Apparatus for forming a reinforcing layer on a sheet-shaped cell culture, method for forming a reinforcing layer on a sheet-shaped cell culture, 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, and a program for forming a reinforcing layer on a sheet-shaped cell culture, which can prevent wrinkles from occurring in the sheet-shaped cell culture when a reinforcing layer is formed on the sheet-shaped cell culture, and which enable stabilization of product quality of the laminate compared to when the sheet-shaped cell culture is stretched manually. [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 on a sheet-shaped cell culture 10, and the processing unit has one or more contact units 220, 230 with lattice-shaped contact surfaces 221a, 231a configured to be able to come into contact with the sheet-shaped cell culture, and the contact units are configured to be able to eject fibrinogen solution and / or thrombin solution onto the sheet-shaped cell culture that has come into contact with the contact surfaces.

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, 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] 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 transplantation into a patient's body.

[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). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent Publication No. 2016-52272 Summary of the Invention [Problem to be solved by the invention]

[0006] The laminate produced by the method of Patent Document 1 has the physical strength of part or all of the sheet-shaped cell culture reinforced by the reinforcing layer, which effectively prevents wrinkling and tearing when handled by workers.

[0007] When forming a reinforcing layer using the method of Patent Document 1 or the like, to ensure that the reinforcing layer has an appropriate thickness and that the shape and quality of the laminate are stable, an operator stretches the sheet-shaped cell culture during fibrin treatment (application of fibrinogen solution and application of thrombin solution) to prevent wrinkles from forming in the sheet-shaped cell culture. However, because this work is performed manually by the operator, it is dependent on the operator's intuition, skill, experience, etc., and therefore may result in variations in the product quality of the laminate.

[0008] 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, and a program that can prevent wrinkles from occurring in the sheet-shaped cell culture when a reinforcing layer is formed on the sheet-shaped cell culture, and that can stabilize the product quality of the laminate compared to when the sheet-shaped cell culture is stretched manually. [Means for solving the problem]

[0009] The present invention is achieved by any one of the following means (1) to (6).

[0010] (1) An apparatus comprising a processing unit for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, the processing section has one or more contact sections each having a lattice-shaped contact surface configured to be able to come into contact with the sheet-shaped cell culture, An apparatus for forming a reinforcing layer on a sheet-shaped cell culture, wherein the contact portion is configured to be able to eject fibrinogen solution and / or thrombin solution onto the sheet-shaped cell culture that has come into contact with the contact surface.

[0011] (2) the processing section has a first contact section and a second contact section on which the sheet-shaped cell culture is placed on the back side of the sheet-shaped cell culture, the first contact portion has a first contact surface and a plurality of first discharge holes capable of discharging the fibrinogen liquid, the second contact portion has a second contact surface and a plurality of second discharge holes capable of discharging the thrombin liquid, An apparatus for forming a reinforcing layer on a sheet-shaped cell culture described in (1), wherein the first discharge hole and the second discharge hole are capable of discharging fibrinogen solution or thrombin solution at positions where they do not overlap each other in the planar direction.

[0012] (3) the processing section has a first contact section and a second contact section that are pressed against the sheet-shaped cell culture from the surface side of the sheet-shaped cell culture, the first contact portion has a first contact surface and a plurality of first discharge holes capable of discharging the fibrinogen liquid, the second contact portion has a second contact surface and a second discharge hole capable of discharging the thrombin solution, An apparatus for forming a reinforcing layer on a sheet-shaped cell culture described in (1), wherein the first discharge hole and the second discharge hole are capable of discharging fibrinogen solution or thrombin solution at positions where they do not overlap each other in the planar direction.

[0013] (4) The device for forming a reinforcing layer on a sheet-shaped cell culture according to (1), wherein the contact part is configured to be able to discharge a thrombin solution.

[0014] (5) A method for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, comprising: a step of bringing the lattice-shaped contact surface of the contact part into contact with one of the front and back surfaces of the sheet-shaped cell culture while the sheet-shaped cell culture is suspended in a predetermined solution; removing the solution; A method for forming a reinforcing layer on a sheet-shaped cell culture, comprising the steps of: contacting the sheet-shaped cell culture with the contact surface and ejecting fibrinogen solution and / or thrombin solution from the contact portion onto the sheet-shaped cell culture.

[0015] (6) 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, a step of bringing the lattice-shaped contact surface of the contact part into contact with one of the front and back surfaces of the sheet-shaped cell culture while the sheet-shaped cell culture is suspended in a predetermined solution; removing the solution; and ejecting fibrinogen solution and / or thrombin solution from the contact portion onto the sheet-shaped cell culture while the sheet-shaped cell culture is in contact with the contact surface. [Effects of the Invention]

[0016] According to 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, and the program of the present invention, a fibrinogen solution and / or a thrombin solution can be ejected onto the sheet-shaped cell culture while the sheet-shaped cell culture is in contact with the contact surface of the contact unit provided in the processing unit. The sheet-shaped cell culture is prevented from wrinkling during fibrin treatment by having the fibrinogen solution and / or thrombin solution sprayed from the contact unit while in contact with the contact surface of the contact unit. Therefore, the product quality of the laminate can be stabilized compared to when an operator manually stretches the sheet-shaped cell culture to prevent wrinkling. [Brief explanation of the drawings]

[0017] [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 plan view for explaining a processing section (first contact section and second contact section) according to the embodiment. [Figure 4]FIG. 2 is a perspective view for explaining a processing section (first contact section and second contact section) according to the embodiment. [Figure 5] FIG. 4 is a cross-sectional view illustrating the internal structure of a first contact portion. [Figure 6] FIG. 4 is a cross-sectional view illustrating the internal structure of a second contact portion. [Figure 7] FIG. 10 is a diagram illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 8] FIG. 10 is a diagram illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to an embodiment. [Figure 9] 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 10] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to Modification 1. [Figure 11] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to Modification 1. [Figure 12] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to Modification 1. [Figure 13] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to Modification 1. [Figure 14] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to Modification 2. [Figure 15] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to Modification 2. [Figure 16] FIG. 10 is a plan view illustrating a method for forming a reinforcing layer on a sheet-shaped cell culture according to Modification 2. [Figure 17] 10 is a cross-sectional view illustrating the internal structure of a first contact portion according to Modification 2. FIG. [Figure 18] 10 is a cross-sectional view illustrating the internal structure of a second contact portion according to Modification 2. FIG. DETAILED DESCRIPTION OF THE INVENTION

[0018] (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.

[0019] 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.

[0020] Fig. 3 is a plan view (viewed from the direction of arrow III in Fig. 1) showing each contact portion 220, 230 of a forming apparatus 200 (hereinafter also referred to as "forming apparatus 200") including a processing unit 210 that forms a reinforcing layer 30 on the sheet-shaped cell culture 10 according to this embodiment, and Fig. 4 is a perspective view showing each contact portion 220, 230. Fig. 5 is a partial cross-sectional view of the first contact portion 220 taken along line VV shown in Fig. 4, and Fig. 6 is a partial cross-sectional view of the second contact portion 230 taken along line VI-VI shown in Fig. 4.

[0021] 7 to 9 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).

[0022] 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).

[0023] <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.

[0024] As shown in Figure 4, the sheet-shaped cell culture 10 can 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.

[0025] 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.

[0026] 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 FIG. 9).

[0027] 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 a solution 330 such as a liquid medium. The sheet-shaped cell culture 10 that is the subject of this embodiment is generally in a state after being detached from the container 320. The front surface 10a of the sheet-shaped cell culture 10 is the surface of the sheet-shaped cell culture 10 that is not adhered to the container 320 such as a culture substrate, and the back surface 10b is the surface of the sheet-shaped cell culture 10 that was adhered to the culture substrate and is the surface that is in contact with the culture substrate when left to stand after detachment.

[0028] The formation method described below can be carried out in a state where the sheet-shaped cell culture 10 is contained in a container 320 (see FIGS. 7 to 9). The formation method can also be started with a solution 330 contained in the container 320, and the solution 330 can be discharged from the container 320 during the formation method. The container 320 can be provided with a mechanism (for example, an openable / closable valve mechanism and a drain tube) that can discharge the solution 330 to the outside of the container 320 under the operational control of the control unit 110.

[0029] As shown in FIG. 7, 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.

[0030] 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. In addition, the opening 321 of the container 320 can be configured to have a size that allows the contact portions 220, 230 to be inserted into and removed from the container 320.

[0031] 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.

[0032] <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).

[0033] To give an overview with reference to Figures 1 and 7 to 9, 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.

[0034] 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.

[0035] 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.

[0036] The detection unit 111 can be used, for example, to detect whether wrinkles have occurred in the sheet-shaped cell culture 10 when the reinforcing layer 30 is formed on the sheet-shaped cell culture 10, based on the image captured by the imaging unit 120. The determination unit 112 can be used, for example, to determine whether wrinkles have occurred, based on the detection result of the detection unit 111, and notify the result to an operator or the like.

[0037] The operational control executed by the control unit 110 includes the operation of forming the reinforcing layer 30 using 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 to execute the following steps: bringing the lattice-shaped first contact surface 221a (or the second contact surface 231a) of the first contact unit 220 (or the second contact unit 230) into contact with one of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10 while the sheet-shaped cell culture 10 is suspended in a predetermined solution 330; removing the solution 330; and discharging a fibrinogen solution and / or a thrombin solution from the first contact unit 220 (or the second contact unit 230) onto the sheet-shaped cell culture 10 while the sheet-shaped cell culture 10 is in contact with the first contact surface 221a (or the second contact surface 231a). Specific steps of the forming method will be described later.

[0038] 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.

[0039] 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.

[0040] The image display unit 130 can be configured, for example, by a known liquid crystal display or the like.

[0041] 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.

[0042] <Forming device 200> As shown in FIGS. 1 and 3 to 9, 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.

[0043] The processing section 210 has one or more contact sections 220, 230 with lattice-shaped contact surfaces 221a, 231a configured to be able to come into contact with the sheet-shaped cell culture 10.

[0044] The contact parts 220, 230 are configured to be able to discharge fibrinogen solution and / or thrombin solution onto the sheet-shaped cell culture 10 in contact with the contact surfaces 221a, 231a.

[0045] In this embodiment, a form will be described in which processing unit 210 is provided with two contact units 220 and 230. However, processing unit 210 can also be configured to have one contact unit 230 configured to be able to eject thrombin liquid, as will be described in modified examples (see FIGS. 10 to 13) below, or to have one contact unit 220 configured to be able to eject fibrinogen liquid.

[0046] As shown in FIGS. 3 and 4, the processing section 210 has a first contact section 220 and a second contact section 230 on which the sheet-shaped cell culture 10 is placed on the rear surface 10b side of the sheet-shaped cell culture 10.

[0047] 4, the first contact portion 220 has a first contact surface 221a and a plurality of first discharge holes 225 capable of discharging the fibrinogen liquid. The first contact surface 221a and the first discharge holes 225 are arranged on the side facing the back surface 10b of the sheet-shaped cell culture 10.

[0048] The second contact portion 230 has a second contact surface 231a and a plurality of second discharge holes 235 capable of discharging the thrombin liquid.

[0049] The first contact portion 220 has a main body portion 221 formed with a first contact surface 221a and a back surface 221b located opposite the first contact surface 221a, and a tube body 227 connected to the main body portion 221 and supplying fibrinogen liquid to the main body portion 221.

[0050] The main body 221 can be configured, for example, to have a shape similar to the planar shape of the sheet-shaped cell culture 10 and to have a first contact surface 221a that is larger than the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10. By configuring the main body 221 in this way, the sheet-shaped cell culture 10 can be stably placed on the first contact portion 220 when forming the reinforcing layer 30 (see FIGS. 8 and 9).

[0051] In this embodiment, the sheet-shaped cell culture 10 has a circular planar shape (see FIG. 4). Therefore, the main body 221 is configured to have a substantially circular planar shape in which the area of ​​the first contact surface 221a in a plan view (corresponding to the projected surface when the outer shape of the main body 221 is circular) is larger than the areas of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10.

[0052] There are no particular limitations on the specific shape or size of the first contact surface 221a, and they can be changed as desired depending on, for example, the planar shape of the sheet-shaped cell culture 10 and the areas of the front surface 10a and back surface 10b.

[0053] The main body 221 has frame portions 222 arranged in a grid pattern on the inner periphery of the main body 221 , and gaps 223 provided between the frame portions 222 .

[0054] The first contact surface 221a is formed by the upper surface of the frame portion 222. The back surface 221b is formed by the lower surface of the frame portion 222 located on the opposite side to the upper surface.

[0055] The frame portion 222 is provided with a plurality of first discharge holes 225 .

[0056] Each first discharge hole 225 opens toward the upper side (upper side of the first contact surface 221a) of the frame part 222. Therefore, as will be described later, by discharging the fibrinogen liquid from the first discharge hole 225 while the sheet-shaped cell culture 10 is placed on the first contact surface 221a side of the first contact part 220, the fibrinogen liquid can be sprayed onto the sheet-shaped cell culture 10 (see FIG. 8).

[0057] Note that the first discharge holes 225 are not limited to opening upward, and may be provided on the side surface of the frame portion 222 so as to open toward the gap 223, for example. Similarly, the second discharge holes 235 are not limited to opening upward, and may be provided on the side surface of the frame portion 232 so as to open toward the gap 233. When these configurations are employed, it is desirable to arrange the first discharge holes 225 and the second discharge holes 235 so as to form a flow path extending from the center position in the surface direction of each of the main body portions 221, 231 toward the peripheral edge portion.

[0058] Each of the first discharge holes 225 may be configured, for example, to have a circular shape.

[0059] As illustrated by the arrow f1 in Figure 4, the first discharge hole 225 can eject (spray) fibrinogen liquid in a radial direction vertically upward from the center position of the first discharge hole 225 (center position when viewed in a plane) and in a 360° radius around the vertically upward.

[0060] 5, a flow path 226 through which the fibrinogen liquid can flow is formed inside the frame 222. The flow path 226 communicates with each of the first discharge holes 225 formed in the frame 222.

[0061] The tubular body 227 connected to the main body 221 is configured to be in fluid communication with a fibrinogen liquid supply source (e.g., a tank, a liquid storage tank, a syringe, etc.) not shown. A flow path 227a through which the fibrinogen liquid can flow is provided inside the tubular body 227. The flow path 227a of the tubular body 227 is in fluid communication with the flow path 226 provided inside the frame 222.

[0062] When discharging fibrinogen liquid onto the sheet-shaped cell culture 10, the forming device 200 sends the fibrinogen liquid from a predetermined supply source to the main body 221 of the first contact part 220. The first contact part 220 can spray the fibrinogen liquid onto the sheet-shaped cell culture 10 through a plurality of first discharge holes 225 formed in the frame part 222.

[0063] As shown in FIG. 4, the second contact portion 230 has a main body portion 231 formed with a second contact surface 231a and a back surface 231b located on the opposite side of the second contact surface 231a, and a tube 237 connected to the main body portion 231 and supplying thrombin liquid to the main body portion 231.

[0064] The main body 231 can be configured to have a shape similar to the planar shape of the sheet-shaped cell culture 10 and a second contact surface 231a larger than the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10. By configuring the main body 231 in this manner, the sheet-shaped cell culture 10 can be stably placed on the second contact surface 230 when forming the reinforcing layer 30.

[0065] In this embodiment, the main body portion 231 is configured to have an approximately circular planar shape in which the area of ​​the second contact surface 231a in a planar view (corresponding to the projected surface when the outer shape of the main body portion 231 is circular) is larger than the area of ​​the front surface 10a and back surface 10b of the sheet-shaped cell culture 10.

[0066] There are no particular limitations on the specific shape or size of the second contact surface 231a, and they can be changed as desired depending on, for example, the planar shape of the sheet-shaped cell culture 10 and the areas of the front surface 10a and back surface 10b.

[0067] The main body 231 has frame portions 232 arranged in a lattice pattern on the inner periphery of the main body 231 , and gaps 233 provided between the frame portions 232 .

[0068] The second contact surface 231a is formed by the upper surface of the frame portion 232. The back surface 231b is formed by the lower surface of the frame portion 222, which is located on the opposite side to the upper surface.

[0069] A plurality of second discharge holes 235 are formed in the frame part 232. Each second discharge hole 235 is open toward the upper side of the frame part 232 (above the second contact surface 231a). Therefore, as will be described later, by discharging the thrombin solution from the second discharge holes 235 while the sheet-shaped cell culture 10 is placed on the second contact surface 231a side of the second contact part 230, the thrombin solution can be sprayed onto the sheet-shaped cell culture 10 (see FIG. 8).

[0070] Each of the second outlet holes 235 may be configured, for example, to have a circular shape.

[0071] As illustrated by arrow f2 in Figure 4, the second outlet hole 235 can eject (spray) thrombin liquid from the center position of the second outlet hole 235 (the center position when viewed from above) in a radial direction vertically upward and 360° around the vertical axis.

[0072] 6, a flow path 236 through which the thrombin solution can flow is formed inside the frame 232. The flow path 236 communicates with each of the second discharge holes 235 formed in the frame 232.

[0073] The tubular body 237 connected to the main body 231 is configured to be in fluid communication with a supply source (such as a tank, a reservoir, or a syringe) of the thrombin liquid (not shown). A flow path 237a through which the thrombin liquid can flow is provided inside the tubular body 237. The flow path 237a of the tubular body 237 is in fluid communication with the flow path 236 provided inside the frame 232.

[0074] When discharging thrombin solution onto the sheet-shaped cell culture 10, the forming device 200 sends the thrombin solution from a predetermined supply source to the main body 231 of the second contact part 230. The second contact part 230 can spray the thrombin solution onto the sheet-shaped cell culture 10 through a plurality of second discharge holes 235 formed in the frame part 232.

[0075] Figure 3 shows the positional relationship between the first contact portion 220 and the second contact portion 230 when ejecting fibrinogen solution and thrombin solution onto the sheet-shaped cell culture 10 (the positional relationship in a plan view as seen from the direction of arrow III in Figure 1).

[0076] As shown in FIG. 4, the first outlet hole 225 of the first contact portion 220 and the second outlet hole 235 of the second contact portion 230 are configured to be able to eject fibrinogen liquid or thrombin liquid at positions that do not overlap with each other in the planar direction.

[0077] In this embodiment, as shown in FIGS. 4 and 7 to 9, when the fibrinogen solution and the thrombin solution are discharged onto the sheet-shaped cell culture 10, the sheet-shaped cell culture 10 is placed on the first contact surface 221a of the first contact section 220, and the second contact section 230 is disposed below the first contact section 220 (toward the bottom 323 of the container 320). Furthermore, as shown in FIG. 3, the first discharge holes 225 formed in the frame section 222 of the first contact section 220 can be disposed at a position that does not overlap with the frame section 232 of the second contact section 230, but overlaps with the gaps 233 of the second contact section 230. Furthermore, the second discharge holes 235 formed in the frame section 232 of the second contact section 230 can be disposed at a position that does not overlap with the frame section 222 of the first contact section 220, but overlaps with the gaps 223 of the first contact section 220. By arranging each discharge hole 225, 235 in this manner, when fibrinogen liquid and thrombin liquid are discharged from each discharge hole 225, 235 onto the sheet-shaped cell culture 10, the spraying of the fibrinogen liquid and thrombin liquid can be prevented from being obstructed by each frame portion 222, 232.

[0078] The back surface 231b of the first contact portion 220 can be disposed so as not to come into contact with the second contact surface 231a of the second contact portion 230. In this case, for example, the frame portion of the first contact portion 220 (the outer frame portion located on the outermost side of the main body portion 221) and the frame portion of the second contact portion 230 (the outer frame portion located on the outermost side of the main body portion 231) can be fitted together with a male-female structure (a structure in which a convex portion and a concave portion are formed on each), and a space can be formed between the first discharge hole 225 and the second discharge hole 235. This also makes it possible to position the first contact portion 220 with respect to the second contact portion 230.

[0079] 5 and 6, the first discharge holes 225 of the first contact portion 220 can be configured to have a larger diameter than the second discharge holes 235 of the second contact portion 230. The fibrinogen liquid used to form the reinforcing layer 30 is generally more viscous than thrombin liquid, which may cause clogging of the first discharge holes 225. Therefore, by increasing the diameter of the first discharge holes 225 that discharge the fibrinogen liquid, clogging of the first discharge holes 225 with the fibrinogen liquid can be prevented. This allows the fibrinogen liquid to be efficiently discharged from the first contact portion 220 onto the sheet-shaped cell culture 10.

[0080] There are no particular restrictions on the number and shape of each discharge hole 225, 235, the arrangement pattern and shape of each frame portion 222, 232, the number and shape of each gap 223, 233, etc., as long as they can prevent wrinkles from occurring in the sheet-shaped cell culture 10 when forming the reinforcing layer 30 on the sheet-shaped cell culture 10, as described below.

[0081] The fibrinogen solution and thrombin solution used to form the reinforcing layer 30 are not particularly limited as long as they can react with each other and solidify to form a fibrin gel. For example, the specific forms of the fibrinogen solution and thrombin solution can be appropriately determined by referring to publicly known information (e.g., JP 2016-52272 A, etc.).

[0082] <Formation method> Next, a forming method according to this embodiment will be described.

[0083] The specific work content of each step will be described below with reference to the flowchart shown in Fig. 2 and Fig. 7 to Fig. 9. 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.

[0084] To start the formation method, a sheet-shaped cell culture 10 is prepared before the formation of the reinforcing layer 30 (step S10). 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).

[0085] 7, before the solution 330 is discharged from the container 320 (a state in which the sheet-shaped cell culture 10 is floating in the solution 330), each contact part 220, 230 is placed inside the container 320. At this time, the sheet-shaped cell culture 10 is placed on the first contact surface 221a of the first contact part 220, and the back surface 10b of the sheet-shaped cell culture 10 is brought into contact with the first contact surface 221a of the first contact part 220 (step S11). The second contact part 230 is placed below the first contact part 220 (towards the bottom part 323 of the container 320).

[0086] Next, as shown in Fig. 8, the solution 330 is removed (discharged) from the container 320 (step S12). Once the solution 330 has been removed, the sheet-shaped cell culture 10 moves toward the bottom 323 under its own weight and is placed on the first contact surface 221a of the first contact part 220. This effectively prevents the sheet-shaped cell culture 10 from becoming twisted or wrinkled during the period from when the solution 330 is removed until when the sheet-shaped cell culture 10 is placed on the first contact surface 221a.

[0087] The reinforcing layer 30 is formed between the sheet-shaped cell culture 10 and the first contact surface 221a. The sheet-shaped cell culture 10 is spaced apart from the first contact surface 221a and placed on the first contact surface 221a via the reinforcing layer 30.

[0088] Next, fibrinogen solution is ejected from the first contact section 220 onto the sheet-shaped cell culture 10, and thrombin solution is ejected from the second contact section 230 onto the sheet-shaped cell culture 10 (step S13). With the sheet-shaped cell culture 10 placed on the first contact surface 221a of the first contact section 220, the processing section 210 sprays the fibrinogen solution onto the sheet-shaped cell culture 10 from each of the first discharge holes 225 that open on the first contact surface 221a side. At this time, even if wrinkles have occurred in the sheet-shaped cell culture 10, the sheet-shaped cell culture 10 can be stretched by spraying the fibrinogen solution, and the wrinkles that have occurred in the sheet-shaped cell culture 10 can be removed. Furthermore, since the thrombin solution can also be sprayed onto the sheet-shaped cell culture 10 from the second contact part 230, which is positioned below the first contact part 220, wrinkles can be efficiently removed even if they occur in the sheet-shaped cell culture 10. In the above-described process, the reinforcing layer 30 is not formed between the first contact part 220 and the second contact part 230. Furthermore, excess fibrinogen solution and thrombin solution can be collected into the container 320 through the gaps 223, 233.

[0089] Next, the process waits until the fibrinogen solution and thrombin dispensed onto the sheet-shaped cell culture 10 solidify and a reinforcing layer 30 made of fibrin gel is formed (step S13). When the solidification of the fibrinogen solution and thrombin solution has progressed sufficiently, the reinforcing layer 30 is formed on the sheet-shaped cell culture 10. At this time, the reinforcing layer 30 is formed between the sheet-shaped cell culture 10 and the first contact surface 221a.

[0090] This embodiment further includes a step of removing the laminate 20 (the sheet-shaped cell culture 10 and the reinforcing layer 30) from the container 320. Because the laminate 20 is in contact with the first contact surface 221a via the reinforcing layer 30 (a portion of the reinforcing layer 30 is adhered to the frame 222), it can be transported as is to a desired location (for example, a transplant device or another container). When peeling the laminate 20 from the first contact surface 221a, peeling is performed from the boundary between the first contact surface 221a and the reinforcing layer 30, allowing the sheet-shaped cell culture 10 to be peeled off without being damaged or wrinkled.

[0091] 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.

[0092] 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 on a sheet-shaped cell culture 10, and the processing unit 210 has one or more first contact portions 220 (or second contact portions 230) that have a lattice-shaped first contact surface 221a (or second contact surface 231a) that is configured to be able to come into contact with the sheet-shaped cell culture 10, and the first contact portion 220 (or second contact portion 230) is configured to be able to eject fibrinogen solution (or thrombin solution) onto the sheet-shaped cell culture 10 that has come into contact with the first contact surface 221a (or second contact surface 231a).

[0093] 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 bringing the lattice-shaped contact surfaces 221a, 231a of the contact portions 220, 230 into contact with one of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10 while the sheet-shaped cell culture 10 is suspended in a predetermined solution 330, removing the solution 330, and ejecting fibrinogen solution and / or thrombin solution from the contact portions 220, 230 onto the sheet-shaped cell culture 10 while the sheet-shaped cell culture 10 is in contact with the contact surfaces 221a, 231a.

[0094] Furthermore, 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 following steps: bringing the lattice-shaped contact surfaces 221a, 231a of the contact parts 220, 230 into contact with one of the front surface 10a and the back surface 10b of the sheet-shaped cell culture 10 while the sheet-shaped cell culture 10 is suspended in a predetermined solution 330; removing the solution 330; and ejecting fibrinogen solution and / or thrombin solution from the contact parts 220, 230 onto the sheet-shaped cell culture 10 while the sheet-shaped cell culture 10 is in contact with the contact surfaces 221a, 231a.

[0095] According to this embodiment, the fibrinogen solution and / or thrombin solution can be ejected onto the sheet-shaped cell culture 10 while the sheet-shaped cell culture 10 is in contact with the first contact surface 221a (or the second contact surface 231a) of the first contact unit 220 (or the second contact unit 230) provided in the processing unit 210. The fibrinogen solution (or the thrombin solution) is sprayed onto the sheet-shaped cell culture 10 from the first contact unit 220 (or the second contact unit 230) while the sheet-shaped cell culture 10 is in contact with the first contact surface 221a (or the second contact surface 231a) of the first contact unit 220 (or the second contact unit 230), thereby preventing wrinkles from forming during the fibrin treatment. Therefore, the product quality of the laminate 20 can be stabilized compared to when an operator manually stretches the sheet-shaped cell culture 10 to prevent wrinkles from forming.

[0096] Although 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, and the program according to the present invention have been described through the embodiments, the present invention is not limited to the configurations described in the embodiments, and can be modified as appropriate based on the claims.

[0097] (Variation 1) For example, the forming device can be configured to include one contact part 230 (corresponding to the second contact part 230 in the above-described embodiment) capable of discharging thrombin liquid.

[0098] A method for forming the reinforcing layer 30 using the forming device according to the first modification having the above configuration will be described below.

[0099] As shown in Figure 10, with the sheet-shaped cell culture 10 suspended in a solution 330 contained in a container 320, the sheet-shaped cell culture 10 is brought into contact with the contact surface 231a of the contact portion 230 (corresponding to the second contact surface 231a in the above-mentioned embodiment).

[0100] 11, the solution 330 is removed from the container 320. When the solution 330 is removed from the container 320, the sheet-shaped cell culture 10 is placed on the contact surface 231a of the contact part 230. This prevents the sheet-shaped cell culture 10 from wrinkling before and after the operation of removing the solution 330.

[0101] Next, as shown in FIG. 12 , a predetermined applicator 250 is used to apply fibrinogen solution to the sheet-shaped cell culture 10. The applicator 250 can be configured to include, for example, a mechanism for dripping the fibrinogen solution onto the sheet-shaped cell culture 10. Examples of mechanisms for dripping the fibrinogen solution include a well-known syringe or pipette. The fibrinogen solution may be applied to the back surface 10b of the sheet-shaped cell culture 10. For example, the applicator 250 may dispense a predetermined amount of fibrinogen solution into a container 320 (culture substrate), store it in the bottom 323 of the container 320, and, with the sheet-shaped cell culture 10 placed on the contact part 230, immerse the back surface 10b of the sheet-shaped cell culture 10 in the fibrinogen solution to apply the fibrinogen solution to the sheet-shaped cell culture 10.

[0102] Next, as shown in Figure 13, with the sheet-shaped cell culture 10 in contact with and placed on the contact surface 231a of the contact section 230, thrombin solution is ejected from the contact section 230 onto the sheet-shaped cell culture 10. As in the previously described embodiment, the sheet-shaped cell culture 10 can be stretched by spraying thrombin solution onto the sheet-shaped cell culture 10 from each ejection hole 235 (corresponding to the second ejection hole 235 in the previously described embodiment) opened on the contact surface 231a side (upper side). Therefore, even if wrinkles have occurred in the sheet-shaped cell culture 10, the wrinkles can be removed.

[0103] After the above steps are completed, the worker waits until the fibrinogen solution and thrombin solidify and a reinforcing layer 30 made of fibrin gel is formed, thereby obtaining a laminate 20 in which the sheet-shaped cell culture 10 is reinforced by the reinforcing layer 30.

[0104] (Variation 2) In the above-described embodiment, an example was described in which the processing unit 210 has the first contact part 220 and the second contact part 230 on which the sheet-shaped cell culture 10 is placed on the back surface 10b side of the sheet-shaped cell culture 10. For example, as will be described in Modification 2 below, the processing unit can also be configured to have the first contact part 220A and the second contact part 230A that are pressed against the sheet-shaped cell culture 10 from the front surface 10a side of the sheet-shaped cell culture 10.

[0105] As shown in FIGS. 14 and 15, the processing section according to the second modification has a first contact part 220A and a second contact part 230A that are pressed against the sheet-shaped cell culture 10 from the surface 10a side of the sheet-shaped cell culture 10.

[0106] The first contact portion 220A has a first contact surface 221b and a plurality of first discharge holes 225 (see FIG. 17) capable of discharging the fibrinogen liquid.

[0107] The first contact part 220A has substantially the same function as the first contact part 220 according to the above-described embodiment (see FIGS. 3 and 4). The main differences between the first contact part 220 and the first contact part 220A are that in the first contact part 220A, the back surface 221b of the main body part 221 forms the first contact surface that is pressed against the sheet-shaped cell culture 10, and that the first discharge holes 225 open toward the back surface 221b (downward) (see FIG. 17).

[0108] The second contact portion 230A has a second contact surface 231b and a plurality of second discharge holes 235 (see FIG. 18) capable of discharging the thrombin liquid.

[0109] The second contact part 230A has substantially the same function as the second contact part 230 according to the above-described embodiment (see FIGS. 3 and 4). The main differences between the second contact part 230 and the second contact part 230A are that in the second contact part 230A, the back surface 231b of the main body part 231 forms a second contact surface that is pressed against the sheet-shaped cell culture 10, and that the second discharge holes 235 open toward the back surface 231b side (downward) (see FIG. 18).

[0110] Furthermore, similar to the above-described embodiment, the first outlet hole 225 of the first contact portion 220A and the second outlet hole 235 of the second contact portion 230A are configured to be able to eject fibrinogen liquid or thrombin liquid at positions that do not overlap with each other in the planar direction.

[0111] In this embodiment, as shown in Figures 15 and 16, when fibrinogen solution and thrombin solution are discharged onto the sheet-shaped cell culture 10, the second contact surface 231b of the second contact portion 230A is disposed so as to be in contact with the surface 10a of the sheet-shaped cell culture 10, and the first contact portion 220A is disposed above the second contact portion 230A (toward the opening 321 of the container 320). As explained in Figure 3 of the above-mentioned embodiment, the first discharge holes 225 formed in the frame portion 222 of the first contact portion 220A can be disposed at a position that does not overlap with the frame portion 232 of the second contact portion 230A but overlaps with the gaps 233 of the second contact portion 230A. Furthermore, the second discharge holes 235 formed in the frame portion 232 of the second contact portion 230A can be disposed at a position that does not overlap with the frame portion 222 of the first contact portion 220A but overlaps with the gaps 223 of the first contact portion 220A. By arranging each discharge hole 225, 235 in this manner, when fibrinogen liquid and thrombin liquid are discharged from each discharge hole 225, 235 onto the sheet-shaped cell culture 10, the spraying of the fibrinogen liquid and thrombin liquid can be prevented from being obstructed by each frame portion 222, 232.

[0112] As in the above-described embodiment, the first outlet hole 225 for ejecting the fibrinogen liquid provided in the first contact portion 220A can be configured to have a larger diameter than the second outlet hole 235 for ejecting the thrombin liquid provided in the second contact portion 230A (see Figures 17 and 18).

[0113] A method for forming the reinforcing layer 30 using the forming device according to the second modification having the above configuration will be described below.

[0114] 14, before the solution 330 is discharged from the container 320 (a state in which the sheet-shaped cell culture 10 is suspended in the solution 330), each contact part 220A, 230A is placed inside the container 320. At this time, the second contact surface 231b of the second contact part 230A is pressed against the sheet-shaped cell culture 10, so that the second contact surface 231b comes into contact with the surface 10a of the sheet-shaped cell culture 10. The first contact part 220A is placed above the second contact part 230A (towards the opening 321 of the container 320).

[0115] 15, the solution 330 is removed (discharged) from the container 320. When the solution 330 is removed from the container 320, the sheet-shaped cell culture 10 is pressed against the bottom 323 of the container 320 by the second contact surface 231b of the second contact portion 230A. This makes it possible to preferably prevent the sheet-shaped cell culture 10 from becoming twisted or wrinkled when the solution 330 is removed.

[0116] Next, fibrinogen solution is discharged from the first contact section 220A onto the sheet-shaped cell culture 10, and thrombin solution is discharged from the second contact section 230A onto the sheet-shaped cell culture 10 (step S13). The processing section can spray thrombin solution onto the sheet-shaped cell culture 10 from each of the second discharge holes 235 opened on the second contact surface 231b side (lower side) while pressing the sheet-shaped cell culture 10 against the bottom 323 of the container 320 by the second contact surface 231b of the second contact section 230A. Therefore, even if wrinkles occur in the sheet-shaped cell culture 10 when the thrombin solution is sprayed, the sheet-shaped cell culture 10 can be straightened by spraying the thrombin solution, and the wrinkles can be removed from the sheet-shaped cell culture 10. In addition, by spraying fibrinogen liquid onto the sheet-shaped cell culture 10 from the first contact portion 220A, which is positioned above the second contact portion 230A, wrinkles formed on the sheet-shaped cell culture 10 can be removed more efficiently.

[0117] After the above steps are completed, the worker waits until the fibrinogen solution and thrombin solidify and a reinforcing layer 30 made of fibrin gel is formed, thereby obtaining a laminate 20 in which the sheet-shaped cell culture 10 is reinforced by the reinforcing layer 30.

[0118] As explained in the embodiments and each modified example, the number of contact parts provided in the processing unit may be one or more, and if the processing unit has multiple contact parts, it is possible to arbitrarily change which contact part's contact surface is configured to contact the front or back surface of the sheet-shaped cell culture.

[0119] 7 to 9, the fibrin treatment may be performed by placing the sheet-shaped cell culture 10 on the second contact surface 231a of the second contact portion 230 capable of discharging thrombin solution, and arranging the first contact portion 220 capable of discharging fibrinogen solution below the second contact portion 230 (toward the bottom 323 of the container 320). Alternatively, in the formation method shown in FIGS. 14 to 16, the fibrin treatment may be performed by pressing the first contact surface 221b of the first contact portion 220A capable of discharging fibrinogen solution against the sheet-shaped cell culture 10, and arranging the second contact portion 230A capable of discharging thrombin solution above the first contact portion 220 (toward the opening 321 of the container 320). Note that the above configuration is more preferable from the viewpoint of enabling direct contact of the first contact surface 221b discharging the fibrinogen solution (first liquid) with the sheet-shaped cell culture 10. [Explanation of symbols]

[0120] 10. Sheet-shaped cell culture 10a Surface of sheet-shaped cell culture 10b Backside of sheet-shaped cell culture 20 laminate 30 Reinforcement layer 100 Formation System 110 control section 200 Forming device 210 Processing section 220 1st contact part 220A First Contact 221 Main body 221a 1st contact surface 221b Back side (1st contact surface) 222 Frame 223 Gap 225 1st discharge hole 230 Second contact part 230A 2nd contact part 231 Main body 231a Second contact surface 231b Back side (second contact surface) 232 Frame 233 Gap 235 2nd discharge hole 250 application part 310 Stage 320 container 330 Solution

Claims

1. An apparatus comprising a processing unit for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, the processing section has one or more contact sections each having a lattice-shaped contact surface configured to be able to come into contact with the sheet-shaped cell culture, An apparatus for forming a reinforcing layer on a sheet-shaped cell culture, wherein the contact portion is configured to be able to eject fibrinogen solution and / or thrombin solution onto the sheet-shaped cell culture that has come into contact with the contact surface.

2. the processing section has a first contact section and a second contact section on which the sheet-shaped cell culture is placed on the back side of the sheet-shaped cell culture, the first contact portion has a first contact surface and a plurality of first discharge holes capable of discharging the fibrinogen liquid, the second contact portion has a second contact surface and a plurality of second discharge holes capable of discharging the thrombin solution, 2. The device for forming a reinforcing layer on a sheet-shaped cell culture according to claim 1, wherein the first discharge hole and the second discharge hole are capable of discharging fibrinogen solution or thrombin solution at positions where they do not overlap each other in the planar direction.

3. the processing section has a first contact section and a second contact section that are pressed against the sheet-shaped cell culture from the surface side of the sheet-shaped cell culture, the first contact portion has a first contact surface and a plurality of first discharge holes capable of discharging the fibrinogen liquid, the second contact portion has a second contact surface and a second discharge hole capable of discharging the thrombin liquid, 2. The device for forming a reinforcing layer on a sheet-shaped cell culture according to claim 1, wherein the first discharge hole and the second discharge hole are capable of discharging fibrinogen solution or thrombin solution at positions where they do not overlap each other in the planar direction.

4. The device for forming a reinforcing layer on a sheet-shaped cell culture according to claim 1 , wherein the contact part is configured to be able to discharge a thrombin solution.

5. A method for forming a reinforcing layer made of fibrin gel on a sheet-shaped cell culture, comprising: a step of bringing the lattice-shaped contact surface of the contact part into contact with one of the front and back surfaces of the sheet-shaped cell culture while the sheet-shaped cell culture is suspended in a predetermined solution; removing the solution; A method for forming a reinforcing layer on a sheet-shaped cell culture, comprising the steps of: contacting the sheet-shaped cell culture with the contact surface and ejecting fibrinogen solution and / or thrombin solution from the contact portion onto the sheet-shaped cell culture.

6. 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, a step of bringing the lattice-shaped contact surface of the contact part into contact with one of the front and back surfaces of the sheet-shaped cell culture while the sheet-shaped cell culture is suspended in a predetermined solution; removing the solution; and ejecting fibrinogen solution and / or thrombin solution from the contact portion onto the sheet-shaped cell culture while the sheet-shaped cell culture is in contact with the contact surface.

Images