Method for transporting laminated body and graft

Abstract

To provide a laminated body capable of reliably protecting a graft to be transported and having high flexibility, and to provide a method for transporting the graft.SOLUTION: A laminated body 10 includes a graft 11 and a support 12 containing a biocompatible compound, the support 12 being foam-like and covers at least one surface of the graft 11. The support 12 has a volume ratio of air to a volume of a material constituting the support 12 of 30% to 200%, and a diameter of the bubble is 10 μm to 100 μm.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a laminate formed for delivery of an implant for implantation in a living body and a method for delivering the implant. [Background technology]

[0002] In recent years, new regenerative medicine has been developed as a solution for treating severe heart failure. One example is a method in which a sheet-shaped cell culture prepared using a temperature-responsive culture dish using tissue engineering is applied to the surface of the heart as a graft in severe myocardial infarction, etc. The method using the sheet-shaped cell culture makes it possible to safely transplant a large amount of cells over a wide area, and is particularly useful for treating, for example, myocardial infarction (including chronic heart failure associated with myocardial infarction), dilated cardiomyopathy, ischemic cardiomyopathy, and heart diseases (e.g., heart failure, especially chronic heart failure) accompanied by systolic dysfunction (e.g., left ventricular systolic dysfunction), etc.

[0003] A sheet-shaped cell culture is formed by culturing cell tissue collected from a patient in a culture facility. To transplant the sheet-shaped cell culture into a patient, the sheet-shaped cell culture is transported from the culture facility to a medical facility. Alternatively, the sheet-shaped cell culture is produced in a cell processing center (CPC) in a medical facility and transported to a surgical room.

[0004] The sheet-shaped cell culture is transported in a sealed culture dish containing the sheet-shaped cell culture and a buffer solution. At this time, the sheet-shaped cell culture is suspended in the buffer solution. Since the sheet-shaped cell culture is fragile, it can easily become wrinkled or folded due to shaking or impact during transportation or the operation of removing the sheet-shaped cell culture from the culture dish, which can cause damage to the cells.

[0005] In order to transport a sheet-shaped cell culture without damaging it, Patent Document 1 discloses covering the sheet-shaped cell culture with a gelled support layer. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] JP 2011-172925 A Summary of the Invention [Problem to be solved by the invention]

[0007] Implants are very fragile, so to transport the cells without damaging them, a laminate and delivery method that is strong and flexible for ease of implantation is desirable.

[0008] The present invention has been made to solve the above-mentioned problems, and has an object to provide a laminate that can reliably protect a graft during transportation and has high flexibility, and a method for transporting a graft. [Means for solving the problem]

[0009] (1) The laminate of the present invention, which achieves the above-mentioned object, is a laminate including a graft and a support containing a biocompatible compound, the support being foam-like and covering at least one surface of the graft.

[0010] (6) A method for transporting a graft according to the present invention, which achieves the above-mentioned object, comprises the steps of placing the graft on a substrate, and spraying a liquid containing a biocompatible compound onto the graft in foam form and solidifying the liquid, thereby coating one side of the graft with a foam support to form a laminate. Effect of the Invention

[0011] The laminate (1) constructed as described above has a high flexibility and high strength during transportation because the support has a large volume due to the foam. Therefore, the laminate can reliably protect the graft during transportation.

[0012] (2) In the laminate of (1) above, the support may have a volume ratio of air to the volume of the material constituting the support of 30% to 200%, which allows the laminate to have a sufficiently large volume of the support and to adequately protect the graft during transportation.

[0013] (3) In the laminate of (1) or (2) above, the support may have bubbles having a diameter of 10 μm to 100 μm, thereby ensuring that the laminate can reliably protect the graft being transported by keeping the bubble size within an appropriate range.

[0014] (4) In any one of the laminates (1) to (3) above, the thickness of the laminate may be 10 μm to 10 mm, thereby ensuring a sufficient thickness for protecting the graft.

[0015] (5) In any of the laminates (1) to (4) above, the biocompatible compound may be fibrin or dextrin. These biocompatible compounds can be laminated in a foamed state on the surface of a graft to form a stable support.

[0016] The (6) method for transporting a graft configured as described above allows the graft to be transported while being protected by the foam support, thereby preventing the graft from wrinkling or folding during transport, and thus preventing damage to the graft.

[0017] (7) The above-mentioned method for transporting a graft according to (6) may further include a step of peeling the laminate from the substrate, and a step of suspending the laminate detached from the substrate in a buffer solution, whereby the graft having a foam-like support can be suspended in the buffer solution, and therefore the graft can be easily removed and transplanted. [Brief description of the drawings]

[0018] [Figure 1] FIG. 2 is a cross-sectional view of a culture dish and a laminate in a state in which the laminate is formed on a substrate in this embodiment. [Diagram 2] FIG. 2 is a cross-sectional view of a culture dish and an explant in a state in which the explant is placed on a substrate. [Diagram 3] FIG. 13 is a cross-sectional view of a culture dish and an explant in a state in which the explant is being sprayed with a support material. [Figure 4] FIG. 2 is a cross-sectional view of a culture dish and a laminate in a state where the laminate is suspended in a buffer solution. [Diagram 5] FIG. 2 is a cross-sectional view of the culture dish and the laminate in a state where the laminate has been removed from the buffer solution with a medical spatula. [Figure 6] Photographs from an example in which a laminate of a sheet-like material and fibrin gel was produced, in which (a) shows the support formed on a dish, (b) shows the laminate lifted up with a spatula, and (c) shows the laminate suspended in a buffer solution. [Figure 7] These are photographs from an example in which a laminate of a sheet-like material and dextrin gel was produced, in which (a) shows the support formed on a dish, (b) shows the laminate being held up with tweezers, and (c) shows the laminate being suspended in a buffer solution. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] Hereinafter, an embodiment of the present invention will be described with reference to the drawings. Note that, for the sake of explanation, the dimensional ratios in the drawings may be exaggerated and may differ from the actual ratios.

[0020] The laminate 10 of this embodiment is formed by laminating a graft 11 and a support 12. The graft 11 is a sheet-shaped cell culture. The sheet-shaped cell culture is formed by cells connecting with each other to form a sheet.

[0021] The sheet-shaped cell culture is formed by culturing tissue collected from a living body. The cells constituting the sheet-shaped cell culture are myoblast cells such as skeletal myoblast cells and fibroblast cells. However, the cells are not limited thereto, and include, for example, somatic stem cells, adult stem cells, mesenchymal stem cells, ES cells (embryonic stem cells), or cardiomyocytes derived from iPS cells (induced pluripotent stem cells). The somatic stem cells preferably include skeletal myoblast cells (myoblast cells). The graft 11 may be a sheet-shaped material other than the sheet-shaped cell culture. The sheet-shaped material includes films made of various materials such as plastic, paper, woven fabric, nonwoven fabric, metal, polymer, and lipid. The sheet-shaped cell culture or sheet-shaped material is a fragile material that shows a breaking load of, for example, less than 0.5 N in a tensile test.

[0022] A sheet-shaped cell culture is formed by suspending cells extracted from tissue collected from a living body and culturing them on a culture substrate. As shown in Fig. 1, a graft 11, which is a sheet-shaped cell culture, is placed on the surface of the bottom surface 20a of a culture dish 20, which serves as the substrate.

[0023] The laminate 10 has a graft 11 arranged on a substrate and a foam-like support 12 covering one side of the graft 11. The support 12 contains a biocompatible compound. Biocompatibility refers to the property of not causing an inflammatory reaction, an immune reaction, or the like to living tissue or cells, being bioinert, or not having biotoxicity. Since the support 12 contains a biocompatible compound, the graft 11 can be transplanted directly to the transplant site without being separated from the laminate 10. Specifically, the biocompatible compound is fibrin or dextrin. However, the biocompatible compound constituting the support 12 may be other than these.

[0024] The material of the support 12 may be any material that contains a biocompatible compound and can be foamed, for example, fibrin gel obtained by mixing a fibrinogen solution with a thrombin solution, an adhesion prevention material obtained by mixing an aqueous solution of NHS-CM dextrin and trehalose with an aqueous solution of sodium carbonate or sodium bicarbonate, etc. However, the material of the support 12 is not limited to these.

[0025] The ratio of the volume of air in the support 12 to the volume of the material constituting the support 12 is 30% to 200%. The bubbles in the support 12 have a diameter of 10 μm to 100 μm. The density of the support 12 is 50 μg / cm 2 ~300mg / cm 2 The thickness of the laminate 10 is 10 μm to 10 mm. In a tensile test, the laminate 10 exhibits a breaking load of 0.1 N to 5 N. The breaking load of the laminate 10 is higher than the breaking load of the graft 11.

[0026] Next, a method for transporting the graft 11 will be described. First, as shown in Fig. 2, the graft 11 is placed on a substrate. The substrate on which the graft 11 is placed may be the surface of a culture dish 20 in which the graft 11 has been cultured, or the surface of a medium formed on the bottom surface 20a of the culture dish 20. The substrate may also be the surface of another container to which the cultured graft 11 has been moved.

[0027] The graft 11 is in a state of being adhered onto the substrate before the laminate 10 is formed. However, the graft 11 may be peeled off from the substrate before the laminate 10 is formed.

[0028] As shown in Fig. 3, the material of the support 12 is sprayed in a foam state from the spray nozzle 30 onto the surface of the graft 11 placed on the substrate. The spray nozzle 30 can mix the solution that is the material of the support 12 with gas near the tip and spray the material in a foam state. The sprayed material of the support 12 solidifies on the surface of the graft 11 to form the foamed support 12. The gas used in the spray nozzle 30 can be air, nitrogen, carbon dioxide, etc., and is not particularly limited.

[0029] In this embodiment, the support 12 is formed to cover the entirety of one side of the graft 11. When the graft 11 is in a state where it is peeled off from the substrate, the support 12 may be formed to cover both sides of the graft 11.

[0030] When the same amount of biocompatible compound is used as in the case of forming a gel-like support by spraying a liquid, the foam-like support 12 formed becomes bulkier than the gel-like support because the foam increases the volume. Therefore, the support 12 has high flexibility and can have higher strength during transportation. In addition, since the support 12 has a large thickness, when the graft 11 is transplanted without separating the support 12, it can be easily sutured with thread to the living body. In addition, since the support 12 is sprayed in a foam state, the range to be sprayed is easily adjusted. Therefore, the laminate 10 can suppress the support 12 from spreading more than necessary, and can facilitate handling during transplantation. In addition, since the support 12 is foam-like, its color becomes opaque, and visibility during transplantation can be improved.

[0031] Once the laminate 10 is formed, the culture dish 20 is filled with the buffer solution 22. By transporting the laminate in this state, the graft 11 can be prevented from wrinkling or folding even if it is shaken or impacted during transportation.

[0032] When transplantation is to be performed, the laminate 10 is peeled off from the substrate. The culture dish 20 is a temperature-responsive culture dish, and a temperature-responsive polymer is immobilized on the surface of the substrate. The temperature-responsive polymer exhibits hydrophobicity to which cells easily adhere at temperatures near those used for culture, and becomes hydrophilic to which cells do not easily adhere at lower temperatures. By utilizing this, the laminate 10 can be easily peeled off from the substrate.

[0033] As shown in Fig. 4, the laminate 10 has a foam-like support 12 and can therefore float in the buffer solution 22. The floating laminate 10 can be easily removed from the buffer solution 22 with, for example, a medical spatula 40, as shown in Fig. 5. EXAMPLES

[0034] (Preparation of sheet-like object) A paper sheet of about 100 μm was cut into a circle of about 40 mm in diameter to prepare a sheet-like object. This was placed on a substrate and immersed in a buffer solution (HBSS(-), Cat No. 14175095, ThermoFisher Scientific). In the buffer solution, the sheet-like object sank to the bottom of the substrate and did not float.

[0035] (Preparation of a laminate of a sheet-like material and fibrin gel) The prepared sheet-like material was soaked in a buffer solution (HBSS(-), Cat No. 14175095, ThermoFisher Scientific), and the wet sheet-like material was placed on a substrate (UpCell (registered trademark)) with a diameter of 9.2 cm, a 10 cm dish (CS3005, CellSeed). 5 ml of fibrinogen solution (contents of vial 1 (lyophilized fibrinogen powder) of Veriplast (registered trademark) for tissue adhesion (CSL Behring) dissolved in contents of vial 2 (fibrinogen dissolving solution), fibrinogen concentration 80 mg / ml) and 5 ml of thrombin solution (Veriplast The contents of vial 3 (lyophilized thrombin powder) of Tissue Adhesive (manufactured by CSL Behring) were dissolved in the contents of vial 4 (thrombin dissolving solution), and a thrombin concentration of 300 units / ml was prepared, which was then filled into the spray syringe of a spray kit (Adspray (registered trademark)) and attached to the nozzle. While supplying carbon dioxide gas under a condition of 0.10 MPa, the fibrinogen solution and the thrombin solution were pushed out of the spray syringe, respectively, and the fibrinogen solution and the thrombin solution were sprayed at the tip of the sprayer. The fibrinogen solution and the thrombin solution were mixed and sprayed in mist form onto the sheet-like material. At this time, the two solutions were sprayed in their entirety evenly so as to cover the top of the sheet-like material and the area around the sheet-like material, which was about 1 cm around the sheet-like material. The fibrin gel formed by mixing the fibrinogen solution and the thrombin solution was layered on the sheet-like material in the form of bubbles (microbubbles) or mist, and was fixed onto the base material, forming a support covering the 1 cm area around the sheet-like material (see FIG. 6(a)). Here, the laminate consisting of the sheet-like material and the fibrin gel had a thickness of 7 mm. The fibrin gel was spread evenly over the sheet-like material, layered in an uneven mountain shape with small bubbles uniformly packed in it. The laminate was opaque white and had a smooth surface. The laminate could be peeled off from the substrate by using a spatula to peel off the part where the support and substrate were in direct contact (see Figure 6(b)). When a buffer solution was added to the substrate, the laminate floated on the buffer solution (see Figure 6(c)). The laminate did not break when held with tweezers or pulled.

[0036] (Preparation of a laminate of a sheet-like material and dextrin gel) The prepared sheet was diluted with a buffer solution (HBSS(-), Cat No. 14175095, ThermoFisher The wetted sheet-like material was placed on a 9.2 cm diameter substrate (UpCell (registered trademark), CellSeed Scientific), and a 10 cm dish (CS3005, CellSeed). Powder A (Adspray (registered trademark), NHS-modified CM dextrin, trehalose hydrate (Terumo Corporation)) and powder B (sodium carbonate, sodium bicarbonate) were dissolved in dissolving water, respectively, and filled into a spray syringe of a spray kit (Adspray (registered trademark)) and attached to a nozzle. Powder A liquid and powder B liquid were pushed out from the spray syringe while supplying carbon dioxide gas under a condition of 0.10 MPa, and the mixed state of powder A liquid and powder B liquid at the tip of the sprayer was sprayed onto the sheet-like material in a mist form. At this time, the entire amount of the two liquids was sprayed evenly so as to cover the top of the sheet-like material and the periphery of the sheet-like material. Powder A liquid and powder B liquid were mixed. The gel formed by combining the two materials was layered on the sheet-like material in the form of bubbles (microbubbles) or mist, and was fixed on the substrate, forming a support covering the entire surface of the substrate including the sheet-like material (see FIG. 7(a)). Here, the laminate consisting of the sheet-like material and the dextrin gel had a thickness of 4 mm, and the dextrin gel was layered flat (flat) on the sheet-like material, with fine bubbles uniformly densely packed. The laminate was pale yellow, mostly translucent, and partially opaque. The surface of the laminate was smooth. The laminate could be peeled off from the substrate by pressing the support around the sheet-like material with a spatula to make an incision and inserting the spatula under the support (see FIG. 7(b)). When a buffer solution was added to the substrate, the laminate floated on the buffer solution (see FIG. 7(c)). The laminate did not break when held with tweezers, but broke when intentionally folded or pulled.

[0037] As described above, the laminate 10 according to the present embodiment (1) is a laminate 10 including a graft 11 and a support 12 containing a biocompatible compound, the support 12 being foam-like and covering at least one surface of the graft 11. The laminate 10 thus configured has high flexibility because the support 12 has a large volume due to the foam, and can have high strength during transportation. Therefore, the laminate 10 can reliably protect the graft 11 during transportation.

[0038] (2) In the laminate 10 of (1) above, the support 12 may have a volume ratio of air of 30% to 200% relative to the volume of the material constituting the support 12. This allows the laminate 10 to have a sufficiently large volume of the support 12, and to fully protect the graft 11 during transportation.

[0039] (3) In the laminate 10 of (1) or (2) above, the support 12 may have bubbles with a diameter of 10 μm to 100 μm. This allows the laminate 10 to reliably protect the graft 11 being transported by keeping the bubble size within an appropriate range.

[0040] (4) In any one of the laminates 10 described above in (1) to (3), the thickness of the laminate 10 may be 10 μm to 10 mm. This allows the laminate 10 to have a sufficient thickness to protect the graft 11.

[0041] (5) In any of the laminates 10 described above in (1) to (4), the biocompatible compound may be fibrin or dextrin. These biocompatible compounds can be laminated in a foamed state on the surface of the graft 11, forming a stable support 12.

[0042] The (6) method for transporting a graft 11 according to this embodiment includes the steps of placing the graft 11 on a substrate, and spraying a liquid containing a biocompatible compound onto the graft 11 in foam form and solidifying the liquid to cover one side of the graft 11 with a foam-like support 12 to form a laminate 10. The method for transporting a graft 11 configured in this manner can transport the graft 11 while protecting it with the foam-like support 12, thereby preventing the graft 11 from wrinkling or folding during transport.

[0043] (7) The above method for transporting the graft 11 in (6) may include a step of peeling the laminate 10 from the substrate, and a step of suspending the laminate 10 peeled from the substrate in the buffer solution 22. In this way, the graft 11 having the foam-like support 12 can be suspended in the buffer solution 22, so that the graft 11 can be easily taken out and transplanted.

[0044] The present invention is not limited to the above-described embodiment, and various modifications can be made by those skilled in the art within the technical concept of the present invention. [Explanation of symbols]

[0045] 10 Laminate 11 Graft 12 Support 20 Culture dish 22 Buffer 30 Spray nozzle 40 Medical Spatula

Claims

1. A laminate comprising an implant and a support containing a biocompatible compound, The support is a foam and a laminate covering at least one surface of the implant.

2. 2. The laminate according to claim 1, wherein the support has a volume ratio of air to the volume of the material constituting the support of 30% to 200%.

3. 3. The laminate according to claim 1, wherein the support has bubbles having a diameter of 10 μm to 100 μm.

4. 3. The laminate according to claim 1, wherein the thickness of the laminate is from 10 μm to 10 mm.

5. 3. The laminate according to claim 1, wherein the biocompatible compound is fibrin or dextrin.

6. 1. A method for delivering a graft, comprising: placing the implant on a substrate; A step of spraying a liquid containing a biocompatible compound onto the implant in the form of a foam and solidifying the foam to cover one side of the implant with a foam support to form a laminate; A method for transporting a graft comprising the steps of:

7. peeling the laminate from the substrate; A step of suspending the laminate peeled off from the substrate in a buffer solution; 7. The method of claim 6, comprising:

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