Compressible bioreactor vessel having gas outlet comprising flow restrictor and method of mixing cell suspension therein

Abstract

A bioreactor is described that includes a compressible bioreactor vessel configured to hold a cell suspension in an interior volume of the compressible bioreactor vessel. The compressible bioreactor vessel includes a gas outlet in fluid communication with an interior volume of the compressible bioreactor vessel and configured to allow gas to flow out of the interior volume of the compressible bioreactor vessel in use. The gas outlet includes a flow restrictor configured such that the flow restrictor restricts gas from flowing out of the gas outlet when the compressible bioreactor vessel is compressed in use.

Description

Technical Field

[0001] The present invention relates to a bioreactor having a compressible bioreactor container, and more particularly to a bioreactor having a compressible bioreactor container with an outlet having a flow restrictor for restricting gas flow out of the bioreactor container. Background Art

[0002] Bioprocessing processes, such as cell and gene therapy (CGT) manufacturing processes, are often complex and include manual steps across several devices. The equipment system used in the various steps or unit operations of cell-based therapeutic product (CTP) manufacturing can include equipment for each unit operation. The unit operations can include, for example, cell collection, cell separation, selection, cell expansion, cell washing, volume reduction, cell storage or transportation. Unit operations can vary greatly based on the manufacturing model (i.e., autologous vs. allogeneic), cell type, intended purpose, and other factors. In addition, cells are "living" entities that are sensitive to even the simplest manipulations (such as differences in cell transfer processes). The role of cell manufacturing equipment in ensuring scalability and reproducibility is an important factor in cell and gene therapy manufacturing.

[0003] In addition, cell-based therapeutic products (CTPs) have gained significant momentum, creating a need for improved cell manufacturing equipment for various cell manufacturing processes, 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, gene modification, incubation / recovery, washing, infusion into patients, and / or freezing.

[0004] The cultivation or treatment of cells often requires the use of a device to hold the cells, for example, in a suitable culture medium when culturing cells. Known devices include shake flasks, roller bottles, T-flasks, and bags. Such bottles or flasks are widely used, but have several disadvantages. During the cell culture period, additional culture medium may be added to the container, and some fluids, such as waste liquids, may be extracted from the container. Summary of the Invention

[0005] According to one aspect of the present disclosure, a bioreactor is provided that includes a compressible bioreactor container configured to hold a cell suspension within an interior volume of the compressible bioreactor container. The compressible bioreactor container includes a gas outlet that is in fluid communication with the interior volume of the compressible bioreactor container and is configured to allow gas to flow out of the interior volume of the compressible bioreactor container during use. The gas outlet includes a flow restrictor that is configured to restrict gas flow out of the gas outlet when the compressible bioreactor container is compressed during use.

[0006] The restrictor limits the flow of gas out of the gas outlet to reduce the flow rate of gas leaving the bioreactor during compression. This allows pressure to build up in the bioreactor, and the compressible bioreactor container can expand to compensate for the pressure buildup. The reduced flow rate out of the bioreactor also minimizes the loss of water vapor from the bioreactor container through the outlet. It is beneficial to keep water vapor (and other gas components) within the bioreactor container in order to maintain an optimal internal environment for cell growth, and the restrictor improves this during the compression of the bioreactor container.

[0007] In one example, the bioreactor may further include an expandable expansion vessel having an internal volume fluidly connected to the internal volume of the compressible bioreactor vessel. In this manner, when the compressible bioreactor vessel is compressed during use, gas within the internal volume of the compressible bioreactor vessel is transferred to the internal volume of the expansion vessel. This increases the gas pressure within the expansion vessel, causing the expansion vessel to expand to compensate for the increase in gas pressure. The expansion vessel also allows for expansion and contraction of the compressible bioreactor vessel without significantly changing the pressure within the compressible bioreactor vessel. This is because gas is transferred between the compressible bioreactor vessel and the expansion vessel as the compressible bioreactor vessel is compressed and expanded.

[0008] In one example, the expansion vessel may include a gas outlet. Thus, as pressure in the expansion vessel increases, gas will flow out of the expansion vessel through the gas outlet. A flow restrictor will restrict the flow of gas out of the gas outlet. The restricted flow of gas out of the gas outlet allows the pressure in the expansion vessel to increase, and the expansion vessel will expand to compensate for the increase in gas pressure. Consequently, less gas escapes through the gas outlet, and the expansion vessel holds more gas due to the expansion under pressure. Gas remaining in the expansion vessel can be returned to the bioreactor vessel, for example, after compression is released.

[0009] In an example, a compressible bioreactor container can include a base section, a top section, and a compressible sidewall extending between the base section and the top section. Thus, the compressible bioreactor container can expand and contract to compensate for pressure therein. The compressible bioreactor container can also be actuated to expand and contract, thereby controlling the internal volume of the compressible bioreactor container or agitating or mixing a cell suspension contained within the internal volume of the compressible bioreactor container.

[0010] In an example, the expansion container may be connected to the top section of the compressible bioreactor container. Thus, the expansion of the expandable container is not restricted by any external surface.

[0011] In an example, the top section of the compressible bioreactor container can include an interface plate having an opening. The expansion vessel can be mounted to the interface plate at the opening such that the interior volume of the expansion vessel is fluidly connected to the interior volume of the compressible bioreactor container via the opening in the interface plate.

[0012] In an example, an expansion vessel may include a base section, a top section, and a compressible sidewall extending between the base section and the top section. The base section may be attached to an interface plate. Thus, the expansion vessel may expand and contract to compensate for pressure therein. The expansion vessel may also be actuated to expand and contract, thereby controlling the volume of the expansion vessel.

[0013] In an example, the compressible bioreactor vessel and / or the expansion vessel may be generally cylindrical, wherein the sidewalls are generally cylindrical.The bioreactor interface plate may be generally circular and planar.

[0014] In one example, the sidewall of the compressible bioreactor vessel and / or expansion vessel can be a bellows wall. The sidewall can include a plurality of inward and outward folds that are staggered with the blade segments. The blade segments can be rigid. The inward and outward folds allow the blade segments to fold relative to each other, thereby compressing the sidewall, or conversely, to extend the sidewall.

[0015] In an example, the bioreactor container may further include one or more ports for introducing materials into the internal volume of the compressible bioreactor container and / or removing materials from the internal volume of the compressible bioreactor container. In an example, one or more ports may be provided in the interface plate. One or more ports may include seals, such as diaphragm seals. Thus, materials may be introduced into the internal volume of the compressible bioreactor container or materials may be extracted from the internal volume of the compressible bioreactor container through the ports. The seals cover and seal the ports to prevent contaminants from being introduced into the internal volume of the compressible bioreactor container and to maintain a sterile environment within the internal volume.

[0016] In some examples, the flow restrictor includes a valve. In some examples, the valve can allow gas to flow into and / or out of the gas outlet when the gas pressure differential at the gas outlet is equal to or greater than a threshold pressure differential. Thus, when the gas pressure differential at the gas outlet is below the threshold pressure differential, gas is prevented from flowing out of the gas outlet, thereby preventing water vapor from being lost from the bioreactor container through the outlet. Furthermore, when the gas pressure differential at the gas outlet is equal to or greater than the threshold pressure differential, gas is allowed to flow into and / or out of the gas outlet. This advantageously prevents large fluctuations in pressure within the bioreactor container during expansion and compression of the bioreactor container and also reduces the force required to expand and compress the bioreactor container.

[0017] In an example, when the gas pressure differential at the gas outlet is equal to or above a threshold pressure differential, the valve may not restrict the flow rate of gas therethrough, thereby reducing the force required to expand and / or compress the bioreactor container.

[0018] In some examples, the valve may be a two-way valve. In some examples, the valve may include a combination of an umbrella valve and a duckbill valve. In other examples, the valve may be a ball valve, particularly a two-way ball valve. In other examples, the valve may be an elastic contraction valve. In other examples, the valve may be a membrane valve having a membrane with one or more slits.

[0019] Thus, during expansion and compression of the bioreactor container, gas is allowed to flow into and out of the gas outlet to compensate for pressure changes in the bioreactor container due to its volume changes during expansion and compression.

[0020] In an example, the flow restrictor may include a restricted flow path having a restricted diameter. Thus, the restricted flow path having a restricted diameter reduces the flow rate of gas through the gas outlet, and thus as the bioreactor container is compressed, the pressure within the bioreactor container increases, which causes the bioreactor container to expand.

[0021] In an example, the restricted diameter can be between about 0.15 mm and about 1.5 mm. The restricted diameter can be between about 0.5 mm and about 1.0 mm. The restricted diameter can be one of about 0.5 mm, about 0.75 mm, or about 1.0 mm.

[0022] In an example, the gas outlet may have an outlet diameter, and the restricted diameter may be smaller than the outlet diameter.Thus, the diameter of the gas outlet is modified to provide a restricted diameter through which gas may flow.

[0023] In an example, the flow restrictor may include a porous structure arranged to restrict the flow of gas out of the outlet. The porous structure may be a sintered material. The porous structure may be a filter. The porous structure may restrict the flow of gas therethrough to reduce the flow rate of the gas through the outlet.

[0024] In some examples, the restrictor may be disposed within the gas outlet. The restrictor may be press-fitted to the gas outlet. In some examples, the restrictor may be disposed above the gas outlet. The restrictor may be press-fitted to an outer surface of the gas outlet. In other examples, the restrictor may be integrally formed with the gas outlet.

[0025] In an example, the flow restrictor may be integrally formed with the gas outlet.

[0026] In an example, the flow restrictor may include a tube fluidly connected to the gas outlet. Thus, when gas flows out of the gas outlet into the tube, the pressure at the gas outlet increases, thereby restricting the flow of gas out of the gas outlet.

[0027] In an example, the inner diameter of the tube may be between about 0.2 mm and about 1 mm. The inner diameter of the tube may be about 0.5 mm.

[0028] In an example, the length of the tube can be at least about 100 mm. The length of the tube can be between about 100 mm and about 700 mm. The length of the tube can be between about 300 mm and about 500 mm. The length of the tube can be one of about 300 mm or about 500 mm.

[0029] In an example, the flow restrictor may include a user-actuable valve. Actuation of the user-actuable valve may be independent of a gas pressure differential across the valve.

[0030] The user-actuable valve may include a spring element operably coupled to a movable plate. The movable plate may be operable to translate to selectively block an inlet or an outlet of the valve.

[0031] The spring element may be arranged to bias the movable plate to the closed position or the open position. The spring element may have a selected spring force such that pressure differential fluctuations during use of the bioreactor do not inadvertently switch the movable plate between the closed position and the open position.

[0032] The user-actuable valve may further include a valve actuator configured to transition the movable plate from the closed valve position to the open valve position and / or vice versa.

[0033] The valve actuator may include a control arm operably connected to the controller. In alternative examples, the user-actuable valve may be actuated by any other suitable valve actuator. For example, the user-actuable valve may be a torsion valve, an electrically actuated valve (e.g., a solenoid valve), or a magnetically actuated valve.

[0034] The user-actuable valve may be configured such that transitioning between open and closed valve positions is independent of a pressure differential across the valve.

[0035] During compression mixing operations, the user-actuated valve may be in a closed valve position. During breathing operations, the user-actuated valve may be in an open valve position to allow maximum air exchange with the contents of the bioreactor vessel and / or expansion vessel.

[0036] The controller may be configured to actuate the valve according to a predefined sequence of operating modes.

[0037] In an example, the bioreactor may further include a filter that is arranged in a gas flow path between the internal volume of the compressible bioreactor container and the gas outlet and / or the restrictor. The filter may be arranged in the gas outlet. The filter may include a hollow body that provides a channel through the filter and a filter element that extends across the channel. The hollow body may be wider at the center of the hollow body, and the filter element may be arranged in the center. The hollow body may have end portions extending from each end of the center. One or both end portions may include a restrictor. Each end portion may be formed as a spigot. The filter element may be a polyethersulfone membrane or a polyvinylidene fluoride membrane. Therefore, the filter can remove pollutants from the gas flowing through it. This prevents pollutants in the surrounding environment from reaching the cell suspension in the internal volume of the compressible bioreactor container, thereby preventing contamination of the cell suspension.

[0038] According to another aspect of the present disclosure, a filter for a gas outlet of a bioreactor is provided. The filter may include a hollow body that provides a passageway. The hollow body includes a central portion and end portions extending from the central portion. The central portion is wider than the end portions. A filter element is arranged within the central portion so as to extend across the passageway. The end portions have a reduced diameter to restrict airflow through the passageway.

[0039] In this way, the filter can be used in the gas outlet of a bioreactor vessel.The reduced diameter of the end portion restricts the flow of gas out of the gas outlet, thereby reducing the flow rate through the filter.

[0040] In an example, the end portion is a first end portion, and the filter may include a second end portion extending from an opposite side of the central portion from the first end portion.

[0041] In an example, a flow restrictor may be provided in the first end portion and / or the second end portion to reduce the diameter and restrict air flow through the passageway.

[0042] In an example, the flow restrictor may be press-fitted to the first end portion and / or the second end portion. In other examples, the flow restrictor may be integrally formed with the first end portion and / or the second end portion.

[0043] In an example, the flow restrictor may be integrally formed with the first end portion and / or the second end portion.

[0044] In an example, the first end portion and / or the second end portion may be formed as a sleeve.

[0045] In an example, the filter element may be a polyethersulfone membrane or a polyvinylidene fluoride membrane.

[0046] According to another aspect of the present disclosure, a method for mixing a cell suspension in a bioreactor is provided. In an example, the method is a method for mixing a cell suspension using the above-mentioned bioreactor container.

[0047] The method includes providing a bioreactor comprising a compressible bioreactor vessel having a gas outlet, the gas outlet comprising a flow restrictor; introducing a cell suspension into an interior volume of the compressible bioreactor vessel; compressing the compressible bioreactor vessel to reduce the interior volume of the compressible bioreactor vessel; and restricting gas flow through the gas outlet by the flow restrictor while compressing the compressible bioreactor vessel.

[0048] In an example, a compressible bioreactor container may include an expansion container having an interior volume fluidly connected to the interior volume of the compressible bioreactor container. The method may include transferring gas from the interior volume of the compressible bioreactor container to the interior volume of the expansion container during compression of the compressible bioreactor container.

[0049] In an example, the flow restrictor may include a valve. Restricting the flow of gas through the gas outlet by the flow restrictor may include blocking the flow of gas through the valve when a gas pressure differential at the gas outlet is below a threshold pressure differential, and allowing the flow of gas through the valve when the gas pressure differential at the gas outlet is equal to or greater than the threshold pressure differential. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Examples of the present invention will now be described hereinafter, by way of example only, with reference to the accompanying drawings, in which:

[0051] Figure 1A and Figure 1B A bioreactor is shown;

[0052] Figure 2 shows a cross-sectional view of the bioreactor of Figure 1;

[0053] Figure 3 shows a cross-sectional perspective view of the bioreactor of Figure 1;

[0054] Figure 4 shows a cross-sectional view of the outlet of the bioreactor of FIG1 with a first example of a flow restrictor;

[0055] Figure 5 (a) and Figure 5 (b) shows Figure 4 The arrangement of the flow restrictor;

[0056] Figure 6 A second example of a current restrictor is shown;

[0057] 7( a ) and 7 ( b ) show the outlet of the bioreactor of FIG. 1 with a third example of a flow restrictor;

[0058] Figure 8(a) to Figure 8(d) A fourth example of a flow restrictor is shown;

[0059] 9( a ) and 9 ( b ) show a fifth example of a current restrictor;

[0060] Figure 10(a) to Figure 10(c) A sixth example of a flow restrictor is shown;

[0061] 11( a ) and 11 ( b ) show a seventh example of a current limiter; and

[0062] Figure 12 Graph showing the load required to compress a bioreactor vessel with different flow restrictors. DETAILED DESCRIPTION

[0063] The described example embodiments relate to assemblies for processing biological materials. In particular, some embodiments relate to aseptic or sterilized assemblies. Note that the terms "aseptic" and "sterile" can be used interchangeably in this disclosure. Reference to fluids in the detailed description is not intended to limit the scope of protection to these materials. As those skilled in the art will recognize, the fluids described herein are merely examples of suitable materials for the described assemblies. Similarly, reference may be made to containers, receptacles, etc., however, such reference is not intended to limit the scope of protection to such containers or receptacles. As those skilled in the art will recognize, containers, receptacles, etc. are described herein merely as examples.

[0064] Certain terms are used in the following description for convenience only and are not limiting. The words "upper" and "lower" indicate directions referenced in the accompanying drawings and are relative to the components being described when assembled and installed. The words "inward," "inwardly," "outwardly," and "outwardly" refer to directions toward and away from, respectively, a designated centerline or geometric center (e.g., a central axis) of the components being described, as will be apparent from the context of the specification. Additionally, the terms "proximal" (i.e., closer) and "distal" (i.e., farther away) refer to positions relative to an axis or point of attachment.

[0065] Furthermore, as used herein, the terms "connected," "attached," "coupled," and the like are intended to include a direct connection between two components without any other components interposed therebetween, as well as an indirect connection between components with one or more other components interposed therebetween. The terminology includes the words specifically mentioned above, derivatives thereof, and words of similar meaning.

[0066] Furthermore, unless otherwise indicated, the use of ordinal adjectives such as "first," "second," "third," etc., merely indicates reference to different instances of similar objects and does not imply that the objects so described are necessarily in a given order temporally, spatially, sequentially, or in any other manner. The same reference numerals are used throughout to describe the same features.

[0067] Figure 1A 、 Figure 1B and Figure 2 A bioreactor 10 of the present invention is shown. The bioreactor 10 comprises a compressible bioreactor vessel 12, an interface plate 13 and an expansion vessel 14, also known as a breathing vessel.

[0068] During use, the compressible bioreactor container 12 has an internal volume for holding a fluid in which cell processing occurs. In particular, the fluid is a cell suspension and includes a cell population present in a liquid culture medium. In various examples, the cell population provided to the compressible bioreactor container 12 in use can include any human or animal cell type, for example: any type of adult stem cells or primary cells, T cells, CAR-T cells, monocytes, leukocytes, erythrocytes, NK cells, γδt cells, tumor infiltrating t cells, mesenchymal stem cells, embryonic stem cells, induced pluripotent stem cells, adipose-derived stem cells, Chinese hamster ovary cells, NS0 mouse myeloma cells, HELA cells, fibroblasts, HEK cells, insect cells, organoid cells, etc. Suitably, the cell population can include T cells. Alternatively, the cell population can include any microbial cell type, for example: bacteria, fungi, archaea, protozoa, algae cells.

[0069] The compressible bioreactor container 12 and the expansion container 14 are compressible, for example, by having bellows walls. The compressible bioreactor container 12 can expand and contract when being filled and emptied. The expansion container 14 can expand and contract when gas is transferred from the compressible bioreactor container 12 into and out of the expansion container 14.

[0070] like Figure 1A 、 Figure 1B and Figure 2 As shown, the compressible bioreactor container 12 has a base section with a bottom wall 15, a top section with an interface plate 13, and compressible side walls 16. The bottom wall 15 is disposed opposite the interface plate 13. The bottom wall 15 is rigid or attached to a rigid plate to provide a bottom surface of the compressible bioreactor container 12. Figure 2As shown, the top portion 17 of the compressible sidewall 16 is attached to the interface plate 13. The top portion 17 may include a rigid ring or the like for attachment to the interface plate 13. The compressible sidewall 16 is compressible so that the bottom wall 15 can move toward and away from the interface plate 13, thereby changing the interior volume of the compressible bioreactor container 12.

[0071] The compressible sidewall 16 can be a bellows wall having an accordion-like arrangement, allowing the compressible sidewall 16 to fold onto itself to compress. Specifically, the compressible sidewall 16 can include a series of alternating inward folds 16a and outward folds 16b, which allow the compressible sidewall 16 to compress like a bellows or accordion. The rigid blade portion extends between the inward folds 16a and the outward folds 16b. The inward folds 16a and the outward folds 16b can be formed by thinned sections in the compressible sidewall 16, wherein the inward folds 16a have thinned sections arranged on the outer surface of the compressible sidewall 16, and the outward folds 16b have thinned sections arranged on the inner surface of the compressible sidewall 16.

[0072] Thus, the compressible bioreactor container 12 can expand and contract, or be expanded and contracted, depending on the material held in the compressible bioreactor container 12. Specifically, the compressible bioreactor container 12 can expand as the cell culture within the compressible bioreactor container 12 grows and / or as additional material is added. The compressible bioreactor container 12 can be contracted and expanded by an actuator (not shown) that is adapted to move (e.g., push and / or pull) the bottom wall 15 and / or the interface plate 13 of the compressible bioreactor container 12 to change the volume of the compressible bioreactor container 12.

[0073] like Figure 2 and Figure 3 As shown, the interface plate 13 has a lower surface that is sealingly connected to the top portion 17 of the compressible side wall 16 of the compressible bioreactor container 12. The interface plate 13 also has an upper surface that is sealingly connected to the lower portion 20 of the compressible side wall 18 of the expansion container 14.

[0074] The interface plate 13 has one or more ports 22 for transferring materials into and out of the interior volume of the compressible bioreactor container 12. External components can be connected to one or more ports 22 to introduce materials through the ports 22. Each port includes a seal 23, such as a septum seal, which maintains a sealed environment within the interior volume of the compressible bioreactor container 12 and also allows a needle to pass through to form a fluid connection into the interior volume of the compressible bioreactor container 12. In an alternative example, each port 22 can have a valve, a cap, or other closure that provides an openable or breakable seal. One or more ports 22 can have a draw tube 24 that extends from the port 22 to the interior volume of the compressible bioreactor container 12. In use, the draw tube 24 can extend into the cell suspension for use in removing material, such as a sample, from the compressible bioreactor container 12.

[0075] The bulkhead 31 is mounted to the interface plate 13 such that the bulkhead 22 is suspended within the interior volume of the compressible bioreactor container 12. The bulkhead 31 can be attached to the interface plate 13 via a threaded connector or via a clip or clamp. The bulkhead 31 is attached to the center of the interface plate 13 such that the bulkhead 31 is centrally positioned within the compressible bioreactor container 12. However, it should be understood that the bulkhead 31 can be positioned off-center within the compressible bioreactor container 12. The bottom surface of the bulkhead 31 is substantially flat and faces the bottom wall 15 of the compressible bioreactor container 12. The bulkhead 31 also has a tapered upper surface that faces the interface plate 13. The bulkhead 31 is circular and sized so as to be spaced apart from the compressible sidewall 16 of the compressible bioreactor container 12. This allows the draw tube 24 to pass between the compressible sidewall 16 and the bulkhead 31 to provide a fluid sampling path from the compressible bioreactor container 12 to the interface plate 13.

[0076] The diaphragm 31 is provided to mix the cell suspension contained in the interior volume of the compressible bioreactor container 12 during use. Specifically, the bottom wall 15 of the compressible bioreactor container 12 can be moved relative to the interface plate 13 and the diaphragm 31 so that the diaphragm 31 contacts and mixes the cell suspension within the interior volume of the compressible bioreactor container 12. In examples, the base wall 15 can be raised and lowered relative to the interface plate 13 (i.e., the distance between the base portion 15 and the interface plate 13 is changed), and / or the base portion 15 can be tilted relative to the interface plate 13, and / or the base portion 15 can be rotated relative to the interface plate 13.

[0077] The interface plate 13 has an orifice 21 extending between the upper and lower surfaces of the interface plate 13 and allowing gas to flow between the interior volume of the compressible bioreactor container 12 and the interior volume of the expansion vessel 14. The orifice 21 may include a filter. The filter may prevent particles (such as cells) from transferring from the compressible bioreactor container 12 to the expansion vessel 14. The filter may also prevent liquid from transferring from the compressible bioreactor container 12 to the expansion vessel 14.

[0078] like Figure 3 As shown, the interface plate 13 has a hollow, annular protrusion 34 extending from the upper side of the interface plate 13. The outer surface of the annular protrusion 34 is connected to the lower portion 20 of the compressible sidewall 18 of the expansion vessel 14. The inner surface of the annular protrusion 34 has a plurality of orifices 21 spaced around the inner surface. The orifices 21 allow fluid communication between the compressible bioreactor container 12 and the expansion vessel 14. A plurality of walls 35 extend inward from the inner surface of the annular protrusion 34 toward the center of the interface plate 13. The walls 35 are positioned approximately at the center of each orifice 21. As gas flows between the compressible bioreactor container 12 and the expansion vessel 14, the walls 35 create a tortuous (non-linear) path for the gas, thereby increasing the surface area that the gas from the compressible bioreactor container 12 contacts as it flows into the expansion vessel 14. When the gas contacts the surfaces surrounding the orifices 21 (including the walls 35), water vapor in the gas condenses on these surfaces. The upper surface of the interface plate 13, within the inner surface of the annular protrusion 34, is sloped toward the orifice 21 to direct liquid droplets back through the orifice 21, thereby returning liquid to the interior volume of the compressible bioreactor vessel. Thus, the increased surface area created by the wall 35 surrounding the orifice 21 reduces the flow of liquid vapor out of the compressible bioreactor vessel 12 while providing an orifice through which gas can flow freely between the compressible bioreactor vessel 12 and the expansion vessel 14.

[0079] The expansion vessel 14 has a bottom section formed by the interface plate 13 , a top section 25 connected to a filter 19 having a gas outlet 26 , and a compressible side wall 18 .

[0080] The lower portion 20 of the compressible sidewall 18 is attached to the interface plate 13. The lower portion 20 may include a rigid ring or the like for attachment to the interface plate 13. The compressible sidewall 18 is compressible so that the top section 25 can move toward and away from the interface plate 13, thereby changing the internal volume of the expansion vessel 14.

[0081] The compressible sidewall 18 can be a bellows wall having an accordion-like arrangement that allows the compressible sidewall 18 to fold onto itself for compression. Specifically, the compressible sidewall 18 can include a series of alternating inward folds 18a and outward folds 18b that allow the compressible sidewall 18 to compress like a bellows or accordion. The rigid blade portion extends between the inward folds 18a and the outward folds 18b. The inward folds 18a and the outward folds 18b can be formed by thinned sections in the compressible sidewall 18, wherein the inward folds 18a have thinned sections arranged on the outer surface of the compressible sidewall 18, and the outward folds 18b have thinned sections arranged on the inner surface of the compressible sidewall 18.

[0082] The expansion vessel 14 allows the compressible bioreactor container 12 to expand and contract without significantly changing the pressure in the compressible bioreactor container 12. Alternatively or additionally, the expansion vessel 14 may be operable, such as by mechanically or manually compressing or expanding, to expand or contract the compressible sidewall 18 of the expansion vessel 14, thereby changing the volume of the compressible bioreactor container 12. Alternatively or additionally, the expansion vessel 14 may be operable, such as by mechanically or manually compressing or expanding, to adjust the pressure within the compressible bioreactor container 12.

[0083] like Figure 1A and Figure 1B As shown, a retainer 29 is provided around the expansion vessel 14 and maintains the expansion vessel in alignment as the expansion vessel 14 expands and contracts. The retainer 29 includes a first retainer portion 29a attached to the interface plate 13 and a second retainer portion 29b slidably mounted to the first retainer portion 29a. The second retainer portion 29b can slide in the direction of expansion and contraction of the expansion vessel 14. A stopper prevents the second retainer portion 29b from detaching from the first retainer portion 29a.

[0084] Thus, the expansion vessel 14 can expand or contract depending on the operating and environmental characteristics of the bioreactor 10. As the expansion vessel 14 expands and contracts, the retainer 29 limits movement of the expansion vessel 14 as the first and second retainer portions 29a, 29b slide relative to each other.

[0085] The holder 29 also includes a clamping feature 30, which in this example is a lip. The clamping feature 30 can be clamped by an actuator. In other examples where the bioreactor 10 does not include an expansion vessel 14, the lip 30 can be provided on the interface plate 13.

[0086] In some examples, a locking element may be provided on the first retainer portion 29a and / or the second retainer portion 29b to lock the second retainer portion 29b in the first retracted position relative to the first retainer portion 29a (see FIG. Figure 1A ) and / or second expanded position (see Figure 1B ). The locking element can be any suitable locking element, such as corresponding recesses and protrusions. In other examples, the second retainer portion 29b can be maintained in the first retracted position and / or the second expanded position by an actuator.

[0087] like Figure 2 、 Figure 4 As shown in FIG7( a), the filter 19 has a hollow body that provides a passage through the filter 19. The passage is the gas outlet 26 of the bioreactor 10. The hollow body has a central portion 32, a first end 27 extending from a first side of the central portion 32, and a second end 28 extending from a second side of the central portion 32. The central portion 32 is wider than the first end 27 and the second end 28 to receive a filter element 33 therein. The filter element 33 extends across the passage so that all gas flowing through the passage passes through the filter element 33. Any suitable filter element 33 that filters particles (such as microorganisms or other contaminants) can be used. For example, the filter element 33 can be a polyethersulfone membrane or a polyvinylidene fluoride membrane. The first end 27 of the filter is positioned within the interior volume of the expansion vessel 14, and the second end 28 of the filter is positioned outside the expansion vessel 14. In some examples, the filter 19 may not include the second end 28 as shown.

[0088] The outlet 26 allows for gas exchange between the interior volume of the expansion vessel 14 and the external environment (e.g., a bioreactor housing). Specifically, as pressure increases within the interior volume of the expansion vessel 14, gas flows out of the expansion vessel 14 through the outlet 26. The pressure within the expansion vessel 14 can increase due to contraction of the expansion vessel 14 and / or contraction of the compressible bioreactor container 12, which forces gas from the compressible bioreactor container 12 into the expansion vessel 14. Additionally, as pressure decreases within the interior volume of the expansion vessel 14 and / or the compressible bioreactor container 12, gas flows into the expansion vessel 14 through the outlet 26. The pressure within the expansion vessel 14 can decrease due to expansion of the expansion vessel 14 and / or expansion of the compressible bioreactor container 12, which draws gas from the expansion vessel 14 into the compressible bioreactor container 12.

[0089] In some embodiments, the outlet 26 may include a one-way valve to only allow gas to flow out of the bioreactor 10. The gas inlet may be provided in one of the compressible bioreactor vessel 12, the interface plate 13, or the expansion vessel 14. The gas inlet may include a one-way valve to only allow gas to flow into the bioreactor 10.

[0090] Figure 4 A first example of a flow restrictor 40 is shown. The flow restrictor 40 is positioned in the second end 28 of the outlet 26. The flow restrictor 40 restricts the flow of gas through the outlet 26. This reduces the rate at which gas leaves the interior volume of the expansion vessel 14, thereby reducing the volume of gas that leaves the interior volume of the expansion vessel 14 during compression and reducing the loss of liquid in the form of vapor leaving the bioreactor 10. Because the gas flow out of the outlet 26 is restricted, the pressure in the interior volume of the expansion vessel 14 increases. Once the pressure in the expansion vessel reaches a certain level, the expansion vessel 14 expands and receives additional gas from the compressible bioreactor container 12.

[0091] Figure 5 (a) and Figure 5 (b) shows Figure 4 An example of a current limiter 40. Figure 5 (a) and Figure 5 As shown in (b), the restrictor 40 has a body having a first end 41 and a second end 42. The body is hollow to provide a gas flow path extending from the first end 41 to the second end 42. The inner diameter (A) of the hollow body is smaller than the outlet 26 (see FIG. Figure 2 ). For example, the diameter (B) of the outlet 26 can be between about 3 mm and about 5 mm, such as about 3.8 mm, and the inner diameter (A) of the hollow body can be between about 0.15 mm and about 1.5 mm, such as about 0.5 mm, about 0.75 mm, or about 1 mm. The restrictor 40 thus restricts the diameter (B) of the outlet 26 to restrict the flow of gas out of the outlet. At the first end 41 of the body, the outer diameter of the body is equal to or less than the diameter (B) of the outlet 26. This allows the first end 41 of the restrictor 40 to be inserted into the second end 28 of the outlet 26.

[0092] In some examples, such as Figure 5 (a) and Figure 5 As shown in (b), the first end 41 of the flow restrictor is inserted into the second end 28 of the outlet 26. The second end 42 of the body has a flange. The flange contacts the outer edge of the second end 28 of the outlet 26. The flange can be used as a stopper to prevent the flow restrictor 40 from moving through the outlet 26. In one example, as shown in FIG. Figure 4 As shown, the first end 41 of the flow restrictor 40 is press-fitted to the second end 28 of the outlet 26. In another example, as shown in FIG. Figure 5 (a) and Figure 5 As shown in FIG. 2( b ), the first end 41 includes an O-ring 43 to provide a seal between the first end 41 of the flow restrictor 40 and the outlet 26 . In another example, the flow restrictor may be integrally formed in the second end 28 of the outlet 26 .

[0093] The inner diameter (A) of the hollow body can be between about 0.15 mm and about 1.5 mm. In some examples, the inner diameter (A) of the hollow body is between about 0.5 mm and about 1.0 mm. In some examples, the inner diameter (A) of the hollow body is about 0.50 mm, or about 0.75 mm, or about 1.00 mm.

[0094] like Figure 5 (a) and Figure 5 As shown in (b), the second end portion 42 of the restrictor 40 may be of different lengths. Figure 5 The length of the second end portion 42 of the flow restrictor 40 shown in (a) is greater than Figure 5 (b) Length of the second end 42 of the restrictor 40 shown. In an example, the length of the second end 42 of the restrictor 40 can be between about 1 mm and about 15 mm. In an example, the length of the second end 42 of the restrictor 40 can be between about 1 mm and about 10 mm. In an example, the length of the second end 42 of the restrictor 40 is about 1.5 mm. In an example, the length of the second end 42 of the restrictor 40 is about 10 mm. It will be understood that the longer the length of the second end 42 of the restrictor 40, the more restricted the flow.

[0095] The flow restrictor 40 can be formed from any suitable material. The flow restrictor 40 can be formed from a polymer that is resistant to gamma radiation. For example, the flow restrictor 40 can be formed from a polymer such as a polyaryletherketone, for example, polyetheretherketone (PEEK). The flow restrictor 40 can be manufactured by machining (turning). Additionally or alternatively, the flow restrictor 40 can be manufactured by an additive manufacturing process or by molding.

[0096] Figure 6 A second example of a flow restrictor 50 is shown. Figure 4 The restrictor 40 is shown positioned in the same manner in the second end 28 of the outlet 26 .

[0097] The flow restrictor 50 has a body having a first end 51 and a second end 52. The first end 51 of the body is hollow to allow gas to enter the flow restrictor 50. The second end 52 of the body has a porous material. In some examples, the second end 52 of the body can be formed of a porous material. In other examples, the flow restrictor 50 is similar to the reference Figures 4 and 5(b) The restrictor described is the same, but with the second end 52 of the body covered with a porous material.

[0098] The porous material allows a restricted flow of gas therethrough. This restricts the flow of gas out of the outlet 26.

[0099] The body of the restrictor 50 can be formed of any suitable material. The body of the restrictor 50 can be formed of a polymer that is resistant to gamma radiation. For example, the body of the restrictor 50 can be formed of a polymer, such as a polyaryletherketone, for example polyetheretherketone (PEEK). The body of the restrictor 50 can be manufactured by machining (turning). Additionally or alternatively, the body of the restrictor 50 can be manufactured by an additive manufacturing process or by molding. The porous material can be a sintered polymer. Any suitable polymer can be used, such as polytetrafluoroethylene (PTFE), polyethylene (PE) or polypropylene (PP).

[0100] Figure 7(a) and 7(b) A third example of a flow restrictor 60 is shown. The flow restrictor 60 has an elongated tubular member 61. A first end portion 62 of the elongated tubular member 61 is positioned in fluid communication with the second end 28 of the outlet 26. The elongated tubular member 61 increases resistance to gas flow out of the outlet 26, thereby restricting gas flow through the outlet.

[0101] The elongated tubular member 61 is secured in fluid communication with the outlet 26 by a tubular member support 63. The tubular member support 63 surrounds the second end 28 of the outlet 26 and the first end portion 62 of the elongated tubular member 61 and maintains the first end portion 62 of the elongated tubular member 61 aligned with the second end 28 of the outlet 26 to allow fluid to flow therebetween.

[0102] The inner diameter of the elongated tubular member 61 is smaller than the diameter (B) of the outlet 26. The inner diameter of the elongated tubular member 61 may be 0.25 mm to 1 mm. In some examples, the inner diameter of the elongated tubular member 61 is 0.5 mm.

[0103] The length of the elongated tubular member 61 is 100 mm to 700 mm. In some examples, the length of the elongated tubular member 61 is 300 mm to 500 mm. In some examples, the length of the elongated tubular member 61 is 300 mm or 500 mm.

[0104] The elongated tube 63 has a bent portion 65 adjacent the tube support 63. The bent portion 65 may be formed of a relatively rigid material compared to the rest of the tube, or the bent portion 65 may be reinforced to prevent the bent portion from kinking and blocking gas flow.

[0105] As shown in FIG. 7( a ), the second retainer portion 29 b covers the tube support 63 and at least a portion of the elongated tube including the second end portion 64 of the elongated tube.

[0106] 7(b), the elongated tubing 63 may be coiled to prevent kinks in the tubing that could impede airflow and to reduce the footprint of the restrictor 60. The tubing support 63 includes a circular base with guides for receiving the elongated tubing 61 in a coiled arrangement.

[0107] According to the illustrated example, the elongated tubular member 61 is held fixed at the second end 28 of the outlet 26 by a tubular member support 63. In other examples, the first end portion 62 of the elongated tubular member 61 can be press-fit into the second end 28 of the outlet 26 or sealed in the second end 28 of the outlet 26 by an O-ring so that the second end 28 maintains the first end portion 62 of the elongated tubular member 61 in fluid communication with the outlet 26.

[0108] Figures 8(a) to 10(c) Examples of valve restrictors 70 , 80 , 90 are shown. Figure 8(a) to Figure 8(d) A fourth example of a flow restrictor 70 in the form of a combination of an umbrella valve and a duckbill valve is shown. Figures 9(a) and 9(b) show a fifth example of a flow restrictor 80 in the form of an elastically contracting valve. Figure 10(a) to Figure 10(c) A sixth example of a flow restrictor 90 in the form of a two-way ball valve is shown. These valves 70, 80, 90 allow gas flow through the valve when the gas pressure differential at the gas outlet 26 is equal to or greater than a threshold pressure differential, and prevent gas flow when the gas pressure differential is below the threshold pressure differential. When the gas pressure differential is below the threshold pressure differential, the valves 70, 80, 90 remain closed, thereby preventing fluid from being lost from the bioreactor vessel 12 in the form of vapor. However, when the gas pressure differential at the gas outlet 26 is equal to or greater than the threshold pressure differential, the valves 70, 80, 90 open.

[0109] When the gas pressure differential increases to a value equal to or above the threshold pressure differential, the valves 70, 80, 90 open. This provides a force profile at the valves 70, 80, 90 that increases at an approximately constant rate. This, in turn, allows the bioreactor container 12 to expand and / or contract without significantly increasing the force required to expand or contract the bioreactor container 12. This also prevents large fluctuations in pressure in the bioreactor container 12 and / or the expansion vessel 14 during expansion and / or contraction.

[0110] Other one-way or two-way pressure-actuated valves (not shown) may be used in the present invention. The valve may be any valve that can be actuated by a gas pressure differential proximal to the valve. For example, the valve may be a membrane valve having one or more slits therein. As the gas pressure differential increases to a pressure differential equal to or above a threshold pressure differential, the membrane of the membrane valve flexes in the direction of the relatively low gas pressure, thereby opening the one or more slits and allowing gas to flow therethrough.

[0111] like Figure 8(a) to Figure 8(d) As shown in FIG. 8 , a fourth example of a combined umbrella valve and duckbill valve 70 of a flow restrictor includes a valve element 71 and a support member 72. As shown in FIG. 8( b ), the valve element 71 has an umbrella portion 73, which is a flange extending from a first upper end portion of the valve element 71. The valve element 71 also has a duckbill portion 74 including a resilient lip.

[0112] The support member 72 has a valve seat 77 at its upper end. The valve seat 77 has at least one opening 75 therein to allow gas to flow therethrough. The valve seat can have multiple openings. In one example, the valve seat has four openings. The thickness of the valve seat can be selected to control the gas pressure differential required to open the valve. In an example, the valve seat has a thickness ranging from approximately 0.6 mm to 1.2 mm. In some examples, the valve seat has a thickness of approximately 0.6 mm, 0.8 mm, 1.0 mm, or 1.2 mm.

[0113] In the rest position, the umbrella portion 73 rests against the valve seat 77, blocking the opening 75 in the valve seat 77. In the rest position, the resilient lips of the duckbill portion 74 rest against each other to block air flow through the central opening 76 of the valve element 71.

[0114] When the gas pressure in the gas outlet 26 is greater than the external pressure, so that the gas pressure difference is equal to or higher than the threshold pressure difference, as shown in FIG8(c), the umbrella portion 73 is lifted away from the support member 72 to allow the air flow G 出 The flow exits the valve 70 through the opening 75 and thus exits the bioreactor container 12 and / or the expansion container 14. 压缩 When the support member 72 is compressed, this increases the pressure within the bioreactor container 12 and / or the expansion vessel 14, and the gas pressure in the gas outlet 26 may increase. When the gas pressure differential at the gas outlet 26 drops below the threshold pressure differential, the umbrella-shaped portion 73 returns to its rest position and rests against the upper end of the support member 72, as shown in Figures 8(b) and 8(d), to prevent gas from flowing through the opening 75.

[0115] When the gas pressure near the central opening 76 of the valve element 71 is greater than the gas pressure in the gas outlet 26, so that the gas pressure difference is equal to or higher than the threshold pressure difference, as shown in FIG8(d), the elastic lips of the duckbill portion 74 separate from each other to allow the gas flow G 进 The bioreactor vessel 12 and / or expansion vessel 14 is accessed through the central opening 76. When the bioreactor vessel 12 and / or expansion vessel 14 is in the direction F 膨胀 8( b ) and 8 ( c ), the resilient lips of the duckbill portion 74 return to their rest positions and rest against each other, as shown in FIG8( b ) and FIG8( c ), to prevent gas from flowing through the central opening 76.

[0116] The support 72 is press-fitted onto an outer surface of the second end 28 of the outlet 26. According to an alternative example, the support 72 may be press-fitted into the second end 28 of the outlet 26. In yet another example, the support 72 may be integrally formed with the second end 28 of the outlet 26.

[0117] As shown in Figures 9(a) and 9(b), a fifth example of a resilient contraction valve 80 of a flow restrictor includes a resilient valve element 81 and a compression ring 82. The resilient valve element 81 is cylindrical and formed of a resilient material. The resilient valve element 81 is biased toward its cylindrical shape. The compression ring 82 is disposed around the outer surface of the resilient valve element 81. The compression ring 82 is made of a resilient material. The compression ring 82 compresses a portion of the resilient valve element 81 in the rest position to reduce the diameter of the resilient valve element 81 at that portion.

[0118] When the gas pressure difference at the gas outlet 26 is equal to or higher than the threshold pressure difference, as shown in FIG9(b), the compression ring 82 expands, causing the diameter of the elastic valve element 81 to expand to allow the gas flow G 出 Gas flows out of or into the bioreactor container 12 and / or the expansion container 14 (not shown) through the resilient valve element 81. When the bioreactor container 12 and / or the expansion container 14 is compressed, this increases the pressure within the bioreactor container 12 and / or the expansion container 14 relative to the external pressure, and the gas pressure differential at the gas outlet 26 may increase. When the bioreactor container 12 and / or the expansion container 14 expands, this decreases the pressure within the bioreactor container 12 and / or the expansion container 14 relative to the external pressure, and the gas pressure differential at the gas outlet 26 may also increase. When the gas pressure differential at the gas outlet 26 drops below a threshold pressure differential, the compression ring compresses back to its rest position and reduces the diameter of the resilient valve element 81 at that portion, as shown in FIG. 9( a), to prevent gas from flowing through the resilient valve element 81.

[0119] The elastically contractile valve 80 may be provided on an inner surface of the gas outlet 26. Alternatively, the elastically contractile valve 80 may be connected to one end of the gas outlet 26.

[0120] like Figure 10(a) to Figure 10(c) As shown, a two-way ball valve 90 of the sixth example of a flow restrictor includes a first ball valve 91 connected to a first spring element 92, a first ball seat 93, a second ball valve 94 connected to a second spring element 95, and a second ball seat 96. The two-way ball valve 90 also has an outlet end 97 and an inlet end 98.

[0121] As shown in FIG. 10( a ), in the rest position, each of the first and second ball valves 91 , 94 is resting in their respective first and second ball seats 93 , 98 .

[0122] A two-way ball valve 90 may be connected to the gas outlet (26, see Figure 2 The two-way ball valve 90 can be incorporated into the gas outlet 26 in any suitable manner.

[0123] When the gas pressure at the outlet end 97 (i.e., the gas pressure in the gas outlet 26) is greater than the gas pressure at the inlet end 98 (i.e., the external pressure) so that the gas pressure difference is equal to or higher than the threshold gas pressure difference, as shown in FIG10( b ), the second ball valve 94 overcomes the bias of the second spring element 95 and lifts away from the second ball seat 96 to allow the gas flow G 出 Gas flows out of the bioreactor vessel 12 and / or expansion vessel 14 through the valve 90. When the gas pressure differential at the gas outlet 26 drops below a threshold pressure differential, the second ball valve 94 is biased by the second spring element 95 to rest in the second ball seat, as shown in Figures 10(a) and 10(c), to prevent gas from flowing through the valve 90.

[0124] When the gas pressure at the inlet end 98 (i.e., the external pressure) is greater than the gas pressure at the outlet end 97 (i.e., the gas pressure in the gas outlet 26) so that the gas pressure difference is equal to or higher than the threshold pressure difference, as shown in FIG10( c ), the first ball valve 91 overcomes the bias of the first spring element 92 and lifts away from the first ball seat 93 to allow the gas flow G 进 Gas enters the bioreactor vessel 12 and / or the expansion vessel 14 through the valve 90. When the gas pressure differential at the gas outlet 26 drops below a threshold pressure differential, the first ball valve 91 is biased by the first spring element 92 to rest in the first ball seat 93, as shown in Figures 10(a) and 10(b), to prevent gas from flowing through the valve 90.

[0125] 11( a )-( b ) show a seventh example of a flow restrictor in the form of a user-actuatable valve 100 . The user-actuatable valve 100 comprises an outlet end 110 and an inlet end 120 .

[0126] A user-actuable valve 100 may be connected to the gas outlet 26 (see Figure 2 ). The user-actuable valve 100 may be incorporated into the gas outlet 26 in any suitable manner.

[0127] The user-actuated valve 100 includes a movable plate 101 and a spring element 102. The movable plate 101 is provided with a seal 103 for sealing the movable plate 101 against the inlet end 120, preventing gas from flowing therethrough. In this example, the seal 103 is an O-ring having an inner diameter that exceeds the diameter of the inlet end 120. In other examples, the seal 103 may be omitted. For example, the movable plate 101 may include a flexible material that is operable to seal against the outer periphery of the inlet end 120.

[0128] When a user actuates the user-actuable valve 100 , the movable plate 101 is operable to translate between a closed position and an open position.

[0129] Figure 11a The user-actuable valve 100 is shown when the movable plate 101 is in a closed position. As shown, when the movable plate 101 is in the closed position, the seal 103 is sandwiched between the outer peripheral wall of the inlet end 120 and the movable plate 101. The movable plate 101 is substantially impermeable to fluid, and thus this configuration prevents gas from exiting or entering the user-actuable valve 100 through the inlet end 120.

[0130] The spring element 102 is operatively connected to the movable plate 101 and is arranged to bias the movable plate 101 to the closed position. The spring element 102 has a selected spring force so that pressure fluctuations during use of the bioreactor 10 do not inadvertently switch the movable plate 101 between the closed and open positions.

[0131] The user-actuated valve 100 also includes a valve actuator for actuating the movable plate 101 between a closed position and an open position. In this example, the valve actuator includes a control arm 104 operatively connected to a controller (not shown). The movable plate 101 includes a connector 105 configured to selectively interact with the control arm 104.

[0132] The controller (not shown) is a button for pressing the control arm 104 toward the movable plate 101. The valve actuator is configured so that when the user actuates the button, the control arm 104 pushes to contact the connector 105, as indicated by the block arrow 106. The force of the control arm 104 overcomes the reaction force of the spring element 102 to separate the movable plate 101 from the inlet end 120. In this manner, the valve actuator is configured to switch the movable plate 101 from the closed valve position to the open valve position.

[0133] In alternative examples, the user-actuable valve 100 may be actuated by any other suitable valve actuator. For example, the user-actuable valve 100 may form a twist valve, an electrically actuated valve (eg, a solenoid valve), or a magnetically actuated valve.

[0134] Figure 11b The user-actuated valve 100 is shown when the movable plate 101 is in an open position. In this position, the inlet end 120 is fluidly connected to the outlet end 110. This configuration allows gas to flow into or out of the bioreactor container 12 and / or the expansion vessel 14 through the user-actuated valve 100. When the movable plate 101 is in the open position, the pressure within the bioreactor container 12 and / or the expansion vessel 14 is balanced relative to the external pressure at the gas outlet 26.

[0135] The controller can be configured such that further actuation of the controller by the user (e.g., releasing a button) transitions the movable plate 101 from the open position to the closed position. Releasing the button lifts the control arm 104 away from the connector 105 of the movable plate 101. Without the force of the control arm 104 on the movable plate 101, the spring element 102 forces the movable plate 101 into the closed position. In this way, the user-actuated valve 100 is configured to enable the user to transition the valve from a closed configuration (in which the movable plate 101 is in the closed position) to an open configuration (in which the movable plate 101 is in the open position), and vice versa.

[0136] The user can close the user-actuated valve 100 during the compression mixing operation. This can reduce the amount of water vapor lost from the inside of the bioreactor 10 during such an operation. As previously described, during such a compression mixing operation, a pressure differential can increase. After the compression mixing operation is complete, the user can open the user-actuated valve 100 to release the pressure from the bioreactor 10. This "breathing operation" can allow for maximum air exchange with the contents of the bioreactor vessel 12 and / or expansion vessel 14 and prevent significant pressure differentials from occurring.

[0137] Actuated by gas pressure difference Figure 8a to Figure 10bCompared to a flow restrictor, the actuation of the user-actuated valve 100 is independent of the gas pressure differential. The user-actuated valve 100 can provide greater control over the environmental parameters experienced by the contents of the bioreactor container 12 and / or expansion container 14.

[0138] In one example, the inlet end 120 of the user-actuable valve 100 can be connected to the end of the gas outlet 26, while the outlet end 110 can be a free end under external pressure. Alternatively, the outlet end 110 can be connected to the end of the gas outlet 26, while the inlet end 120 can be a free end under external pressure.

[0139] In an alternative embodiment, the valve actuator can be configured such that pressing a button transitions the movable plate 101 from an open position to a closed position. For example, the control arm 104 can be weighted so that the weight of the control arm 104 on the movable plate 101 overcomes the spring force of the spring element 102. The controller can be configured such that pressing a button lifts the control arm 104 away from the connector 105, thereby transitioning the movable plate from an open position to a closed position. Although the controller of the user-actuated valve 100 is described as a button, any suitable controller, such as a lever, a dial, or an electronic controller, may be used instead. The user-actuated valve 100 described herein may be provided in the bioreactor 10 in place of, or in addition to, the flow restrictors 40, 50, or 60 or valves 70, 80, or 90 of the aforementioned embodiments.

[0140] The use of the bioreactor 10 of the present invention is described with reference to Figures 1 to 10(b). During use, the bottom wall 15 of the compressible bioreactor container 12 is tilted and / or moved in the axial direction of the compressible bioreactor container 12 by an actuator. This movement of the bottom wall 15 compresses and / or expands the compressible bioreactor container 12. This is done to mix or agitate the fluid within the internal volume of the compressible bioreactor container 12, to control the volume of the compressible bioreactor container 12 before adding material to or extracting a sample from the compressible bioreactor container 12, or to perform one or more cell processing steps.

[0141] As the compressible bioreactor container 12 is compressed, the pressure within the compressible bioreactor container 12 increases, forcing gas to flow from the compressible bioreactor container 12 to the expansion container 14 to equalize the pressure within the compressible bioreactor container 12 and the expansion container 14. The restrictor 40, 50, or 60 or valve 70, 80, 90, or 100 according to any of the above-described embodiments prevents gas from freely flowing out of the expansion container 14. Because the flow of gas out of the expansion container 14 is restricted, the expansion container 14 expands to compensate for the increased pressure of the introduced gas. Thus, the restrictor 40, 50, or 60 causes expansion of the expansion container 14 and reduces gas escape through the gas outlet 26, thereby reducing vapor loss through the gas outlet 26. Similarly, when the valve 70, 80, 90, or 100 is closed, the valve 70, 80, 90, or 100 blocks gas from flowing out of the expansion container, thereby causing the expansion container 14 to expand, which prevents gas escape and vapor loss through the gas outlet 26.

[0142] As the compressible bioreactor container 12 expands, the pressure in the compressible bioreactor container 12 decreases, thereby drawing gas from the expansion vessel 14 back into the compressible bioreactor container 12 to equalize the pressures in the compressible bioreactor container 12 and the expansion vessel 14. The restrictor 40, 50, or 60 or the valve 70, 80, 90, or 100 according to any of the above-described embodiments prevents gas from flowing freely from outside the bioreactor 10 into the expansion vessel 14. As the flow of gas into the expansion vessel 14 is restricted, the expansion vessel 14 will contract to compensate for the decrease in pressure caused by the gas moving out of the expansion vessel 14.

[0143] The expansion vessel 14 can also be locked or held in the retracted position by the holder 29 to allow gas exchange in the bioreactor 10. The expansion vessel 14 is first actuated to the retracted position by actuating the second holder portion 29b relative to the first holder portion 29a to a first retracted position, as shown in FIG. Figure 1AAs shown, the second retainer portion 29b is locked or held in place relative to the first retainer portion 29a to prevent expansion of the expansion vessel 14. The bottom wall 15 of the compressible bioreactor container 12 is moved upwardly, toward the expansion vessel 14, by the actuator to compress the compressible bioreactor container 12, thereby increasing the gas pressure at the outlet 26 and venting gas from the headspace above the fluid in the interior volume of the bioreactor container 12 through the contracted expansion vessel 14 and the outlet 26. The bottom wall 15 of the compressible bioreactor container 12 is then moved downwardly, away from the expansion vessel 14, by the actuator to expand the compressible bioreactor container 12, thereby decreasing the gas pressure at the outlet 26 and drawing ambient gas into the interior volume of the compressible bioreactor container 12 through the outlet 26 and the contracted expansion vessel 14. The second retainer portion 29b can then be released from the first contracted position, allowing the second retainer portion 29b to freely move relative to the first retainer portion 29a and allowing the expansion vessel 14 to freely expand and contract.

[0144] Stress test data

[0145] Figure 12 The results of compression test data for three flow restrictors are shown: a combination of an umbrella valve and a duckbill valve (combination valve) with a valve seat having a thickness of 0.8 mm, a membrane valve having two slits (i.e., a cross-shaped slit), a restrictor having a restricted diameter of 0.5 mm, and a control with no flow restrictor. The graph shows the load required to compress the bioreactor vessel for each flow restrictor.

[0146] The first test was to compress the bioreactor container 12 at a speed of 2.5 mm / s, as shown in solid lines. The second test was to compress the bioreactor container 12 at a speed of 1.0 mm / s, as shown in dashed lines. During this test, the expansion container 14 remained locked in the retracted position.

[0147] The control without a flow restrictor required the least force to compress the bioreactor vessel. As bioreactor displacement increased, the load increased at a relatively slow rate.

[0148] The force required to compress the bioreactor container with the membrane valve was slightly higher than that required for the control. As the displacement of the bioreactor container increased, the rate of load increase was comparable to that of the control.

[0149] The force required to compress the bioreactor vessel with the combination valve was higher than the force required for the control. In one test, the force required to compress the combination valve was higher than that required for the membrane valve at lower displacement levels and comparable to that required for the membrane valve at higher displacement levels. In another test, the force required to compress the combination valve was higher than that required for the membrane valve at all displacement levels. The rate of load increase with increasing bioreactor vessel displacement was slower than that for the control and membrane valves.

[0150] The force required to compress the valve assembly is the greatest. As the displacement of the bioreactor vessel increases, the rate of load increase is also the highest.

[0151] Therefore, the membrane valve provides the minimum force required to compress the bioreactor vessel. This reduces the load on the actuator compressing the bioreactor vessel. Compared to other flow restrictors, the combination valve allows the bioreactor to be compressed at a more stable rate. This allows a more stable load to be applied by the actuator when compressing the bioreactor vessel and also reduces the load required to compress the bioreactor vessel compared to a flow restrictor with a reduced diameter. The flow restrictor with a reduced diameter provides the maximum force required to compress the bioreactor.

[0152] According to the illustrated example, a filter is disposed in the gas outlet 26 of the expansion vessel 14. In other examples, the filter may be disposed at any suitable location between the flow restrictor 40, 50, or 60 or the valve 70, 80, or 90 and the fluid contained in the compressible bioreactor container 12 to prevent the introduction of contaminants into the fluid. For example, the filter may be disposed in the flow restrictor 40, 50, or 60 or the valve 70, 80, or 90, the filter may be disposed in the interface plate 13 between the expansion vessel 12 and the compressible bioreactor container 12, or the filter may be disposed in the compressible bioreactor container 12 above the level of the fluid during use.

[0153] According to the example shown, the outlet 26 is provided in the expansion vessel 14. In other examples, the bioreactor does not comprise an expansion vessel, and the outlet 26 is provided in the compressible bioreactor vessel 12, for example in the interface plate 13.

[0154] In general, it will be understood by those skilled in the art that the above embodiments have been described by way of example only and without any limiting meaning, and that various substitutions and modifications are possible without departing from the scope of the invention as defined by the appended claims. Various modifications to the above detailed designs are possible, for example, there may be changes in shape, size, arrangement, assembly, sequence, etc. For example, any of the housing, planar interface, component retaining elements, etc. may be used in any suitable combination. In addition, although the present invention has been described with respect to an automated process, it will be understood by those skilled in the art that a user may perform one or more of the above process steps manually or semi-automatically.

Claims

1. A bioreactor comprising: a compressible bioreactor container configured to hold a cell suspension within an interior volume of the compressible bioreactor container, the compressible bioreactor container comprising a gas outlet in fluid communication with the interior volume of the compressible bioreactor container and configured to allow, in use, gas to flow out of the interior volume of the compressible bioreactor container; Wherein the gas outlet comprises a flow restrictor, the flow restrictor being configured such that when the compressible bioreactor container is compressed in use, the flow restrictor restricts the flow of gas out of the gas outlet.

2. The bioreactor according to claim 1, wherein The bioreactor further comprises an expandable expansion vessel having an interior volume fluidly connected to the interior volume of the compressible bioreactor vessel.

3. The bioreactor according to claim 2, wherein The expansion vessel includes the gas outlet.

4. The bioreactor according to claim 2 or claim 3, wherein The compressible bioreactor container comprises a base section, a top section, and a compressible sidewall extending between the base section and the top section, and wherein the expansion vessel is connected to the top section of the compressible bioreactor container.

5. The bioreactor according to claim 4, wherein The top section of the compressible bioreactor container comprises an interface plate having an opening, and wherein the expansion vessel is mounted to the interface plate at the opening such that the interior volume of the expansion vessel is fluidly connected to the interior volume of the compressible bioreactor container via the opening in the interface plate.

6. The bioreactor according to claim 5, wherein The expansion vessel includes a base section, a top section, and a compressible sidewall extending between the base section and the top section, and wherein the base section is attached to the interface plate.

7. The bioreactor according to claim 5 or claim 6, wherein The one or more ports are provided in the interface board.

8. A bioreactor according to any one of the preceding claims, wherein The flow restrictor includes a valve.

9. The bioreactor according to claim 8, wherein The valve consists of a user-actuable valve, and actuation of the valve is independent of the gas pressure differential across the valve.

10. The bioreactor according to claim 8, wherein When the gas pressure differential at the gas outlet is equal to or higher than a threshold pressure differential, the valve allows gas to flow into and / or out of the gas outlet.

11. The bioreactor according to any one of claims 8 to 10, wherein The valve is a two-way valve.

12. The bioreactor according to claim 11, wherein The valve comprises a combination of an umbrella valve and a duckbill valve.

13. The bioreactor according to any one of claims 1 to 7, wherein The flow restrictor includes a flow restriction path having a restricted diameter.

14. The bioreactor according to claim 13, wherein The restricted diameter is between about 0.15 mm and about 1.5 mm.

15. The bioreactor according to claim 14, wherein The restricted diameter is between about 0.5 mm and about 1.0 mm.

16. The bioreactor according to any one of claims 13 to 15, wherein The gas outlet has an outlet diameter, and wherein the restricted diameter is smaller than the outlet diameter.

17. A bioreactor according to any one of the preceding claims, wherein The flow restrictor comprises a porous structure arranged to restrict the flow of gas out of the outlet.

18. The bioreactor according to claim 17, wherein The porous structure is a sintered material.

19. A bioreactor according to any one of the preceding claims, wherein The flow restrictor is disposed in the gas outlet, for example press-fitted to the gas outlet.

20. A bioreactor according to any one of the preceding claims, wherein The flow restrictor is disposed above the gas outlet, for example, press-fitted onto an outer surface of the gas outlet.

21. The bioreactor according to any one of claims 1 to 18, wherein The restrictor is integrally formed with the gas outlet.

22. The bioreactor according to any one of claims 1 to 7 or 13 to 15, wherein The flow restrictor includes a tube fluidly connected to the gas outlet.

23. The bioreactor according to claim 22, wherein The inner diameter of the tube is between about 0.2 mm and about 1 mm.

24. The bioreactor according to claim 23, wherein The inner diameter of the tube is approximately 0.5 mm.

25. The bioreactor according to any one of claims 22 to 24, wherein The length of the tube is at least about 100 mm, for example between about 100 mm and about 700 mm.

26. The bioreactor according to claim 25, wherein The length of the tube is between about 300 mm and about 500 mm.

27. The bioreactor of any preceding claim, further comprising a filter disposed in the gas flow path between the interior volume of the compressible bioreactor vessel and the gas outlet and / or the flow restrictor.

28. The bioreactor according to claim 27, wherein The filter is disposed in the gas outlet.

29. The bioreactor of any preceding claim, further comprising one or more ports for introducing material into and / or removing material from the interior volume of the compressible bioreactor container.

30. A method of mixing a cell suspension in a bioreactor, the method comprising: providing a bioreactor comprising a compressible bioreactor vessel having a gas outlet, the gas outlet comprising a flow restrictor; introducing a cell suspension into the interior volume of the compressible bioreactor vessel; compressing the compressible bioreactor container to reduce an interior volume of the compressible bioreactor container; and While the compressible bioreactor container is being compressed, gas flow through the gas outlet is restricted by the flow restrictor.

31. The method according to claim 30, wherein The compressible bioreactor container comprises an expansion vessel having an interior volume fluidly connected to the interior volume of the compressible bioreactor container, and wherein the method comprises transferring gas from the interior volume of the compressible bioreactor container to the interior volume of the expansion vessel during compression of the compressible bioreactor container.

32. The method according to claim 30 or 31, wherein The flow restrictor comprises a valve, and wherein, Restricting the flow of gas through the gas outlet by the flow restrictor includes blocking gas flow through the valve when the gas pressure differential at the gas outlet is below a threshold pressure differential, and allowing gas flow through the valve when the gas pressure differential at the gas outlet is equal to or above the threshold pressure differential.

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