Tension Measuring Device

Abstract

To provide a tension force measuring device capable of reducing vibration which applies impact to tension force detecting means, for correctly measuring tension force.SOLUTION: A tension force measuring device 1 comprises: a first container 40 in which a cell structure CS is arranged and in which a medium is filled so as to immerse the cell structure; a second container 50 into which medicine administered into the first container, is injected; a first tube 60 for liquid tightly coupling the first and second containers; and a pump 70 which is arranged on the first tube, and causes liquid in the second container to flow into the first container.SELECTED DRAWING: Figure 6

Description

[Technical field]

[0001] The present invention relates to a device for measuring tension in a cellular structure including a muscle cell. [Background technology]

[0002] In recent years, drug discovery screening methods have been developed that utilize pluripotent stem cells, such as iPS cells, which have the ability to differentiate into various functional cells. However, the evaluation systems that have been used so far use individual cells and do not reflect the state of living tissues. Therefore, there is a demand to develop an evaluation system that mimics living tissues from somatic cells induced to differentiate from pluripotent stem cells.

[0003] Attempts to construct cells three-dimensionally have included, for example, a method of seeding cells onto a three-dimensional structure called a scaffold, a method of decellularizing organs and tissues and seeding cells into the remaining matrix to create a three-dimensional structure, and a method of stacking cell sheets that have been detached into sheets in a three-dimensional manner (e.g., Patent Document 1 and Non-Patent Document 1).

[0004] Using these techniques, research and development of evaluation systems for use in drug discovery screening is being conducted. In this regard, for example, Patent Document 2 listed below discloses a tension measuring device, system, and kit for sheet-like tissue containing cardiomyocytes. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 02-211865 [Patent Document 2] JP 2019-76046 A [Non-patent literature]

[0006] [Non-Patent Document 1] Haraguchi Y.,et al.,Scaffold-free tissue engineering using cell sheet technology.RSC Adv.,2012;2:2184-2190 Summary of the Invention [Problem to be solved by the invention]

[0007] The tension measuring device according to Patent Document 2 is provided with a highly sensitive tension detecting means for measuring minute tensions resulting from the pulsation of cardiomyocytes. In addition, in the tension measuring device according to Patent Document 2, a drug is directly introduced into a culture medium tank (first container) in which the cardiomyocytes are placed. For this reason, there is a risk that vibrations caused by the introduction of the drug may affect the tension detecting means, making it difficult to measure tension accurately.

[0008] The present invention has been invented to solve the above-mentioned problems, and has an object to provide a tension measuring device that can reduce vibrations that affect the tension detection means and accurately measure tension. [Means for solving the problem]

[0009] The tension measuring device according to the present invention, which achieves the above object, is a tension measuring device for a cell structure including muscle cells. The tension measuring device includes a gel adapter holder assembly for holding a gel and the cell structure, a first container in which the cell structure and the gel adapter holder assembly are disposed and which is filled with a culture medium so that the cell structure is immersed, a second container into which a drug to be administered is injected into the first container, a first tube for fluid-tightly connecting the first container and the second container, and a pump disposed on the first tube for causing a liquid in the second container to flow into the first container. When the first container is viewed from above in a vertical direction, The first tube is At a position offset from a center line passing through the center of the first container in a direction perpendicular to the vertical direction of the first container It is coupled to a side of the first container. Effect of the Invention

[0010] According to the above-mentioned tension measuring device, the drug and culture medium in the second container are flowed into the first container through the first tube by the pump. Therefore, it is possible to reduce vibrations that affect the tension detecting means compared to the case where the drug is directly flowed into the first container. Therefore, it is possible to provide a tension measuring device that can reduce vibrations that affect the tension detecting means and accurately measure tension. [Brief description of the drawings]

[0011] [Figure 1] 1 is a schematic perspective view showing a tension measuring device according to a first embodiment of the present invention. [Diagram 2] FIG. 2 is a schematic diagram showing a gel adapter holder assembly. [Diagram 3] FIG. 2 is a schematic diagram showing a first gel adapter holder. [Figure 4] FIG. 13 is a schematic diagram showing a second gel adapter holder. [Diagram 5] FIG. 13 is a diagram for explaining a method for placing a gel and a cell structure in a first gel adaptor holder and a second gel adaptor holder. [Figure 6] FIG. 1 is a schematic diagram showing a part of a tension measuring device according to a first embodiment. [Figure 7] FIG. 4 is a schematic diagram showing a part of a tension measuring device according to a second embodiment of the present invention. [Figure 8] FIG. 11 is a schematic diagram showing a part of a tension measuring device according to a third embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] First Embodiment Hereinafter, a first embodiment of the present invention will be described with reference to the drawings. In the description of the drawings, the same elements are given the same reference numerals, and duplicated descriptions will be omitted. The dimensional ratios in the drawings are exaggerated for the convenience of explanation, and may differ from the actual ratios.

[0013] FIG. 1 is a schematic perspective view showing a tension measuring device 1 according to a first embodiment of the present invention. FIG. 2 is a schematic view showing a gel adapter holder assembly 10. FIG. 3 is a schematic view showing a first gel adapter holder 11. FIG. 4 is a schematic view showing a second gel adapter holder 12. FIG. 5 is a diagram for explaining a method of arranging a gel G and a cell structure CS in the first gel adapter holder 11 and the second gel adapter holder 12. FIG. 6 is a schematic view showing a part of the tension measuring device 1 according to the first embodiment.

[0014] Before describing the configuration of the tension measuring device 1 according to this embodiment, a cell structure CS including muscle cells that is the measurement target of the tension measuring device 1 will be described below.

[0015] <Cell structure containing muscle cells> In this specification, the term "cell structure containing muscle cells" refers to biological tissue containing muscle cells collected from a living body (e.g., cardiac muscle tissue, skeletal muscle tissue, smooth muscle tissue, etc.) or a structure containing muscle cells. The cell structure containing muscle cells that can be applied to the present invention may be biological tissue itself collected from a living body, or biological tissue obtained by processing biological tissue collected from a living body, such as cardiac muscle tissue using human iPS cell-derived cardiac muscle cells. In addition, the cell structure containing muscle cells that can be applied to the present invention may be a cell structure formed by mixing a suspension containing muscle cells with a gel solution or a gelling agent, or may be a cell sheet. Furthermore, the cell structure containing muscle cells that can be applied to the present invention may be one formed by directly seeding a group of cells containing muscle cells on a gel and culturing it.

[0016] In one embodiment, the cell structure containing muscle cells that can be applied to the present invention may be at least one selected from the group consisting of a sheet-like, rod-like, and string-like shape, and is preferably a sheet-like cell structure.

[0017] In this specification, the term "sheet-like cell structure" refers to a membrane-like cell structure having an average thickness of, for example, about 10 μm (e.g., the thickness of one cell) to about 2 mm and a length that can be applied between a first gel holding part and a second gel holding part described below. The width of the "sheet-like cell structure" is not limited as long as it is a width that can be applied to the first gel holding part and the second gel holding part. One "sheet-like cell structure" may be applied to the tension measuring device, or multiple "sheet-like cell structures" may be applied to the tension measuring device. Multiple "sheet-like cell structures" may be applied in parallel between the first gel holding part and the second gel holding part, or may be applied in a stacked manner.

[0018] In this specification, the term "rod-shaped cell structure" refers to a cell structure having, for example, an average diameter of about 100 μm or more and about 5 mm or less, and a length that can be applied between a first gel holding part and a second gel holding part described below. A single "rod-shaped cell structure" may be applied to the tension measuring device, or multiple "sheet-shaped cell structures" may be applied to the tension measuring device. Multiple "rod-shaped cell structures" may be applied in parallel between the first gel holding part and the second gel holding part, or may be applied bundled together.

[0019] In this specification, the term "string-like cell structure" refers to a three-dimensional cell structure having, for example, an average diameter of about 10 μm or more and less than about 100 μm, and a length that can be applied between the first gel holding part and the second gel holding part described below. One "string-like cell structure" may be applied to the tension measuring device, or multiple "string-like cell structures" may be applied to the tension measuring device. Multiple "string-like cell structures" may be applied in parallel between the first gel holding part and the second gel holding part, or may be applied bundled together.

[0020] Note that, for convenience, "rod-shaped cell structures" and "string-shaped cell structures" are distinguished as above based on their diameter, but both refer to cell structures with an elongated shape, and the terms may be used interchangeably.

[0021] A "rod-shaped cell structure" or "string-shaped cell structure" that can be applied to the present invention may be, for example, a sheet-shaped cell structure formed into a rod or string shape (e.g., by rolling up, twisting, shrinking, or cutting), or may be formed by pouring a suspension containing cells and any gel into a rod-shaped or string-shaped mold (see, for example, Zhao Y, Cell. 2019 Feb 7; 176(4): 913-927).

[0022] As used herein, the term "cell structure containing muscle cells" refers to a cell structure containing at least 10% muscle cells, for example, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, or 99% or more. As used herein, "muscle cells" refer to cells having contractility that form muscle tissue in an animal body, and examples of such cells include cardiac muscle cells, skeletal muscle cells, and smooth muscle cells. In one embodiment, the "muscle cells" used in the present invention are at least one selected from the group consisting of cardiac muscle cells, skeletal muscle cells, and smooth muscle cells. The muscle cells that can be used in the present invention may be derived from animals, and for example, mammalian, avian, amphibian, reptile, and fish muscle cells can be used. Preferably, the muscle cells are derived from a mammal, and for example, muscle cells derived from a mammal such as mouse, rat, human, monkey, pig, dog, sheep, cat, or goat can be used.

[0023] The muscle cells that can be used in the present invention may be primary cells collected from biological tissue, or may be established cell lines, or may be cells induced to differentiate from pluripotent stem cells or tissue stem cells.

[0024] As used herein, the term "pluripotent stem cells" is intended to collectively refer to stem cells that have the ability to differentiate into any tissue cell (differentiation pluripotency). Pluripotent stem cells include, but are not limited to, embryonic stem cells (ES cells), embryonic carcinoma cells (EC cells), trophoblast stem cells (TS cells), epiblast stem cells (EpiS cells), embryonic germ cells (EG cells), multipotent germline stem cells (mGS cells), induced pluripotent stem cells (iPS cells), Muse cells, and the like. ES cells or iPS cells are preferred. Any known pluripotent stem cells can be used, and for example, the pluripotent stem cells described in International Publication No. 2009 / 123349 (PCT / JP2009 / 057041) can be used.

[0025] The muscle cells that can be used in the present invention may be cells induced to differentiate from pluripotent stem cells. A known method can be used to differentiate pluripotent cells into muscle cells (see, for example, Matsuura K., et al. Creation of human cardiac cell sheets using pluripotent stem cells. Biochem. Biophys. Res. Commun. 2012 Aug. 24; 425(2): 321-327, etc.).

[0026] The cell structure containing muscle cells may contain cells other than muscle cells, such as cardiac myoblasts, myoblasts, mesenchymal stem cells, vascular endothelial cells, vascular endothelial precursor cells, and fibroblasts.

[0027] The cell structure CS including muscle cells has been described above. Next, the configuration of the tension measuring device 1 according to this embodiment will be described.

[0028] As shown in FIG. 1, the tension measuring device 1 comprises a gel adapter holder assembly 10 in which a gel G and a cell structure CS are held, a tension detection means 20 for detecting tension caused by the pulsation of the cell structure CS, a connecting member 30 for connecting the gel adapter holder assembly 10 and the tension detection means 20, a first container 40 in which the cell structure CS and the gel adapter holder assembly 10 are placed, a second container 50 into which a drug to be administered is injected into the first container 40, a first tube 60 for liquid-tightly connecting the first container 40 and the second container 50, a pump 70 placed on the first tube 60, and a second tube 80 for liquid-tightly connecting the first container 40 and the second container 50.

[0029] As shown in FIGS. 2 to 5, the gel adapter holder assembly 10 holds the gel G and the cell structure CS.

[0030] The gel that can be used in the present invention is any gel that (1) can adhere to a cell structure including muscle cells, (2) has the strength to maintain the sheet shape, and (3) does not adversely affect cell growth, function expression, etc., that is, is biocompatible. An example of a gel that can be used in the present invention is a hydrogel. Examples of hydrogels that can be used in the present invention include hydrogels that are chemically crosslinked with water-soluble, water-affinity, or water-absorbent synthetic polymers such as polyacrylamide, polyacrylic acid, polyhydroxyethyl methacrylate, polyvinyl alcohol, polylactic acid, and polyglycolic acid, polysaccharides, proteins, and nucleic acids. Examples of polysaccharides include glycosaminoglycans such as hyaluronic acid and chondroitin sulfate, starch, glycogen, agarose, pectin, and cellulose. Examples of proteins include collagen and its hydrolyzates such as gelatin, proteoglycan, fibronectin, vitronectin, laminin, entactin, tenascin, thrombospondin, von Willebrand factor, osteopontin, and fibrinogen (for example, fibrin gel obtained by reacting fibrinogen with thrombin). These hydrogels may be crosslinked using a known method to increase their strength before use. A preferred gel that can be applied to the present invention is a fibrin gel. In one embodiment of the present invention, the gel may be formed by mixing it with cells in advance.

[0031] An example of the configuration of the gel adapter holder assembly 10 will be described below, but the configuration of the gel adapter holder assembly 10 is not limited to the following configuration as long as it is capable of holding the gel G and the cell structure CS.

[0032] The gel adapter holder assembly 10 may be that described in Japanese Patent Application No. 2019-036677. For this reason, a detailed description of the gel adapter holder assembly 10 will be omitted here, and only the general configuration will be described.

[0033] The gel adapter holder assembly 10 has a first gel adapter holder 11 and a second gel adapter holder 12, as shown in FIGS.

[0034] The first gel adapter holder 11 and the second gel adapter holder 12 are assembled together to form a gel adapter holder assembly 10 as shown in FIG.

[0035] As shown in Figures 2 and 3, the first gel adapter holder 11 has a frame member 110, a first gel holding portion 111 for fixing one end of the gel G, and a pair of claw portions 117 fixed to the lid body 41 of the first container 40.

[0036] By providing the frame member 110, the gel adapter holder assembly 10 can be attached to a lid 41 (described below) of the first container 40 while maintaining the shapes of the gel G and the cell structure CS.

[0037] The material constituting the first gel adapter holder 11 is not particularly limited, but examples include polyethylene, polypropylene, polycarbonate, polystyrene, polyvinyl chloride, nylon, polyurethane, polyurea, polylactic acid, polyglycolic acid, polyvinyl alcohol, polyvinyl acetate, poly(meth)acrylic acid, poly(meth)acrylic acid derivatives, polyacrylonitrile, poly(meth)acrylamide, poly(meth)acrylamide derivatives, polysulfone, cellulose, cellulose derivatives, polysilicone, metal, and the like.

[0038] As shown in Figures 2 and 4, the second gel adapter holder 12 has a connection portion 120 to which the connecting member 30 is connected, a second gel holding portion 121 for fixing the other end of the gel G, and a connecting portion 124 that connects the connection portion 120 and the second gel holding portion 121.

[0039] The connection portion 120 is provided with a connection port 126 for connection to the connecting member 30 .

[0040] The material constituting the second gel adapter holder 12 can be the same as the material constituting the first gel adapter holder 11 .

[0041] Hereinafter, a method for arranging the gel G and the cell structure CS in the first gel adapter holder 11 and the second gel adapter holder 12 will be described with reference to Fig. 5. Note that the method can be the method described in Japanese Patent Application No. 2019-036677. For this reason, a detailed description of the method for arranging the gel G and the cell structure CS will be omitted here, and only an outline will be described.

[0042] First, the first gel adapter holder 11 and the second gel adapter holder 12 are set on the substrate 13, and a pre-hardened gelling agent (e.g., a mixture of fibrinogen (SIGMA bovine plasma-derived Type IS), thrombin (SIGMA bovine plasma-derived T4648), CaCl2 solution (8 mM), and Factor XIII (CSL Behring Fibrogammin P for intravenous injection)) is injected using a pipette P (Figure 5(A)).

[0043] Next, the first gel holding portion 111 and the second gel holding portion 121 are covered with the gel-forming lid 14 (FIG. 5(B)).

[0044] Next, after the gel has hardened, the gel-forming lid 14 and the substrate 13 are removed from the first gel adapter holder 11 and the second gel adapter holder 12 (FIG. 5(C)).

[0045] Separately from the above, a group of cells including muscle cells is seeded on a temperature-responsive culture dish D1 (for example, UpCell (registered trademark) (CellSeed, Tokyo, Japan)) and cultured in advance at 37° C. until the cells become confluent.

[0046] Then, the first gel adaptor holder 11 and the second gel adaptor holder 12 equipped with the gel obtained above are placed on the cell construct CS containing muscle cells in the temperature-responsive culture dish D1 (FIG. 5(C)).

[0047] Then, the temperature-responsive culture dish D1 is kept below the lower critical solution temperature, for example at 20°C, and the cell structure CS containing muscle cells is detached from the temperature-responsive culture dish D1, while at the same time, the cell structure CS containing muscle cells is adhered to the underside of the gel G (Figure 5(D)).

[0048] The description of the configuration of the tension measuring device 1 will now be returned to.

[0049] The tension detection means 20 measures the tension generated by the pulsation of the cell structure CS containing muscle cells. The tension detection means 20 is connected to the gel adapter holder assembly 10 via a connecting member 30, as shown in Fig. 1. When the cell structure CS containing muscle cells contracts, the second gel adapter holder 12 is pulled vertically downward, and the load is detected by the tension detection means 20 via the connecting member 30. The tension detection means 20 can be, for example, a known load cell.

[0050] 1, the connecting member 30 connects the gel adapter holder assembly 10 and the tension detection means 20. The connecting member 30 has a hook 31 and a connection portion 32.

[0051] The hook 31 is inserted into the connection port 126 of the second gel adapter holder 12. The connection part 32 is connected to the tension detection means 20.

[0052] The connecting member 30 is not limited to the above configuration as long as it is capable of connecting the second gel adapter holder 12 and the tension detection means 20 .

[0053] The gel adapter holder assembly 10 having the gel G and the cell structure CS is housed in the first container 40. A lid 41 is attached to the first container 40.

[0054] A pair of claws 117 of the first gel adapter holder 11 are fitted into the cover 41 .

[0055] The second container 50 is configured to have approximately the same height as the first container 40. The drug to be administered into the first container 40 is injected into the second container 50. The second container 50 has a smaller surface area than the first container 40. With this configuration, the drug and medium can be sent from the second container 50 to the first container 40 using a small amount of the drug and medium. In the following description, the drug and / or medium may be referred to as a liquid.

[0056] As shown in Fig. 6, a heater 51 is provided below the second container 50. Here, for example, if a heater is provided below the first container 40, a local temperature rise will cause medium evaporation in the first container 40. In contrast, in this embodiment, the heater 51 is provided below the second container 50, so that medium evaporation due to a local temperature rise in the first container 40 can be prevented. Note that the present invention also includes a configuration in which a heater is provided below the first container 40.

[0057] The first tube 60 connects the first container 40 and the second container 50 in a liquid-tight manner. As shown in FIG. 6, when viewed from above in the vertical direction, the first tube 60 is connected to the first container 40 so as to be offset from a center line L1 passing through the center of the first container 40 in a direction perpendicular to the vertical direction of the first container 40. With this configuration, a rotating flow F is generated in the first container 40, promoting mixing. Therefore, the drug can be mixed in the culture medium more quickly. Furthermore, the concentration gradient in which the oxygen concentration is high in the upper part of the first container 40 can be flattened.

[0058] 6, the first tube 60 is branched into two in the vertical direction at a point 61 where the first tube 60 is connected to the first container 40. With this configuration, the flow rate of the liquid flowing into the first container 40 can be reduced and the medicine can be mixed more quickly, compared to a configuration in which the first tube 60 is not branched into two in the vertical direction but is connected as a single tube.

[0059] 6, the pump 70 is disposed on the first tube 60. Although there is no particular limitation on the pump 70, a tube pump 70 can be used. By using the tube pump 70 as the pump 70 in this manner, the movement of the liquid from the second container 50 to the first container 40 can be easily controlled.

[0060] As shown in FIG. 6, the tube pump 70 has three rollers 71 arranged at 120 degree intervals in the circumferential direction, a rotating part 72 that rotates the three rollers 71 in the circumferential direction, and a tube 73 connected to the first tube 60.

[0061] When the three rollers 71 of the tube pump 70 rotate in the circumferential direction, the tube 73 is pressed, and the liquid in the second container 50 is moved into the first container 40.

[0062] 6, the second tube 80 connects the first container 40 and the second container 50 in a liquid-tight manner. No pump is disposed on the second tube 80.

[0063] By providing the second tube 80 in this manner, when the tube pump 70 moves liquid from the second container 50 to the first container 40 and the height of the liquid in the first container 40 becomes higher than the height of the liquid in the second container 50, the liquid in the first container 40 moves to the second container 50 via the second tube 80. This makes it possible to continuously maintain the heights of the liquid in the first container 40 and the second container 50 at the same level.

[0064] Next, a method of using the tension measuring device 1 according to this embodiment will be described.

[0065] First, the first container 40 and the second container 50 are connected by the first tube 60 and the second tube 80, and a tube pump 70 is installed on the first tube 60. The gel adapter holder assembly 10 to which the gel G and the cell structure CS are attached is set in the first container 40. At this time, for example, 45 ml of medium is placed in the first container 40, and 5 ml of medium is placed in the second container 50.

[0066] Then, the liquids in the first container 40 and the second container 50 are circulated by the tube pump 70 and the second tube 80. In this state, for example, 5 μl of a drug is administered to the second container 50 at any timing.

[0067] Here, the tube pump 70 moves the liquid in the second container 50 to the first container 40. As a result, the liquid in the second container 50 decreases and the liquid in the first container 40 increases. Here, the liquid moves from the first container 40 to the second container 50 by the second tube 80 so that the heights of the first container 40 and the second container 50 are the same.

[0068] As described above, the tension measuring device 1 according to this embodiment is a tension measuring device 1 for a cell structure CS including muscle cells. The tension measuring device 1 includes a first container 40 in which the cell structure CS is placed and in which a culture medium is filled so that the cell structure CS is immersed, a second container 50 in which a drug to be administered is injected into the first container 40, a first tube 60 that connects the first container 40 and the second container 50 in a liquid-tight manner, and a pump 70 that is placed on the first tube 60 and causes the liquid in the second container 50 to flow into the first container. According to the tension measuring device 1 configured in this manner, the drug and culture medium in the second container 50 are caused to flow into the first container 40 via the first tube 60 by the pump 70. Therefore, compared to the case where the drug is directly flowed into the first container 40, it is possible to reduce vibrations that affect the tension detecting means 20. Therefore, it is possible to reduce vibrations that affect the tension detecting means 20 and accurately measure tension.

[0069] Furthermore, since the tube pump 70 is disposed on the first tube 60, away from the first container 40, it is possible to suitably prevent the accuracy of the tension measurement from being reduced due to vibration of the tube pump 70.

[0070] Furthermore, for example, when a drug is administered directly into the first container 40, the drug is administered near the cell structure CS containing muscle cells, so there is a possibility that the cell structure CS will be suddenly exposed to a high concentration of the drug. In contrast, according to the tension measuring device 1 of this embodiment, the drug diluted with the culture medium in the second container 50 is administered into the first container 40, so it is possible to prevent the cell structure CS from being suddenly exposed to a high concentration of the drug.

[0071] Moreover, the tension measuring device 1 further includes a second tube 80 that is separate from the first tube 60 and liquid-tightly connects the first container 40 and the second container 50. With this configuration, the liquid can be moved from the first container 40 to the second container 50 without reversely rotating the tube pump 70. Therefore, the liquid in the first container 40 and the second container 50 can be easily circulated.

[0072] Moreover, when the first container 40 is viewed from above in the vertical direction, the first tube 60 is connected to the first container 40 at a position offset from the center line L1. With this configuration, a rotating flow F can be generated in the first container 40, enabling mixing. This allows the drug to be mixed into the culture medium more quickly. Furthermore, the concentration gradient in which the oxygen concentration is high in the upper part of the first container 40 can be flattened.

[0073] In addition, the first tube 60 is branched into two in the vertical direction at the point where it is connected to the first container 40. With this configuration, the flow rate of the liquid flowing into the first container 40 can be reduced and the medicine can be mixed more quickly, compared to a configuration in which the first tube 60 is not branched into two in the vertical direction but is connected as a single tube.

[0074] Furthermore, the second container 50 has a smaller surface area than the first container 40. With this configuration, the drug and medium can be transferred from the second container 50 to the first container 40 using small amounts of the drug and medium.

[0075] <Second embodiment> Next, a second embodiment of the present invention will be described with reference to Fig. 7. Descriptions of parts common to the first embodiment will be omitted, and only features unique to the second embodiment will be described.

[0076] FIG. 7 is a schematic diagram showing a part of a tension measuring device 2 according to a second embodiment of the present invention.

[0077] As shown in FIG. 7, the tension measuring device 2 according to the second embodiment differs from the first embodiment in that a memory M is marked on the second container 50, a third tube 190 is connected to the second container 50, and an opening / closing valve 81 is provided on the second tube 80.

[0078] In the tension measuring device 2 according to the second embodiment, the second container 50 is provided with a scale M as shown in Fig. 7. The interval at which the scale M is provided is not particularly limited, but is, for example, 1 mm along the vertical direction.

[0079] The on-off valve 81 is provided on the second tube 80. The on-off valve 81 is always open during tension measurement.

[0080] The third tube 190 is liquid-tightly connected to the second container 50. An on-off valve 191 is disposed on the third tube 190.

[0081] The third tube 190 is a waste tube. In the tension measuring device 2 according to the second embodiment, the culture medium in the first container 40 and the second container 50 can be replaced by opening the on-off valve 191. Then, fresh culture medium can be introduced from the second container 50 to the first container 40 by driving the tube pump 70.

[0082] In the tension measuring device 2, when the culture medium in the first container 40 evaporates, it is necessary to add an amount of culture medium equal to the amount that evaporated, so that the amount of culture medium in the first container 40 remains constant.

[0083] In the tension measuring device 2 according to the second embodiment, when the medium in the first container 40 evaporates, the on-off valve 81 of the second tube 80 is closed. Then, the tube pump 70 supplies liquid from the second container 50 to the first container 40 so that the inside of the first container 40 becomes 45 ml. Then, the amount of the medium in the first container 40 can be kept constant by finding the amount of the liquid reduced in the second container 50 from the memory M and introducing the medium into the second container 50. Here, since the surface area of ​​the second container 50 is set smaller than the surface area of ​​the first container 40, the amount of the medium reduced in the first container 40 can be found in the second container 50 in an appropriate manner.

[0084] In addition, when the culture medium in the first container 40 evaporates, the height of the first container 40 can be lowered so that 45 ml remains in the first container 40, and the amount of liquid lost in the second container 50 at this time can be determined from the memory M.

[0085] <Third embodiment> Next, a third embodiment of the present invention will be described with reference to Fig. 8. Descriptions of parts common to the first embodiment will be omitted, and only features unique to the third embodiment will be described.

[0086] FIG. 8 is a schematic diagram showing a part of a tension measuring device 3 according to a third embodiment of the present invention.

[0087] The tension measuring device 3 according to the third embodiment is different in that a fourth tube 290 is provided in a system separate from the first tube 60 and the second tube 80, as shown in FIG.

[0088] The tension measuring device 3 according to the third embodiment has a fourth tube 290 that liquid-tightly connects the first container 40 and the second container 50. A tube pump 291 (corresponding to an inflow pump) is disposed on the fourth tube 290.

[0089] The tube pump 291 moves the liquid in the first container 40 to the second container 50. The liquid in the second container 50 is moved to the first container 40 by the tube pump 70, and the liquid in the first container 40 is moved to the second container 50 by the tube pump 291, thereby circulating the liquid in the first container 40 and the second container 50.

[0090] Furthermore, even if the flow rates of the tube pump 70 and the tube pump 291 are different, the second tube 80 allows the liquid levels in the first container 40 and the second container 50 to be the same.

[0091] The present invention is not limited to the above-described embodiment, and various modifications can be made within the scope of the claims.

[0092] For example, in the first embodiment described above, the heater 51 is disposed below the second container 50. However, the heater may be provided on the outer periphery of the first tube 60 or the second tube 80.

[0093] In the first embodiment described above, the tension measuring device 1 has the second tube 80. However, the tension measuring device does not have to have the second tube 80. In this case, the liquid is circulated between the first container 40 and the second container 50 by rotating the tube pump 70 forward and backward.

[0094] In the first embodiment described above, the first tube 60 is connected to the first container 40 at a position offset from the center line L1. However, for example, when the gel G and the cell structure CS held by the gel adapter holder assembly 10 are offset from the center line L1 passing through the center of the first container 40 in a direction perpendicular to the vertical direction of the first container 40, the first tube may be connected to the first container 40 at a position that is not offset from the center line L1.

[0095] In the first embodiment described above, the first tube 60 is branched into two in the vertical direction at the point where it is connected to the first container 40. However, the number of branches may be three or more when the tube inner diameter is small or the flow rate in the tube is fast. Furthermore, the first tube 60 may be branched not only in the vertical direction at the point where it is connected to the first container 40, but also in the up-down direction, left-right direction, etc. Furthermore, the first tube does not have to be branched at the point where it is connected to the first container 40.

[0096] In the first embodiment described above, the second container 50 is configured to have a smaller surface area than the first container 40. However, the second container may be configured to have a larger or equal surface area than the first container 40.

[0097] In the first embodiment described above, the tube pump 70 disposed on the first tube 60 is configured to have three rollers 71 arranged at 120 degree intervals in the circumferential direction. However, the number of rollers 71 is not limited to three, and may be three or more or three or less.

[0098] In the second embodiment described above, the second container 50 is configured to have a scale M. However, the scale M is not limited to the second container 50, and may be configured to be attached to the first container 40. In this case, the interval at which the scale M is attached is not particularly limited, but is, for example, 1 mm along the vertical direction. [Explanation of symbols]

[0099] 1, 2, 3 tension measuring device, 10 Gel adapter holder assembly, 11 first gel adapter holder, 12 second gel adapter holder, 40 first container, 50 second container, 51 Heater, 60 1st tube, 70 Tube pump (pump), 80 2nd tube, 190 3rd tube, 290 4th tube, 291 Tube pump (inflow pump), CS cell structure, G Gel, L1 centerline, M memory.

Claims

1. A device for measuring tension of a cell structure including muscle cells, comprising: a gel adapter holder assembly in which the gel and the cell structure are held; a first container in which the cell structure and a gel adapter holder assembly are placed and which is filled with a medium so that the cell structure is submerged; a second container into which the drug to be administered is injected; a first tube that connects the first container and the second container in a liquid-tight manner; a pump disposed on the first tube for causing liquid in the second container to flow into the first container; A tension measuring device in which, when the first container is viewed from above in the vertical direction, the first tube is connected to the side of the first container at a position offset from a center line passing through the center of the first container in a direction perpendicular to the vertical direction of the first container.

2. The tension measuring device according to claim 1 , further comprising a second tube that is separate from the first tube and that connects the first container and the second container in a liquid-tight manner.

3. The tension measuring device according to claim 1 , wherein the first tube is branched into a plurality of tubes in a vertical direction at a location where the first tube is connected to the first container.

4. The tension measuring device according to any one of claims 1 to 3, wherein the second container has a smaller surface area than the first container.

5. 5. The tension measuring device according to claim 1, wherein the second container is marked with a mark indicating the remaining amount of liquid.

6. 3. The tension measuring device of claim 2, further comprising a third tube connected to the second container for disposing of liquid in the second container.

7. a fourth tube that is separate from the first tube and that connects the first container and the second container in a liquid-tight manner; The tension measuring device of claim 6 , further comprising an inflow pump disposed on the fourth tube for causing liquid in the first container to flow into the second container.

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