System for forming a reinforcing layer, method for forming a reinforcing layer, and program

Abstract

A system for forming a reinforcing layer is provided that allows for uniform formation of the laminate. [Solution] The formation system 100 includes a first application unit 111 that applies fibrinogen liquid L1 to a sheet-shaped cell culture 10, and a second application unit 112 that applies thrombin liquid L2 to the sheet-shaped cell culture, and is further equipped with a processing unit 110 for forming a reinforcing layer made of fibrin gel on the sheet-shaped cell culture, an imaging unit 120 that images the sheet-shaped cell culture to which the fibrinogen liquid has been applied, a detection unit 131 that detects the thickness of the fibrinogen liquid applied by the first application unit based on image information acquired by the imaging unit, a determination unit 132 that determines whether the thickness distribution of the fibrinogen liquid is uniform based on the thickness distribution of the fibrinogen liquid detected by the detection unit, and a control unit 130 that controls the application speed of the second application unit that applies thrombin liquid to each region.

Description

[Technical Field]

[0001] The present invention relates to a system for forming a reinforcing layer, a method for forming a reinforcing layer, 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 allowing them to adhere and aggregate into a sheet.

[0003] Because the sheet-shaped cell culture is constructed in the form of a thin film sheet, it is physically fragile and therefore requires careful handling, as it is prone to wrinkling and tearing during isolation from the culture substrate, transportation and storage of the sheet-shaped cell culture, and actual transplantation into a patient's body.

[0004] To solve the above problems, a method (fibrin treatment) 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). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent Publication No. 2014-179151 Summary of the Invention [Problem to be solved by the invention]

[0006] During fibrin treatment, some areas may contain a large amount of fibrinogen solution and some areas may contain less. If thrombin solution is supplied in such a situation where some areas contain a large amount of fibrinogen solution and some areas contain less, the thickness of the reinforcing layer that is finally formed may become uneven. As a result, the thickness of the laminate consisting of the sheet-shaped cell culture and the reinforcing layer may become unintentionally uneven.

[0007] When there are regions where the thickness of the laminate is uneven, the laminate is difficult to handle, and the sheet-shaped cell culture may be damaged or may be difficult to transplant into the target location.

[0008] An object of the present invention is to provide a system for forming a reinforcing layer, a method for forming a reinforcing layer, and a program for forming a reinforcing layer, which enable a laminate to be formed uniformly. [Means for solving the problem]

[0009] The present invention is achieved by any one of the following means (1) to (5).

[0010] (1) a processing unit for forming a reinforcing layer made of fibrin gel on the sheet-shaped cell culture, the processing unit comprising a first application unit for applying a fibrinogen solution to the sheet-shaped cell culture and a second application unit for applying a thrombin solution to the sheet-shaped cell culture; an imaging unit that images the sheet-shaped cell culture on which the fibrinogen solution has been applied; a detection unit that detects the thickness of the fibrinogen solution applied by the first application unit based on the image information acquired by the imaging unit; a determination unit that determines whether the thickness distribution of the fibrinogen liquid is uniform based on the thickness distribution of the fibrinogen liquid detected by the detection unit; and a control unit that controls the application speed of the second application unit that applies the thrombin solution in each region based on the thickness distribution of the fibrinogen solution when the thickness distribution of the fibrinogen solution is not uniform.

[0011] (2) The system for forming a reinforcing layer according to (1) further comprises a vibration unit for vibrating a container in which the sheet-shaped cell culture is placed.

[0012] (3) a step in which a first applicator applies a fibrinogen solution to the sheet-shaped cell culture; an imaging unit imaging the sheet-shaped cell culture on which the fibrinogen solution has been applied; a step in which a detection unit detects the thickness of the fibrinogen solution based on image information acquired by the imaging unit; a step in which a determination unit determines whether or not the thickness distribution of the fibrinogen liquid is uniform; If it is determined that the thickness distribution of the fibrinogen liquid is not uniform, the control unit controls the second application unit to control the application speed of the thrombin liquid in each region based on the thickness distribution of the fibrinogen liquid.

[0013] (4) The method for forming a reinforcing layer described in (3) further comprises a step of making the thickness of the fibrinogen liquid uniform if it is determined in the step of determining whether the thickness distribution of the fibrinogen liquid is uniform that the thickness distribution of the fibrinogen liquid is not uniform.

[0014] (5) A system having a processing unit for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, applying a fibrinogen solution to the sheet-shaped cell culture; taking an image of the sheet-shaped cell culture on which the fibrinogen solution has been applied; detecting the thickness of the applied fibrinogen solution; determining whether the thickness distribution of the applied fibrinogen solution is uniform; If it is determined that the thickness distribution of the fibrinogen liquid is not uniform, the program executes a step of controlling the application speed of the thrombin liquid in each region based on the thickness distribution of the fibrinogen liquid. [Effects of the Invention]

[0015] According to the system, method, and program for forming a reinforcing layer of the present invention, the first applicator applies fibrinogen liquid to a sheet-shaped cell culture, the imager captures an image of the sheet-shaped cell culture coated with the fibrinogen liquid, the detector detects the thickness of the fibrinogen liquid based on the image information acquired by the imager, the determiner determines whether the thickness distribution of the fibrinogen liquid is uniform, and if it is determined that the thickness distribution of the fibrinogen liquid is not uniform, the controller controls the second applicator to control the application speed of thrombin liquid in each region based on the thickness distribution of the fibrinogen liquid. Therefore, in areas where the fibrinogen liquid is thick, thrombin liquid is sprayed at a high speed, diffusing the fibrinogen liquid in the plane direction and thinning the thickness of the fibrinogen liquid. This allows the formation of a uniform laminate. [Brief explanation of the drawings]

[0016] [Figure 1] FIG. 1 is a block diagram illustrating a system for forming a reinforcement layer, according to an embodiment. [Figure 2] 4 is a flowchart illustrating steps in a method for forming a reinforcement layer, according to an embodiment. [Figure 3] FIG. 10 is a cross-sectional view for explaining a method for forming a reinforcing layer according to an embodiment, showing a state before a homogenization process is performed. [Figure 4] 10A and 10B are cross-sectional views for explaining a method for forming a reinforcing layer according to an embodiment, showing a state after a homogenization process has been performed. [Figure 5]This is a cross-sectional view for explaining a method of forming a reinforcing layer in an embodiment, and shows how, after a homogenization process, the thickness of the fibrinogen solution is still uneven and thrombin solution is supplied at varying speeds. [Figure 6] 10 is a flowchart showing steps of a method for forming a reinforcing layer according to Modification 1. [Figure 7] 10 is a flowchart showing steps of a method for forming a reinforcing layer according to a second modification. [Figure 8] 10 is a flowchart showing steps of a method for forming a reinforcing layer according to a third modification. DETAILED DESCRIPTION OF THE INVENTION

[0017] 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.

[0018] FIG. 1 is a simplified block diagram showing the overall configuration of a system 100 for forming a reinforcing layer according to the present embodiment (hereinafter also referred to as "forming system 100"). FIG. 2 is a flowchart showing each step of a method for forming a reinforcing layer according to the present embodiment (hereinafter also referred to as "forming method"). FIG. 3 is a cross-sectional view for explaining the method for forming a reinforcing layer according to the present embodiment, showing the state before a homogenization process is performed. FIG. 4 is a cross-sectional view for explaining the method for forming a reinforcing layer according to the present embodiment, showing the state after a homogenization process is performed.

[0019] Arrows X1 and X2 in each figure indicate directions parallel to the surface direction of the sheet-shaped cell culture 10, and arrows Z1 and Z2 indicate the thickness direction perpendicular to the surface direction of the sheet-shaped cell culture 10 and the laminate.

[0020] <Sheet-shaped cell culture 10> The sheet-shaped cell culture 10 used in this embodiment can contain any cells capable of forming the sheet-shaped cell culture 10. The specific types of such cells are not particularly limited, but include, for example, somatic stem cells (adult stem cells), mesenchymal stem cells, or iPS cell (induced pluripotent stem cell)-derived cardiomyocytes, fibroblasts, synovial cells, epithelial cells, and endothelial cells. Somatic stem cells preferably include skeletal myoblasts (myoblast cells). Furthermore, the term "sheet-shaped cell culture" as used herein refers to a sheet-like (thin film-like) structure in which cells are interconnected.

[0021] 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.

[0022] As shown in Figure 3, the sheet-shaped cell culture 10 has a surface 10a, a back surface 10b located opposite the surface 10a and facing the bottom 223 of the container 220, and a peripheral portion 10c having a circular outer shape.

[0023] 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 220 .

[0024] The container 220 is, for example, a culture substrate used for culturing the sheet-shaped cell culture 10. When the container 220 is a culture substrate, the container 220 can be made of a material that is impermeable to liquids such as liquid culture media. When the container 220 is a culture substrate, the solution (liquid culture media, etc.) used for culturing is discharged from the container 220 before starting the formation method.

[0025] As shown in Figures 3 and 4, the container 220 has an opening 221 that opens upward, a bottom 223 on which the back surface 10b of the sheet-shaped cell culture 10 is placed, and a side wall 225 that surrounds the bottom 223.

[0026] The container 220 has a substantially circular planar shape. The bottom 223 of the container 220 has a size (inner diameter) that is the same as or larger than that of the sheet-shaped cell culture 10.

[0027] 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.

[0028] <Forming system 100> 1, the formation system 100 includes a processing unit 110 that performs processing to form a reinforcing layer made of fibrin gel on the sheet-shaped cell culture 10, a first application unit 111 that applies fibrinogen liquid to the sheet-shaped cell culture 10, and a second application unit 112 that applies thrombin liquid to the sheet-shaped cell culture 10, an imaging unit 120 that images the sheet-shaped cell culture 10 to which the fibrinogen liquid has been applied, a detection unit 131 that detects the thickness of the fibrinogen liquid applied by the first application unit 111 based on image information acquired by the imaging unit 120, a determination unit 132 that determines whether the thickness distribution of the fibrinogen liquid is uniform based on the thickness distribution of the fibrinogen liquid detected by the detection unit 131, and a control unit 130 that controls the application speed of the second application unit 112 that applies thrombin liquid to each region based on the thickness distribution of the fibrinogen liquid if the thickness distribution of the fibrinogen liquid is not uniform.

[0029] As shown in FIG. 1, the formation system 100 may further include a stage 210 on which a container 220 containing a sheet-shaped cell culture 10 can be placed, a vibration unit 140 arranged above the stage 210 and vibrating the container 220, and an image display unit 150 capable of displaying an image generated from image information (image data) acquired by the imaging unit 120.

[0030] As shown in Figure 1, the processing unit 110 can be configured to have a first application unit 111 that applies fibrinogen liquid (liquid containing fibrinogen) L1 to the sheet-shaped cell culture 10, and a second application unit 112 that applies thrombin liquid (liquid containing thrombin) L2 to the sheet-shaped cell culture 10.

[0031] The first applicator 111 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.

[0032] The second applicator 112 can be configured to include, for example, a mechanism for spraying the thrombin solution L2 onto the sheet-shaped cell culture 10. The mechanism for spraying the thrombin solution L2 can be, for example, a known spray.

[0033] 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 and volume ratios of the respective solutions L1 and L2, the amount applied per unit time, the application pressure, etc. The specific forms of the respective solutions L1 and L2 used in this embodiment can be determined by referring to known information (e.g., JP 2014-179151 A, etc.).

[0034] The imaging unit 120 can be configured with a known camera device capable of acquiring still image data and / or video data. As shown in Fig. 1, the imaging unit 120 is disposed on the front side of the sheet-shaped cell culture 10 in order to detect the thickness of the fibrinogen solution L1 applied by the first applicator 111.

[0035] The control unit 130 is configured to comprehensively control each unit of the forming system 100. The control unit 130 is configured with a CPU (Central Processing Unit), memories such as RAM (Random Access Memory) and ROM (Read Only Memory), and a storage unit, and controls each of the above units and performs various arithmetic processing according to a program.

[0036] The storage unit is configured with an HDD (Hard Disc 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 130 to function as a detection unit 131 and a determination unit 132.

[0037] The operational control executed by the control unit 130 includes the operation of forming a reinforcing layer using the forming system 100. In carrying out the above operations, the control unit 130 controls the operation of the forming system 100 to execute the following steps: applying fibrinogen liquid L1 to the sheet-shaped cell culture 10; imaging the sheet-shaped cell culture 10 to which the fibrinogen liquid L1 has been applied; detecting the thickness of the applied fibrinogen liquid L1; determining whether the thickness distribution of the applied fibrinogen liquid L1 is uniform; and, if the thickness distribution of the fibrinogen liquid L1 is not uniform, controlling the application speed for applying the thrombin liquid L2 in each region based on the thickness distribution of the fibrinogen liquid L1. Specific steps of the forming method will be described later.

[0038] The detection unit 131 is configured to be able to detect the thickness of the fibrinogen solution L1 based on the image information acquired by the imaging unit 120. For this reason, as shown in Fig. 1, the imaging unit 120 is placed on the front side of the sheet-shaped cell culture 10, and images are taken along the horizontal direction. In this case, it is also possible to capture images of the 360° periphery of the sheet-shaped cell culture 10.

[0039] The determination unit 132 determines whether the thickness distribution of the fibrinogen liquid L1 is uniform or not based on the thickness distribution of the fibrinogen liquid L1 detected by the detection unit 131. Specifically, if the thickness of the fibrinogen liquid L1 is constant in the horizontal direction in the region directly above the sheet-shaped cell culture 10, it is determined to be uniform, and if the thickness of the fibrinogen liquid L1 is wavy, ridged, or uneven, it is determined to be non-uniform.

[0040] In the step of controlling the second application unit 112 to determine whether the thickness distribution of the fibrinogen liquid L1 is uniform, if it is determined that the thickness distribution of the fibrinogen liquid L1 is not uniform, the control unit 130 controls the application speed of the thrombin liquid L2 in each region based on the thickness distribution of the fibrinogen liquid L1.

[0041] The vibrating unit 140 is disposed above the stage 210. The vibrating unit 140 vibrates the container 220 in which the sheet-shaped cell culture 10 is placed. The vibrating unit 140 may have any configuration as long as it can vibrate the container 220. This allows the thickness distribution of the fibrinogen solution L1 to be suitably uniform.

[0042] The image display unit 150 can be configured, for example, by a known liquid crystal display or the like.

[0043] The forming system 100 can also be configured as a portable or non-portable information terminal device in which the imaging unit 120, the control unit 130, and the image display unit 150 are integrally incorporated.

[0044] <Formation method> Next, a forming method according to this embodiment will be described.

[0045] The specific work content of each step will be described below with reference to the flowchart shown in Fig. 2 and Fig. 3 to Fig. 5. The following description also describes an example in which the forming system 100 shown in Fig. 1 automatically performs steps S10 to S18 of the forming method.

[0046] To start the formation method, a container 220 is prepared, which contains a sheet-shaped cell culture 10 before a reinforcing layer made of fibrin gel is formed. The container 220 is placed on a stage 210 (see FIG. 1).

[0047] Next, the fibrinogen solution L1 is supplied to the sheet-shaped cell culture 10 contained in the container 220 by the first applicator 111 (step S10).

[0048] Next, the imaging unit 120 starts to capture an image of the sheet-shaped cell culture 10 to which the fibrinogen liquid L1 has been applied (step S11).

[0049] Next, the detection unit 131 detects the distribution of the thickness of the fibrinogen liquid L1 (step S12). The thickness of the fibrinogen liquid L1 is detected based on image information captured by the imaging unit 120. Alternatively, the thickness of the fibrinogen liquid L1 may be detected by irradiating the fibrinogen liquid L1 with a laser of a specific wavelength and detecting the thickness based on the amount of light that depends on the amount of fibrinogen liquid L1.

[0050] Next, the determination unit 132 determines whether the thickness distribution of the fibrinogen liquid L1 is uniform (step S13). Specifically, the determination unit 132 makes the determination based on the thickness distribution of the fibrinogen liquid L1 detected by the detection unit 131. If the determination unit 132 determines that the thickness distribution of the fibrinogen liquid L1 is uniform (step S13: YES), the second application unit 112 supplies the thrombin liquid L2 to the sheet-shaped cell culture 10 to which the fibrinogen liquid L1 has been applied (step S14). At this time, the application rate of the thrombin liquid L2 applied to the sheet-shaped cell culture 10 is uniform in each region in a planar view. When the thrombin liquid L2 is supplied to the sheet-shaped cell culture 10, the fibrinogen liquid L1 and the thrombin liquid L2 applied to the sheet-shaped cell culture 10 start to react, and solidification progresses as a result, forming a reinforcing layer.

[0051] On the other hand, as shown in FIG. 3, if the determining unit 132 determines that the thickness distribution of the fibrinogen liquid L1 is not uniform (step S13: NO), a process for uniforming the thickness of the fibrinogen liquid L1 is performed (step S15).

[0052] In the process of uniforming the thickness of the fibrinogen liquid L1, a predetermined waiting time is performed and / or the container 220 is vibrated by the vibrating unit 140. This causes the thickness of the fibrinogen liquid L1 to be uniform, as shown in FIG.

[0053] Next, the sheet-shaped cell culture 10 in which the thickness of the fibrinogen liquid L1 has been made uniform is photographed again by the imaging unit 120 (step S16).

[0054] Next, the determination unit 132 again determines whether the thickness distribution of the fibrinogen liquid L1 is uniform (step S17). If the determination unit 132 determines that the thickness distribution of the fibrinogen liquid L1 is uniform (step S17: YES), the process proceeds to step S14, and the second applicator 112 supplies the thrombin liquid L2 (step S14). At this time, the application speed of the thrombin liquid L2 applied to the sheet-shaped cell culture 10 is uniform in each region in a planar view.

[0055] Whether the thickness distribution of the fibrinogen solution L1 is uniform or not can be determined, for example, by the following method. When the difference between the maximum thickness length and the minimum thickness length of the reinforcing layer is equal to or less than the maximum thickness length of the reinforcing layer multiplied by a tolerance, the distribution is determined to be uniform. In this case, the tolerance can be set, for example, between 0 and 10%. For example, if the thickness of the sheet-shaped cell culture 10 is 100 to 1000 μm, the maximum thickness length of the reinforcing layer is 5000 μm, and the minimum thickness length is 0 μm, and the tolerance for the difference is set to 10%, the thickness distribution of the fibrinogen solution L1 is determined to be uniform when the thickness of the reinforcing layer is 4500 μm to 5000 μm, and is determined to be inhomogeneous when it is less than 4500 μm.

[0056] On the other hand, if the determination unit 132 still determines that the thickness distribution of the fibrinogen liquid L1 is not uniform (step S17: NO), as shown in FIG. 5, the second applicator 112 supplies the thrombin liquid L2 at varying speeds (step S18). Specifically, as shown in FIG. 5, at the thicker portions LH of the fibrinogen liquid L1, the thrombin liquid L2 is sprayed at a high speed toward the apex of the thickness so that the thicker portions are evenly distributed in the planar direction. This spreads the fibrinogen liquid L1 in the planar direction, thinning the thickness of the fibrinogen liquid L1. The thrombin liquid L2 is then supplied at a constant speed to areas other than the thicker portions of the fibrinogen liquid L1. Note that in FIG. 5, thick arrows indicate a high speed, and thin arrows indicate a low speed. Note that after the thrombin liquid L2 is supplied at a high speed to the thicker portions LH, the thrombin liquid L2 may be supplied at a constant speed to the entire upper surface of the sheet-shaped cell culture 10. When step S18 is completed, the forming method ends.

[0057] As described above, the formation system 100 according to this embodiment includes a first application unit 111 that applies fibrinogen liquid L1 to the sheet-shaped cell culture 10, and a second application unit 112 that applies thrombin liquid L2 to the sheet-shaped cell culture 10, a processing unit 110 that forms a reinforcing layer made of fibrin gel on the sheet-shaped cell culture 10, an imaging unit 120 that images the sheet-shaped cell culture 10 to which the fibrinogen liquid L1 has been applied, and a first application unit 112 that applies thrombin liquid L2 to the sheet-shaped cell culture 10 based on image information acquired by the imaging unit 120. The device includes a detection unit 131 that detects the thickness of the fibrinogen liquid L1 applied by the application unit 111, a determination unit 132 that determines whether the thickness distribution of the fibrinogen liquid L1 is uniform or not based on the thickness distribution of the fibrinogen liquid L1 detected by the detection unit 131, and a control unit 130 that, if the thickness distribution of the fibrinogen liquid L1 is not uniform, controls the application speed of the second application unit 112 that applies thrombin liquid to each region based on the thickness distribution of the fibrinogen liquid L1.

[0058] Furthermore, the forming method according to this embodiment includes the steps of: a first application unit 111 applying fibrinogen liquid L1 to the sheet-shaped cell culture 10; an imaging unit 120 imaging the sheet-shaped cell culture 10 to which the fibrinogen liquid L1 has been applied; a detection unit 131 detecting the thickness of the fibrinogen liquid L1 based on the image information acquired by the imaging unit 120; a determination unit 132 determining whether the thickness distribution of the fibrinogen liquid L1 is uniform or not; and, if it is determined that the thickness distribution of the fibrinogen liquid L1 is not uniform, a control unit 130 controlling the second application unit 112 to control the application speed of the thrombin liquid L2 in each region based on the thickness distribution of the fibrinogen liquid L1.

[0059] Furthermore, the program for executing the forming method according to this embodiment causes a system having a processing unit 110 for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture 10 to execute the following steps: applying fibrinogen liquid L1 to the sheet-shaped cell culture 10; imaging the sheet-shaped cell culture 10 to which the fibrinogen liquid L1 has been applied; detecting the thickness of the applied fibrinogen liquid L1; determining whether the thickness distribution of the applied fibrinogen liquid L1 is uniform; and, if it is determined that the thickness distribution of the fibrinogen liquid L1 is not uniform, controlling the application speed of the thrombin liquid L2 in each region based on the thickness distribution of the fibrinogen liquid L1.

[0060] According to this embodiment, the first applicator 111 applies fibrinogen liquid L1 to the sheet-shaped cell culture 10, the imager 120 captures an image of the sheet-shaped cell culture 10 to which the fibrinogen liquid L1 has been applied, the detector 131 detects the thickness of the fibrinogen liquid L1 based on the image information acquired by the imager 120, the determiner 132 determines whether the thickness distribution of the fibrinogen liquid L1 is uniform, and if it is determined that the thickness distribution of the fibrinogen liquid L1 is not uniform, the controller 130 controls the second applicator 112 to control the application speed of the thrombin liquid L2 in each region based on the thickness distribution of the fibrinogen liquid L1. Therefore, in areas where the fibrinogen liquid L1 is thick, the thrombin liquid L2 is sprayed at a high speed to diffuse the fibrinogen liquid L1 in the planar direction, thereby thinning the thickness of the fibrinogen liquid L1. This allows the formation of a uniform laminate. Here, "uniformly formed" does not necessarily mean that the thickness is completely uniform.

[0061] Next, methods of forming a reinforcing layer according to Modifications 1 to 3 will be described with reference to Fig. 6 to Fig. 8. Fig. 6 is a flowchart showing the steps of the method of forming a reinforcing layer according to Modification 1. Fig. 7 is a flowchart showing the steps of the method of forming a reinforcing layer according to Modification 2. Fig. 8 is a flowchart showing the steps of the method of forming a reinforcing layer according to Modification 3.

[0062] In the above-described embodiment, after the homogenization process in step S15, it is determined again whether the thickness distribution of the fibrinogen liquid L1 is uniform (step S17), and if the thickness distribution of the fibrinogen liquid L1 is not uniform (step S17: NO), the thrombin liquid L2 is supplied at a variable speed. However, in the formation methods according to Modifications 1 and 2, steps S16 and onward can be omitted. That is, as shown in FIGS. 6 and 7, in step S15, after waiting for a predetermined time (for example, calculated based on the dispersion rate based on the viscosity of the fibrinogen liquid L1 and the elapsed time) or vibrating the container 220 with the vibration unit 140, the process may return to the image capturing step of step S11, and after confirming that the thickness distribution of the fibrinogen liquid has become uniform, the thrombin liquid may be supplied at a constant speed in step S14. In addition, after the judgment unit 132 judges that the thickness distribution of the fibrinogen liquid L1 is not uniform (step S13: NO), steps S15 to S17 may be omitted and the process may proceed to supplying the thrombin liquid L2 (with speed change) in step S18.

[0063] Furthermore, in the above-described embodiment, if it was determined in step S13 that the thickness distribution of the fibrinogen liquid L1 was not uniform (step S13: NO), a process for uniforming the thickness of the fibrinogen liquid L1 was performed (step S15). However, in the forming method according to Modification 3, steps S15, S16, and S17 can be omitted. That is, as shown in Fig. 8, if it was determined that the thickness distribution of the fibrinogen liquid L1 was not uniform (step S13: NO), the second applicator 112 supplies the thrombin liquid L2 at a variable speed (step S18).

[0064] The system for forming a reinforcing layer, the method for forming a reinforcing layer, and the program according to the present invention have been described through embodiments and modified examples, but the present invention is not limited to the configurations described in the embodiments, and can be modified as appropriate based on the claims. [Explanation of symbols]

[0065] 10. Sheet-shaped cell culture 100 Formation System 110 Processing section 111 First application section 112 Second application section 120 Imaging unit 130 control section 131 Detection unit 132 Judgment section 140 Vibration unit 150 Image display unit 210 Stages 220 Container L1 fibrinogen liquid L2 thrombin solution

Claims

1. a processing unit for forming a reinforcing layer made of fibrin gel on the sheet-shaped cell culture, the processing unit comprising a first applying unit for applying a fibrinogen solution to the sheet-shaped cell culture and a second applying unit for applying a thrombin solution to the sheet-shaped cell culture; an imaging unit that images the sheet-shaped cell culture coated with the fibrinogen solution; a detection unit that detects the thickness of the fibrinogen solution applied by the first application unit based on the image information acquired by the imaging unit; a determination unit that determines whether the thickness distribution of the fibrinogen liquid is uniform based on the thickness distribution of the fibrinogen liquid detected by the detection unit; A system for forming a reinforcing layer having a control unit that controls the application speed of the second application unit that applies the thrombin solution in each region based on the thickness distribution of the fibrinogen solution when the thickness distribution of the fibrinogen solution is not uniform.

2. The system for forming a reinforcing layer according to claim 1 , further comprising a vibration unit for vibrating a container in which the sheet-shaped cell culture is placed.

3. a step in which a first applicator applies a fibrinogen solution to the sheet-shaped cell culture; an imaging unit imaging the sheet-shaped cell culture on which the fibrinogen solution has been applied; a step in which a detection unit detects the thickness of the fibrinogen solution based on image information acquired by the imaging unit; a step in which a determination unit determines whether or not the thickness distribution of the fibrinogen liquid is uniform; A method for forming a reinforcing layer, comprising: if it is determined that the thickness distribution of the fibrinogen liquid is not uniform, the control unit controls the second application unit to control the application speed of the thrombin liquid in each region based on the thickness distribution of the fibrinogen liquid.

4. A method for forming a reinforcing layer as described in claim 3, further comprising a step of making the thickness of the fibrinogen liquid uniform if it is determined in the step of determining whether the thickness distribution of the fibrinogen liquid is uniform that the thickness distribution of the fibrinogen liquid is not uniform.

5. A system having a processing unit for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, applying a fibrinogen solution to the sheet-shaped cell culture; taking an image of the sheet-shaped cell culture on which the fibrinogen solution has been applied; detecting the thickness of the applied fibrinogen solution; determining whether the thickness distribution of the applied fibrinogen solution is uniform; If it is determined that the thickness distribution of the fibrinogen liquid is not uniform, the program executes a step of controlling the application speed of the thrombin liquid in each region based on the thickness distribution of the fibrinogen liquid.

Images