Apparatus for forming a reinforcing layer on a sheet-shaped cell culture, method for forming a reinforcing layer on a sheet-shaped cell culture, laminate, and program

Abstract

Provided are an apparatus for forming a reinforcing layer on a sheet-shaped cell culture, a method for forming a reinforcing layer on a sheet-shaped cell culture, a laminate, and a program, which enable a laminate formed by forming a reinforcing layer on a sheet-shaped cell culture to be smoothly attached to a transplantation device and enable the laminate to be smoothly delivered to a transplantation site using the transplantation device. [Solution] The forming device 200 is an apparatus equipped with a processing unit 210 that performs processing to form a reinforcing layer 30 made of fibrin gel that extends linearly on a sheet-shaped cell culture 10, and the processing unit has a spray unit 230 that can be positioned to face one of the front and back surfaces 10a and 10b of the sheet-shaped cell culture 10 and can spray thrombin solution L2 onto each of multiple different locations on one surface.

Description

[Technical Field]

[0001] The present invention relates to an apparatus for forming a reinforcing layer on a sheet-shaped cell culture, a method for forming a reinforcing layer on a sheet-shaped cell culture, a laminate, and a program. [Background technology]

[0002] Sheet-shaped cell cultures (cell sheets) are widely known for use in fields such as regenerative medicine. Sheet-shaped cell cultures are produced by storing and culturing cells collected from humans or non-human animals in a culture substrate (culture vessel) containing a culture medium, and then allowing them to adhere and aggregate into a sheet.

[0003] Sheet-shaped cell cultures are constructed in the form of a thin sheet (e.g., a circular sheet). Therefore, they are prone to wrinkling and tearing during isolation from the culture substrate, transportation and storage, and transplantation into the patient's body, and therefore require careful handling.

[0004] To solve the above problems, a method has been proposed in which a fibrinogen solution and a thrombin solution are applied to a sheet-shaped cell culture, a reinforcing layer made of fibrin gel is formed on the sheet-shaped cell culture, and a laminate consisting of the sheet-shaped cell culture and the reinforcing layer is produced (see, for example, Patent Document 1).

[0005] Various methods have been considered for transplanting a laminate with a reinforcing layer into the human body, and for example, a body cavity scope (e.g., a thoracoscope) may be used depending on the transplant site in the human body. In addition, in a procedure using a body cavity scope, a predetermined transfer device that can be introduced into the human body together with the body cavity scope may be used through a relatively small incision made in the patient's body.

[0006] As the above-mentioned transfer device, for example, one has been proposed that includes a cylindrical member (outer cylinder portion) with a circular tubular structure that allows the laminate formed into a circular sheet to be stored and deployed (extracted) to the outside (see, for example, Patent Document 2).

[0007] In the method of transplanting a laminate using a transfer device disclosed in Patent Document 2, the laminate is housed in the inner cavity of an outer tube while being supported in a curved state (deformed into a cylindrical shape) relative to a predetermined support. After delivering the laminate to the transplant site in the human body, the laminate is unfolded from the outer tube, thereby maintaining the laminate in a compact shape until delivery. Therefore, by using the transfer device described above, the laminate can be smoothly delivered to the transplant site. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Patent Publication No. 2016-52272 [Patent Document 2] WO2023 / 234380 Summary of the Invention [Problem to be solved by the invention]

[0009] As described above, when using a transplant device such as that proposed in Patent Document 2, the laminate needs to be deformed into a curved (cylindrical) shape when attached to the outer tube. However, if a reinforcing layer of uniform thickness is formed on the front (or back) surface of the sheet-shaped cell culture, the reinforcing layer may hinder easy deformation of the laminate. This may result in the work of setting the laminate in the transplant device and delivering the laminate to the transplant site being time-consuming.

[0010] The present invention aims to provide an apparatus for forming a reinforcing layer on a sheet-shaped cell culture, a method for forming a reinforcing layer on a sheet-shaped cell culture, a laminate, and a program that enable a laminate formed by forming a reinforcing layer on a sheet-shaped cell culture to be smoothly attached to a transplant device and that enables the laminate to be smoothly delivered to the transplant site using the transplant device. [Means for solving the problem]

[0011] The present invention is achieved by any one of the following means (1) to (7).

[0012] (1) An apparatus including a processing unit that performs a process for forming a linearly extending reinforcing layer made of fibrin gel on a sheet-shaped cell culture, The processing unit can be positioned to face one of the front and back surfaces of the sheet-shaped cell culture, and has a spray unit capable of spraying thrombin solution onto each of multiple different locations on the one surface, in an apparatus for forming a reinforcing layer on a sheet-shaped cell culture.

[0013] (2) the spray unit has a plurality of spray holes for spraying the thrombin liquid, An apparatus for forming a reinforcing layer on a sheet-shaped cell culture described in (1), wherein the plurality of nozzles have portions that increase and portions that decrease the amount of thrombin solution sprayed per specified area on one side of the sheet-shaped cell culture.

[0014] (3) The apparatus for forming a reinforcing layer on a sheet-shaped cell culture according to (2), wherein the plurality of nozzles are arranged in a plurality of parallel straight lines.

[0015] (4) An apparatus for forming a reinforcing layer on a sheet-shaped cell culture according to (2) or (3), wherein the plurality of nozzles are arranged in a predetermined pattern including dense and sparsely spaced areas along the surface direction of one of the surfaces of the sheet-shaped cell culture.

[0016] (5) An apparatus for forming a reinforcing layer on a sheet-shaped cell culture described in (2) or (3), wherein the plurality of nozzles include a first nozzle that sprays a larger amount of the thrombin solution per unit time, and a second nozzle that sprays a smaller amount of the thrombin solution per unit time than the first nozzle.

[0017] (6) A method for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, comprising: a step in which a processing unit applies a fibrinogen solution to one of the front and back surfaces of the sheet-shaped cell culture; A method for forming a reinforcing layer on a sheet-shaped cell culture, comprising the step of: the processing unit spraying thrombin solution onto one surface of the sheet-shaped cell culture to form a reinforcing layer consisting of fibrin gel extending linearly on the one surface.

[0018] (7) A laminate having a sheet-shaped cell culture and a reinforcing layer made of fibrin gel formed on the sheet-shaped cell culture, The reinforcing layer extends linearly on one of the front and back surfaces of the sheet-shaped cell culture.

[0019] (8) An apparatus including a processing unit for performing a process to form a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, applying a fibrinogen solution to one of the front and back surfaces of the sheet-shaped cell culture; and spraying a thrombin solution onto the one surface of the sheet-shaped cell culture to form the reinforcing layer made of fibrin gel extending linearly on the one surface. [Effects of the Invention]

[0020] The device for forming a reinforcing layer on a sheet-shaped cell culture, the method for forming a reinforcing layer on a sheet-shaped cell culture, the laminate, and the program according to the present invention make it possible to produce a laminate having a linearly extending reinforcing layer on one of the front and back surfaces of a sheet-shaped cell culture. When the laminate is bent in a direction intersecting the surface direction, the thinner portions of the reinforcing layer become the origin of deformation, allowing for easy bending. This allows the surgeon to smoothly load the laminate, consisting of a sheet-shaped cell culture with a reinforcing layer formed thereon, into a transplantation device, and to smoothly deliver the laminate to the transplantation site using the transplantation device. [Brief explanation of the drawings]

[0021] [Figure 1] FIG. 1 is a block diagram showing a system for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 2] 1 is a flowchart showing the steps of a method for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 3] FIG. 2 is a perspective view illustrating a spray unit according to the embodiment. [Figure 4] FIG. 10 is a diagram illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 5] FIG. 10 is a diagram illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 6] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 7] FIG. 2 is a plan view of a laminate manufactured using the processing unit according to the embodiment. [Figure 8] FIG. 8 is a cross-sectional view of the laminate taken along line VIII-VIII shown in FIG. 7. [Figure 9] FIG. 8 is a cross-sectional view of the laminate taken along line IX-IX shown in FIG. 7. [Figure 10] FIG. 8 is a cross-sectional view of the laminate taken along line XX shown in FIG. 7. [Figure 11]FIG. 10 is a cross-sectional view illustrating the state when the laminate is attached to a transplantation device. [Figure 12] FIG. 10 is a perspective view illustrating a spray unit according to a modified example. [Figure 13] FIG. 10 is a plan view of a laminate manufactured using a spray unit according to a modified example. [Figure 14] 14 is a cross-sectional view of the laminate taken along line XIV-XIV shown in FIG. 13. FIG. [Figure 15] 14 is a cross-sectional view of the laminate taken along line XV-XV shown in FIG. 13. FIG. [Figure 16] 16 is a cross-sectional view of the laminate taken along line XVI-XVI shown in FIG. 13. DETAILED DESCRIPTION OF THE INVENTION

[0022] (Embodiment) Hereinafter, embodiments of the present invention will be described with reference to the drawings. Note that the dimensional proportions in the drawings are exaggerated for the sake of explanation and may differ from the actual proportions.

[0023] FIG. 1 is a simplified block diagram showing the overall configuration of a system 100 (hereinafter also referred to as "forming system 100") for forming a reinforcing layer 30 on a sheet-shaped cell culture 10 according to this embodiment. FIG. 2 is a flowchart showing each step of a method (hereinafter also referred to as "forming method") for forming a reinforcing layer 30 on a sheet-shaped cell culture 10 according to this embodiment. FIG. 3 is a perspective view showing a spraying unit 230 of a forming device 200 (hereinafter also referred to as "forming device 200") equipped with a processing unit 210 for forming a reinforcing layer 30 on a sheet-shaped cell culture 10 according to this embodiment.

[0024] 4 to 6 are diagrams for explaining the formation method, and show partial cross-sectional views along the height direction of the container 320 (the same direction as the thickness direction of the sheet-shaped cell culture 10 and the laminate 20).

[0025] 7 to 11 are diagrams illustrating the laminate 20 according to this embodiment. Fig. 7 is a plan view of the laminate 20 as viewed from the direction of arrow VII shown in Fig. 1. Fig. 8 is a cross-sectional view of the laminate 20 taken along line VIII-VIII shown in Fig. 7, Fig. 9 is a cross-sectional view of the laminate 20 taken along line IX-IX shown in Fig. 7, and Fig. 10 is a cross-sectional view of the laminate 20 taken along line XX shown in Fig. 7. Fig. 11 is a cross-sectional view illustrating the laminate 20 according to this embodiment housed in a tubular portion 400 of a predetermined transplant device.

[0026] The arrows X1, X2, Y1, and Y2 in each figure indicate directions parallel to the surface direction of the sheet-shaped cell culture 10 (directions parallel to the front surface 10a and back surface 10b), and the arrows Z1 and Z2 indicate the thickness direction perpendicular to the surface direction of the sheet-shaped cell culture 10 (laminated body 20).

[0027] <Sheet-shaped cell culture 10> The sheet-shaped cell culture 10 used in this embodiment can contain any cells capable of constituting the sheet-shaped cell culture 10. Cells constituting the sheet-shaped cell culture 10 include, for example, somatic stem cells (adult stem cells), mesenchymal stem cells, or cardiomyocytes derived from iPS cells (induced pluripotent stem cells). Somatic stem cells preferably include skeletal myoblasts (myoblast cells), fibroblasts, synovial cells, epithelial cells, endothelial cells, etc. Furthermore, the term "sheet-shaped cell culture" as used herein refers to a sheet-like (thin film-like) structure in which cells are interconnected.

[0028] As shown in Figure 3, the sheet-shaped cell culture 10 may be configured to have, for example, a substantially circular planar shape in its natural state when no external force is applied. The planar shape of the sheet-shaped cell culture 10 is not limited to a circle, and may be, for example, a rectangle, a triangle, an oval, or any other geometric shape. There are also no particular limitations on the specific size or thickness of the sheet-shaped cell culture 10.

[0029] The sheet-shaped cell culture 10 has a surface 10a, a back surface 10b located opposite the surface 10a and arranged to face the bottom 323 of the container 320, and a peripheral edge 13 having a circular outer shape.

[0030] In this embodiment, the laminate in which the reinforcing layer 30 is formed in a position overlapping at least a portion of the sheet-shaped cell culture 10 is defined as a laminate 20 (see FIGS. 7 to 11).

[0031] The formation method described below can be carried out in a state where the sheet-shaped cell culture 10 is housed in a predetermined container 320 (see FIGS. 4 to 6).

[0032] The container 320 is, for example, a culture substrate used for culturing the sheet-shaped cell culture 10. When the container 320 is a culture substrate, the container 320 can be made of a material that is impermeable to solutions such as liquid culture media. When the container 320 is a culture substrate, before starting the formation method, an operation is appropriately performed to drain the solution (liquid culture media, etc.) used for the culture from the container 320.

[0033] As shown in FIG. 4, the container 320 has an opening 321 that opens upward, a bottom 323 on which the back surface 10b of the sheet-shaped cell culture 10 is placed, and a side wall 325 that surrounds the periphery of the bottom 323.

[0034] The container 320 can be configured to have a substantially circular shape. The bottom 323 of the container 320 has a size (inner diameter) larger than the outer shape of the sheet-shaped cell culture 10 before the reinforcing layer 30 is formed.

[0035] There are no particular restrictions on the specific form of the sheet-shaped cell culture 10 (e.g., manufacturing method, physical properties, function, etc.) and the specific form of the culture substrate (e.g., material, function, structure and shape, type of liquid medium used, etc.), and publicly known content (e.g., Patent Publication No. 2011-155869, Patent Publication No. 2022-106986, Patent Publication No. 2021-106609, etc.) can be taken into consideration as appropriate.

[0036] <Forming system 100> 1, the forming device 200 according to this embodiment can be used in a state where it is incorporated into the forming system 100. Note that the forming device 200 can also be used for the operation of forming the reinforcing layer 30 (the operation of manufacturing the laminate 20) in a state where it is not incorporated into the forming system 100 (a state where the forming device 200 can operate independently).

[0037] To give an overview with reference to Figures 1 and 3 to 6, the formation system 100 comprises a formation device 200, a control unit 110 that performs overall operational control of the formation system 100 and the formation device 200, an imaging unit 120 that photographs the sheet-shaped cell culture 10 and the laminate 20, an image display unit 130 that can display an image generated from image information (image data) acquired by the imaging unit 120, and a stage 310 on which a container 320 containing the sheet-shaped cell culture 10 and the laminate 20 can be placed.

[0038] The control unit 110 is composed of, for example, a CPU (Central Processing Unit), memories such as RAM (Random Access Memory) and ROM (Read Only Memory), and a storage unit, and controls each part of the forming system 100 and performs various arithmetic processing according to a program.

[0039] The storage unit is configured with an HDD (Hard Disk Drive) or an SSD (Solid State Drive) and stores various programs and data. The storage unit can store a series of programs necessary for controlling the operation of the forming system 100. The above programs can incorporate algorithms for causing the control unit 110 to function as the detection unit 111 and the determination unit 112.

[0040] The detection unit 111 can be used, for example, to detect whether the reinforcing layer 30 is formed to have rib portions 35 (see FIG. 10 ), which will be described later, based on an image captured by the imaging unit 120. The determination unit 112 can be used, for example, to determine whether the rib portions 35 are properly formed based on the detection result of the detection unit 111, and to notify the worker of the determination result.

[0041] The operational control executed by the control unit 110 includes the operation of forming the reinforcing layer 30 by the forming device 200. In carrying out the above operation, the control unit 110 controls the operation of the forming system 100 and the forming device 200 so that the processing unit 210 of the forming device 200 applies fibrinogen solution L1 to one of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10, and sprays thrombin solution L2 onto one surface of the sheet-shaped cell culture 10 to form a reinforcing layer 30 made of fibrin gel extending linearly on one surface. Specific steps of the forming method will be described later.

[0042] The image capturing unit 120 can be configured, for example, by a known camera device capable of acquiring still image data and / or video data.

[0043] 1, the imaging unit 120 can be laid out above the stage 310 on which the container 320 containing the sheet-shaped cell culture 10 is placed. By arranging the imaging unit 120 in this manner, it becomes possible to capture the entire sheet-shaped cell culture 10 and the laminate 20 within the imaging field of view from above the stage 310. The placement of the imaging unit 120 is not particularly limited as long as it is possible to photograph the sheet-shaped cell culture 10 and the laminate 20, and it is also possible to lay out the imaging unit 120 below the container 320, for example.

[0044] The image display unit 130 can be configured, for example, by a known liquid crystal display or the like.

[0045] The forming system 100 can also be configured as a portable or non-portable information terminal device in which the control unit 110, the imaging unit 120, and the image display unit 130 are integrally incorporated.

[0046] <Forming device 200> As shown in FIGS. 1 and 3 to 5, the forming device 200 includes a processing unit 210 that performs processing on the sheet-shaped cell culture 10 to form a reinforcing layer 30 made of fibrin gel that extends linearly.

[0047] The processing unit 210 has an application unit 220 (see Figure 4) that applies fibrinogen liquid (liquid containing fibrinogen) L1 to one side of the sheet-shaped cell culture 10, and a spray unit 230 that can be positioned opposite one side of the sheet-shaped cell culture 10 and can spray thrombin liquid (liquid containing thrombin) L2 onto each of multiple different locations on one side.

[0048] In this embodiment, an example will be described in which the processing unit 210 selects the "front surface 10a" as one surface of the sheet-shaped cell culture 10 on which the reinforcing layer 30 is to be formed. However, the processing unit 210 may also select the "rear surface 10b" as one surface of the sheet-shaped cell culture 10 on which the reinforcing layer 30 is to be formed.

[0049] The applicator 220 can be configured to include, for example, a mechanism for dripping the fibrinogen liquid L1 onto the sheet-shaped cell culture 10. Examples of the mechanism for dripping the fibrinogen liquid L1 include a known syringe or pipette.

[0050] As shown in Figure 3, the spraying section 230 has a main body 231 with an opposing surface 233 that is positioned to face the surface 10a of the sheet-shaped cell culture 10 when spraying the thrombin solution L2 onto the surface 10a of the sheet-shaped cell culture 10, and a tubular body 232 that is fluidly connected to the inside of the main body 231 and supplies the thrombin solution L2 to the main body 231.

[0051] The main body 231 can be configured, for example, to have a shape similar to the planar shape of the sheet-shaped cell culture 10 and to have an opposing surface 233 larger than the surface 10a of the sheet-shaped cell culture 10.

[0052] In this embodiment, the sheet-shaped cell culture 10 is configured to have a circular planar shape, and therefore the main body 231 can be configured to have a circular opposing surface 233 with an area that is approximately the same as or larger than the surface 10a of the sheet-shaped cell culture 10. There are no particular restrictions on the specific shapes of the main body 231 and the opposing surface 233, and they can be arbitrarily changed depending on, for example, the planar shape of the sheet-shaped cell culture 10 and the size of the surface 10a.

[0053] The tubular body 232 is configured to be in fluid communication with a supply source (a tank or a liquid storage tank) of the thrombin liquid L2 (not shown). Inside the tubular body 232, a flow path 232a is provided through which the thrombin liquid L2 can flow.

[0054] When spraying the thrombin solution L2 onto the sheet-shaped cell culture 10, the spraying unit 230 delivers the thrombin solution L2 from a predetermined supply source to the main body 231. The spraying unit 230 can spray the thrombin solution L2 onto the surface 10a of the sheet-shaped cell culture 10 through a plurality of ejection holes 235 formed in the main body 231. Note that, inside the main body 231, a flow path is formed for moving the thrombin solution L2 delivered via the tubular body 232 to each of the ejection holes 235.

[0055] 3, the spray unit 230 has a plurality of nozzles 235 that spray the thrombin solution L2. The plurality of nozzles 235 are arranged in parallel linear fashion (in the form of first portions 235A and second portions 235B, which will be described later).

[0056] Each of the ejection holes 235 is open so as to communicate with the outside on the opposing surface 233 side of the main body 231. Each of the ejection holes 235 can be configured, for example, in a circular shape.

[0057] As shown by arrow f in FIG. 3, the ejection hole 235 can eject the thrombin liquid L2 in a radial direction vertically downward and in a 360° radius from the center position of the ejection hole 235 (center position in a plan view).

[0058] As shown in Figure 3, the multiple nozzles 235 have a portion (hereinafter referred to as "first portion 235A") that increases the amount of thrombin solution sprayed per given area of ​​the surface 10a of the sheet-shaped cell culture 10, and a portion (hereinafter referred to as "second portion 235B") that decreases the amount of thrombin solution sprayed per given area of ​​the surface 10a of the sheet-shaped cell culture 10.

[0059] The first portion 235A (region) indicated by the dashed line in FIG. 3 is arranged in the vertical direction along the surface of the main body 231 (the direction corresponding to the arrows Y1-Y2, which is the surface direction of the sheet-shaped cell culture 10). Similarly, the second portion 235B (region) indicated by the dashed line is arranged parallel to the first portion 235A in the vertical direction along the surface of the main body 231. The first portion 235A and the second portion 235B are arranged alternately in the horizontal direction (the direction corresponding to the arrows X1-X2, which is the surface direction of the sheet-shaped cell culture 10) perpendicular to the vertical direction. The multiple first portions 235A and the multiple second portions 235B may be arranged alternately and parallel to each other. The ejection holes 235 are arranged at equal intervals in each of the first portion 235A and the second portion 235B.

[0060] 7, when the thrombin solution L2 is sprayed from the spray unit 230 configured as described above, a greater amount of thrombin solution L2 is sprayed in the region of the sheet-shaped cell culture 10 arranged to face the first portion 235A (hereinafter referred to as the "first region A1") than in the region of the sheet-shaped cell culture 10 arranged to face the second portion 235B (hereinafter referred to as the "second region A2") Therefore, in the laminate 20, the thickness of the reinforcing layer 30 in the portion corresponding to the first region A1 is greater than the thickness of the reinforcing layer 30 in the portion corresponding to the second region A2.

[0061] In FIG. 7, the positions where the nozzle holes 235 are disposed in the laminate 20 when the thrombin solution L2 is sprayed onto the sheet-shaped cell culture 10 are shown by dashed lines.

[0062] Third regions A3 are formed near both side portions of the sheet-shaped cell culture 10 that are located further laterally than the first region A1, which is located most laterally (in the direction indicated by the arrows X1-X2). The third region A3 is a region where the nozzles 235 of the sprayer 230 do not overlap in the planar direction when spraying the thrombin solution L2 from the sprayer 230. Therefore, a portion of the thrombin solution L2 sprayed from the sprayer 230 located at the first portion 235A disposed adjacent to the third region A3 is sprayed onto the third region A3. Therefore, the thickness of the reinforcing layer 30 formed in the third region A3 is thinner than that of the first region A1 and substantially the same as that of the second region A2.

[0063] As described above, the first portion 235A extends in the vertical direction, and therefore the first region A1, where the amount of sprayed is large, is formed continuously in the vertical direction. Therefore, the portion where the reinforcing layer 30 is formed thick due to the large amount of sprayed thrombin solution L2 has a substantially constant thickness in the vertical direction (the direction of the arrows Y1-Y2), as shown in the cross-sectional view of FIG.

[0064] Furthermore, because the second portion 235B extends in the vertical direction as described above, the second region A2, where the amount of sprayed thrombin solution L2 is small, is formed continuously in the vertical direction. Therefore, the portion of the reinforcing layer 30 where the amount of sprayed thrombin solution L2 is small and thus formed thinner than the first region A1, has a substantially constant thickness in the vertical direction (the direction of arrows Y1-Y2), as shown in the cross-sectional view of FIG. 8. The thickness of the reinforcing layer 30 in the first region A1 formed by the first portion 235A is greater than the thickness of the reinforcing layer 30 in the second region A2 formed by the second portion 235B. As a result, there is a difference in the thickness of the reinforcing layer 30 between the first region A1 and the second region A2.

[0065] 10, in the horizontal direction (arrow X1-X2 direction) perpendicular to the vertical direction (arrow Y1-Y2 direction), the reinforcing layer 30 forms ridge portions 35 in which thick convex portions 35a (convex cross-sectional portions) and thin concave portions 35b (concave cross-sectional portions) are alternately repeated corresponding to the first region A1 and the second region A2. The ridge portions 35 are in a state in which thick convex portions 35a of the reinforcing layer 30 and thin concave portions 35b of the reinforcing layer 30 are connected in parallel.

[0066] As described above, the laminate 20 is configured to have a reinforcing layer 30 of a predetermined pattern having convex portions 35a extending linearly with a constant thickness in the vertical direction (arrow Y1-Y2 direction) and concave portions 35b adjacent to the convex portions 35a in the horizontal direction (arrow X1-X2 direction) and extending linearly with a thickness smaller than that of the convex portions 35a, by spraying the thrombin solution L2 onto the sheet-shaped cell culture 10 from the spray section 230 having the first portion 235A and the second portion 235B.

[0067] The thrombin solution L2 used to form the reinforcing layer 30 made of fibrin gel generally has a lower viscosity than the fibrinogen solution L1. Therefore, when forming the linearly extending reinforcing layer 30 having the ridged portions 35, the spraying position and spray amount of the thrombin solution L2 are easier to control than with the fibrinogen solution L1. Therefore, by spraying the thrombin solution L2 onto the sheet-shaped cell culture 10 using the spray unit 230, it is possible to appropriately and easily form the linearly extending reinforcing layer 30 in a predetermined pattern.

[0068] The laminate 20 can exhibit the following effects by forming the reinforcing layer 30 extending linearly on the surface 10a of the sheet-shaped cell culture 10.

[0069] 11 shows a cross-sectional view of the laminate 20 in a curved state housed in a lumen 410 of a tubular portion (outer tubular portion) 400 of a transplantation device used to deliver the laminate 20 to a transplantation site in the human body. As shown in this cross-sectional view, when delivering the laminate 20 to a transplantation site in the human body, the entire laminate 20 is deformed into a curved shape having an arc-shaped cross section based on the central axis c1 of the tubular portion 400, thereby making it possible to deliver the laminate 20 to the transplantation site in a compact state. A carrier 420 on which the laminate 20 is placed is disposed on the inner circumferential side of the tubular portion 400.

[0070] When the surgeon deforms the laminate 20 and accommodates it in the tubular section 400 as described above, the surgeon bends the laminate 20 so that the surface 10a of the sheet-shaped cell culture 10 on which the rib portions 35 are formed faces inward (toward the central axis c1). When the surgeon bends the laminate 20 as described above, the depressions 35b of the rib portions 35, where the thickness of the reinforcing layer 30 is small, become the starting point of deformation. Therefore, compared to when the reinforcing layer 30 is formed with a uniform thickness over the entire surface 10a, the laminate 20 can be easily deformed to have an arc-shaped cross section as shown in FIG. 11. At this time, the distance between the vertices of the multiple protrusions 35a becomes shorter, allowing the laminate 20 to be accommodated along the inner circumferential surface of the tubular section 400. Therefore, cracks or the like can be prevented from occurring in the reinforcing layer 30 when it is folded. Note that multiple adjacent protrusions 35a may come into contact with each other when accommodated. As described above, the surgeon can smoothly set the laminate 20 into the tubular portion 400 of the transplantation device, and can smoothly deliver the laminate 20 to the transplantation site using the transplantation device.

[0071] The specific shape of the linear pattern of the reinforcing layer 30 formed in the sheet-shaped cell culture 10 is not particularly limited as long as it can achieve the effects described above. For example, the cross-sectional shape of the ridge portion 35, the number of convex portions 35a and concave portions 35b forming the ridge portion 35, the thickness of each portion, etc. can be changed as desired. However, preferably, there are at least two to six convex portions 35a, and at least three to seven concave portions 35b.

[0072] As shown in FIG. 3, the plurality of orifices 235 are arranged in a predetermined pattern including dense and sparsely spaced portions along the surface direction of the front surface 10a of the sheet-shaped cell culture 10.

[0073] Specifically, the number of the plurality of ejection holes 235 arranged in the first portion 235A per given area is greater than the number of the plurality of ejection holes 235 arranged in the second portion 235B. In other words, the ejection holes 235 are arranged more densely in the first portion 235A than in the second portion 235B. Furthermore, the plurality of ejection holes 235 have different opening areas per unit area in the first portion 235A and the second portion 235B. The opening area of ​​the ejection holes 235 per unit area in the first portion 235A is larger than the opening area of ​​the ejection holes 235 per unit area in the second portion 235B.

[0074] As described above, the thrombin solution L2 discharged from the nozzle holes 235 is sprayed vertically downward and in radial directions extending 360° from the center of the nozzle hole 235 (the center in a plan view). In this embodiment, the nozzle holes 235 all have the same shape and opening area. Therefore, when the thrombin solution L2 is sprayed simultaneously from multiple nozzle holes 235, the amount of thrombin solution L2 sprayed onto the sheet-shaped cell culture 10 is greater in the first portion 235A where the distance between adjacent nozzle holes 235 is short in the vertical direction (the direction corresponding to the arrows Y1-Y2 on the sheet-shaped cell culture 10) and the horizontal direction (the direction corresponding to the arrows X1-X2 on the sheet-shaped cell culture 10) than in the second portion 235B where the distance between adjacent nozzle holes 235 is long. Therefore, by spraying thrombin solution L2 from each of the nozzles 235 onto the sheet-shaped cell culture 10, the spray unit 230 makes it possible to more easily and reliably manufacture the laminate 20 having the linearly extending reinforcing layer 30 shown in Figure 10.

[0075] In this embodiment, the nozzle holes 235 are configured to have the same shape and the same opening area, but the specific form of the nozzle holes 235 is not particularly limited as long as a linearly extending reinforcing layer 30 can be formed in the sheet-shaped cell culture 10. For example, as will be described in a modified example below, the opening areas of the nozzle holes 235 may be configured to be different between the first portion 235A and the second portion 235B, or each nozzle hole 235 may be configured to have a different shape or opening area. Furthermore, the degree of density of the nozzle holes 235 in the first portion 235A and the second portion 235B (such as the distance between the nozzle holes 235) is not particularly limited as long as a linearly extending reinforcing layer 30 can be formed in the sheet-shaped cell culture 10. For example, the nozzle holes 235 may be configured as linear openings.

[0076] The fibrinogen solution L1 and the thrombin solution L2 are not particularly limited as long as they react with each other and solidify to form a fibrin gel. There are also no particular limitations on the concentrations of the solutions L1 and L2, the amounts applied per unit time, the volume ratio of the solutions L1 and L2, the application pressure, etc. The specific forms of the solutions L1 and L2 used in this embodiment can be determined by referring to known information (e.g., JP 2016-52272 A, etc.).

[0077] <Formation method> Next, a forming method according to this embodiment will be described.

[0078] The specific work content of each step will be described below with reference to the flowchart shown in Fig. 2 and Fig. 4 to Fig. 6. The following description will also explain an example in which the forming system 100 and forming device 200 shown in Fig. 1 automatically perform steps S11 to S14 of the forming method.

[0079] To start the formation method, a sheet-shaped cell culture 10 is prepared before the formation of the reinforcing layer 30 (step S10), as shown in Fig. 3. Specifically, a container 320 containing the sheet-shaped cell culture 10 is prepared, and the container 320 is placed on a stage 310 (see Fig. 1).

[0080] 4, the fibrinogen liquid L1 is applied to the sheet-shaped cell culture 10 (step S11). The control unit 110 controls the operation of the application unit 220 of the processing unit 210 to apply the fibrinogen liquid L1 to the surface 10a (corresponding to "one side") of the sheet-shaped cell culture 10.

[0081] 5, the thrombin solution L2 is sprayed onto the sheet-shaped cell culture 10 (step S12). The control unit 110 controls the operation of the spray unit 230 of the processing unit 210 to spray the thrombin solution L2 onto the surface 10a of the sheet-shaped cell culture 10 on which the fibrinogen solution L1 has been applied.

[0082] Next, the process waits until the fibrinogen solution L1 and the thrombin solution L2 applied to the surface 10a of the sheet-shaped cell culture 10 solidify to form a reinforcing layer 30 made of fibrin gel (step S13). When the solidification of the fibrinogen solution L1 and the thrombin solution L2 has progressed sufficiently, the reinforcing layer 30 extending linearly as described above is formed on the surface 10a of the sheet-shaped cell culture 10 (see FIGS. 7 and 10).

[0083] By the above operations, a laminate 20 in which the sheet-shaped cell culture 10 is reinforced by the reinforcing layer 30 can be obtained.

[0084] As described above, the forming device 200 of this embodiment is an apparatus equipped with a processing unit 210 that performs processing to form a reinforcing layer 30 made of fibrin gel that extends linearly on the sheet-shaped cell culture 10, and the processing unit 210 has a spray unit 230 that can be positioned to face one of the front and back surfaces 10a and 10b of the sheet-shaped cell culture 10 and can spray thrombin solution L2 onto each of multiple different locations on one surface.

[0085] Furthermore, the formation method according to this embodiment is a method for forming a reinforcing layer 30 made of fibrin gel on a sheet-shaped cell culture 10, and includes the steps of a processing unit 210 applying a fibrinogen liquid L1 to one of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10, and a processing unit 210 spraying a thrombin liquid L2 onto one surface of the sheet-shaped cell culture 10 to form a reinforcing layer 30 made of fibrin gel extending linearly on one surface.

[0086] Furthermore, the laminate 20 of this embodiment is a laminate 20 having a sheet-shaped cell culture 10 and a reinforcing layer 30 consisting of a fibrin gel formed on the sheet-shaped cell culture 10, and the reinforcing layer 30 extends linearly on one of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10.

[0087] In addition, the program according to this embodiment causes a forming device 200 equipped with a processing unit 210 that performs processing to form a reinforcing layer 30 made of fibrin gel on a sheet-shaped cell culture 10 to execute the steps of applying a fibrinogen solution L1 to one of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10, and spraying a thrombin solution L2 onto one surface of the sheet-shaped cell culture 10 to form a reinforcing layer 30 made of fibrin gel extending linearly on one surface.

[0088] According to this embodiment, it is possible to produce a laminate 20 having a reinforcing layer 30 extending linearly on one of the front surface 10a and back surface 10b of the sheet-shaped cell culture 10. When the laminate 20 is bent in a direction intersecting the planar direction, the thinner portions of the reinforcing layer 30 become the starting points of deformation, making it possible to easily achieve bending. Therefore, the surgeon can smoothly set the laminate 20, which is formed by forming the reinforcing layer 30 on the sheet-shaped cell culture 10, into a transplantation instrument, and can smoothly deliver the laminate 20 to the transplantation site using the transplantation instrument.

[0089] Through the embodiments, the device for forming a reinforcing layer on a sheet-shaped cell culture, the method for forming a reinforcing layer on a sheet-shaped cell culture, the laminate, and the program according to the present invention have been described. However, the present invention is not limited to the configurations described in the embodiments, and can be modified as appropriate based on the claims.

[0090] For example, the processing unit of the forming device may be configured to include at least a spray unit, i.e., the processing unit may not include an application unit for applying fibrinogen liquid, and the spray unit may be incorporated alone into an apparatus or system for manufacturing a laminate.

[0091] (Variation) Furthermore, for example, the forming device may employ the configuration of spray section 230A according to a modified example shown in FIG.

[0092] In the above-described embodiment, an example of a configuration of the spray section 230 has been described in which a plurality of nozzles 235 are arranged in a predetermined pattern including densely spaced portions and sparsely spaced portions along the planar direction of one surface (e.g., surface 10a) of the sheet-shaped cell culture 10 (see FIG. 3). However, there are no particular limitations on the shape and arrangement of the plurality of nozzles formed in the spray section, as long as they can form a reinforcing layer 30 that extends linearly relative to the sheet-shaped cell culture 10.

[0093] In the spray section 230A shown in Figure 12, the multiple nozzles 235 include a first nozzle 235a that sprays a larger amount of thrombin liquid L2 per unit time, and a second nozzle 235b that sprays a smaller amount of thrombin liquid L2 per unit time than the first nozzle 235a.

[0094] First nozzle hole 235a and second nozzle hole 235b have a circular shape. The opening area of ​​first nozzle hole 235a is larger than the opening area of ​​second nozzle hole 235b. Therefore, when thrombin liquid L2 is sprayed from spray part 230A, the amount of thrombin liquid L2 sprayed per unit time from first nozzle hole 235a is greater than that from second nozzle hole 235b.

[0095] 12, the first portion 235A of the spray unit 230A is configured with a region in which a plurality of first nozzle holes 235a are arranged in the vertical direction. The second portion 235B of the spray unit 230A is configured with a region in which a plurality of second nozzle holes 235b are arranged in the vertical direction. The first portion 235A and the second portion 235B are arranged alternately in the horizontal direction perpendicular to the vertical direction.

[0096] 13, when thrombin solution L2 is sprayed from the spray unit 230 having the portions 235A and 235B, a greater amount of thrombin solution L2 is sprayed in the first region A1 arranged to face the first portion 235A in the sheet-shaped cell culture 10 than in the second region A2 arranged to face the second portion 235B in the sheet-shaped cell culture 10. Therefore, in the laminate 20, the thickness of the reinforcing layer 30 in the portion corresponding to the first region A1 is greater than the thickness of the reinforcing layer 30 in the portion corresponding to the second region A2.

[0097] Furthermore, a third region A3 is formed near both sides of the first region A1, which is located most laterally (in the direction indicated by the arrows X1-X2) in the sheet-shaped cell culture 10. A portion of the thrombin solution L2 sprayed from the first portion 235A disposed adjacent to the third region A3 is sprayed onto the third region A3. Therefore, the thickness of the reinforcing layer 30 formed in the third region A3 is thinner than that of the first region A1 and substantially the same as that of the second region A2.

[0098] In FIG. 13, the positions where the nozzle holes 235a and 235b are disposed in the laminate 20 when the thrombin solution L2 is sprayed onto the sheet-shaped cell culture 10 are shown by dashed lines.

[0099] Because the first portion 235A extends in the vertical direction, the first region A1, where the amount of sprayed is large, is formed continuously in the vertical direction. Therefore, the portion where the reinforcing layer 30 is formed thick due to the large amount of sprayed thrombin solution L2 has a substantially constant thickness in the vertical direction (the direction of arrows Y1-Y2), as shown in the cross-sectional view of FIG.

[0100] Furthermore, because the second portion 235B extends in the vertical direction, the second region A2, where the amount of sprayed is small, is formed continuously in the vertical direction. Therefore, the portion where the reinforcing layer 30 is formed thinner than the first region A1 due to the small amount of sprayed thrombin solution L2 has a substantially constant thickness in the vertical direction (the direction of the arrows Y1-Y2), as shown in the cross-sectional view of FIG.

[0101] Also, as shown in Figure 16, in the horizontal direction (arrow X1-X2 direction) perpendicular to the vertical direction (arrow Y1-Y2 direction), the reinforcing layer 30 forms ridge portions 35 in which thick convex portions 35a (convex cross-sectional portions) and thin concave portions 35b (concave cross-sectional portions) are alternately repeated corresponding to the first region A1 and the second region A2.

[0102] As described above, the laminate 20 is configured to have a reinforcing layer 30 of a predetermined pattern having convex portions 35a extending linearly with a constant thickness in the vertical direction (arrow Y1-Y2 direction) and concave portions 35b adjacent to the convex portions 35a in the horizontal direction (arrow X1-X2 direction) and extending linearly with a thickness smaller than that of the convex portions 35a, by spraying the thrombin solution L2 onto the sheet-shaped cell culture 10 from the spray section 230 having the first portion 235A and the second portion 235B.

[0103] As explained in the modified example, the spray unit according to the present invention is not particularly limited in its specific configuration, as long as it can form a reinforcing layer that extends linearly in a predetermined pattern on the sheet-shaped cell culture 10. [Explanation of symbols]

[0104] 10. Sheet-shaped cell culture 10a Surface of sheet-shaped cell culture 10b Backside of sheet-shaped cell culture 20 laminate 30 Reinforcement layer 35 Ridged part 35a Convex part 35b recess 100 Formation System 200 Forming device 210 Processing section 220 Application section 230 Spray section 230A spray section 231 Main body of spray unit 232 Tube 232a Channel 233 Opposite surface of main body 235 Spout hole 235A Part 1 235B 2nd part 235a 1st vent 235b 2nd vent 310 Stage 320 container 400 Cylindrical part 410 Lumen of cylindrical part A1: First region of the stack A2 Second region of the stack A3 Third region of the laminate L1 fibrinogen liquid L2 thrombin solution c1 Central axis of the cylindrical part

Claims

1. An apparatus including a processing unit that performs a process for forming a linearly extending reinforcing layer made of fibrin gel on a sheet-shaped cell culture, The processing unit can be positioned to face one of the front and back surfaces of the sheet-shaped cell culture, and has a spray unit capable of spraying thrombin solution onto each of multiple different locations on the one surface, in an apparatus for forming a reinforcing layer on a sheet-shaped cell culture.

2. the spray unit has a plurality of spray holes for spraying the thrombin liquid, An apparatus for forming a reinforcing layer on a sheet-shaped cell culture as described in claim 1, wherein the plurality of nozzles have portions where the amount of thrombin solution sprayed per specified area on one side of the sheet-shaped cell culture is increased and portions where it is decreased.

3. The device for forming a reinforcing layer on a sheet-shaped cell culture according to claim 2 , wherein the plurality of nozzles are arranged in a plurality of parallel straight lines.

4. 4. An apparatus for forming a reinforcing layer on a sheet-shaped cell culture as described in claim 2 or claim 3, wherein the plurality of nozzles are arranged in a predetermined pattern including densely spaced portions and sparsely spaced portions along the surface direction of the one surface of the sheet-shaped cell culture.

5. 4. An apparatus for forming a reinforcing layer on a sheet-shaped cell culture as described in claim 2 or claim 3, wherein the plurality of nozzles include a first nozzle that sprays a larger amount of the thrombin liquid per unit time and a second nozzle that sprays a smaller amount of the thrombin liquid per unit time than the first nozzle.

6. A method for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, comprising: a step in which a processing unit applies a fibrinogen solution to one of the front and back surfaces of the sheet-shaped cell culture; A method for forming a reinforcing layer on a sheet-shaped cell culture, comprising the step of: the processing unit spraying thrombin solution onto one surface of the sheet-shaped cell culture to form a reinforcing layer consisting of fibrin gel extending linearly on the one surface.

7. A laminate having a sheet-shaped cell culture and a reinforcing layer made of fibrin gel formed on the sheet-shaped cell culture, The reinforcing layer extends linearly on one of the front and back surfaces of the sheet-shaped cell culture.

8. An apparatus including a processing unit for performing a process to form a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, applying a fibrinogen solution to one of the front and back surfaces of the sheet-shaped cell culture; and spraying a thrombin solution onto the one surface of the sheet-shaped cell culture to form the reinforcing layer made of fibrin gel extending linearly on the one surface.

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