Cell culture container

Abstract

There is a need for compact, automated cell processing devices (e.g., multi-stage cell processing devices) that can perform unit operations without contamination. [Solution] This disclosure provides a cell culture vessel (2) comprising a base portion (7) and a compressible wall element (6). The compressible wall element (6) extends axially from the base portion (7) and defines the internal volume of the cell culture vessel (2). The compressible wall element (6) is compressible in the axial direction. The base portion (7) comprises a substantially planar rigid base plate (12). A bioreactor (1) comprising the cell culture vessel (2) is also disclosed.

Description

Technical Field

[0001] The present invention relates to a cell culture container, such as a cell culture container of a bioreactor for use in cell culture processing, for example.

Background Art

[0002] Cell therapy manufacturing processes and gene therapy manufacturing processes are often complex and include manual or semi-automatic steps across several devices. The equipment systems used in various steps (i.e., unit operations) of the manufacture of cell-based therapeutic products (CTPs) can include devices for cell collection, cell separation / selection, cell expansion, cell washing and volume reduction, cell storage, and transportation. Unit operations can vary greatly based on, among several factors, the manufacturing model (i.e., autologous vs. allogeneic), cell type, and intended purpose. Also, cells are "living" entities that are sensitive even to the most simple operations, such as differences in the procedure for transferring cells. The role of cell manufacturing equipment to ensure scalability and reproducibility is an important factor in cell therapy manufacturing and gene therapy manufacturing.

[0003] Also, cell-based therapeutic products (CTPs) are gaining significant momentum, and thus, improved cell manufacturing equipment is required for various cell manufacturing procedures, such as, but not limited to, stem cell enrichment, generation of chimeric antigen receptor (CAR) T cells, and various cell manufacturing processes such as collection, purification, genetic recombination, culture / harvest, washing, injection into patients, and / or freezing.

[0004] Cell culture or processing typically requires the use of a device to hold the cells when culturing them, for example, in a suitable culture medium. Known devices include shake flasks, roller bottles, T-flasks, and bags.

[0005] A significant limiting factor in the production of cell or gene therapies for pharmaceutical use is the lack of compact, automated, and closed systems for performing unit operations without contamination. For example, during cell culture, upstream or subsequent cell handling poses a risk of contamination when adding cells to incubators, removing cells, or removing liquid samples. Operating systems are largely manual and therefore expensive to operate. Multiple instruments are typically required to cover all non-cell culture steps, which involve many transfers, each of which presents an opportunity for operator error and contamination. Furthermore, the increased manual actions increase the risk of manual error, and therefore, current labor-intensive processes lack the robustness required for the production of clinical-grade therapies. [Overview of the project] [Problems that the invention aims to solve]

[0006] Therefore, there is a need for cell processing devices (e.g., multi-stage cell processing devices) that allow such processing and enable such processing, avoiding the requirement of constant cell transfer to a new device. [Means for solving the problem]

[0007] According to an aspect of the present invention, a cell culture vessel is provided comprising a base portion and a compressible wall element. The compressible wall element extends axially from the base portion and defines the internal volume of the cell culture vessel. The compressible wall element is compressible in the axial direction. The base portion comprises a substantially planar rigid base plate.

[0008] A planar, rigid base plate provides a substantially planar, or flat, bottom to the internal volume of the cell culture vessel. Advantageously, a planar bottom to a cell culture vessel can provide improved cell culture, such as enhanced mixing and control across the cell culture. A planar bottom to a cell culture vessel can help ensure that cells are distributed substantially uniformly across the cross-section of the vessel as they settle to the bottom. In contrast, if the base portion is not planar, cells become concentrated in a smaller volume, which can be detrimental to cell culture. A planar, rigid base plate to a cell culture vessel also helps prevent fluids from becoming trapped in the vessel when cells are harvested or extracted at the end of the cell culture process.

[0009] In the example, the compressible wall element may be bonded or welded to the rigid base plate. For example, the compressible wall element may be hot-plate welded or ultrasonically welded to the rigid base plate. In the example, the compressible wall element may be fastened or clipped to the rigid base plate. For example, the compressible wall element may have a protruding edge that is clipped into a groove formed in the rigid base plate. In the example, the compressible wall element may be molded integrally with the rigid base plate. For example, the rigid base plate may be overmolded onto a portion of the compressible wall element. Specifically, the rigid base plate may be overmolded onto the hem of the compressible wall element. In the example, the compressible wall element is attached to the rigid base plate by sealing.

[0010] In the example, the compressible wall element may have a deformable portion located at the joint between the compressible wall element and the rigid base plate, or immediately adjacent to that joint. Such an arrangement ensures that the compressible wall element can be fully compressed, thereby preventing or reducing fatigue stress in the compressible wall.

[0011] In the example, the cell culture vessel may further comprise a base sheet extending across a rigid base plate within the internal volume of the cell culture vessel. The base sheet may define the bottom surface of the internal volume of the cell culture vessel. The base sheet may extend from a compressible wall element. Specifically, the base sheet may be molded integrally with the compressible wall element, for example, the compressible wall element and the base sheet may be formed integrally by blow molding. Alternatively, the base sheet may be attached to the compressible wall element and / or rigid base plate, for example, by adhesive or welding.

[0012] In the example, the base sheet may be gas permeable. Specifically, the base sheet may be oxygen permeable. In the example, the base sheet may contain silicone.

[0013] In this example, the rigid base plate may have one or more gas-permeable openings. Each of these gas-permeable openings may be a hole or opening in the rigid base plate. Thus, a gas flow, such as airflow, is supplied to the outer surface of the base sheet through one or more openings in the rigid base plate.

[0014] In the example, the rigid base plate may include one or more spacers adapted to separate the base sheet from the rigid base plate. Thus, a gas flow, such as airflow, can be provided to a larger surface area on the outer surface of the base sheet.

[0015] In this example, the compressible wall element may comprise an inwardly deformable portion, an outwardly deformable portion, and a leaf portion extending between the inwardly and outwardly deformable portions. In this configuration, the deformation of the inwardly and outwardly deformable portions compresses the compressible wall element. Specifically, the leaf portions can overlap each other to reduce the height of the compressible wall element in the axial direction. It should be understood that the compressible wall element is also expandable by the same or a similar mechanism.

[0016] In this example, either the inwardly deformable or outwardly deformable portion is located at the joint between the compressible wall element and the rigid base plate, or immediately adjacent to that joint. Such arrangement ensures that the compressible wall element can be fully compressed, thereby preventing or reducing fatigue stress in the compressible wall.

[0017] In one example, the rigid base plate may have a transparent or translucent sensor window. In another example, the rigid base plate is transparent or translucent, or a portion of the rigid base plate has a sensor window. In yet another example, the rigid base plate is opaque, and the rigid base plate has a transparent or translucent sensor window that is attached, inserted, or molded integrally with it. In yet another example, the rigid base plate has a sensor window made of opaque high-density polyethylene and transparent polycarbonate that is attached to the rigid base plate or molded integrally with the rigid base plate. In an example where the cell culture vessel has a base sheet, the base sheet may contain a transparent or translucent material. Alternatively, the base sheet may have an opening corresponding to the sensor window. In this example, the base sheet may be sealed to the rigid base plate around the opening.

[0018] In the example, the cell culture vessel comprises one or more sensor elements positioned in a sensor window. The one or more sensor elements may be positioned on the inner surface of the sensor window within the internal volume of the cell culture vessel. The one or more sensor elements may include optical dots, such as optical dots for use with an optical fluorescence sensor.

[0019] In the example, the cell culture vessel may further comprise one or more sensors, specifically optical sensors, mounted in a sensor window. One or more optical sensors may transmit and receive light through the sensor window to detect parameters of the fluid in the cell culture vessel. One or more optical sensors may cooperate with one or more sensor elements, specifically optical dots. The optical sensors and / or optical dots may measure the dissolved oxygen concentration of the fluid in the cell culture vessel.

[0020] In one example, the compressible wall element may contain silicone. In another example, the compressible wall element may contain low-density polyethylene. In yet another example, the compressible wall element may contain a thermoplastic elastomer. In one example, the compressible wall element may comprise an outer portion and a lining or insert. For example, the outer portion may contain a thermoplastic elastomer, and the lining may be blow-molded onto the outer portion. In yet another example, the compressible wall element may comprise an inner portion and a jacket. The jacket may be overmolded onto the inner portion. In one example, at least a portion of the compressible wall element may have a coating, such as a gas-impermeable coating.

[0021] In the example, the rigid base plate may include high-density polyethylene or polycarbonate. In the example, the rigid base plate may be transparent or translucent, or may have a transparent or translucent sensor window.

[0022] A further aspect of the present invention provides a bioreactor for cell culture processing. The bioreactor comprises the cell culture vessel described above. The bioreactor may further comprise a joint plate that can be attached to a compressible wall element opposite the base portion for closing the cell culture vessel. The joint plate can act as a lid or closure for the cell culture vessel. The joint plate can seal the internal volume of the cell culture vessel.

[0023] In the example, the connecting plate may be equipped with a connector joint. For example, the connector joint can facilitate the fluid connection of a container for introducing fluid into a cell culture vessel, or the fluid connection of a sampler for sampling fluid from a cell culture vessel.

[0024] In an example, the bioreactor may further comprise one or more sensors, specifically one or more optical sensors, disposed in a sensor window in a rigid base plate. The one or more optical sensors can send and receive light through the sensor window to detect parameters of the fluid in the cell culture vessel. The one or more optical sensors can cooperate with one or more sensor elements, specifically optical dots, disposed in the sensor window. The optical sensors and / or the optical dots can measure the dissolved oxygen concentration of the fluid in the cell culture vessel.

[0025] According to a further aspect of the present invention, there is provided a cell processing system comprising a bioreactor as described above.

[0026] In an example, the cell processing system may further comprise a stirrer configured to move a base portion to stir the fluid in the cell culture vessel. The stirrer may be configured to move the base portion relative to a joining plate by at least partially compressing or expanding a compressible wall.

[0027] Embodiments of the present invention will be described hereinafter with reference to the accompanying drawings.

Brief Description of the Drawings

[0028] [Figure 1] It is a view of a bioreactor having a cell culture vessel. [Figure 2a] It is a cross-sectional view of an example cell culture vessel. [Figure 2b] It is an exploded assembly view of the cell culture vessel of FIG. 2a. [Figure 3a] It is a view of an example cell culture vessel. [Figure 3b] It is a cross-sectional view of the cell culture vessel of FIG. 3a. [Figure 3c] It is a detailed cross-sectional view of an example joint between a compressible wall element and a rigid base plate of the cell culture vessel of FIGS. 3a and 3b. [Figure 3d]Figures 3a and 3b show detailed cross-sectional views of an example of the joint between a compressible wall element and a rigid base plate in a cell culture vessel. [Figure 4] This is a cross-sectional view of the cell culture vessel in question. [Figure 5a] This is a diagram showing an example of a cell culture vessel. [Figure 5b] This is a diagram showing an example of a cell culture vessel. [Figure 5c] This is a diagram showing an example of a cell culture vessel. [Figure 6a] This is a cross-sectional view of the cell culture vessel in question. [Figure 6b] Figure 6a is a disassembled diagram of a cell culture vessel. [Figure 7a] This is a cross-sectional view of the cell culture vessel in question. [Figure 7b] Figure 7a is a bottom view of the cell culture vessel. [Figure 7c] Figures 7a and 7b are detailed cross-sectional views of the joint between the compressible wall element and the rigid base plate of the cell culture vessel. [Figure 8] This is a cross-sectional view of the cell culture vessel in question. [Figure 9a] This is a cross-sectional view of the cell culture vessel in question. [Figure 9b] Figure 9a is a detailed cross-sectional view of the mounting portion between the compressible wall element and the rigid base plate of the cell culture vessel. [Figure 9c] Figure 9a is a detailed cross-sectional view of the mounting portion between the compressible wall element and the rigid base plate of the cell culture vessel. [Figure 10] This is a schematic cross-sectional view of a cell culture vessel showing a channel for gas permeability. [Figure 11] This is a diagram of a compressible wall element in an example of a cell culture vessel. [Figure 12] This is a diagram of a compressible wall element in an example of a cell culture vessel. [Figure 13] This is a diagram of a compressible wall element in an example of a cell culture vessel. [Figure 14] This is a diagram of a support ring, an example of a compressible wall element for a cell culture vessel. [Figure 15a] This is a diagram of the exit of an example cell culture vessel. [Figure 15b] This is a diagram of the exit of an example cell culture vessel. [Modes for carrying out the invention]

[0029] The bioreactor 1 shown in Figure 1 comprises a cell culture vessel 2 and a connecting plate 3. During use, the cell culture vessel 2 holds a fluid 4 in which cell processing takes place. Specifically, the fluid 4 is a cell suspension containing a population of cells present in a liquid medium. The cells can be cultured for reproduction or processed to produce cell-based therapeutic products.

[0030] The junction plate 3 is attached to the top of the cell culture vessel 2 and acts, for example, as a lid or closure. The junction plate 3 includes at least one connector joint 5 for connecting to external elements, such as consumables for delivering fluid to or from the cell culture vessel 2. Thus, the junction plate 3 provides for adding media and other fluids to the cell culture vessel 2 during cell processing, and / or for removing fluid from the cell culture vessel 2 during processing, for example, to remove sample or waste fluid.

[0031] The cell culture vessel 2 may be expandable and / or compressible. Specifically, the cell culture vessel 2 has compressible wall elements 6, such as bellows walls. The cell culture vessel 2 has a base portion 7 positioned opposite the connecting plate 3 and compressible wall elements 6 defining the side walls of the cell culture vessel 2. The upper part of the compressible wall elements 6 is attached to the connecting plate 3. The upper part of the compressible wall elements 6 may have a rigid ring 8 or similar for attachment to the connecting plate 3. The compressible wall elements 6 are compressible and / or expandable so that the base portion 7 can move toward or away from the connecting plate 3, thereby changing the internal volume of the cell culture vessel 2. The base portion 7 can be moved relative to the connecting plate 3 to agitate or mix the fluid 4 in the cell culture vessel 2.

[0032] The compressible wall element 6 may be a bellows-like wall having a concertina-like arrangement that causes the compressible wall element 6 to fold in itself for compression. Specifically, as shown, the compressible wall element 6 comprises a series of alternately arranged deformable portions 9a, 9b, and more specifically, may comprise inwardly deformable portions 9a and outwardly deformable portions 9b. A leaf portion 10 extends between the deformable portions 9a, 9b. The leaf portion 10 is more rigid than the deformable portions 9a, 9b. The deformable portions 9a, 9b act as hinges that allow the compressible wall element 6 to fold like a bellows or concertina while the leaf portion 10 remains substantially undeformed.

[0033] The compressible wall element 6 may comprise, for example, at least one inwardly deformable portion 9a and at least one outwardly deformable portion 9b, such as at least two inwardly deformable portions 9a and at least two outwardly deformable portions 9b. The compressible wall element 6 may comprise three, four, or more inwardly deformable portions 9a and three, four, or more outwardly deformable portions 9b.

[0034] The inwardly deformable portion 9a and the outwardly deformable portion 9b may be formed by thinned portions of the compressible wall element 6. The inwardly deformable portion 9a may include a thinned portion located on the outer surface of the compressible wall element 6 so as to be deformable inward. The outwardly deformable portion 9b may include a thinned portion located on the inner surface of the compressible wall element 6 so as to be deformable outward.

[0035] In one example, the compressible wall element 6 comprises silicone, specifically liquid silicone rubber. In another example, the compressible wall element 6 comprises low-density polyethylene (LDPE). In yet another example, the compressible wall element 6 comprises thermoplastic elastomer (TPE). In the example, as will be further described later, the compressible wall element 6 may be coated, laminated, or otherwise treated to reduce the gas permeability of the compressible wall element 6, or to make the compressible wall element 6 impermeable to gases, particularly oxygen. In one example, the compressible wall element 6 comprises layers and an outer sheath, jacket, or coating. For example, the compressible wall element 6 may comprise an inner portion and a jacket overmolded onto the LDPE inner portion. The inner portion may comprise LDPE, and the jacket may comprise TPE. In another example, the compressible wall element 6 may comprise an elastomer outer portion, such as the outside of TPE, and a lining. For example, an LDPE lining may be blown into the inner surface of the outside of the elastomer to form the lining. In other examples, the lining may be an insert, such as an LDPE insert that is received inside the outer elastomer but is not co-molded with the outer elastomer. In such examples, it may be considered preferable that the lining comprises a base sheet and defines (excluding the top) a sealed container for holding cell cultures.

[0036] Therefore, the cell culture vessel 2 can expand and contract, or can be expanded and contracted depending on the material held in the cell culture vessel 2. Specifically, the cell culture vessel 2 can expand as the volume of fluid 4 inside the cell culture vessel 2 increases and / or as additional material is added.

[0037] As illustrated, the junction plate 3 also includes an expansion vessel 11, also called a breathing vessel. The expansion vessel 11 can expand and contract the cell culture vessel 2 without significantly changing the pressure in the cell culture vessel 2. Alternatively or additionally, the expansion vessel 11 may be operable by compressing or expanding, for example mechanically or manually, to change the volume of the cell culture vessel 2 by expanding or retracting the compressible walls 6 of the cell culture vessel 2. Alternatively or additionally, the expansion vessel 11 may be operable by compressing or expanding, for example mechanically or manually, to change the pressure inside the cell culture vessel 2.

[0038] In the various examples described later, the base portion 7 comprises a rigid base plate 12. The rigid base plate 12 is generally planar, or flat. The rigid base plate 12 is attached to or molded with the compressible wall element 6, as will be further described later.

[0039] The rigid base plate 12 is substantially planar, thereby defining the substantially flat bottom of the rigid cell culture vessel 2. The planar bottom of the cell culture vessel 2 can provide improved cell culture, specifically in terms of mixing and control over the cell culture. The flat bottom of the cell culture vessel 2 helps ensure that the cells are substantially uniformly dispersed across the cross-section of the cell culture vessel 2 when they settle to the bottom of the vessel, whereas if the base portion 7 is not flat, the cells will be concentrated in a smaller volume, which can be detrimental to cell culture. The flat bottom of the cell culture vessel 2 also helps prevent the fluid 4 from becoming trapped in the cell culture vessel 2 when the cells are harvested or extracted at the end of the cell culture process.

[0040] In various examples, the rigid base plate 12 includes a thermoplastic polymer such as high-density polyethylene (HDPE) or polycarbonate, or other rigid polymers. As will be further described later, the rigid base plate 12 may be opaque, transparent, or translucent.

[0041] In various examples as described later, the base portion 7, specifically the rigid base plate 12, has a sensor window. The sensor window is transparent or translucent and provides an optical path to the cell culture vessel. Thus, the optical sensor can send light to the cell culture in the cell culture vessel and receive light from the cell culture.

[0042] In various examples, the sensor window may be centrally located on the rigid base plate 12. The central position of the sensor window ensures that the fluid 4 is located in the sensor window during mixing and stirring so that the sensor operating through the sensor window can function.

[0043] In the illustrated example, the cell culture vessel 2 is generally cylindrical, comprising a generally circular base portion 7 and generally cylindrical compressible wall elements 6. Thus, the axial direction is defined between the base portion 7 and the ends of the compressible wall elements 6 to which the connecting plate 3 is attached. However, it should be understood that the cell culture vessel 2 may take on alternative forms, such as a generally triangular or square cross-sectional shape.

[0044] As shown in the example Figures 2a and 2b, the rigid base plate 12 comprises a planar central portion 13 corresponding to the internal volume of the cell culture vessel 2 and an edge portion 14. The compressible wall element 6 is attached to the edge portion 14. The compressible wall element 6 may be additionally or alternatively attached to a peripheral portion 15 outside the edge portion 14, outside the internal volume of the cell culture vessel 2. Specifically, the compressible wall element 6 may comprise a base portion 16 attached to the edge portion 14 and / or the peripheral portion 15. Alternatively, the bottom leaf portion 10a of the compressible wall element 6 may be attached to the edge portion 14 and / or the peripheral portion 15.

[0045] The compressible wall element 6, specifically the hem portion 16 or bottom leaf portion 10a, may be bonded or welded to the rigid base plate 12, specifically the edge portion 14 and / or surrounding portion 15. In the example, the compressible wall element 6, specifically the hem portion 16 or bottom leaf portion 10a, is ultrasonically welded to the rigid base plate 12, specifically the edge portion 14 and / or surrounding portion 15. In the example, the compressible wall element 6, specifically the hem portion 16 or bottom leaf portion 10a, is thermally welded to the rigid base plate 12, specifically the edge portion 14 and / or surrounding portion 15, for example, by hot plate welding. The compressible wall element 6 is sealed to the rigid base plate 12.

[0046] As shown in the figure, the compressible wall element 6 is attached to the rigid base plate 12, for example by adhesive or welding, such that the deformable portion 9c is positioned at or near the tip 18 of the edge 14. In this illustrated example, the deformable portion 9c positioned at the tip 18 of the edge 14 is a portion that can be deformed inward.

[0047] In this example, the base portion 7, specifically the rigid base plate 12, includes an opaque HDPE material. The rigid base plate 12 can be molded, for example, by injection molding.

[0048] As shown in Figures 2a and 2b, the base portion 7 also includes a sensor window 19. In this example, the sensor window 19 is transparent or translucent. For example, the sensor window 19 may be a polycarbonate window that is mounted or molded into an opening in the rigid base plate 12. The sensor window 19 may be an insert in the opening in the rigid base plate 12.

[0049] One or more sensor elements 20 may be attached to or molded into the sensor window 19. The sensor elements 20 may be, for example, optical dots for use with an optical sensor to detect the amount of dissolved oxygen in the fluid in the cell culture vessel 2 during use.

[0050] In other examples, the rigid base plate 12 may be made of a transparent or translucent material such as polycarbonate (PC), and the sensor window 19 may be defined as part of the planar central portion 13.

[0051] As schematically shown, the rigid base plate 12 may be further equipped with a valve 24 for the extraction of fluid from the cell culture vessel 2, for example, to collect cells from the cell culture vessel 2 at the end of the cell culture process.

[0052] As shown in Figures 3a and 3b of the example, the rigid base plate 12 has a planar central portion 13 corresponding to the internal volume of the cell culture vessel 2. In this example, the rigid base plate 12 has shoulders 21 such that the peripheral portion 22 is stepped relative to the planar central portion 13. Specifically, as shown in Figures 3c and 3d, the compressible wall element 6 is molded into the rigid base plate 12 at the shoulders 21. In the illustrated example, the shoulders 21 provide an outer circumferential surface to which the compressible wall element 6 is attached. However, it should be understood that the shoulders 21 may extend in the opposite direction to provide a recess having an inner circumferential surface to which the compressible wall element 6 is attached.

[0053] In the examples shown in Figures 3a to 3d, the hem portion 23 at the end of the compressible wall element 6 is molded into the rigid base plate 12, specifically embedded in the rigid base plate 12, thereby providing a sealed connection between the compressible wall element 6 and the rigid base plate 12.

[0054] In the example shown in Figure 3c, the base portion 23 of the compressible wall element 6 is molded into the rigid base plate 12 at the shoulder portion 21. The base portion 23 extends from the inwardly deformable portion 9a and extends radially inward toward the planar central portion 13. The base portion 23 can be molded into the rigid base plate 12 by a two-stage molding process, specifically a two-stage injection molding process.

[0055] In the example shown in Figure 3d, the base portion 23 of the compressible wall element 6 is molded to the rigid base plate 12 at the shoulder portion 21. The base portion 23 has a first portion 23a that extends radially inward from an inwardly deformable portion 9a toward a planar central portion 13. The first portion 23a is located in part of the planar central portion 13 and may or may not be molded to the rigid base plate 12. The base portion 23 also has a second portion 23b that extends toward the rigid base plate 12 in a direction generally perpendicular to the first portion 23a in the axial direction. The second portion 23b of the base is molded to the rigid base plate 12. The second portion 23b of the base can be molded to the rigid base plate 12 by a two-stage molding process, specifically a two-stage injection molding process.

[0056] In the examples shown in Figures 3a to 3d, the inwardly deformable portion 9a is positioned on the shoulder portion 21, so that the compressible wall element 6 can be folded onto the rigid base plate 12, specifically the central planar portion 13.

[0057] In this example, the base portion 7, specifically the rigid base plate 12, includes an opaque HDPE material.

[0058] As shown in Figure 3b, the base portion 7 also includes a sensor window 19. In this example, the sensor window 19 is transparent or translucent. For example, the sensor window 19 may be a polycarbonate window that is mounted or molded into an opening in the rigid base plate 12. The sensor window 19 may be an insert in the opening in the rigid base plate 12.

[0059] One or more sensor elements 20 may be attached to or molded into the sensor window. The sensor elements 20 may be, for example, optical dots for use with an optical sensor to detect the amount of dissolved oxygen in the fluid in the cell culture vessel 2 during use.

[0060] In other examples, the rigid base plate 12 may be made of a transparent or translucent material such as polycarbonate, and the sensor window 19 may be defined as part of the planar central portion 13.

[0061] Similar to the examples in Figures 2a and 2b, the rigid base plate 12 may be further equipped with a valve (reference numeral 24, see Figure 2a) for the extraction of fluid from the cell culture vessel 2, for example, to collect cells from the cell culture vessel 2 at the end of the cell culture process.

[0062] In the example shown in Figure 4, the rigid base plate 12 includes a planar central portion 13 corresponding to the internal volume of the cell culture vessel 2. The rigid base plate 12 also includes a peripheral portion 25 radially outward from the planar central portion 13. The bottom leaf portion 10a of the compressible wall element 6 is attached to the peripheral portion 25. For example, the bottom leaf portion 10a is bonded or welded to the peripheral portion 25. The bottom leaf portion 10a can be hot-plate welded or ultrasonically welded to the peripheral portion 25. The compressible wall element 6 is sealed to the rigid base plate 12.

[0063] As shown in the diagram, the bottom leaf portion 10a is attached to the surrounding portion 25 such that the portion 9a that is deformable inward of the compressible wall element 6 is positioned on the rigid base plate 12 or immediately adjacent to the rigid base plate 12.

[0064] In this example, the base portion 7, specifically the rigid base plate 12, comprises a transparent polycarbonate (PC) material. The rigid base plate 12 includes a number of reinforcing ribs 26 molded onto the surface of the rigid base plate 12 opposite the compressible wall element 6 in order to improve the strength and rigidity of the rigid base plate 12 and to reduce the risk of shattering.

[0065] As shown in Figure 4, the base portion 7 also includes a sensor window 19. In this example, the sensor window 19 comprises a planar portion of the rigid base plate 12 where the reinforcing ribs 26 do not extend.

[0066] One or more sensor elements 20 may be attached to or molded onto the rigid base plate 12 in the sensor window 19. The sensor elements 20 may be, for example, optical dots for use with an optical sensor to detect the amount of dissolved oxygen in the fluid in the cell culture vessel 2 during use.

[0067] Alternatively, as in the examples in Figures 2a to 3d, the rigid base plate 12 may include an opaque material such as opaque HDPE, and the sensor window 19 may include a transparent material that is integrally molded, attached, or inserted, such as a polycarbonate insert.

[0068] Similar to the examples in Figures 2a and 2b, the rigid base plate 12 may be further equipped with a valve (reference numeral 24, see Figure 2a) for the extraction of fluid from the cell culture vessel 2, for example, to collect cells from the cell culture vessel 2 at the end of the cell culture process.

[0069] In the example shown in Figures 5a and 5c, the cell culture vessel 2 comprises a rigid base plate 12 with a planar central portion 13 corresponding to the internal volume of the cell culture vessel 2. The cell culture vessel 2 also has a compressible wall element 6 attached to the rigid base plate 12 by a ring 38 in this example. The rigid base plate 12 comprises a peripheral portion 25 radially outward from the planar central portion 13. The bottom leaf portion 10a of the compressible wall element 6 is attached to the ring 38, for example, by adhesive or by welding such as ultrasonic welding or hot plate welding 39. The compressible wall element 6 is sealed to the ring 38. The ring 38 can be attached to the peripheral portion 25 of the rigid base plate 12, for example, by one or more fasteners. A seal, for example, an O-ring 40, may be provided between the ring 38 and the rigid base plate 12 to provide a sealed attachment of the ring 38 to the rigid base plate 12.

[0070] As shown in Figure 5c, the bottom leaf portion 10a is attached to the ring 38 such that the portion 9a that is deformable inward of the compressible wall element 6 is positioned on the rigid base plate 12 or immediately adjacent to the rigid base plate 12.

[0071] In this example, the base portion 7, specifically the rigid base plate 12, includes a transparent polycarbonate (PC) material.

[0072] As shown in Figure 4, the base portion 7 also includes a sensor window 19. One or more sensor elements 20 may be attached to or molded into the rigid base plate 12 in the sensor window 19. The sensor elements 20 may be, for example, optical dots for use with an optical sensor to detect the amount of dissolved oxygen in the fluid in the cell culture vessel 2 during use.

[0073] Alternatively, as in the examples in Figures 2a to 3d, the rigid base plate 12 may include an opaque material such as opaque HDPE, and the sensor window 19 may include a transparent material that is integrally molded, attached, or inserted, such as a polycarbonate insert.

[0074] Similar to the examples in Figures 2a and 2b, the rigid base plate 12 may be further equipped with a valve (reference numeral 24, see Figure 2a) for the extraction of fluid from the cell culture vessel 2, for example, to collect cells from the cell culture vessel 2 at the end of the cell culture process.

[0075] In the example shown in Figures 6a and 6b, the cell culture vessel 2 comprises compressible wall elements 6 and a rigid base plate 12 that can be attached to each other in any of the methods described with reference to Figures 2a to 5c. In this example, the cell culture vessel 2 further comprises a base sheet 27, which extends across the rigid base plate 12 within the cell culture vessel 2.

[0076] In this example, the base sheet 27 is attached to the compressible wall element 6, specifically by sealing. This seals the internal volume of the cell culture vessel 2 between the compressible wall element 6 and the base sheet 27, preventing fluid from contacting the rigid base plate 12. In this example, the base sheet 27 is attached to the compressible wall element 6 by sealing, for example, by welding such as hot plate welding or ultrasonic welding.

[0077] In addition or alternatively, the base sheet 27 is attached to the rigid base plate 12, specifically by sealing. The base sheet 27 may be attached to the rigid base plate 12 at the joint between the compressible wall element 6 and the rigid base plate 12, or adjacent to that joint, around the base sheet 27. Thereafter, the internal volume of the cell culture vessel 2 is sealed between the compressible wall element 6 and the base sheet 27. In this example, the base sheet 27 is attached to the rigid base plate 12 by sealing, for example, by welding such as hot plate welding or ultrasonic welding.

[0078] In this example, the base sheet 27 may be permeable to gases. Specifically, the base sheet 27 may be permeable to oxygen. The base sheet 27 may contain silicone, and more specifically, it may contain liquid silicone rubber.

[0079] The rigid base plate 12 is provided with one or more openings, specifically holes 28. Thus, the base sheet 27 is exposed to the atmosphere through the holes 28, and gases such as oxygen can permeate through the base sheet 27.

[0080] In this example, the compressible wall element 6 can be made gas permeable, specifically oxygen permeable, or gas impermeable, specifically oxygen impermeable. In this example, the compressible wall element 6 can be coated or laminated to make it gas impermeable, specifically oxygen impermeable. In one example, the compressible wall element 6 comprises an inner silicone layer and an outer LDPE sheath or coating. The inner silicone layer may be a lining inside the outer LDPE sheath.

[0081] Similar to the previous example, the rigid base plate 12 may have a sensor window. The base sheet 27 may be transparent or translucent. The sensor window may be a transparent or translucent window that is attached to or molded into an opening in the rigid base plate 12. The sensor window may be an insert in the opening in the rigid base plate 12. The rigid base plate 12 may be transparent, and the sensor window may be a part of the rigid base plate 12.

[0082] One or more sensor elements may be attached to or molded onto the rigid base plate 12 in the sensor window. The sensor elements may be, for example, optical dots for use with an optical sensor to detect the amount of dissolved oxygen in the fluid in the cell culture vessel 2 during use.

[0083] Similar to the examples in Figures 2a and 2b, the rigid base plate 12 may be further equipped with a valve (reference numeral 24, see Figure 2a) for the extraction of fluid from the cell culture vessel 2, for example, to collect cells from the cell culture vessel 2 at the end of the cell culture process.

[0084] In the example shown in Figures 7a–7c, the cell culture vessel 2 comprises compressible wall elements 6 and a rigid base plate 12 that can be attached to each other in any of the methods described with reference to Figures 2a–5c. In this example, the cell culture vessel 2 further comprises a base sheet 27, which extends across the rigid base plate 12 within the cell culture vessel 2, as is most clearly shown in Figures 7a and 7c.

[0085] In this example, the base sheet 27 is part of the compressible wall element 6. Specifically, the base sheet 27 is formed as part of the same mold forming as the compressible wall element 6, for example by blow molding. As shown in Figure 7c, the compressible wall element 6 may have a hem 23 similar to those described with reference to Figures 3a-3d, which is attached to the rigid base plate 12. In other examples, the compressible wall element 6 may be attached to the rigid base plate 12 similar to those described with reference to Figures 2a, 2b, 4, and 5a-5c, for example by being glued or welded.

[0086] As a result, in the examples shown in Figures 7a and 7c, the internal volume of the cell culture vessel 2 is defined solely within the compressible wall element 6, and there is no sealing joint between the compressible wall element 6 and the rigid base plate 12. Therefore, the fluid does not come into contact with the rigid base plate 12.

[0087] In one example, a portion of the base sheet 27 is attached to the rigid base plate 12, for example, by adhesive or welding, specifically by spot welding. The base sheet 27 may be attached to the rigid base plate 12 around the periphery of the base sheet 27.

[0088] In the example, the base sheet 27 may be gas permeable. Specifically, the base sheet 27 may be oxygen permeable. The base sheet 27 may contain silicone, such as liquid silicone rubber. The base sheet 27 is made from the same material as the compressible wall element 6. All or part of the compressible wall element 6 may be coated or laminated to make all or part of it gas impermeable, specifically oxygen impermeable. In one example, the compressible wall element 6 comprises an inner silicone layer and an outer LDPE sheath or coating. The inner silicone layer may be a lining for the LDPE sheath.

[0089] In this example, as shown in Figures 7b and 7c, the rigid base plate 12 has one or more openings 29. Thus, the base sheet 27 is exposed to the atmosphere through the openings 29, and gases such as oxygen can permeate through the base sheet 27.

[0090] Similar to the previous example, the rigid base plate 12 may include a sensor window 19, as shown in Figure 7b. The base sheet 27 may be transparent or translucent. The sensor window 19 may consist of a transparent or translucent window that is mounted or molded into an opening in the rigid base plate 12. The sensor window 19 may be an insert in the opening in the rigid base plate 12. The rigid base plate 12 may be transparent, and the sensor window 19 may be part of the rigid base plate 12.

[0091] One or more sensor elements 20 may be attached to or molded onto the rigid base plate 12 in the sensor window 19. The sensor elements 20 may be, for example, optical dots for use with an optical sensor to detect the amount of dissolved oxygen in the fluid in the cell culture vessel 2 during use.

[0092] Similar to the examples in Figures 2a and 2b, the rigid base plate 12 may be further equipped with a valve (reference numeral 24, see Figure 2a) for the extraction of fluid from the cell culture vessel 2, for example, to collect cells from the cell culture vessel 2 at the end of the cell culture process.

[0093] In the example of Figure 8, similar to the examples in Figures 6a to 7c, the compressible wall element 6 comprises a base sheet 27. In this example, the base sheet 27 is opaque. As shown, the base sheet 27 comprises an opening 30 that aligns with the sensor window 19 in the rigid base plate 12. The base sheet 27 is attached to the rigid base plate 12 by sealing around the opening 30. For example, the base sheet 27 is welded or bonded to the rigid base plate 12 around the opening 30.

[0094] In the examples of Figures 9a to 9c, the compressible wall element 6 is attached to the rigid base plate 12 by clip fastening. The rigid base plate 12 and the compressible wall element 6 may be as described in the examples of Figures 6a to 8. Specifically, as illustrated, in this example, the compressible wall element 6 comprises a single base sheet 27 such that the internal volume of the cell culture vessel 2 is defined within the compressible wall element 6.

[0095] The compressible wall element 6 generally comprises a radially extending protrusion 31. The protrusion 31 may be flexible or deformable and / or may have increased thickness to have greater rigidity. The protrusion 31 is received in a groove 33 formed in the peripheral portion 32 of the rigid base plate 12. The groove 33 is molded to receive the protrusion 31 of the compressible wall element 6 so that the protrusion 31 is held in the groove 33. The groove 33 may comprise one or more fan-shaped sections 34 to facilitate the insertion of the protrusion 31 into the groove 33. Thus, the compressible wall element 6 can be clipped to the rigid base plate 12 by clipping the protrusion 31 into the groove 33.

[0096] Figure 10 shows an example of a cell culture vessel 2 having a base sheet 27, as described with reference to Figures 6a to 9c, for example. The base sheet 27 may be integrated with the compressible wall element 6, or it may be separate, as shown in Figure 10. In the illustrated example, the base sheet 27 is sealed to the rigid base plate 12 within the internal volume of the cell culture vessel 2.

[0097] In this example, the base sheet 27 is gas permeable, specifically oxygen permeable. The rigid base plate 12 comprises one or more openings 37 and one or more separating ribs 35 extending from the rigid base plate 12 toward the internal volume of the cell culture vessel 2, specifically toward the base sheet 27. In this example, the separating ribs 35 are positioned to separate the base sheet 27 from the rigid base plate 12 to create a fluid passage 36 for gas circulation, specifically air circulation. Thus, air can reach the underside of the base sheet 27 and permeate into the cell culture vessel 2 during use.

[0098] Figures 11 to 13 show alternative compressible wall elements 6 that may be used with any of the cell culture vessels 2 described with reference to Figures 2a to 10. Specifically, each of Figures 11 to 13 shows a compressible wall element 6 that is attached to a rigid base plate 12 to form the cell culture vessel 2.

[0099] In the example shown in Figure 11, the compressible wall element comprises an inner portion 41 and a jacket 42. In this example, the jacket 42 is overmolded onto the inner portion 41, so that they are formed as a single unit. In this example, the inner portion 41 is made of LDPE and the jacket 42 is made of thermoplastic elastomer (TPE).

[0100] In the example shown in Figure 12, the compressible wall element 6 may comprise an elastomer outer 43, such as the outer casing of TPE, and a lining 44. For example, an LDPE lining 44 may be blown into the inner surface of the elastomer outer casing 43 to form the lining 44. As shown, the lining 44 comprises a base sheet 27 as previously described.

[0101] In the example of Figure 13, the compressible wall element may comprise an elastomer outer 43, such as the outer casing of TPE, and an insert 45. The insert 45 may include LDPE. The insert 45 is received within the elastomer outer casing 43 but is not co-molded with the elastomer outer casing 43. As shown, the insert 45 comprises a base sheet 27 as previously described.

[0102] Figure 14 shows a cell culture vessel in which a support ring 46 is provided on the top of a compressible wall element 6. The support ring 46 is attached to the uppermost leaf portion 10b of the compressible wall element 6, for example, by adhesive, welding, or fasteners. The support ring 46 is rigid and engages with another component of the bioreactor 1 shown in Figure 1, specifically the joint plate 3. It should be understood that the support ring 46 may also be provided for example cell culture vessels 2 of any other examples described herein.

[0103] Figures 15a and 15b illustrate an example of providing an outlet 47 for the extraction of fluid from a cell culture vessel, for example, to harvest cells from the cell culture vessel at the end of a cell culture process. The outlet 47 is formed in a rigid base plate 12 and is therefore formed at the lower end of the cell culture vessel, so that gravity can assist in harvesting cells through the outlet 47. The outlet 47 may be equipped with a valve or an openable closure.

[0104] In the example of Figure 15a, the exit 47 is formed in the opening in the rigid base plate 12 within an insert 48 that seals the opening. The insert 48 may be a thermoplastic elastomer. As shown, the insert 48 may be part of a compressible wall element 6 that extends from the compressible wall element 6 to the rigid base plate 12 towards the opening. The insert 48 may be bonded to the rigid base plate 12 or otherwise attached to provide a seal between the insert 48 and the rigid base plate 12.

[0105] In the example shown in Figure 15b, the exit 47 is specifically formed as part of the rigid base plate 12 at the protruding portion 49 of the rigid base plate 12.

[0106] Throughout this specification, the words “equipped with” and “including,” and their variations, mean “including but not limited to,” and are not intended (and do not) to exclude other components, integers, or steps. Throughout this specification, the singular encompasses the plural unless the context otherwise requires. Specifically, where the indefinite article is used, this specification is understood to consider the singular in addition to the plural unless the context otherwise requires.

[0107] Any features, integers, characteristics, or groups described in conjunction with specific aspects, embodiments, or examples of the present invention are understood to be applicable to any other aspects, embodiments, or examples described herein, insofar as they do not conflict. All features disclosed herein (including the appended claims, abstract, and drawings), and / or all steps of the methods or processes so disclosed herein, can be combined in any combination, except for any combination in which at least some of such features and / or steps are mutually exclusive. The present invention is not limited to the details of the embodiments described herein. The present invention extends to any novel one or any novel combination of features disclosed herein (including the appended claims, abstract, and drawings), or to any novel one or any novel combination of the steps of the methods or processes so disclosed herein.

[0108] [Section 1] The base part, A compressible wall element extending axially from the base portion and defining the internal volume of the cell culture vessel, wherein the compressible wall element is compressible in the axial direction Equipped with, The aforementioned base portion comprises a rigid base plate that is substantially planar, and is used as a cell culture vessel. [Section 2] The cell culture vessel according to paragraph 1, wherein the compressible wall element is bonded or welded to the rigid base plate. [Section 3] The cell culture vessel according to paragraph 1, wherein the compressible wall element is fastened or clipped to the rigid base plate. [Section 4] The cell culture vessel according to paragraph 1, wherein the compressible wall element is molded integrally with the rigid base plate, for example, the rigid base plate is overmolded onto a portion of the compressible wall element. [Section 5] The cell culture vessel according to any one of the first to fourth claims, wherein the compressible wall element comprises a deformable portion located at the joint between the compressible wall element and the rigid base plate, or immediately adjacent to that joint. [Section 6] A cell culture vessel according to any one of the first to fifth claims, comprising a base sheet extending over the rigid base plate within the internal volume of the cell culture vessel. [Section 7] The cell culture vessel according to paragraph 6, wherein the base sheet extends from the compressible wall element, and in particular, the base sheet is molded integrally with the compressible wall element. [Section 8] The cell culture vessel according to item 6 or 7, wherein the base sheet is permeable to gas, and in particular to oxygen. [Section 9] The cell culture vessel according to paragraph 8, wherein the rigid base plate is provided with one or more gas-permeable openings. [Section 10] The cell culture vessel according to paragraph 8 or 9, wherein the rigid base plate comprises one or more spacers fitted to space the base sheet away from the rigid base plate. [Section 11] The cell culture vessel according to any one of the first to tenth paragraphs, wherein the compressible wall element comprises an inwardly deformable portion, an outwardly deformable portion, and a leaf portion extending between the inwardly deformable portion and the outwardly deformable portion such that the deformation of the inwardly deformable portion and the deformation of the outwardly deformable portion compress the compressible wall element. [Section 12] The cell culture vessel according to paragraph 11, wherein one of the inwardly deformable portion or the outwardly deformable portion is located at the joint between the compressible wall element and the rigid base plate, or immediately adjacent to that joint. [Section 13] The cell culture vessel according to any one of claims 1 to 12, wherein the rigid base plate is provided with a transparent or translucent sensor window. [Section 14] The cell culture vessel according to any one of claims 1 to 13, wherein the compressible wall element comprises silicone, low-density polyethylene, or a thermoplastic elastomer. [Section 15] The cell culture vessel according to any one of claims 1 to 14, wherein the compressible wall element comprises an outer portion and a lining or insert. [Section 16] The cell culture vessel according to any one of the first to fifth paragraphs, wherein the compressible wall element comprises an inner portion and a jacket. [Section 17] A cell culture vessel according to any one of the first to sixth paragraphs, wherein at least a portion of the compressible wall element is provided with a gas-impermeable coating. [Section 18] The cell culture vessel according to any one of claims 1 to 17, wherein the rigid base plate comprises high-density polyethylene or polycarbonate. [Section 19] The cell culture vessel according to paragraph 18, wherein the rigid base plate is transparent or translucent, or has a transparent or translucent sensor window. [Section 20] A bioreactor for cell culture processing, comprising any of the cell culture vessels described in paragraphs 1 to 19. [Section 21] The bioreactor according to paragraph 20, further comprising a joint plate that can be attached to the compressible wall element opposite the base portion for closing the cell culture vessel. [Section 22] The bioreactor according to paragraph 21, wherein the connecting plate is provided with a connector connecting portion. [Section 23] A cell processing system comprising the bioreactor described in any of paragraphs 20 to 22. [Section 24] The cell processing system according to paragraph 23, further comprising a stirrer configured to move the base portion in order to agitate the fluid in the cell culture vessel. [Explanation of symbols]

[0109] 1. Biological reactor 2 Cell culture vessel 3 Joint plate 4 fluid 5. Connector joint 6 Compressible wall elements 7 Base part 8 Rigid ring 9a Inwardly deformable portion 9b Outwardly deformable portion 9c Deformable part 10 Leaf section 10a Bottom leaf section 10b The uppermost leaf section 11 Expandable container 12 Rigid base plate 13 Center part 14 Edge 15 Surrounding area 16 Hem 18 Tip 19 Sensor window 20 Sensor Elements 21 Shoulder 22 Peripheral area 23 Hem 23a Part 1 23b Second part 24 valves 25 Peripheral area 26 Reinforcement Ribs 27 Base Sheet 28 holes 29 Opening of rigid base plate 12 30 Opening of base sheet 27 31 Edge 32 Peripheral area 33 Groove 34 sector area 35 Separating ribs 36 Fluid passage 37 Aperture 38 ring 39. Hot plate welding 40 O-rings 41 Inner part 42 Jacket 43 Outside 44 Lining 45 Insertion 46 Support ring 47 Exit 48 Insertion body 49 Protrusion

Claims

[Claim 1] The base part, A compressible wall element extending axially from the base portion and defining the internal volume of the cell culture vessel, wherein the compressible wall element is compressible in the axial direction Equipped with, The aforementioned base portion comprises a rigid base plate that is substantially planar, and is used as a cell culture vessel.

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