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

Abstract

A system for forming a reinforcing layer on a sheet-shaped cell culture is provided, which can form a reinforcing layer at a position where a tear has occurred in the sheet-shaped cell culture. [Solution] The forming system 100 includes an imaging unit 110 that photographs the sheet-like cell culture 10, a processing unit 120 that performs processing to form a reinforcing layer 30 made of fibrin gel on the sheet-like cell culture 10, a detection unit 131 that detects tears 10t in the sheet-like cell culture 10 based on image information acquired by the imaging unit 110, and a control unit 130 that controls the operation of the processing unit 120 so as to form the reinforcing layer 30 at the position where the tears 10t in the sheet-like cell culture 10 are pressing down.

Description

[Technical Field]

[0001] The present invention relates to a system for forming a reinforcing layer on a sheet-like cell culture, a method for forming a reinforcing layer, and a program for doing so. [Background technology]

[0002] Sheet-shaped cell cultures (cell sheets) are widely known for use in fields such as regenerative medicine. Generally, 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] Because sheet-shaped cell cultures are constructed in the form of a thin film sheet, they are generally physically fragile and prone to wrinkling and tearing during isolation from the culture substrate and subsequent handling, making them extremely difficult to handle when transporting and storing the sheet-shaped cell cultures, as well as when actually transplanting them 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 Patent Document 1). [Prior art documents] [Patent documents]

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

[0006] On the other hand, the condition of the sheet-shaped cell culture varies from patient to patient, and if the sheet-shaped cell culture is torn and fragile, even if a reinforcing layer is formed, the sheet-shaped cell culture may be difficult to handle when transplanted into the patient's body, or the tearing of the sheet-shaped cell culture may progress.

[0007] The present invention aims to provide a system for forming a reinforcing layer on a sheet-shaped cell culture, a method for forming a reinforcing layer, and a program for forming a reinforcing layer at locations where tears have occurred in the sheet-shaped cell culture. [Means for solving the problem]

[0008] The present invention is achieved by any one of the following means (1) to (3).

[0009] (1) an imaging unit that images the sheet-shaped cell culture; a processing unit that performs a process to form a reinforcing layer made of fibrin gel on the sheet-shaped cell culture; a detection unit that detects tears in the sheet-shaped cell culture based on image information acquired by the imaging unit; a control unit that controls the operation of the processing unit so as to form the reinforcing layer at the location where the tear has occurred in the sheet-shaped cell culture.

[0010] (2) an imaging unit capturing an image of the sheet-shaped cell culture; a step in which a detection unit detects tears in the sheet-shaped cell culture based on image information of the sheet-shaped cell culture that has been photographed; a step in which a processing unit forms a reinforcing layer made of fibrin gel at the location where the tear has occurred in the sheet-shaped cell culture.

[0011] (3) A system having a processing unit for performing a process to form a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, taking an image of the sheet-shaped cell culture; detecting tears in the sheet-shaped cell culture based on image information of the sheet-shaped cell culture; and a program for executing a step of controlling the operation of the processing unit so as to form the reinforcing layer at the location where the tear has occurred in the sheet-shaped cell culture. [Effects of the Invention]

[0012] According to the system, method, and program for forming a reinforcing layer on a sheet-shaped cell culture of the present invention, the detection unit detects tears in the sheet-shaped cell culture, and based on the results of the detection unit, the processing unit performs processing (application of fibrinogen solution and application of thrombin solution) to form a reinforcing layer at the location of the tear in the sheet-shaped cell culture. Therefore, according to the present invention, a reinforcing layer made of fibrin gel can be appropriately and simply formed at the location of the tear in the sheet-shaped cell culture. [Brief explanation of the drawings]

[0013] [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 each step of a method for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 3] FIG. 10 is a plan view showing a state in which a tear has occurred in the sheet-shaped cell culture according to the embodiment. [Figure 4] This is a plan view showing how a reinforcing layer is formed against tears occurring in a sheet-like cell culture according to the embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0014] 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.

[0015] Figure 1 is a simplified block diagram showing the overall configuration of the system 100 (hereinafter also referred to as "formation system 100") for forming a reinforcing layer 30 on a sheet-like cell culture 10 according to this embodiment. Figure 2 is a flowchart showing each step of the method (hereinafter also referred to as "formation method") for forming a reinforcing layer 30 on a sheet-like cell culture 10 according to this embodiment. Figure 3 is a plan view showing a tear 10t occurring in the sheet-like cell culture 10 according to this embodiment. Figure 4 is a plan view showing the formation of a reinforcing layer on the tear 10t occurring in the sheet-like cell culture 10 according to this embodiment.

[0016] The arrows X1, X2, Y1, and Y2 in each figure indicate directions parallel to the plane direction of the sheet-like cell culture 10, while the arrows Z1 and Z2 indicate the thickness direction perpendicular to the plane direction of the sheet-like cell culture 10.

[0017] <Sheet-shaped cell culture 10> The sheet-like cell culture 10 used in this embodiment may contain any cells capable of forming the sheet-like cell culture 10. There are no particular restrictions on the specific types of such cells, but examples include 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, "sheet-like cell culture" as used herein means a material in which cells are interconnected to form a sheet-like (thin-film) structure.

[0018] As shown in Figure 3, the sheet-shaped cell culture 10 can be configured to have, for example, a substantially circular planar shape in its natural state without the application of external force. 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.

[0019] The sheet-like cell culture 10 has a surface 10a and a back surface located opposite to the surface 10a and positioned facing the bottom 223 of the container 220. In this embodiment, the sheet-like cell culture 10 is assumed to have a tear 10t, as shown in Figures 3 and 4.

[0020] In this embodiment, the tear 10t reaches the peripheral edge 10c of the sheet-shaped cell culture 10 in plan view. However, the tear 10t does not have to reach the peripheral edge 10c of the sheet-shaped cell culture 10 in plan view. The shape of the tear 10t is not limited to the shape shown in Fig. 3, and the tear may overlap, turn up, or have a gap in the torn portion.

[0021] The formation method described below can be carried out with the sheet-shaped cell culture 10 contained in a predetermined container 220. In this embodiment, the target sheet-shaped cell culture 10 is generally in a state after it has been detached from the container 220. The sheet-shaped cell culture 10 may be in a state before it is detached from the container 220, and particularly when tears and curling are already present before detachment, a reinforcing layer may be formed to prevent the tears and curling before detachment.

[0022] 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.

[0023] As shown in Figure 3, the container 220 comprises an opening 221 that opens upward, a bottom 223 on which the back surface of the sheet-like cell culture 10 is placed, and side walls 225 surrounding the bottom 223.

[0024] The container 220 has a roughly circular shape in the plan view shown in Figure 3. The bottom 223 of the container 220 has the same size (inner diameter) as or larger than the sheet-like cell culture 10.

[0025] 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.

[0026] <Forming system 100> 1, 3, and 4, the formation system 100 comprises an imaging unit 110 that photographs the sheet-shaped cell culture 10, a processing unit 120 that performs processing to form a reinforcing layer 30 made of fibrin gel on the sheet-shaped cell culture 10, a detection unit 131 that detects tears 10t in the sheet-shaped cell culture 10, and a control unit 130 that controls the operation of the processing unit 120 so as to form a reinforcing layer at the location where the tear 10t has occurred in the sheet-shaped cell culture 10.

[0027] As shown in Figure 1, the formation system 100 may further include a stage 210 on which a container 220 containing a sheet-like cell culture 10 can be placed, and an image display unit 140 capable of displaying an image generated from image information (image data) acquired by the imaging unit 110.

[0028] The imaging unit 110 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 110 can be laid out above a stage 210 on which a container 220 containing a sheet-shaped cell culture 10 is placed. By arranging the imaging unit 110 in this manner, it becomes possible to capture the entire surface 10a of the sheet-shaped cell culture 10 within the imaging field of view from above the stage 210. The imaging unit 110 may also capture images of the sheet-shaped cell culture 10 from below.

[0029] As shown in Figure 1, the processing unit 120 can be configured to include a first coating unit 121 for coating the sheet-like cell culture 10 with fibrinogen solution (a liquid containing fibrinogen), and a second coating unit 122 for coating the sheet-like cell culture 10 with thrombin solution (a liquid containing thrombin).

[0030] The first coating unit 121 can be configured to include, for example, a mechanism for dropping fibrinogen solution onto the sheet-like cell culture 10. Examples of mechanisms for dropping fibrinogen solution include known syringes and pipettes.

[0031] The second coating unit 122 can be configured to include, for example, a mechanism for spraying thrombin solution onto the sheet-like cell culture 10. Examples of the mechanism for spraying thrombin solution include a known spray.

[0032] The fibrinogen solution and the thrombin solution 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 concentration of each solution, the amount applied per unit time, the volume ratio of each solution, the application pressure, etc. The specific form of each solution used in this embodiment can be determined by referring to known information (e.g., JP 2014-179151 A, etc.).

[0033] 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.

[0034] 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 130 to function as the detection unit 131 and the determination unit 132.

[0035] The operational control executed by the control unit 130 includes the operation of forming the reinforcing layer 30 by the formation system 100. In carrying out the above operation, the control unit 130 controls the operation of the formation system 100 to execute the following steps: the imaging unit 110 photographs the sheet-shaped cell culture 10; the detection unit 131 detects tears 10t in the sheet-shaped cell culture 10 based on image information of the photographed sheet-shaped cell culture 10; and the processing unit 120 forms the reinforcing layer 30 made of fibrin gel at the location of the tear 10t in the sheet-shaped cell culture 10. Specific steps of the formation method will be described later.

[0036] The image display unit 140 can be configured, for example, by a known liquid crystal display or the like.

[0037] The forming system 100 can also be configured as a portable or non-portable information terminal device in which the imaging unit 110, the control unit 130, and the image display unit 140 are integrally incorporated.

[0038] <Formation method> Next, a forming method according to this embodiment will be described.

[0039] The specific work content of each step will be described below with reference to the flowchart shown in Fig. 2 and Fig. 3 and Fig. 4. The following description will also explain an example in which the forming system 100 shown in Fig. 1 automatically performs steps S10 to S16 of the forming method.

[0040] To start the formation method, a container 220 containing the sheet-shaped cell culture 10 before the formation of the reinforcing layer 30 is prepared as shown in Fig. 3. The container 220 is placed on a stage 210 (see Fig. 1).

[0041] Next, the imaging unit 110 starts imaging the sheet-like cell culture 10 (step S10).

[0042] Next, the detection unit 131 detects tears 10t in the sheet-shaped cell culture 10 (step S11). The detection unit 131 detects tears 10t in the sheet-shaped cell culture 10 based on the image information acquired by the imaging unit 110. Specifically, a tear 10t is detected when the outline of the tear 10t is visible, and when a tear 10t has occurred, the transparent portion of the container 220 becomes visible.

[0043] Next, the control unit 130 (determination unit 132) determines whether or not a rupture 10t has occurred in the sheet-like cell culture 10 (step S12).

[0044] If the determining unit 132 determines that no tears 10t have occurred in the sheet-shaped cell culture 10 (step S12: NO), fibrinogen liquid is supplied to the entire sheet-shaped cell culture 10 (step S13), and then thrombin liquid is supplied (step S14) in order to improve operability. Note that, in this example, fibrinogen liquid is applied, and then thrombin liquid is applied to the area where fibrinogen liquid was applied, but the fibrinogen liquid and thrombin liquid may be applied to the entire sheet-shaped cell culture 10 at the same time.

[0045] On the other hand, as shown in Figure 3, if the determination unit 132 determines that a tear 10t has occurred in the sheet-like cell culture 10 (step S12: YES), the detection unit 131 detects the location of the tear 10t in the sheet-like cell culture 10 based on the image information captured by the imaging unit 110 (step S15).

[0046] Next, the control unit 130 controls the processing unit 120 to perform fibrin treatment at the position where the tear 10t occurred in the sheet-shaped cell culture 10 (step S16). Specifically, the control unit 130 controls the first applicator 121 to supply fibrinogen liquid to the position where the tear 10t occurred, and controls the second applicator 122 to supply thrombin liquid to the position where the fibrinogen liquid was supplied, thereby forming the reinforcing layer 30 as shown in Fig. 4. At this time, the fibrin treatment may be performed from directly above the position where the tear 10t occurred, or from the peripheral portion of the position where the tear 10t occurred, or the peripheral portion of the position where the tear 10t occurred may be bordered with fibrinogen liquid and thrombin liquid may be supplied inside the border. Then, after fibrin treatment at the location where the sheet-like cell culture 10 has ruptured 10t, the process proceeds to step S10, and the operation is repeated in step S12 until it is determined that no rupture 10t has occurred in the sheet-like cell culture 10.

[0047] After fibrin treatment at the location where the tear 10t has occurred, the fibrin treatment may be performed over the entire upper surface of the sheet-shaped cell culture 10, including the previously treated location (which has the effect of making the work easier), or may be performed on parts of the upper surface of the sheet-shaped cell culture 10 other than the previously treated location (which has the effect of making the thickness uniform).

[0048] By controlling the operation of the processing unit 120 as described above, the control unit 130 can form a reinforcing layer 30 at the position where the tear 10t occurred in the sheet-shaped cell culture 10, as shown in Figure 4. The formation of the reinforcing layer 30 improves the operability of the sheet-shaped cell culture 10. Then, by performing steps S13 and S14 and subjecting the entire sheet-shaped cell culture 10 to fibrin treatment, the handleability of the sheet-shaped cell culture 10 when transplanted into a patient's body is improved.

[0049] As described above, the formation system 100 of this embodiment includes an imaging unit 110 that photographs the sheet-shaped cell culture 10, a processing unit 120 that performs processing to form a reinforcing layer 30 made of fibrin gel on the sheet-shaped cell culture 10, a detection unit 131 that detects tears 10t in the sheet-shaped cell culture 10 based on image information acquired by the imaging unit 110, and a control unit 130 that controls the operation of the processing unit 120 so as to form the reinforcing layer 30 at the position dominated by the tear 10t in the sheet-shaped cell culture 10.

[0050] Furthermore, the forming method according to this embodiment includes the steps of: an imaging unit 110 photographing the sheet-shaped cell culture 10; a detection unit 131 detecting tears 10t in the sheet-shaped cell culture 10 based on image information of the photographed sheet-shaped cell culture 10; and a processing unit 120 forming a reinforcing layer 30 made of fibrin gel at the position where the tear 10t has occurred in the sheet-shaped cell culture 10.

[0051] Furthermore, the program for executing the formation method according to this embodiment is a program for causing a formation system 100 having a processing unit 120 that performs processing to form a reinforcing layer 30 made of fibrin gel on a sheet-shaped cell culture 10 to execute the following steps: imaging the sheet-shaped cell culture; detecting tears 10t in the sheet-shaped cell culture 10 based on image information of the sheet-shaped cell culture 10; and controlling the operation of the processing unit 120 so as to form a reinforcing layer 30 at the position where the tear 10t in the sheet-shaped cell culture 10 has occurred.

[0052] According to this embodiment, the detection unit 131 detects tears 10t in the sheet-shaped cell culture 10 based on image information of the sheet-shaped cell culture 10 captured by the imaging unit 110, and the processing unit 120 performs processing (application of fibrinogen solution L1 and application of thrombin solution L2) to form a reinforcing layer 30 at the position where the tear 10t has occurred in the sheet-shaped cell culture 10 based on the results of detection by the detection unit 131. Therefore, according to this embodiment, the reinforcing layer 30 made of fibrin gel can be appropriately and simply formed at the position where the tear 10t has occurred in the sheet-shaped cell culture 10.

[0053] The system for forming a reinforcing layer on a sheet-shaped cell culture, the method for forming a reinforcing layer on a sheet-shaped cell culture, and the program according to the present invention have been described through the embodiments, 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]

[0054] 10. Sheet-shaped cell culture 30 Reinforcement layer 100 Formation System 110 Imaging unit 120 Processing section 121 First application section 122 Second application section 130 Control Unit 131 Detection unit 140 Image display unit

Claims

1. an imaging unit that images the sheet-shaped cell culture; a processing unit that performs a process to form a reinforcing layer made of fibrin gel on the sheet-shaped cell culture; a detection unit that detects tears in the sheet-shaped cell culture based on image information acquired by the imaging unit; a control unit that controls the operation of the processing unit so as to form the reinforcing layer at the location where the tear has occurred in the sheet-shaped cell culture.

2. an imaging unit capturing an image of the sheet-shaped cell culture; a step in which a detection unit detects tears in the sheet-shaped cell culture based on image information of the sheet-shaped cell culture that has been photographed; a step in which a processing unit forms a reinforcing layer made of fibrin gel at the location where the tear has occurred in the sheet-shaped cell culture.

3. A system having a processing unit for performing a process to form a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, taking an image of the sheet-shaped cell culture; detecting tears in the sheet-shaped cell culture based on image information of the sheet-shaped cell culture; and a program for executing a step of controlling the operation of the processing unit so as to form the reinforcing layer at the location where the tear has occurred in the sheet-shaped cell culture.

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