be described. The following embodiment shows one aspect of the present invention, does not limit the present invention, and can be arbitrarily modified within the scope of the technical idea of ​​the present invention. In addition, in the following drawings, the scale and number of each structure are different from the actual structure in order to make each configuration easier to understand.

[0014] [First embodiment] FIG. 1 is a cross-sectional view of a manufacturing apparatus 1 for an artificial three-dimensional tissue according to the first embodiment (hereinafter, referred to as the manufacturing apparatus 1). As shown in FIG. 1, the manufacturing apparatus 1 includes a mold 10, an annular member 20, a case 30, and a cap portion 40.

[0015] The mold 10 forms the inner surface of a bag-shaped artificial three-dimensional tissue with a closed tip (one end). The mold 10 has a mold body (shaft-shaped portion) 11 and a support base 16. The mold body 11 is cylindrical and extends along an axis J that extends in the vertical direction. The mold body 11 has a protrusion 12 at its upper end. The protrusion 12 in this embodiment is in the shape of a hemispherical dome.

[0016] The support base 16 is provided at the lower end of the mold body 11. The support base 16 is formed in a disk shape that is coaxial with the mold body 11 and has a diameter larger than that of the mold body 11. The mold 10 is manufactured using, for example, a lithography method, injection molding, a 3D printer, or the like. The material of the mold 10 is not particularly limited as long as it does not adversely affect the artificial three-dimensional tissue, and an appropriate material can be used depending on the selected manufacturing method.

[0017] In the following description, the direction in which the axis J extends will be referred to as the up-down direction (predetermined direction), and the horizontal direction perpendicular to the axis J will be referred to as the radial direction. Furthermore, the up-down direction, upper side (one side of the predetermined direction), lower side (the other side of the predetermined direction), and horizontal direction are names simply used to describe the relative positions of the various parts, and the actual relative positions or directions of use may be other than those indicated by these names.

[0018] The annular member 20 is formed in a circular ring shape. The outer diameter of the annular member 20 is approximately the same as the outer diameter of the support base 16. The annular member 20 has a through hole 21 and an anchor structure 22. The diameter of the through hole 21 is approximately the same as the diameter of the mold body 11. FIG. 2 is an external perspective view of the mold 10 and the annular member 20. As shown in FIG. 2, the annular member 20 is provided in the through hole 21 so as to be freely insertable (insertable and detachable) into the mold body 11 from above.

[0019] As shown in Fig. 1, a seal member 13 (not shown in Fig. 2) provided on the outer peripheral surface of the mold body 11 comes into contact with the through hole 21, thereby liquid-tightly sealing the gap between the mold body 11 and the annular member 20. The annular member 20 is inserted into the mold body 11 from above in the through hole 21, and its underside comes into contact with the upper surface of the support base 16. When the underside of the annular member 20 comes into contact with the upper surface of the support base 16, the part of the mold body 11 that protrudes upward from the annular member 20 is a protruding portion 14 that forms the inner surface of the artificial three-dimensional tissue.

[0020] The anchor structure 22 is disposed on the upper surface of the annular member 20 near the mold body 11. As shown in FIG. 2, a plurality of anchor structures 22 (eight in FIG. 2) are provided at intervals in the circumferential direction centered on the axis J. Each anchor structure 22 has a column portion 23 and a flange portion 24. The column portion 23 is columnar and extends upward from the upper surface of the annular member 20. The flange portion 24 is disk-shaped and extends radially outward from the tip of the column portion 23 over the entire circumference. The outer diameter of the flange portion 24 is larger than the outer diameter of the column portion 23. The side surface of the flange portion 24 may be in contact with the protruding portion 14. The lower surface of the flange portion 24 is an anchor portion 25 that engages with the artificial three-dimensional tissue from above. The anchor portion 25 is located below the protrusion 12. The anchor portions 25 are located on both sides of the protrusion 12 in the radial direction.

[0021] The annular member 20 and the anchor structure 22 are manufactured, for example, by lithography, injection molding, a 3D printer, or the like, in the same manner as the mold 10. The material of the annular member 20 and the anchor structure 22 is not particularly limited as long as it does not adversely affect the above-mentioned artificial three-dimensional tissue, and an appropriate material can be used depending on the selected manufacturing method.

[0022] The annular member 20, whose lower surface is in contact with the upper surface of the support base 16, the anchor structure 22 and the mold 10 constitute a holder for holding the artificial three-dimensional tissue.

[0023] The case 30 has an annular member 20, a bottom plate portion 31, and a tubular portion 32. The bottom plate portion 31 is disk-shaped and located below the support base 16. The tubular portion 32 is cylindrical and extends upward from the periphery of the bottom plate portion 31. The tubular portion 32 fits liquid-tightly into the outer circumferential surface of the support base 16 and the outer circumferential surface of the annular member 20. The tubular portion 32 surrounds the mold 10 (projection portion 14), the annular member 20, and the radial outside of the anchor structure 22 to form a culture space 33 for an artificial three-dimensional tissue. The culture space 33 is formed above the annular member 20.

[0024] The case 30 is preferably made of a material with high oxygen permeability. In this embodiment, the case 30 is made of PDMS (polydimethylsiloxane). Although oxygen is consumed during cell culture, by forming the case 30 from PDMS with high oxygen permeability, oxygen can be supplied through the case 30, and an artificial three-dimensional tissue can be formed without causing oxygen shortage.

[0025] The cap portion 40 is detachably attached to the upper end side of the tube portion 32. The cap portion 40 has a molding surface 41 and a supply port 42. The molding surface 41 is provided facing the lower culture space 33. The molding surface 41 molds the upper side of the artificial three-dimensional tissue. Specifically, the molding surface 41 has a cross-sectional contour such that the shape of the outer diameter side of the culture space 33 is offset by a certain amount from the outer contour of the protrusion portion 14, and the width of the culture space (i.e., the thickness of the artificial three-dimensional tissue) is constant. The supply port 42 penetrates the cap portion 40 coaxially with the axis J.

[0026] Next, a method for producing an artificial three-dimensional tissue using the above-mentioned production apparatus 1 will be described with reference to Fig. 3 to Fig. 7. Fig. 3 is a cross-sectional view of the production apparatus 1 in which a solution of extracellular matrix M containing cells C is filled into the culture space 33. Fig. 4 is a cross-sectional view of the production apparatus 1 in which an artificial three-dimensional tissue T is formed after culturing the solution of extracellular matrix M containing cells C for a predetermined period of time.

[0027] The method for producing an artificial three-dimensional tissue of this embodiment includes preparing the above-mentioned production apparatus 1, filling the culture space 33 in the production apparatus 1 with a solution of extracellular matrix M containing cells C through the supply port 42, and culturing the extracellular matrix M containing cells C for a predetermined period of time to form an artificial three-dimensional tissue T. Thereafter, by removing the cap portion 40 and the case 30 in the production apparatus 1, a holder is obtained that is an artificial three-dimensional tissue model in which the upper side covers the protrusion 12 and is folded back to cover the outer circumferential surface of the protruding portion 14, and the lower opening side is engaged with and fixed to the anchor portion 25 of the anchor structure 22 from below, and the bag-shaped artificial three-dimensional tissue T formed in a U-shape in cross section is held. That is, the artificial three-dimensional tissue T has a bulging portion 91 that bulges upward and a side portion 92 that extends downward from the bulging portion 91 and is formed around the entire axis J, and when the artificial three-dimensional tissue T is removed from the manufacturing apparatus 1, a bag-shaped artificial three-dimensional tissue T is formed having an anchor mark 93 at the end (part) of the side portion 92.

[0028] In this embodiment, an example will be described in which dermal cells (hereinafter, appropriately referred to as dermal cells C) are used as cells C, collagen (hereinafter, appropriately referred to as collagen M) is used as extracellular matrix M, and dermal tissue (hereinafter, appropriately referred to as dermal tissue T) is formed as artificial three-dimensional tissue T.

[0029] As the dermal cells C, for example, fibroblasts can be used. Fibroblasts include cells derived from mammals such as humans, mice, and rats, and are preferably derived from humans. Human fibroblasts include normal human dermal fibroblasts (NHDF), human pulmonary fibroblasts (HPF), human cardiac fibroblasts (HCF), human aortic adventitial fibroblasts (HAoAF), human uterine fibroblasts (HUF), and human villous mesenchymal fibroblasts (HVMF), and are preferably derived from humans.

[0030] Examples of collagen include, but are not limited to, collagen (type I, type II, type III, type V, type XI, etc.) and mouse EHS tumor extract (type IV collagen). Examples of extracellular matrix M other than collagen include basement membrane components (product name: Matrigel) reconstituted from mouse EHS tumor extract (containing laminin, heparan sulfate proteoglycan, etc.), gelatin, agar, agarose, fibrin, glycosaminoglycan, hyaluronic acid, proteoglycan, etc.

[0031] (Preparation of manufacturing equipment 1) As shown in FIG. 2, the preparation of the manufacturing apparatus 1 is performed by first inserting the through hole 21 of the annular member 20 into the mold body 11 from above, and bringing the lower surface of the annular member 20 into contact with the upper surface of the support base 16. Next, the assembled annular member 20 and the mold body 11 are inserted into the inside of the cylindrical portion 32 of the case 30 from above, and bringing the lower surface of the support base 16 into contact with the upper surface of the bottom plate portion 31. Alternatively, the mold body 11 alone may be inserted into the inside of the cylindrical portion 32 of the case 30 from above, bringing the lower surface of the support base 16 into contact with the upper surface of the bottom plate portion 31, and then inserting the through hole 21 of the annular member 20 into the mold body 11 from above. Next, the cap portion 40 is attached to the upper end side of the cylindrical portion 32. The position of the cap portion 40 in the direction of the axis J is a position where the distance between the molding surface 41 and the protrusion 12 is approximately the same as the distance between the mold body 11 and the inner surface of the cylindrical portion 32. This completes the preparation of the manufacturing apparatus 1 having the culture space 33.

[0032] (Solution filling of extracellular matrix containing cells) When preparation of the manufacturing apparatus 1 is completed, the culture space 33 is filled with a solution of collagen M containing dermal cells C. As the solution of collagen M containing dermal cells C, a solution containing the above-mentioned NHDF at a concentration of 4.0×106 cells / ml was used. The solution of collagen M containing dermal cells C is injected into the culture space 33 via the supply port 42 using a syringe, pipette, or the like, as shown in FIG.

[0033] (Formation of dermal tissue T) Once the mixture of dermal cells C and collagen M is injected into the culture space 33, it is cultured under predetermined conditions. The culture conditions are such that the density of the dermal tissue T is equivalent to that of human dermis. As an example, the culture is performed at a temperature of 37°C for 4 days (96 hours). After about one day of culture, the collagen M solution gels, so the cap portion 40 may be detached from the tube portion 32.

[0034] As a result of the above-mentioned culture, the collagen M solution gels and is contracted by the dermal cells C, forming dermal tissue T covering the protrusion 14, as shown in Fig. 4. The gel contracted radially when the dermal tissue T is formed is held on the outer circumferential surface of the protrusion 14. The upper gel contracted in the direction of axis J engages with the projection 12 from above and is held there. The lower gel contracted in the direction of axis J engages with the anchor portion 25 from below and is held there. Therefore, the artificial dermal tissue T produced by culturing using this manufacturing apparatus 1 has a three-dimensional structure that protrudes upward, and is formed into a bag-like shape that is closed at the top and open at the bottom when subjected to gravity in the shear direction.

[0035] Fig. 5 is a photograph of dermal tissue produced by the above-mentioned production apparatus 1. Fig. 6 is a photograph of dermal tissue produced by the above-mentioned production apparatus 1 without using the anchor structure 22 (anchor portion 25). Fig. 7 is a photograph of dermal tissue produced by the above-mentioned production apparatus 1 without using the anchor structure 22 (anchor portion 25) and the cap portion 40.

[0036] The dermal tissue shown in FIG. 5 was formed without rolling up because the lower side was engaged and held by the anchor portion 25. The dermal tissue shown in FIG. 5 was formed with a hemispherical (dome-shaped) upper end along the protrusion 12. The dermal tissue shown in FIG. 6 was formed with a hemispherical upper end along the protrusion 12, but the lower side of the free end not having the anchor portion rolled up. The dermal tissue shown in FIG. 7 was formed without rolling up because the lower side was engaged and held by the anchor portion 25, but the gel was not formed by the molding surface 41 of the cap portion 40, and the gel spread more in the radial direction than the lower side above the position of the protrusion 12, where the gel is thicker than the radial thickness below the protrusion 12, and the shape did not follow the outer shape of the protruding portion 14.

[0037] As described above, in the manufacturing apparatus 1 of this embodiment, by providing the anchor parts 25 located below the protrusions 12 and located on both sides of the protrusions 12 in the radial direction, the upper side of the dermal tissue T is restrained from contracting downward by the protrusions 12, and the lower side of the dermal tissue T is restrained from contracting upward by the anchor parts 25 arranged on both sides of the protrusions 12 in the radial direction. Therefore, in this embodiment, the dermal tissue T having a three-dimensional structure in which the upper side is covered with a hemisphere along the protrusions 12 without the lower side being rolled up can be formed without providing the anchor parts 25 facing each other in the vertical direction. In addition, in the manufacturing apparatus 1 of this embodiment, by using the case 30 and the cap part 40, it is possible to easily form the dermal tissue T having a shape that follows the outer shape of the protrusions 14 and a uniform thickness.

[0038] [Second embodiment] Next, a second embodiment of the manufacturing apparatus 1 will be described with reference to FIGS. In these figures, the same elements as those in the first embodiment shown in FIGS. 1 to 7 are designated by the same reference numerals, and the description thereof will be omitted.

[0039] The manufacturing apparatus 1 of the second embodiment differs from that of the first embodiment in the configuration of the mold 10, and therefore in the following description, the configuration of the mold 10 that differs from that of the first embodiment will be described. Fig. 8 is a cross-sectional view of the manufacturing apparatus 1 for artificial three-dimensional tissue according to the second embodiment. Fig. 9 is a perspective view of the appearance of the mold 10 according to the second embodiment.

[0040] As shown in Fig. 8, the mold 10 in the manufacturing apparatus 1 of this embodiment has an anchor structure 22 provided on the upper surface of a support base 16, and the protruding portion 14 of the mold body 11 is formed of a plurality of divided bodies 14A to 14C that are divided in the vertical direction. As shown in Fig. 9, a plurality of anchor structures 22 (eight in Fig. 9) are provided on the upper surface of the support base 16 at intervals in the circumferential direction centered on the axis J.

[0041] As shown in Figs. 8 and 9, the protruding portion 14 constitutes a three-joint finger-type model. The protruding portion 14 has divided bodies 14A, 14B, and 14C provided in order from the bottom. Of the adjacent divided bodies 14A, 14B, and 14C, the upper divided body can swing around a rotation axis extending in the radial direction relative to the divided body on the lower side 2. Specifically, the divided body 14A is connected to the upper side of the mold main body 11 extending upward from the upper surface of the support base 16 so as to be able to swing around the rotation axis 15A. The rotation axis 15A extends in the radial direction (the direction perpendicular to the paper surface of Fig. 8). The divided body 14A can swing between an upright position shown in Fig. 8 and a tilted position tilted clockwise in Fig. 8 from the upright position around the rotation axis 15A relative to the mold main body 11. The divided body 14B is connected to the upper side of the divided body 14A so as to be able to swing around the rotation axis 15B. The rotation shaft 15B extends in the radial direction (the direction perpendicular to the paper surface of FIG. 8). The divided body 14B can swing between an upright position shown in FIG. 8 and a tilted position tilted clockwise in FIG. 8 around the rotation shaft 15B from the upright position with respect to the divided body 14A. The divided body 14C is connected to the upper side of the divided body 14C so as to be able to swing around the rotation shaft 15C. The rotation shaft 15C extends in the radial direction (the direction perpendicular to the paper surface of FIG. 8). The divided body 14C has a fingertip-shaped protrusion 12. The divided body 14C can swing between an upright position shown in FIG. 8 and a tilted position tilted clockwise in FIG. 8 around the rotation shaft 15C from the upright position with respect to the divided body 14C.

[0042] One end of wire 17A and one end of wire 17B are connected to segment 14C. As shown in Fig. 8, one end of wire 17A is connected to segment 14C on the opposite side to the swing direction from axis J (left side in Fig. 8). The other end of wire 17A, one end of which is connected to segment 14C, extends downward and passes through segment 14B, segment 14A, mold main body 11, and support base 16 in this order, and is drawn out to the bottom of support base 16. One end of wire 17B is connected to segment 14C in the swing direction (right side in FIG. 8) from axis J. The other end of wire 17B, which is connected to segment 14C at one end, extends downward and passes through segment 14B, segment 14A, mold body 11, and support base 16 in that order, and is drawn out to the bottom of support base 16.

[0043] The segments 14A, 14B, and 14C constituting the three-jointed finger-type model are placed in the upright position shown in FIG. 8 by pulling wire 17A downward, and in the tilted position shown in FIG. 9 by pulling wire 17B downward. The other configurations are similar to those of the first embodiment.

[0044] When forming dermal tissue T using the manufacturing apparatus 1 configured as above, as shown in Fig. 10, the mold 10 including the divided bodies 14A, 14B, 14C in an upright position is inserted from above into the inside of the tube part 32 of the case 30, and the lower surface of the support base 16 is brought into contact with the upper surface of the bottom plate part 31. Next, the cap part 40 is attached to the upper end side of the tube part 32. This completes the preparation of the manufacturing apparatus 1 having the culture space 33.

[0045] When preparation of the manufacturing apparatus 1 is completed, the culture space 33 is filled with a solution of collagen M containing dermal cells C, as shown in FIG. 10. The solution of collagen M containing dermal cells C is injected into the culture space 33 through the supply port 42 using a syringe, pipette, or the like. Once the mixture of dermal cells C and collagen M is injected into the culture space 33, it is cultured under predetermined conditions. The culture conditions are the same as those in the first embodiment. Note that the wires 17A and 17B are not shown in FIG. 10.

[0046] The collagen M solution gels as a result of the above-mentioned culturing, and is contracted by the dermal cells C, forming dermal tissue T that covers the segments 14A, 14B, and 14C that make up the three-jointed finger-type model, as shown in Fig. 11. The gel that contracts in the radial direction when the dermal tissue T is formed is held on the outer circumferential surface of the protruding portion 14. The upper gel that contracts in the direction of axis J engages with the protrusion 12 from above and is held there. The lower gel that contracts in the direction of axis J engages with the anchor portion 25 from below and is held there. Therefore, the artificial dermal tissue T produced by culturing using the present production apparatus 1 is formed into a bag-like shape covering the model of the three-joint finger-shaped divided bodies 14A, 14B, and 14C, and constitutes a three-joint finger-shaped artificial three-dimensional tissue model. That is, a bag-like artificial three-dimensional tissue T is formed having a bulging portion 91 that bulges out upward and a side portion 92 that extends downward from the bulging portion 91 and is formed all around the axis J, and has an anchor mark 93 at the end (part) of the side portion 92 when the artificial three-dimensional tissue T is removed from the production apparatus 1.

[0047] FIG. 12 is a cross-sectional view showing the movement of the dermis tissue T swinging between an upright position and an inclined position. 12, the dermal tissue T of the three-joint finger model is brought to an upright position by pulling wire 17A downward while being held by a holder constituted by mold 10 having anchor portions 25. The dermal tissue T is brought to a tilted position by pulling wire 17B downward. That is, the dermal tissue T can be swung between the upright position and the tilted position by switching between wire 17A and wire 17B and pulling them downward.

[0048] Fig. 13 is a photograph showing the dermal tissue T in an upright position, and Fig. 14 is a photograph showing the dermal tissue T in an inclined position. 13 and 14, it was confirmed that the dermal tissue T of the three-joint finger model can perform joint movement without hindrance in a state where the lower end is engaged and restrained by the anchor portion 25. This confirmed the practical use of the artificial dermal tissue T as, for example, a covering material for a robot.

[0049] [Third embodiment] Next, a third embodiment of the manufacturing apparatus 1 will be described with reference to FIGS. In these figures, the same elements as those in the first embodiment shown in FIGS. 1 to 8 are designated by the same reference numerals, and the description thereof will be omitted.

[0050] The manufacturing apparatus 1 of the third embodiment differs from that of the first embodiment in the configuration of the mold 10, and therefore, in the following description, the configuration of the mold 10 that differs from that of the first embodiment will be described. 15 and 16 are cross-sectional views of the manufacturing apparatus 1 for artificial three-dimensional tissue according to the third embodiment.

[0051] 15, the mold 10 in the manufacturing apparatus 1 of this embodiment is not provided with a support stand, and the lower surface of the mold body 11 contacts the upper surface of the bottom plate portion 31. The lower side of the mold body 11 fits into the through hole 21 of the annular member 20.

[0052] The protruding portion 14 of the mold body 11 has a human face shape facing upward. The protrusion 12 in this embodiment is, for example, a nose portion of the human face shape. The other configurations are similar to those of the first embodiment.

[0053] 15, when forming dermal tissue T using the manufacturing apparatus 1 having the above configuration, first, the through hole 21 of the annular member 20 is inserted into the mold body 11 from above, and then the assembled annular member 20 and mold body 11 are inserted into the inside of the tube portion 32 of the case 30 from above, and the lower surface of the annular member 20 and the lower surface of the mold body 11 are brought into contact with the upper surface of the bottom plate portion 31. Alternatively, the procedure may be such that only the annular member 20 is inserted into the inside of the tube portion 32 of the case 30 from above, and the lower surface of the annular member 20 is brought into contact with the upper surface of the bottom plate portion 31, and then the mold body 11 is inserted into the through hole 21 of the annular member 20 from above.

[0054] Next, the cap part 40 is attached to the upper end side of the cylindrical part 32. The position of the cap part 40 in the direction of the axis J is a position where the distance between the molding surface 41 and the protruding part 14 is approximately the same as the distance between the mold body 11 and the inner surface of the cylindrical part 32. This completes the preparation of the manufacturing device 1 having the culture space 33.

[0055] When preparation of the manufacturing apparatus 1 is completed, the culture space 33 is filled with a solution of collagen M containing dermal cells C. The solution of collagen M containing dermal cells C is injected into the culture space 33 via the supply port 42 using the above-mentioned syringe, pipette, or the like, to fill the culture space 33.

[0056] When the mixture of dermal cells C and collagen M is injected into the culture space 33, it is cultured under the above-mentioned culture conditions. In this embodiment, after about one day has passed since the culture and the collagen M solution has gelled, the cap part 40 is detached from the tube part 32. Then, as shown in FIG. 16, it is cultured for three days with the human face shape aligned in the up-down direction. This makes it possible to form an artificial three-dimensional tissue model provided with dermal tissue T having a human face shape of a face part. That is, an artificial three-dimensional tissue T having a human face shape is formed, which has a bulging part 91 bulging upward and a side part 92 extending downward from the bulging part 91 and formed around the entire circumference around the axis J, and has an anchor mark 93 at the end (part) of the side part 92 when the artificial three-dimensional tissue T is removed from the manufacturing device 1.

[0057] In this embodiment, in addition to obtaining the same actions and effects as those of the first embodiment, it becomes possible to easily produce dermal tissue T in a human face shape having many uneven parts. In addition, in this embodiment, culture can be easily performed in a state where the surface direction is the gravity direction.

[0058] [Fourth embodiment] Next, a fourth embodiment of the manufacturing apparatus 1 will be described with reference to FIGS. In these figures, the same elements as those in the third embodiment shown in FIGS. 15 and 16 are designated by the same reference numerals, and the description thereof will be omitted.

[0059] In the third embodiment, the anchor portion 25 is provided at a position separated from the mold body 11. In the present embodiment, however, a configuration in which the anchor portion 25 is provided on the protruding portion 14 will be described.

[0060] Fig. 17 is a cross-sectional view of the manufacturing apparatus 1 for artificial three-dimensional tissue according to the fourth embodiment. Note that in Fig. 17, dermal cells C are not shown. 17, the protruding portion 14 has a plurality of flow paths 18 with a circular cross section that open below the projection 12. The protruding portion 14 also has a recess 50 that faces upward and is recessed downward to form an eye socket or the like.

[0061] FIG. 18 is a cross-sectional view taken at the center position of the flow passage 18 and perpendicular to the direction of the axis J. As shown in Fig. 18, a plurality of flow paths 18 (eight in Fig. 18) are provided at both longitudinal ends of the protruding portion 14, both lateral ends, and in the circumferential direction between the flow paths at the longitudinal ends and the flow paths in the lateral direction in the plan view. Each flow path 18 opens to the side surface of the protruding portion 14 at both ends, and is formed in a U-shape that extends in the radial direction and curves in an arc before connecting. Each flow path 18 is formed along a plane perpendicular to the axis J. The inner circumferential surface of the flow path 18 that is located on the upper side and faces the lower side forms an anchor portion 19.

[0062] FIG. 19 is a cross-sectional view taken along a plane including the axis J of the recess 50. As shown in FIG. As shown in Fig. 19, the protrusion 14 has a plurality of flow paths 52 (three in Fig. 19) that open to the inner circumferential surface 51 of the recess 50. The flow paths 52 are respectively disposed at the bottom of the recess 50 and at the opposing side portions sandwiching the recess 50. Both ends of each flow path 52 open to the inner circumferential surface 51, and are formed into a U-shape that extends in the normal direction of the inner circumferential surface 51 and is curved in an arc to join the ends.

[0063] As an example, each flow passage 52 is formed along a plane including the direction of the axis J. Of the inner circumferential surfaces of the flow passages 52, the inner circumferential surface located on the upper side and facing the lower side forms an anchor portion 53. Note that the flow passages 52 disposed on the sides may be formed along a plane perpendicular to the direction of the axis J, similar to the flow passages 18.

[0064] 17, when forming dermal tissue T using the manufacturing apparatus 1 having the above configuration, first, the through hole 21 of the annular member 20 is inserted into the mold body 11 from above, and then the assembled annular member 20 and mold body 11 are inserted into the inside of the tubular portion 32 of the case 30 from above, and the lower surface of the annular member 20 and the lower surface of the mold body 11 are brought into contact with the upper surface of the bottom plate portion 31. Alternatively, the procedure may be such that only the annular member 20 is inserted into the inside of the tubular portion 32 of the case 30 from above, and the lower surface of the annular member 20 is brought into contact with the upper surface of the bottom plate portion 31, and then the mold body 11 is inserted into the through hole 21 of the annular member 20 from above.

[0065] Next, the cap portion 40 is attached to the upper end side of the tube portion 32, and when preparation of the manufacturing apparatus 1 having the culture space 33 is completed, the culture space 33 is filled with a solution of collagen M containing dermal cells C. The solution of collagen M containing dermal cells C is injected into the culture space 33 through the supply port 42 using the above-mentioned syringe, pipette, or the like, and filled therein. The solution of collagen M containing dermal cells C injected into the culture space 33 flows into and fills the flow channel 18, and also fills the flow channel 52 through the recess 50.

[0066] Once the mixture of dermal cells C and collagen M is injected into culture space 33, it is cultured under the culture conditions described above. This allows for the formation of dermal tissue T having a human face shape, as shown in Fig. 20. The upper gel that has contracted in the direction of axis J when dermal tissue T is formed is engaged and held by protrusions 12 from above. Meanwhile, the lower gel that has contracted in the direction of axis J is engaged and held by anchor portions 19 from below in flow channel 18, and is engaged and held by anchor portions 53 from below in flow channel 52.

[0067] 16, when the anchor structure 22 is used, the end of the dermal tissue T that engages with the anchor structure 22 is formed by rolling up, resulting in a shape that differs from the contour of an actual human face. In contrast, in this embodiment, the dermal tissue T engages with the anchor portion 19 in the flow path 18 provided inside the protruding portion 14, making it possible to form the dermal tissue T in a shape that follows the contour of an actual human face.

[0068] 21 , when a recess 50 is provided in the protrusion 12 and anchor structures 22 are provided on both sides of the protrusion 12 in the radial direction below the protrusion 12, it is assumed that it is difficult to cover the inner circumferential surface of the recess 50 with the dermal tissue T because the dermal tissue T is restricted from contracting downward by the protrusion 12. In contrast, in this embodiment, by providing a flow path 52 opening into the inner circumferential surface 51, it is possible to engage the dermal tissue T with the anchor portion 53 close to the inner circumferential surface 51. Therefore, the inner circumferential surface of the recess 50 provided in the protrusion 12 can also be covered with the dermal tissue T.

[0069] Thus, in the manufacturing apparatus 1 of this embodiment, in addition to obtaining the same functions and effects as those of the third embodiment, it becomes possible to form dermal tissue T in a shape that follows the contour of an actual human face, and to easily form dermal tissue T that covers recesses such as the eye sockets, the mouth, the ears, and the nostrils.

[0070] [Modification of the fourth embodiment] In the fourth embodiment, a configuration is exemplified in which a solution of collagen M containing dermal cells C is injected into the culture space 33 through the supply port 42 of the cap portion 40, but this configuration is not limited to this, and for example, a configuration in which a solution of collagen M containing dermal cells C is injected using the above-mentioned flow path 18 may be used.

[0071] 18, the protrusion 14 has one end connected to the flow path 18 on the radially outer side and the other end extending toward the center of the protrusion 14 in a plan view, and each of the flow paths 18 has a communication path 60. The other end of each communication path 60 is connected to a solution supply unit 61 provided in the center of the protrusion 14. The solution supply unit 61 opens to the bottom surface of the mold body 11, and can be configured to be able to supply a solution of collagen M containing dermal cells C from the outside. By supplying the solution of collagen M containing dermal cells C from inside the protrusion 14 in this manner, it becomes unnecessary to provide the cap portion 40 with a supply port 42. The molding surface 41 of the cap portion 40 can be made into an offset surface aligned with the upper surface of the protrusion 14 including the projections 12 over its entirety. Therefore, by making the width of the culture space 33 more constant, the cultured dermal tissue T can be formed with high precision following the uneven shape of the protrusions 14 including the projections 12.

[0072] [Fifth embodiment] Next, a fifth embodiment of the manufacturing apparatus 1 will be described with reference to FIG. In this figure, the same elements as those in the first embodiment shown in FIGS. 1 to 7 are designated by the same reference numerals, and the description thereof will be omitted.

[0073] FIG. 22 is a cross-sectional view of the manufacturing apparatus 1 for artificial three-dimensional tissue according to the fifth embodiment. 22, the manufacturing apparatus 1 of this embodiment has a shaft-shaped mold body 11 extending upward from the top surface of the bottom plate portion 31 of the case 30, and a bowl-shaped protrusion 14 that expands radially from the upper end of the mold body 11 toward the upper side and opens upward. The bowl-shaped protrusion 14 that opens upward has an opening end 71 located on the upper side, a concave surface 72 surrounded by the opening end 71 facing the upper side and recessed downward, and a convex surface 73 located below the concave surface 72 and bulging toward the lower side. The molding surface 41 of the cap portion 40 protrudes downward in a shape that is spaced from the concave surface 72 by a certain distance.

[0074] In this embodiment, the open end 71 constitutes the protrusion 12. In this embodiment, the convex surface 73 constitutes the anchor portion 74. Note that Fig. 22 illustrates a state in which the culture space 33 is filled with a solution of collagen M containing dermal cells C.

[0075] In the manufacturing apparatus 1 configured as above, a solution of collagen M containing dermal cells C is cultured under the culture conditions described above. When dermal tissue is formed by culture, the upper gel that has contracted downward in the direction of axis J is engaged from above with open end 71 that constitutes protrusion 12 and held thereon. Meanwhile, the lower gel that has contracted upward in the direction of axis J is engaged from below with convex surface 73 that constitutes anchor portion 74 and held thereon.

[0076] Therefore, in the manufacturing apparatus 1 of this embodiment, dermal tissue is formed that covers the convex surface 73 from the concave surface 72 of the bowl-shaped protrusion 14 through the open end 71. In this way, the manufacturing apparatus 1 of this embodiment makes it possible to easily form dermal tissue that covers recesses such as the eye sockets, mouth, ears, and nostrils.

[0077] [Wound repair model] Next, a wound repair model of dermal tissue formed by the above-mentioned manufacturing apparatus 1 will be described with reference to FIGS.

[0078] The wound repair model of the present invention is a wound repair model having an artificial three-dimensional tissue T, at least a portion of which protrudes in a predetermined direction (direction of axis J), and includes a protruding portion 14 having at least a protrusion 12 protruding on one side of the predetermined direction, a plurality of anchor portions 25 located on the other side of the protrusion 12 in the predetermined direction and located on both sides of the protrusion 12 in a radial direction perpendicular to the predetermined direction and facing the other side of the predetermined direction, an artificial three-dimensional tissue T having a wound portion, whose end portion is folded back by the protrusion 12 to cover the protruding portion 14 and engages with the anchor portion 25, and an extracellular matrix CS that is attached to the wound portion to repair the wound.

[0079] In human wound treatment, cultured dermis made from the patient's cells may be transplanted to treat deep wounds that reach the dermis. As an example, a deep defect penetrating the dermal tissue produced in the three-joint finger model described in the second embodiment is created, and a repair test is then performed to confirm the practicality of the cultured dermis.

[0080] Specifically, we carried out the following: "evaluation of cell migration to the transplant site," "evaluation of strength recovery after repair," and "repair of the dermis covering the above-mentioned mold."

[0081] "Evaluation of cell migration to transplant site" A circular wound with a diameter of 6 mm was created in a planar dermal tissue, and a collagen sheet was grafted into this wound. An experiment was conducted to observe cell migration into the grafted collagen sheet. Figure 23 is a stained photograph of the collagen sheet on the third day after grafting. Figure 24 is a stained photograph of the collagen sheet on the seventh day after grafting. As shown in Figures 23 and 24, these are stained photographs of the collagen sheet on the seventh day after grafting. It was confirmed that the cells gradually migrated into the collagen sheet, and reached the center by the seventh day.

[0082] "Evaluation of strength recovery after repair" Next, a tensile test was performed to confirm the change in the strength of the dermal tissue during repair. Using a culture device designed to be attached to a tensile tester, sample A without a wound hole, sample B with an unrepaired wound hole, and sample C in which the wound hole was filled with collagen and then cultured for 7 days to repair the wound hole were created. Each sample had a rectangular cross section with a width of approximately 4 mm and a thickness of approximately 10 mm. For samples B and C, a cylindrical cavity with a diameter of approximately 2 mm and a depth of approximately 8 mm was created at a location 4 mm wide as the wound hole. For sample C, the cavity was filled with collagen to repair the cavity.

[0083] FIG. 25 is a diagram showing the relationship between the tensile length and the tensile force of Samples A to C in the tensile test. As shown in Figure 25, the dermal tissue of sample B, which has an unrepaired wound hole, broke at a shorter tensile length and with a smaller tensile force than sample A, which does not have a wound hole. On the other hand, the dermal tissue of sample C, in which the wound hole was filled with collagen and then cultured for 7 days to repair the wound hole, broke at the same tensile length and tensile force as sample A, which does not have a wound hole. From these results, it was confirmed that sample C, in which the wound hole was filled with collagen and then cultured for 7 days to repair the wound hole, was able to recover strength to the same extent as sample A, which does not have a wound hole.

[0084] "Repair of the dermis covering the mold" In the dermal tissue produced in the three-joint finger model described in the second embodiment, a wound was made (wounded) using a knife to penetrate the dermal tissue on the back (nail side) of the second joint. Fig. 26 is a perspective view of the appearance of the dermal tissue produced in the three-joint finger model. As shown in Fig. 26, a collagen sheet CS was grafted into the wound in the dermal tissue of the second joint in the three-joint finger model.

[0085] The three-joint finger model in which the collagen sheet CS was grafted into the wound of the dermal tissue was cultured for 7 days. FIG. 27 is a photograph of the three-joint finger model in the upright position described above. FIG. 28 is a photograph of the three-joint finger model in the inclined position described above. As shown in FIG. 27 and FIG. 28, it was confirmed that the dermal tissue of the second joint in which the collagen sheet CS was grafted into the wound was repaired, and that the joint movement could be performed without any problems. Therefore, from the above experiments, a wound repair model SM was obtained that has the practicality of artificial dermis as a robot covering material, as an example of a repairable wound.

[0086] [Epidermal tissue formation] In the above embodiment, a configuration for forming dermal tissue as an artificial three-dimensional tissue has been illustrated, but a configuration for laminating epidermal tissue on the formed dermal tissue will be described with reference to Figures 29 and 30. In these figures, the same elements as those in the first embodiment shown in Figures 1 to 7 are denoted by the same reference numerals, and description thereof will be omitted.

[0087] Epidermal cells that form epidermal tissue are seeded on dermal tissue and cultured for several hours to become fixed to the dermal tissue. On the other hand, as shown in Figures 4 and 11, when epidermal cells are seeded on dermal tissue that covers the approximately cylindrical protruding portion 14, the epidermal cells slide off the surface of the dermal tissue due to gravity, making it difficult to fix the epidermal cells to the dermal tissue.

[0088] Therefore, in this embodiment, the above-mentioned mold 10 for forming dermal tissue and an auxiliary mold for forming epidermal tissue after forming the dermal tissue are used in combination. Here, an example using the mold 10 described in the first embodiment will be described.

[0089] 29 is an external perspective view of the auxiliary mold 10A. The auxiliary mold 10A has a mold body 11 and a support stand 16. The diameter, thickness, length in the direction of the axis J, etc. of the mold body 11 and the support stand 16 in the auxiliary mold 10A are the same as those of the mold 10 described in the first embodiment above.

[0090] The mold body 11 of the auxiliary mold 10A is formed in a semicylindrical shape with a flat surface 81 including the axis J cut out above a position below the position of the top surface of the annular member 20 shown in Fig. 4. The mold body 11 has a recess 82 that opens into the flat surface 81. The inner surface of the recess 82 is shaped to be a certain distance (thickness) from the outer circumferential surfaces of the projection 12 and the protruding portion 14. In other words, the recess 82 has a semicircular cross section perpendicular to the axis J and is recessed in a hemispherical shape on the upper side, forming a container that opens into the flat surface 81.

[0091] When forming epidermal tissue using the auxiliary mold 10A, first, as shown in FIG. 4, the mold 10 is used to form a bag-shaped dermal tissue T that is engaged and held by the projections 12 from above and engaged and held by the anchor portions 25 from below. Next, the mold 10 and the annular member 20 are detached and removed from the case 30. After this, the mold body 11 is pulled out from the through hole 21 of the annular member 20 in a state of being turned upside down, and the mold body 11 of the auxiliary mold 10A is inserted into the through hole 21. This replaces the mold 10 with the auxiliary mold 10A. In addition, in the auxiliary mold 10A, the dermal tissue T engages with the projections 12 from above and contacts the outer circumferential surface of the semi-cylindrical mold body 11. That is, the dermal tissue T is held by a holder composed of the auxiliary mold 10A having the projections 12 and the annular member 20 having the anchor structure 22.

[0092] Next, the holder is tilted so that axis J extends horizontally and the opening of recess 82 faces upward. Fig. 30 is a cross-sectional view perpendicular to axis J of the holder tilted so that the opening of recess 82 faces upward. As shown in Fig. 30, the upper side of dermal tissue T, which has a substantially circular cross section, is deflected into recess 82 due to its own weight and becomes a concave shape recessed downward.

[0093] Epidermal cells E are seeded on the surface of this concave dermal tissue T. At this time, since the dermal tissue T is bent into the recess 82 and has a concave shape, the seeded epidermal cells E remain on the surface of the dermal tissue T without sliding down.

[0094] Examples of epidermal cells E include cells derived from mammals such as humans, mice, and rats, with human-derived epidermal cells being preferred. Examples of human-derived epidermal cells include epidermal keratinocytes and epidermal melanocytes, with epidermal keratinocytes being preferred. Examples of epidermal keratinocytes include normal human epidermal keratinocytes (NHEK). Examples of epidermal melanocytes include normal human epidermal melanocytes (NHEM).

[0095] The epidermal cells E seeded on the surface of the dermal tissue T are fixed to the dermal tissue T in about three hours, so when three hours have passed since seeding, the dermal tissue T is rotated 90° in the circumferential direction relative to the auxiliary mold 10A, and the dermal tissue T including the portion where the epidermal cells E have not been seeded is positioned in the recess 82. Then, as described above, the epidermal cells E are seeded on the surface of the dermal tissue T that has become a recess. In this way, the epidermal cells E can be fixed to the surface of the dermal tissue T over the entire circumference by repeating four times the steps of seeding the epidermal cells E on the surface of the dermal tissue T, allowing three hours to pass after seeding the epidermal cells E, and rotating the dermal tissue T 90° in the circumferential direction after the three hours have passed.

[0096] Thereafter, the epidermal cells E are cultured, whereby the epidermal cells E are induced to differentiate and form epidermal tissue. As an example of the culture conditions for forming epidermal tissue, the cells are cultured at 37° C. for 9 days.

[0097] Although the preferred embodiment of the present invention has been described above with reference to the accompanying drawings, it goes without saying that the present invention is not limited to the examples. The shapes and combinations of the components shown in the above examples are merely examples, and various modifications can be made based on design requirements, etc., without departing from the spirit of the present invention.

[0098] For example, by forming epidermal tissue to protect dermal tissue from drying and constructing a perfusion route for nutritional supply, it becomes possible to use artificial skin as a robot covering material for a long period of time. In addition, by covering a robot using biological tissue such as muscle with the present invention, it becomes possible to create a biohybrid robot that can operate in air.

[0099] In the above embodiment, the artificial three-dimensional tissue is exemplified as an artificial skin tissue, but is not limited thereto. The artificial three-dimensional tissue may be, for example, muscle tissue using skeletal muscle cells or cardiac muscle cells instead of the above-mentioned fibroblasts. Furthermore, the artificial three-dimensional tissue may be digestive system tissue such as liver tissue using liver cells or pancreatic tissue using pancreatic cells, urinary system tissue such as kidney tissue using kidney cells, or nerve tissue using nerve cells. When the present invention is applied to such tissues, the extracellular matrix used in the above embodiment does not necessarily need to be present, and may be an accumulation of only various types of cells. [Explanation of symbols]

[0100] 1...Manufacturing apparatus (manufacturing apparatus for artificial three-dimensional tissue), 11...Mold body (shaft-shaped portion), 12...Protrusion, 14...Protruding portion, 14A, 14B, 14C...Divided body, 15A, 15B, 15C...Rotating shaft, 18, 52...Flow path, 19, 25, 53, 74...Anchor portion, 20...Annular member, 23...Column portion, 24...Flange portion, 30...Case, 32...Cylinder portion, 40...Cap portion, 41...Molding surface, 50...Depression, 60...Communicating passage, 61...Solution supply portion, 71...Open end, 72...Concave surface, C...Cells (dermal cells), CS...Collagen sheet (extracellular matrix), J...Axis, M...Extracellular matrix (dermal cells), SM...Wound repair model, T...Artificial three-dimensional tissue (dermal tissue)

Claims

1. An apparatus for manufacturing an artificial three-dimensional tissue having at least a portion protruding in a predetermined direction, a protruding portion having at least a protrusion protruding to one side in the predetermined direction; a plurality of anchor portions located on the other side of the protrusion in the predetermined direction, and located on both sides of the protrusion in a radial direction perpendicular to the predetermined direction, facing the other side in the predetermined direction; a cap portion disposed on one side of the protrusion in the predetermined direction; having The cap portion has a molding surface provided on the other side in the predetermined direction facing the culture space of the artificial three-dimensional tissue, The molding surface molds the one end of the artificial three-dimensional tissue,

2. a case having a cylindrical portion extending in the predetermined direction to surround the radial outer side of the protrusion portion and the anchor portion to form the culture space; The cap portion is detachably attached to one end portion of the cylindrical portion in the predetermined direction. The apparatus for producing an artificial three-dimensional tissue according to claim 1 .

3. a shaft-shaped portion extending in the predetermined direction and having the protruding portion on the one side; an annular member that is provided on the shaft-shaped portion so as to be freely inserted and removed from the one side and is located on the other side of the protruding portion of the shaft-shaped portion; having The one side of the annular member includes a plurality of column portions extending to the one side and a plurality of flange portions extending from a tip of the column portion to an outer side in the radial direction of the column portion over an entire circumference, The anchor portion is the other side surface of the flange portion.

3. An apparatus for producing an artificial three-dimensional tissue according to claim 1 or 2.

4. The protrusion is hemispherical dome shaped. An apparatus for producing an artificial three-dimensional tissue according to any one of claims 1 to 3.

5. The protrusion portion has a plurality of divided bodies divided in the predetermined direction, Among adjacent divided bodies, the divided body on the one side is swingable around a rotation axis extending in the radial direction relative to the divided body on the other side.

3. An apparatus for producing an artificial three-dimensional tissue according to claim 1 or 2.

6. The protruding portion is formed in a bowl shape that widens in the radial direction toward the one side and opens to the one side, the bowl-shaped protruding portion opening to the one side has an open end on the one side, a concave surface surrounded by the open end, facing the one side, and recessed toward the other side, and a convex surface located on the other side of the concave surface in the predetermined direction and bulging toward the other side, the protrusion is the open end, The anchor portion is the convex surface.

3. An apparatus for producing an artificial three-dimensional tissue according to claim 1 or 2.

7. The protruding portion has a human face shape including the protrusion, a plurality of flow paths extending in the radial direction and opening at a position on the other side of the protrusion; The anchor portion is an inner circumferential surface located on the one side of the inner circumferential surface facing the flow path.

3. An apparatus for producing an artificial three-dimensional tissue according to claim 1 or 2.

8. Both ends of the plurality of flow paths are located on the other side of the protrusion and open to the protruding portion. The apparatus for producing an artificial three-dimensional tissue according to claim 7.

9. The plurality of flow paths have one end open at a position on the other side of the protrusion, and communicate with a solution supply unit inside the protruding portion. The apparatus for producing an artificial three-dimensional tissue according to claim 7.

10. The protruding portion has a recess recessed on the other side, The recess has an inner periphery, The flow path opens to the inner circumferential surface. The apparatus for producing an artificial three-dimensional tissue according to claim 7.

11. A wound repair model having an artificial three-dimensional tissue, at least a portion of which protrudes in a predetermined direction, a protruding portion having at least a protrusion protruding to one side in the predetermined direction; a plurality of anchor portions located on the other side of the protrusion in the predetermined direction, and located on both sides of the protrusion in a radial direction perpendicular to the predetermined direction, facing the other side in the predetermined direction; An artificial three-dimensional tissue having a wound portion, the end of which is folded back by the protrusion to cover the protruding portion and engaged with the anchor portion; An extracellular matrix that is applied to the wound area to repair the wound area; A wound repair model having

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