Bioreactor chamber for growing and / or maturing tissue and method for growing and / or maturing tissue in a bioreactor chamber - Patent Application 20070122999

Abstract

A bioreactor chamber for growing and / or maturing tissues comprises a container (1) for containing a culture medium and a fixing means (2) configured to fix a flexible support (S) permeable to the culture medium on which cells forming the tissue are placed, inside the container. The chamber further comprises a mechanical stimulation means (3) for generating a periodic movement of the support (S) by a pressure surface (4) that presses against the support (S) by intermittent contact with the support (S). A method for growing and / or maturing tissues in a bioreactor chamber is also provided.

Description

[Technical Field]

[0001] The present invention relates to a bioreactor chamber for growing and / or maturing tissue and a method for growing and / or maturing tissue in a bioreactor chamber. [Background technology]

[0002] Bioreactor chambers are known for growing and / or maturing tissues and for maintaining tissue-forming cells in a culture medium under controlled conditions. In many cases, maintaining cells in a culture medium is not sufficient, and stimuli are required to induce vital functions in the tissue-forming cells.

[0003] Bioreactor chambers for growing and / or maturing tissues, including mechanical stimulation to induce vital functions in cells forming the tissue, for example cardiac or pulmonary tissue, are known.

[0004] For example, WO2015108869 discloses a bioreactor chamber for growing cardiac tissue, the bioreactor chamber comprising a container for containing a culture medium, a fixing means configured to fix a flexible support permeable to the culture medium on which cells forming the tissue are disposed, inside the container, and a mechanical stimulation means for generating a cyclic motion of the support. The chamber is configured such that the mechanical stimulation means acts on the culture medium to generate the cyclic motion of the support. It also discloses a method for growing cardiac tissue in the chamber. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2015108869 Brochure Summary of the Invention

[0006] It is an object of the present invention to provide a bioreactor chamber for growing and / or maturing tissue, as well as a method for growing and / or maturing tissue in a bioreactor chamber, as claimed.

[0007] A first aspect of the present invention relates to a bioreactor chamber for growing and / or maturing tissues, comprising a container for containing a culture medium and fixation means configured to fix a flexible support permeable to the culture medium on which the cells that will form the tissue are placed inside the container, the chamber further comprising mechanical stimulation means for generating a periodic movement of the support by means of a pressure surface that presses against the support by intermittent contact with the support.

[0008] A second aspect of the invention relates to a bioreactor comprising a chamber as described above.

[0009] A third aspect of the invention relates to a method for growing and / or maturing a cellular tissue in a bioreactor chamber, the method comprising the steps of fixing a flexible support permeable to a culture medium in which cells forming the tissue are disposed in the chamber, and generating a cyclical movement of the support by mechanical stimulation means, the cyclical movement being performed by pressing the support with a pressure surface that intermittently contacts the support.

[0010] The support is thereby acted upon by contact with a pressure surface, allowing for more precise control over the deformations exerted on the cells forming the tissue, while maintaining the cells in the culture medium within one and the same container.

[0011] These and other advantages and features of the present invention will become apparent in view of the drawings and detailed description of the invention. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 is an exploded view of a bioreactor chamber according to one embodiment of the present invention. [Figure 2]FIG. 2 is a cross-sectional perspective view of the chamber of FIG. 1 in a standby position. [Figure 3] FIG. 2 is a schematic diagram of cyclic motion within the chamber of FIG. 1. [Figure 4] 2 is a cross-sectional front view of the container and lid of the chamber of FIG. 1. [Figure 5] FIG. 2 is a cross-sectional perspective view of a fixing means of the chamber of FIG. 1; [Figure 6] FIG. 10 is a cross-sectional perspective view of a fixing means for a bioreactor chamber according to another embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0013] 1 to 3 show a bioreactor chamber 100 for growing and / or maturing tissues, comprising a container 1 for containing a culture medium and fixation means 2 configured to fix inside the container a flexible support S, which is permeable to the culture medium, on which the cells for growing and forming the tissue are placed. The chamber further comprises mechanical stimulation means 3 for generating a periodic movement of the support S by means of a pressure surface 4 that presses against the support S by intermittently contacting it.

[0014] The chamber of the present invention thereby allows cells deposited on a support to better mimic the physiological conditions experienced by tissues in vivo. By pressing the support with intermittent contact of a pressure surface, the deformation exerted on the cells forming the tissue can be more precisely controlled while the cells remain immersed in culture medium in one and the same container.

[0015] In the context of the present invention, the term "support" refers to a scaffold, construct, or membrane on which deposited cells are supported to grow and form tissue. Supports can be made using naturally occurring materials, such as collagen, alginate, hyaluronic acid, gelatin, fibrin, biopolymers, such as PCL, PLA, PGA, PLA-PEG, PLGA, decellularized matrices, or combinations thereof. They can be made with structures of different pore sizes or structures that can be adapted to each tissue. Supports are usually biocompatible, inert to culture media, gradually degradable with cell growth and tissue formation, have a porosity suitable for cell attachment, and their derivatives are non-toxic.

[0016] The support S of the present invention is flexible and permeable to the culture medium, so that when the pressure surface 4 presses the support S while the fixing means 2 fixes the support S, the support S is deformed. As shown in Figures 5 and 6, the support S comprises a regeneration region S1 where cells are deposited and pressed by the pressure surface 4, and a fixing region S2 that is at least partially fixed by the fixing means 2.

[0017] In the context of the present invention, the cyclic motion of the support refers to a motion that is repeated periodically, in which the support is deformed and returned to its initial position in each cycle, and thus the cells arranged on the support are also cyclically deformed. This cyclic motion allows the cells that form tissue, such as cardiac tissue, to undergo deformations intended to simulate the actual motion of tissue in a living organ. Thus, the cyclic motion makes it possible to mimic the physiological conditions that the tissue will experience in future implantation in vivo.

[0018] 3 shows a schematic representation of the periodic motion generated in the bioreactor chamber 100 shown in FIGS. 1 and 2. In the standby position X0, the mechanical stimulation means 3 is not in operation, and the pressure surface 4 is not in contact with the support S. The mechanical stimulation means 3 moves from the standby position X0 and back to the standby position X0 in each cycle, and as the mechanical stimulation means 3 moves, the pressure surface 4 presses the support S by intermittently contacting the support S. In order to actually press the support S with the pressure surface 4 in each cycle, the mechanical stimulation means 3 moves the pressure surface 4 beyond a threshold position X1, which is the position at which the pressure surface 4 actually contacts the support S.

[0019] The culture medium provides the nutrients necessary for the cells to grow as they remain immersed in the culture medium.

[0020] In the context of the present invention, intermittent contact refers to the fact that in each cycle the pressure surface is not in continuous contact but is in and out of contact with the support, which on the one hand allows nutrients to reach all cells more easily and on the other hand allows gas exchange so that the cells have a sufficient oxygen supply, thereby reducing the risk of tissue necrosis due to lack of nutrients and oxygen.

[0021] The fixation means 2 keeps the support S immersed in the culture medium even with cyclical movements so that the cells placed on the support S can remain viable and grow during the tissue formation and stimulation process, which can last for several months depending on the type of cells used.

[0022] In a preferred embodiment, the support S is a membrane. In this manner, the regeneration area of ​​the support is a substantially two-dimensional area, making it easier for the pressure surface to act across the entire regeneration area of ​​the support, thereby providing a more uniform mechanical stimulus to the cells disposed on the support.

[0023] In a preferred embodiment, the pressure surface 4 is a deformable wall of the container 1, which is arranged between the mechanical stimulation means 3 and the fixing means 2, as can be seen in the preferred embodiment shown in Figures 1 to 3. In this way, the mechanical stimulation means is arranged outside the container, thereby avoiding the introduction of foreign elements into the container that could affect the sterility of the culture medium and the cells arranged on the support, and the need for additional components to ensure leaktightness.

[0024] However, in another embodiment not shown, the pressure surface can be the surface of a mechanical stimulation means arranged inside the container.

[0025] In a preferred embodiment, the container 1 is formed by a single deformable part, which results in a leak-proof container with a simple container structure that is easy to manufacture and assemble.

[0026] The container 1 is preferably made of an elastomeric material, more preferably silicone. The high elasticity of the elastomeric material allows the deformable wall of the container to deform and move, contacting and pressing against the support, only elastically deforming when the mechanical stimulation means is applied, and recovering its original shape and ceasing contact with the support when the mechanical stimulation means is no longer applied. Silicone stands out among the most elastic elastomeric materials, but the container can also be made of other elastomeric materials such as TPU, TPE or EVA.

[0027] In a preferred embodiment, the container 1 is covered by a rigid casing 5 that partially covers the container 1 and immobilizes the fixing means 2 inside the container 1. The casing 5 can be made of metal, composite or rigid polymer. In this way, the fixing means remains immobile inside the container even when parts of the container, e.g. the deformable wall, are deformed.

[0028] The casing 5 can be formed by several parts which, when assembled, immobilize the fastening means 2 inside the container 1. To that end, the parts can have a shape complementary to the part of the container 1 that is covered by these parts.

[0029] In a preferred embodiment, the fixation means 2 is configured to generate a loading stimulus of the support S. The loading stimulus refers to a type of contraction in which the cells or tissues generated on the flexible and permeable support attempt to overcome the resistance of the elastomeric material to radial elongation.

[0030] In a preferred embodiment, the fixing means 2 comprises a first fixing element 21 and a second fixing element 22 between which the peripheral edge of the support S is fixed, the first fixing element 21 being preferably a ring. This allows the peripheral edge of the support to be fixed, but the central region of the support can be used to position cells without fixing means that may damage the cells, and can also be used to press the support by intermittently contacting the pressure surface with the support. That is, the fixing region S2 of the support S is located at the peripheral edge, and the regeneration region S1 of the support S is located in the central region of the support S.

[0031] In a preferred embodiment, the second fixation element 22 is a second ring or a wall of the container 1. This provides better fixation against mechanical stimuli.

[0032] In another preferred embodiment, the second fixation element 22 is a coating made of an elastomeric material that covers the first fixation element 21, as shown in Figure 6. The fixation means 2 is particularly advantageous for generating a tonic stimulus in the cells forming the tissue arranged on the support S.

[0033] In the fixation means 2 shown in FIG. 6 , the first fixation element 21 is a ring made of a rigid material, and the second fixation element 22 is a coating made of an elastomeric material such as silicone that covers the first fixation element 21 and the peripheral edge of the support S. The fixation means 2 is particularly advantageous for generating a stress stimulus in tissue-forming cells placed on the support S by fixing the previously radially stretched support S with the silicone coating. The support S attempts to overcome the resistance to the radial extension exerted by the fixation means 2, generating a contractile force on the support S and the cells. In this way, the fixation means allows for the application of a stress stimulus to the tissue formed by the cells, in addition to allowing for the application of a mechanical stimulus.

[0034] The fixation means 2 can be removably placed inside the container 1. In this way, the fixation means of the container can be easily removed together with the support and put back into place as many times as desired, allowing the tissue to be analysed at any time during tissue growth without the need to touch the tissue, or after tissue growth is complete, the tissue can be removed from the container without destroying it so that it can be implanted in a patient.

[0035] In a preferred embodiment, the mechanical stimulation means 3 are configured to apply different contact pressures to the support S by means of the pressure surface 4. In this way, it becomes possible to adjust the degree of deformation of the support S according to the degree of deformation required throughout cell growth.

[0036] The mechanical stimulation means 3 may comprise an actuator 31 with a set of interchangeable heads, each head having a different surface contour. When the mechanical stimulation means 3 is acted upon, the actuator 31 adopts in each case the surface contour of the head used and presses against the pressure surface 4 which is deformed in such a way that different contact pressures can be applied by each head used to the support S depending on the degree of deformation required throughout the cell growth.

[0037] The mechanical stimulation means 3 preferably comprises a linear actuator 31 which moves vertically and is arranged below the container 1 as shown in Figures 1 to 3 .

[0038] Chamber 100 may include at least one medium inlet port 61 and one medium outlet port 62, which are connected to the interior of vessel 1 to recirculate the culture medium. In this way, continuous and controlled supply of nutrients for cell growth and tissue formation over long periods of time can be ensured without the need for scheduled shutdowns to refresh the culture medium in the vessel.

[0039] In a preferred embodiment, the chamber 100 comprises a first culture medium inlet port 61 a and a second culture medium inlet port 61 b, and a first culture medium outlet port 62 a and a second culture medium outlet port 62 b. The first inlet port 61 a and the first outlet port 62 a are arranged above the immobilization means 2, and the second inlet port 61 b ​​and the second outlet port 62 b are arranged below the immobilization means 2, so that the culture medium can be recirculated above and below the support, thereby reducing the risk of cell necrosis due to a lack of nutrient regeneration on one side of the support.

[0040] In a preferred embodiment, chamber 100 includes at least one electrode port 7 through which an electrode is inserted into vessel 1 for electrical stimulation of cells. In this way, cells can be stimulated mechanically and electrically within one and the same chamber. Vessel 1 may include guide rails 142 that slide along to guide and secure the electrode within vessel 1 when the electrode is inserted through electrode port 7.

[0041] Additionally, chamber 100 may include at least one gas inlet port 8a and at least one gas outlet port 8b connected to the interior of vessel 1 for recirculating gas within vessel 1. This allows for gas supply and control outside the chamber at optimal conditions for continuous gas supply during long-term growth without the need to place the chamber inside another device, such as an incubator, that controls optimal conditions of gas concentration, temperature, and pressure inside vessel 1 for tissue growth.

[0042] In addition to the vessel 1, the chamber 100 may also include a lid 9 for closing the vessel 1. This makes it easier to reach and maintain the optimal conditions of gas concentration, temperature and pressure within the vessel 1 for tissue growth.

[0043] The lid 9 may be deformable in the same way as the container 1. In this way, the lid 9 may also be deformed so that no additional pressure is generated inside the container 1 when the pressure surface 4 is deformed.

[0044] Similarly, the lid 9 can be made of an elastomeric material like the container 1. In this way, the lid 9 is easily deformable and, once deformed, can return to its original shape in the absence of external forces.

[0045] The lid 9 is preferably a transparent optical grade silicone, which thus facilitates the study of tissue growth and / or maturation within the chamber, as it is not necessary to open the lid 9 or remove the fixation means 2 to measure applied deformation or to analyze tissue growth and / or maturation parameters.

[0046] In a preferred embodiment, the lid 9 includes at least one electrode port 7 and / or a gas inlet port 8 a and a gas outlet port 8 b, which allows the entry of several external elements into the chamber from above the vessel to be concentrated, simplifying the vessel structure and reducing the potential impact on such external elements caused by the movement of the pressure surface.

[0047] In the preferred embodiment shown in the figures, the lid 9 includes two electrode ports 7 and a gas inlet port 8a and a gas outlet port 8b. Electrodes can be inserted from above through each electrode port 7 in the lid 9 until they contact the support on which the cells are placed, allowing electrical stimulation to be applied directly to the cells or formed tissue. The gas ports 8a and 8b allow gas trapped within the container 1 to be recirculated between the lid 9 and the culture medium.

[0048] One preferred embodiment of the chamber 100 shown in Figures 1-3 is described below.

[0049] Regarding the container 1, in this preferred embodiment the container 1 comprises a base 13, the inner wall of which comprises a pressure surface 4 and a side wall 14 on which the fastening means 2 are arranged, as shown in FIG.

[0050] The base 13 may comprise side folds 131 and a flat central portion 132 whose inner wall is the pressure surface 4. In this way, the side folds allow the flat central portion of the base to move vertically so as to press against the support when the mechanical stimulation means acts on the base.

[0051] In this preferred embodiment, the container 1 is formed by a single deformable cylindrical section made of silicone, the base 13 being circular and the side wall 14 constituting a single wall.

[0052] However, in other embodiments not shown, the container 1 and base 13 can have other shapes, such as oval, square or rectangular.

[0053] With regard to the casing 5, in a preferred embodiment, the casing 5 is formed from three parts 5a, 5b, and 5c. Parts 5a and 5b of the casing 5 cover the side wall 14 of the container 1, and part 5c of the casing 5 covers the edges of the container 1 and the lid 9. In this way, the entire base and most of the lid can be deformed, but the side wall remains immobile, ensuring a tight seal of the container.

[0054] With regard to the fixing means 2, in a preferred embodiment the first fixing element 21 and the second fixing element 22 of the fixing means 2 are rings (see Figure 5), and the periphery of the support S is fixed between these rings, which are preferably made of a rigid material.

[0055] The side wall 14 of the container 1 may be provided with a groove 141 for detachably locating the fixing means 2, this groove 141 being in a plane parallel to the plane of the flat central part 132 of the base 13. To connect them, it is sufficient to lift the lid 9 and insert the fixing means 2 from above the container 1 until they fit into the groove 141 by deforming the side wall 14, and they can be easily removed again by pulling them to release the connection. With this configuration, the container 1 can be divided into two compartments and supports can be fixed on the side wall 14 immersed in the culture medium.

[0056] With regard to the mechanical stimulation means 3, in a preferred embodiment the mechanical stimulation means 3 is linear and comprises an actuator 31 arranged outside the container 1, below the base 13 of the container 1. A flat central part 132 of the base 13 is supported on the actuator 31, close to but spaced from the support S, in a standby position X0, in which the mechanical stimulation means 3 is inactive.

[0057] However, in other embodiments not shown, the mechanical stimulation means may comprise an actuator that, in addition to allowing the pressure surface to intermittently contact the support, also allows a twist to be applied to the support.

[0058] With regard to the medium inlet ports 61 a and 61 b ​​and the medium outlet ports 62 a and 62 b, in a preferred embodiment these are located in the side wall 14 of the vessel 1, and the parts 5 a and 5 b of the casing 5 covering the side wall 14 include openings complementary to the medium ports 61 a, 61 b, 62 a and 62 b for passing through these ports. In this way, possible damage to the medium ports due to movement of the pressure surfaces is reduced.

[0059] The present invention also relates to a bioreactor for growing and / or maturing tissue, comprising the chamber 100 described.

[0060] The bioreactor is preferably a bioreactor for growing and / or maturing cardiac tissue. The use of the chamber of the present invention in a bioreactor for growing and / or maturing cardiac tissue allows for better mimicking the physiological conditions experienced by tissue in vivo in terms of myocardial deformation. In this way, the tissue acquires structural and functional properties very similar to those of native myocardial tissue, thereby enabling successful transplantation into a patient.

[0061] Although a bioreactor for growing and / or maturing cardiac tissue is specifically described, a bioreactor according to the invention may be for growing and / or maturing other types of tissue, such as skeletal muscle tissue, skin tissue, lung tissue, bladder tissue, cartilage tissue or bone tissue.

[0062] Preferably, the container 1 and / or the fixing means 2 are removable. The container 1 and / or the fixing means 2 can be connected to the remaining components so that they can be easily removed from the bioreactor. In this way, it is possible to take the container with only the fixing means for fixing the support or together with the fixing means for fixing the support to the operating room where the tissue will be transplanted, simplifying the transport and preventing the tissue from being contaminated or degraded during transport.

[0063] Another aspect of the invention relates to a method for growing and / or maturing tissue in a bioreactor chamber, the method comprising: a step of fixing a flexible support, which is permeable to a culture medium on which cells that form a tissue are placed, surrounded by the culture medium in a bioreactor chamber; generating a periodic motion of the support by mechanical stimulation means, the periodic motion being carried out by pressing the support with a pressure surface that intermittently contacts the support; Includes:

[0064] In this way, the deformations applied to the cells that form the tissue can be more precisely controlled while the cells are kept in culture medium in one and the same container.

[0065] In a preferred embodiment of the method, a loading stimulus is generated in the support during the fixation step. Loading stimulus refers to a type of contraction in which cells or tissues generated on the flexible and permeable support attempt to overcome the resistance to radial elongation of an elastomeric material, such as the fixation element 22 of the support S in FIG. 6. In this way, the tissue acquires a higher contractile force that is more similar to natural tissue, thereby enabling successful implantation into a patient.

[0066] Optionally, the stress stimulation of the support during the immobilization step can be combined with electrical stimulation.

[0067] The method can be used with the chamber 100 as described above.

Claims

1. 1. A bioreactor chamber for growing and / or maturing a tissue, comprising: a container (1) for containing a culture medium; fixation means (2) configured to fix a flexible support (S) permeable to the culture medium inside the container (1) on which cells that form the tissue are placed; and mechanical stimulation means (3) for generating a periodic movement of the support (S), characterized in that the mechanical stimulation means (3) generates the periodic movement by means of a pressure surface (4) that presses against the support (S) by intermittently contacting the support (S).

2. 2. A bioreactor chamber according to claim 1, wherein the support (S) is a membrane.

3. 3. A bioreactor chamber according to claim 1 or 2, wherein the pressure surface (4) is a deformable wall of the container (1) arranged between the mechanical stimulation means (3) and the fixing means (2).

4. Bioreactor chamber according to any one of claims 1 to 3, wherein the container (1) is formed by a single deformable part.

5. Bioreactor chamber according to any one of claims 1 to 4, wherein the container (1) is made of an elastomeric material, preferably silicone.

6. 6. A bioreactor chamber according to any one of claims 1 to 5, wherein the vessel (1) is covered by a rigid casing (5) that partially covers the vessel (1) and immobilizes the fixing means (2) inside the vessel (1).

7. 7. A bioreactor chamber according to any one of claims 1 to 6, wherein the fixing means (2) are adapted to generate a load-inducing stimulus of the support (S).

8. 8. A bioreactor chamber according to any one of claims 1 to 7, wherein the fixing means (2) comprises a first fixing element (21) and a second fixing element (22) between which the periphery of the support (S) is fixed, the first fixing element (21) being preferably a ring.

9. 9. Bioreactor chamber according to claim 8, wherein the second fixing element (22) is a second ring or a wall of the container (1).

10. 10. A bioreactor chamber according to claim 9, wherein the second fixing element (22) is a coating made of an elastomeric material covering the first fixing element (21).

11. 11. Bioreactor chamber according to any one of claims 1 to 10, wherein the fixing means (2) are detachably arranged inside the container (1).

12. 12. A bioreactor chamber according to any one of claims 1 to 11, wherein the mechanical stimulation means (3) are configured to apply different contact pressures to the support (S) by means of the pressure surfaces (4).

13. 13. The bioreactor chamber according to any one of claims 1 to 12, comprising at least one medium inlet port (61) and one medium outlet port (62) connected to the interior of the vessel (1) for recirculating the culture medium.

14. 14. The bioreactor chamber according to claim 13, comprising a first culture medium inlet port (61 a) and a second culture medium inlet port (61 b) and a first culture medium outlet port (62 a) and a second culture medium outlet port (62 b), wherein the first inlet port (61 a) and the first outlet port (62 a) are arranged above the fixing means (2) and the second inlet port (61 b) and the second outlet port (62 b) are arranged below the fixing means (2), whereby the culture medium can be recirculated above and below the support (S).

15. 15. A bioreactor chamber according to any one of claims 1 to 14, comprising at least one electrode port (7) through which an electrode is inserted into the vessel (1) for electrical stimulation of the cells.

16. 16. A bioreactor chamber according to any one of claims 1 to 15, comprising at least one gas inlet port (8a) and at least one gas outlet port (8b) connected to the interior of the vessel (1) for recirculating gas within the vessel (1).

17. 17. Bioreactor chamber according to any one of claims 1 to 16, comprising a lid (9) for closing the vessel (1).

18. 18. Bioreactor chamber according to claim 17, wherein the lid (9) comprises the at least one electrode port (7) and / or the gas inlet port (8a) and the gas outlet port (8b).

19. 19. Bioreactor chamber according to claim 17 or 18, wherein the lid (9) is deformable.

20. 20. Bioreactor chamber according to any one of claims 17 to 19, wherein the lid (9) is made of an elastomeric material, preferably transparent optical grade silicone.

21. 21. A bioreactor chamber according to any one of claims 1 to 20, wherein the vessel (1) comprises a base (13) whose inner wall comprises the pressure surface (4) and a side wall (14) on which the fixing means (2) are arranged.

22. A bioreactor for growing and / or maturing tissue, comprising a chamber (100) according to any one of claims 1 to 21.

23. 23. The bioreactor of claim 22, wherein the tissue is cardiac tissue.

24. 24. Bioreactor according to claim 22 or 23, wherein the fixing means (2) for fixing the vessel (1) and / or the support (S) are removable.

25. 1. A method for growing and / or maturing tissue in a bioreactor chamber, comprising: a step of fixing a flexible support, which is permeable to a culture medium in which cells forming the tissue are placed, surrounded by the culture medium in the bioreactor chamber; generating a periodic motion of said support by mechanical stimulation means; wherein the cyclical movement is performed by pressing the support with a pressure surface that intermittently contacts the support.

26. 26. The method for growing and / or maturing tissue according to claim 25, wherein a stress stimulus is generated in the support during the fixing step.

27. 27. A method for growing and / or maturing tissue according to claim 25 or 26, wherein said chamber is a chamber (100) according to any one of claims 1 to 21.

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