Cell culture chamber for increasing cell yield

Abstract

The present disclosure provides cell culture chambers for use in automated cell engineering systems, and in particular cell culture chambers that provide increased cell yield. The cell culture chamber may include a first body forming a first portion of the cell culture chamber. The cell culture chamber may include a second body forming a second portion of the cell culture chamber, the first body and the second body configured to be coupled together to form a closed volume with a first non-porous breathable material disposed on the first body; and wherein the first non-porous breathable material and the first body are formed together. A second non-porous breathable material may be disposed on the second body, and the second non-porous breathable material and the second body may be formed together. The cell culture chamber may further include at least one bubble trap integrally formed with the first body and / or the second body.

Description

Technical Field

[0001] The present disclosure provides cell culture chambers for use in automated cell engineering systems, and in particular provides cell culture chambers that provide increased cell yields. Background Art

[0002] As accelerated clinical adoption of advanced cell therapies is expected to be established, more attention is turning to the fundamental manufacturing strategies that will make these therapies available to patients worldwide. While cell therapies hold great promise in the clinic, high manufacturing costs relative to reimbursement are a huge barrier to commercialization. Therefore, the need for cost-effectiveness, process efficiency, and product consistency is driving efforts towards automation in many areas of cell therapy, and specifically T cell therapy.

[0003] Integrating cell activation, transduction, and expansion into commercial manufacturing platforms is critical to translating these important immunotherapies to a broad patient population. In order to make these life-saving treatments available to a global patient population, a shift in manufacturing technology must be implemented to support personalized medicine. The benefits of automation include labor time savings associated with the use of automation, as well as improved product consistency, reduced room classification, reduced cleanroom footprint, reduced training complexity, and improved scale-up and tracking logistics. In addition, software can be used to simplify the record-keeping process by providing a historical record of all processing equipment, reagents, patient identification, operator identification, sensor data in the process, etc. using automatically generated electronic batch records.

[0004] What is needed to advance these therapies and automated systems are components of cell expansion systems, such as cell culture chambers, that can increase cell output or provide desired cell properties. The present application meets these needs. Summary of the invention

[0005] In some aspects, the technology described herein relates to a cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: a first body, the first body forming a first portion of the cell culture chamber; a second body, the second body forming a second portion of the cell culture chamber, the first body and the second body being configured to be coupled together to thereby form a closed volume, wherein a first non-porous, gas-permeable material is disposed on the first body; and wherein the first non-porous, gas-permeable material and the first body are formed together.

[0006] In some aspects, the technology described herein relates to a cell culture chamber, further comprising a second non-porous gas-permeable material disposed on the second body; and wherein the second non-porous gas-permeable material and the second body are formed together.

[0007] In some aspects, the technology described herein relates to a cell culture chamber, the cell culture chamber further comprising at least one bubble trap, the at least one bubble trap being integrally formed with the cell culture chamber.

[0008] In some aspects, the technology described herein relates to a cell culture chamber further comprising one or more structural islands.

[0009] In some aspects, the technology described herein relates to a cell culture chamber, wherein the first body and the second body are coupled together using mating elements.

[0010] In some aspects, the technology described herein relates to a cell culture chamber, wherein the first body includes a plurality of openings configured to allow gas to be exposed to the external environment through an outer surface of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

[0011] In some aspects, the technology described herein relates to a cell culture chamber, wherein the plurality of openings comprises a geometric shape, a hexagonal shape, a heptagonal shape, an octagonal shape, a symmetrical shape, a honeycomb shape, a circular shape, a ring shape, a rectangular shape, an elliptical shape, or a square shape.

[0012] In some aspects, the technology described herein relates to a cell culture chamber, wherein the first non-porous gas-permeable material is positioned on an inner surface of the first body, the first non-porous gas-permeable material being configured to cover the plurality of openings of the first body.

[0013] In some aspects, the technology described herein relates to a cell culture chamber, wherein the structural section of the second body includes multiple openings, and the multiple openings are configured to allow gas to be exposed to the external environment through the outer surface of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

[0014] In some aspects, the technology described herein relates to a cell culture chamber, wherein the plurality of openings comprises a geometric shape, a hexagonal shape, a heptagonal shape, an octagonal shape, a symmetrical shape, a honeycomb shape, a circular shape, a ring shape, a rectangular shape, an elliptical shape, or a square shape.

[0015] In some aspects, the technology described herein relates to a cell culture chamber, wherein the second non-porous gas-permeable material is positioned on an inner surface of the first body, the second non-porous gas-permeable material being configured to cover the plurality of openings of the second body.

[0016] In some aspects, the technology described herein relates to a cell culture chamber, wherein the at least one bubble trap is integrally formed with the first body of the cell culture chamber, and the cell culture chamber further comprises a bubble trap track, which is integrally formed with the first body and fluidly connected to the at least one bubble trap.

[0017] In some aspects, the technology described herein relates to a cell culture chamber, wherein at least two bubble traps are integrally formed with the first body of the cell culture chamber, and the bubble trap track is fluidly connected to the at least two bubble traps.

[0018] In some aspects, the technology described herein relates to a cell culture chamber in which the width of the bubble trap track tapers.

[0019] In some aspects, the technology described herein relates to a cell culture chamber wherein the bubble trap track tapers in height.

[0020] In some aspects, the technology described herein relates to a cell culture chamber, wherein at least two bubble traps are integrally formed with the first body of the cell culture chamber, and at least two corresponding bubble trap tracks are fluidly connected to each of the at least two bubble traps such that the at least two corresponding bubble trap tracks are disconnected from each other.

[0021] In some aspects, the technology described herein relates to an automated cell engineering system, comprising: a box, the box comprising: a cell culture chamber, the cell culture chamber having at least one bubble trap disposed thereon and further comprising: a first body, the first body forming a first part of the cell culture chamber; a second body, the second body forming a second part of the cell culture chamber, the first body and the second body being configured to be coupled together to thereby form a closed volume; wherein a first non-porous gas-permeable material is disposed on the first body and / or a second non-porous gas-permeable material is disposed on the second body; and wherein the first non-porous gas-permeable material and the first body are formed together, and the second non-porous gas-permeable material and the second body are formed together.

[0022] In some aspects, the technology described herein relates to an automated cell engineering system wherein the cell culture chamber is in a horizontal orientation.

[0023] In some aspects, the technology described herein relates to an automated cell engineering system, wherein the cell culture chamber includes a plurality of bubble traps disposed thereon.

[0024] In some aspects, the technology described herein relates to an automated cell engineering system, wherein the first body comprises a plurality of openings and the second body comprises a plurality of openings.

[0025] In some aspects, the technology described herein relates to an automated cell engineering system, wherein the first non-porous gas-permeable material is positioned on an inner surface of the first body, the first non-porous gas-permeable material being configured to cover the plurality of openings of the first body.

[0026] In some aspects, the technology described herein relates to an automated cell engineering system, wherein the second non-porous gas-permeable material is positioned on an inner surface of the second body, the second non-porous gas-permeable material being configured to cover the plurality of openings of the second body.

[0027] In some aspects, the technology described herein relates to an automated cell engineering system, comprising: a cell culture chamber, the cell culture chamber comprising: a first body, the first body forming a first part of the cell culture chamber and having a plurality of bubble traps disposed thereon, and a bubble trap track, the bubble trap track disposed between and connected to the plurality of bubble traps; a second body, the second body forming a second part of the cell culture chamber, the first body and the second body being configured to be coupled together to thereby form a closed volume; wherein a first non-porous gas-permeable material is disposed on the first body and a second non-porous gas-permeable material is disposed on the second body; and wherein the first non-porous gas-permeable material and the first body are formed together, and the second non-porous gas-permeable material and the second body are formed together.

[0028] In some aspects, the technology described herein relates to an automated cell engineering system wherein the cell culture chamber is in a horizontal orientation.

[0029] In some aspects, the technology described herein relates to an automated cell engineering system wherein the bubble trap track tapers in height and / or width.

[0030] In some aspects, the technology described herein relates to an automated cell engineering system, wherein each of the plurality of bubble traps is formed at opposite ends of the first body, respectively.

[0031] In some aspects, the technology described herein relates to an automated cell engineering system, wherein the first non-porous gas-permeable material is positioned on an inner surface of the first body, the first non-porous gas-permeable material being configured to cover a plurality of openings of the first body.

[0032] In some aspects, the technology described herein relates to an automated cell engineering system, wherein the second non-porous gas-permeable material is positioned on an inner surface of the second body, the second non-porous gas-permeable material being configured to cover the plurality of openings of the second body.

[0033] In some aspects, the technology described herein relates to a cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: at least one body, the at least one body forming a closed volume; and a non-porous gas-permeable material, the non-porous gas-permeable material being disposed on the at least one body such that the non-porous gas-permeable material and the at least one body are formed together; wherein no structure is disposed within the closed volume. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 A cassette for use in an automated cell engineering system as described herein is shown.

[0035] Figure 2 A first view of a cell culture chamber as described herein is shown.

[0036] Figure 3 Shown is a second view of a cell culture chamber as described herein.

[0037] Figure 4 Shown is an interior view of a cell culture chamber as described herein.

[0038] Figure 5 Components of an automated cell engineering system as described herein are shown.

[0039] Figure 6 Shown is a satellite chamber of an automated cell engineering system as described herein.

[0040] Figure 7 Shown is a cross-sectional view of another embodiment of a cell culture chamber for use in an automated cell engineering system as described herein.

[0041] Figure 8 Shown is a second cross-sectional view of a cell culture chamber as described herein.

[0042] Fig. 9 Shown is a third view of a cell culture chamber as described herein.

[0043] Fig.10 Shown is a fourth view of a cell culture chamber as described herein.

[0044] Fig.11 Shown is a fifth view of a cell culture chamber as described herein.

[0045] Fig.12 Shown is a sixth view of a cell culture chamber as described herein.

[0046] Figures 13A-13C Describes the Figure 7 Various embodiments of a cell culture chamber integrally formed with at least one bubble trap and a bubble trap track. DETAILED DESCRIPTION

[0047] It should be understood that the various aspects disclosed herein may be combined in different combinations than those specifically presented in the specification and drawings. It should also be understood that, depending on the example, certain actions or events in any of the processes or methods described herein may be performed in a different order, may be added, combined, or omitted entirely (e.g., all described actions or events may not be necessary to perform the technique).

[0048] The published patents, patent applications, websites, company names and scientific literature mentioned herein are hereby incorporated by reference in their entirety, and the extent of incorporation is as if each is accurately and individually indicated to be incorporated by reference. Any conflict between any reference cited herein and the specific teachings of this specification should be resolved in a manner that is favorable to the latter. Similarly, any conflict between the definition of a word or phrase understood in the art and the definition of a word or phrase specifically taught in this specification should be resolved in a manner that is favorable to the latter.

[0049] As used in this specification, the singular forms "a / an" and "the" specifically also encompass the plural forms of the terms to which they refer, unless the context clearly indicates otherwise. The term "about" is used herein to mean approximately, within a certain range, roughly, or around. When the term "about" is used in conjunction with a numerical range, the term modifies the range by extending the limits above and below the stated numerical values. The term "about" is generally used herein to modify a numerical value above and below the stated value by a variation of 20%.

[0050] The automated cell engineering system 102 as described herein is a fully enclosed automated system for performing activation, transduction, expansion, concentration and / or harvesting steps of cell cultures. The automated cell engineering system (also referred to throughout as an automated bioprocessing unit) provides automated production of cell cultures. As used herein, "cell culture" refers to any suitable cell type, including single cells, as well as multiple cells or cells that can form tissue structures. Exemplary cell cultures include blood cells, skin cells, muscle cells, bone cells, cells from various tissues and organs, and the like. In an embodiment, genetically modified immune cells as described herein can be produced, including CAR T cells. Exemplary automated cell engineering systems are also referred to as or full name System (see, e.g., U.S. Published Patent Application No. 2019 / 0169572, the contents of which are incorporated herein by reference in their entirety).

[0051] The cell culture chamber 101 for use in an automated cell engineering system includes a first body 110 and a second body 120, which are configured to be coupled together, e.g. Figure 1 and Figure 4 As shown. The first body 110 forms the first portion 105 of the cell culture chamber 101, and the second body 120 forms the second portion 115 of the cell culture chamber 101. When coupled together, the first body 110 and the second body 120 form an enclosed volume 325. At least one structural member 130 extends from the first body 110 through the enclosed volume 325 to the second body 120, see Figure 1 and Figure 2-3 The structural member 130 includes a series of cavities 135 that span from the first body 110 to the second body 120 through the enclosed volume 325. The first non-porous breathable material 140 is disposed on the first body 110, and the second non-porous breathable material 145 is disposed on the second body 120, see Figure 4 In one embodiment, the first non-porous air-permeable material 140 may be formed together with the first body 110, and the second non-porous air-permeable material 145 may be formed together with the second body 120, the forming procedure being described in more detail below.

[0052] In another embodiment, the cell culture chamber includes at least one body 110', the at least one body forming an enclosed volume 325'. The at least one body can be formed as a single solid element, the single solid element forming the enclosed volume 325', the enclosed volume further containing a gas permeable material 140' disposed thereon. At least one structural member 130' extends from the at least one body 110' through the enclosed volume 325'. A series of lumens 135' further spans within the enclosed volume of the at least one body. As described herein, the enclosed volume 325' and the gas permeable material 140' are suitable for preparation using a two-shot molding technique.

[0053] As described above, the enclosed volume 325 is formed in the area between the first body 110 and the second body 120. The enclosed volume 325 is configured to contain a cell culture and is configured to be fluid-tight (not allowing liquid to flow into or out of the enclosed volume, but allowing gas flow, as described herein). Figure 4As shown, the first body 110 and the second body 120 are coupled together using a mating element, such as a snap fit, a form fit, or other methods known in the art, for example, at the coupling portion 321. Alternatively, it is contemplated that the first body 110 and the second body 120 may be formed as a unified body without coupling the first body 110 and the second body 120 together.

[0054] The structural member 130 extends from the first body 110 through the closed volume 325 to the second body 120, and includes a first side 132 and a second side 134. The first side 132 is positioned at the first body 110 and formed as a part of the first body, and the second side 134 is positioned at the second body 120 and formed as a part of the second body. The structural member 130 is configured to maintain the integrity of the closed volume 325 while also defining an inner cavity 135 through the closed volume 325. In some embodiments, the inner cavity 135 is configured to receive a valve 136 or a connector, which can be configured to be connected to an automated cell engineering system. Such components allow external machinery such as pumps, pipes, or other machinery to interact with the culture medium and cells in the closed volume 325 and the components of the automated cell engineering system, and provide additional support for them. It is conceivable that a series of inner cavities 135 are configured to receive the valves or connectors. For example, valve 136 can be a trumpet valve, ie, a valve having two positions (open "0" or closed "1") and capable of compressing or pinching the flow path. The lumen can also include a coupler (not shown) allowing connection of various syringes, sampling devices, etc.

[0055] like Figure 2 and 3 As shown, in some embodiments, the cell culture chamber 101 includes at least one or more structural islands 150, which extend from the first body 110 into the enclosed volume 325 to the second body 120. In one embodiment, each of the first body 110 and the second body 120 can include a structural island 150, which extends into the enclosed volume 325. Each structural island 150 provides additional structural support for the cell culture chamber 101 and allows a larger volume to be positioned in the cell culture chamber 101 without including structural integrity. In addition, the structural island 150 also provides a larger surface area for cells to stay in the cell culture chamber, allowing for an increase in growth. The structural island 150 can also provide increased turbulent fluid flow (i.e., non-laminar flow) during filling and moving fluids in the cell culture chamber to help cells mix and / or remove from adherent surfaces. In an embodiment, a lumen can be positioned within the structural island 150 to allow air to flow around and through the cell culture chamber and the box, thereby increasing the potential cooling and temperature management of the box.

[0056] In order to allow gas exchange between the enclosed volume 325 and the external environment, the first body 110 and / or the second body 120 may include a plurality of openings 155, which are configured to allow gas to be exposed to the external environment through the outer surface of the first non-porous gas-permeable material 140 and / or the second non-porous gas-permeable material 145 disposed in the cell culture chamber 101. The plurality of openings 155 can be in a variety of shapes, including geometric, symmetrical, honeycomb, circular, annular, rectangular, oval, square, etc. Additionally, the design of the plurality of openings 155 can be used to provide additional structural support and strength for the first body 110 and the second body 120. For example, the use of a honeycomb structure allows a high ratio of open space to structure, allowing a higher volume of gas to be exchanged while reducing the amount of structure required to support the first body 110 and / or the second body 120.

[0057] The inner surface 311 of the first body 110 of the cell culture chamber 101, also referred to as the culture medium contact surface, and the inner surface 316 or culture medium contact surface of the second body 120 are suitably lined or covered with a non-porous gas-permeable material to prevent water or other liquids from evaporating from the cell culture chamber 101. As used herein, "non-porous gas-permeable material" means any composition, film or material for a gas-permeable cell culture device that allows gas to pass through and enter the cell culture chamber 101, but does not contain holes or holes that allow liquid (i.e., cell culture medium) to pass through or leak. Exemplary non-porous gas-permeable materials include, but are not limited to, silicone, fluoroethylene polypropylene (FEP), polyolefins, ethyl vinyl olefin (EVO) and ethylene vinyl acetate copolymers. As described herein, non-porous gas-permeable materials are suitably helpful in delivering one or more gases including oxygen, nitrogen, CO2, etc. to the cells in the cell culture chamber 101. Additionally, the width of the non-porous gas-permeable material can be reduced or increased to increase or decrease gas exchange through the material.

[0058] like Figure 4As shown, in an embodiment, a first non-porous gas-permeable layer or material 140 is positioned at the inner surface 311 of the first body 110, and a second non-porous gas-permeable material 145 is positioned at the inner surface 316 of the second body 120. During the formation of the cell culture chamber 101, for example, by using molding, the first and second non-porous gas-permeable layers 140 / 145 can be simultaneously produced at the inner surface 311 / 316 by a double shot molding technique. In one embodiment, such a molding technique includes an injection molding technique. For example, the first body 110 and the second body 120 of the cell culture chamber 101 are formed by the first injection of the double shot molding technique. After a short delay allowing the thermoplastic to harden, the core portion of the mold is retracted with a small volume to create space for the second shot of silicone (or any other suitable gas-permeable material) to form the gas-permeable material 140 / 145 in the first body 110 and the second body 120, respectively. The measured thickness of the gas-permeable material 140 / 145 depends on the amount of mold retracted from the first injection, and can therefore be adjusted according to the needs of the end user. Additionally, the non-porous breathable material can be cast onto or into the cell culture chamber 101 to provide the breathable properties described herein. The casting of the non-porous breathable material can be performed using various methods known in the art. By casting the non-porous breathable material onto or into the cell culture chamber 101, the structural strength and integrity of the breathable membrane can be increased while maintaining the breathable properties. The non-porous breathable material can also be coated, sprayed, painted, layered or otherwise arranged on the inner surface 311 / 316 of the first 110 body and the second 120 body. The first and second non-porous breathable layers 140 / 145 can be made of the same or different materials, and in an embodiment, each layer 140 / 145 can include a variety of different materials.

[0059] In an exemplary embodiment, a "portion" of the inner surface, such as 311, comprises a non-porous gas-permeable material. As used herein, "portion" refers to at least about 20% of the cell contact surface or non-cell contact surface being composed of a non-porous gas-permeable material. In an embodiment, both the culture medium contact surfaces 311 and 316 comprise a non-porous gas-permeable material. In such embodiments in which less than 100% of the culture medium contact surface is made of a non-porous gas-permeable material, the remainder of the culture medium contact surface 311 may include other suitable materials, including various plastics (e.g., polypropylene, polystyrene, polystyrene, etc.) that promote adhesion and promote cell adhesion and growth. In embodiments in which less than 100% of the culture medium contact surface is made of a non-porous gas-permeable material, the remainder of the culture medium contact surface 316 may include other suitable materials, including various plastics (e.g., polypropylene, polystyrene, polystyrene, etc.) that provide structural support. In one embodiment, the culture medium contact surface or inner surface 311 of the first body 110 may include a thermoplastic suitable for cell culture, in which case the first body 110 provides a structure and closes the chamber volume without using a gas-permeable material. The first body 110 of the cell culture chamber 101 may suitably be made of a transparent material that allows visual inspection or imaging of the cell culture within the cell culture chamber 101 .

[0060] In an embodiment, at least about 30% of the culture medium contact surface is made of a non-porous, breathable material, more suitably at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or at least about 100% of the culture medium contact surface (i.e., the entire culture medium contact surface) comprises a non-porous, breathable material.

[0061] In an embodiment, the inner surface of the first body 110 and the second body 120 may further include a surface coating, such as a surface coating that activates cells; a surface coating that regulates biological pathways in cells; a surface coating that enhances the growth of cells; a surface coating that increases the transduction efficiency of cells; a surface coating that improves the selection of a certain type of cells; a surface coating that improves the adhesion of cells; a surface coating that inhibits cells; a surface coating with controlled solubility. Therefore, in an embodiment, the non-porous, gas-permeable material that suitably constitutes part or all of the culture medium contact surface may further include a surface coating as described herein. In other embodiments, the remainder of the culture medium contact surface that does not include a non-porous, gas-permeable material may also include a surface coating as described herein. In other embodiments, the entire culture medium contact surface does not include a non-porous, gas-permeable material, but still includes a surface coating as described herein. The adhesion of cells can also provide an opportunity for cell selection to promote the growth of a desired population while not allowing adhesion of an undesirable population.

[0062] As used herein, a "cell-activating surface coating" refers to a material, substrate or composition that induces cell proliferation and / or differentiation.

[0063] As used herein, a "surface coating that modulates a biological pathway in a cell" refers to a material, substrate, or component that causes one or more interactions between molecules in a cell, thereby producing a certain product or change in the cell. For example, such a surface coating can trigger the assembly of new molecules such as fats or proteins, turn genes on and off, or cause cells to move.

[0064] As used herein, a "cell growth enhancing surface coating" refers to a material, substrate, or composition that causes cells to grow faster or in greater numbers than in the absence of the material.

[0065] As used herein, "surface coating that improves cell adhesion" refers to a material, substrate or composition that enables cells to better interact with and attach to a surface, and when adhered to a surface, also interact with other cells.

[0066] As used herein, a "cytostatic surface coating" refers to a material, substrate or composition that causes cells not to grow and / or adhere to a surface in contact with culture medium.

[0067] As used herein, a "surface coating that responds to culture medium conditions" refers to a material, substrate, or component that changes when culture medium conditions change. Exemplary changes include changes in temperature, pH, oxygen levels or concentrations, toxic gas levels, the presence of toxic substances, and include, for example, color changes as a monitoring aid.

[0068] As used herein, "surface coating with controlled solubility" refers to a material, substrate or component that is released from a surface at a specific time or in response to a specific temperature or pH, for example, to obtain controlled release of the contents of the coating.

[0069] Surface treatments (e.g., corona, plasma, etching, etc.) can be used to promote binding (or repulsion) of the coating. This can allow selective surface modification to enable two different cell types to be grown in the same cell culture chamber, or selective unit operations to be performed in the same chamber.

[0070] like Figure 1 As shown, in some embodiments, the cell culture chamber 101 is configured to be positioned as part of a box 100 for use in an automated cell engineering system 102 (see Figure 5As used herein, a "box" refers to a highly independent, removable and replaceable element of the cell engineering system 102, which includes one or more chambers for performing various elements of cell production, and suitably also includes one or more of cell culture medium, activation reagents, carriers, etc.

[0071] Box 100 includes a cryogenic chamber suitable for storing cell culture medium, and a cryogenic chamber suitable for activation, transduction and / or amplification of cell culture including immune cell culture. The cryogenic chamber can be separated from the cryogenic chamber by a thermal barrier. As used herein, "cryogenic chamber" refers to a chamber that is appropriately maintained below room temperature and more appropriately maintained at about 4°C. to about 8°C. to maintain cell culture medium, etc. at a refrigerated temperature. The cryogenic chamber may include a bag or other holder for culture medium, including about 1L, about 2L, about 3L, about 4L or about 5L of fluid. Additional culture medium bags or other fluid sources can be connected to the box externally and connected to the box through an access port. The box further includes one or more fluid passages, which are connected to the cell culture chamber, wherein the fluid passage provides recirculation, waste removal and homogenous gas exchange and distribution of nutrients to the cell culture chamber without interfering with the cells in the cell culture chamber. Box 100 further includes a pump system, which includes one or more pumps for driving fluid through the box as described herein, including a peristaltic pump and one or more valves for controlling the flow through various fluid passages.

[0072] In an embodiment, the cell culture chamber 101 is a non-flexible, contained container. The use of a non-flexible chamber allows the cells to remain in a substantially undisturbed state. As described above, the orientation of the cell culture chamber 101 can allow the immune cell culture to spread throughout the cell culture chamber. Figure 5As shown, cell culture chamber 101 is suitably maintained in a horizontal position parallel to the floor or table, so as to maintain the cell culture in an undisturbed state, allowing the cell culture to spread over a large area of ​​the cell culture chamber. In an embodiment, the total thickness of the cell culture chamber 101 is on the order of about 0.5cm to about 5cm. Suitably, the volume of the cell culture chamber can be between about 0.50ml and about 1L, more suitably about 0.5 to about 900ml, or the volume of the cell culture chamber is about 825ml. Suitably, the volume of the cell culture chamber can be between about 0.5ml and about 500ml, between about 10ml and about 300ml, or more suitably about 50ml and about 200ml, or the volume of the cell culture chamber is about 180ml. Use lower chamber height (less than 5cm, suitably less than 4cm, less than 3cm or less than 2cm) to allow effective culture medium and gas exchange near the cell. Port is configured to allow mixing without disturbing cells by the recirculation of fluid. Static vessels of greater height can create concentration gradients, so that oxygen and fresh nutrients are limited in the vicinity of cells. Through controlled flow dynamics, medium exchange can be performed without cell disturbance. Medium can be removed from another chamber (where cells are not present) without risk of cell loss.

[0073] As described herein, in exemplary embodiments, the cartridge is pre-filled with one or more of cell culture, culture medium, activation reagents and / or carriers, including any combination of these. In further embodiments, these various elements may be added later via a suitable injection port or the like.

[0074] The cell engineering system 102 may include components such as gas control seals, warm zones, actuators, pivots for rocking or tilting the cell engineering system as needed, and low temperature zones for maintaining a low temperature chamber. The cell engineering system 102 may also include a user interface, which may include a barcode reader and the ability to receive input using a touch pad or other similar device.

[0075] like Figure 1 As shown, the box 100 can further include additional chamber volumes or satellite volumes 165 for increasing the working volume of the cell culture chamber by providing additional volume for culture medium and other working fluids, or can be used to contain the cell culture or a portion of the cell culture during activation, transduction, expansion, concentration and / or harvesting steps of the cell culture. Suitably, the satellite volume 165 is oriented substantially perpendicular to the cell culture chamber 101 and is fluidly connected to the cell culture chamber so that culture medium is exchanged with the culture chamber without disturbing the cell culture. In exemplary embodiments, the satellite volume is a bag, and in other embodiments, the satellite volume is a non-curved chamber, such as Figure 1As shown. In an embodiment, the satellite volume is between about 0.50ml and about 1L, or between about 10ml and about 800ml, or between about 100ml and about 500ml, or between about 100ml and about 300ml, more preferably between about 150ml and about 200ml. In an embodiment, the satellite volume is further configured to allow culture medium to be removed without losing the cells of the immune cell culture. That is, when the culture medium exchange between the satellite volume and the cell culture chamber is carried out in such a way that the cell is not disturbed and is not removed from the cell culture chamber. In certain embodiments, the satellite volume can additionally include a port, an inlet, an incision, an inner cavity or a passage that allows airflow to be transmitted from the outside of the box to the inside of the box.

[0076] like Figure 1 As shown, the satellite volume 165 suitably includes a satellite volume lumen 170 , which is a passageway of the satellite volume 165 that allows gas to flow through the satellite volume 165 while still maintaining the fluid integrity of the satellite volume 165 . Figure 5 The orientation of the cell culture chamber 101 within the cell engineering system 102 is shown, wherein the cell culture chamber is oriented substantially horizontally and the satellite volume 165 is oriented substantially vertically, wherein the housing 500 of the cell engineering system is a removable cover surrounding the system. As shown, the satellite volume lumen 170 is positioned so that air 502 can move through the lumen to the center of the cell engineering system 102 and the cassette 100 to provide increased airflow and temperature control and regulation.

[0077] The horizontal position of the cell culture chamber 101 within the cell engineering system 102 is positioned by rotating the entire cell engineering system 102 by approximately 90 degrees, so that the originally vertically oriented cell culture chamber 101 (see Figure 1 , the position on the box 100) becomes a horizontal position, such as Figure 5 This allows the cell culture chamber 101 to utilize a larger volume because the surface of the cassette 100 at the location where the cell culture chamber 101 is located is larger than the top section of the cassette.

[0078] Figure 6 A temperature gradient diagram of satellite volume 165 is shown, wherein satellite volume interior cavity 170 is shown.

[0079] In an embodiment, a cell culture chamber 601 for use in an automated cell engineering system includes a first body 610 and a second body 620, which are configured to be coupled together, for example, Figure 7-12The first body 610 forms a first portion 612 of the cell culture chamber 601, and the second body 620 forms a second portion 622 of the cell culture chamber 601. When coupled together, the first body 610 and the second body 620 form a closed volume 625, as shown in FIG. Figure 7 and 8 The cell culture chamber 601 can be formed without placing any structure in the closed volume 625 to allow for an increased volume of culture medium in the closed volume 625, such as Figure 7 and 8 The first non-porous breathable material 640 may be disposed on the first body 610, and the second non-porous breathable material 645 may be disposed on the second body 620, see Figure 7 In one embodiment, using the formation procedures previously described herein, a first non-porous gas-permeable material 640 may be formed with the first body 610, and a second non-porous gas-permeable material 645 may be formed with the second body 620. In an embodiment, the first body 610 may not include the first non-porous gas-permeable material 640, while the second body 620 includes the second non-porous gas-permeable material 645. Alternatively, the first body 610 includes the first non-porous gas-permeable material 640, while the second body 620 may not include the second non-porous gas-permeable material 645. In an embodiment where only one of the first body 610 or the second body 620 includes the non-porous gas-permeable material 640 / 645, the advantage of maximizing the oxygen level within the cell culture chamber 601 may be achieved. In another embodiment, the first body 610 may include the first non-porous gas-permeable material 640, while the second body 620 also includes the second non-porous gas-permeable material 645.

[0080] As described above, the enclosed volume 625 is formed in the area between the first body 610 and the second body 620. The enclosed volume 625 can be configured to contain a cell culture and can be configured to be liquid-tight (not allowing liquid to flow into or out of the enclosed volume, but allowing gas flow, as described herein), unless intended through a designated valve, port, etc. For example, the input / output port 636 can be connected to other elements of the cartridge 100 or cell engineering system 102 to exchange culture medium or other reagents with the enclosed volume 625 and to allow harvesting of the cell culture within the enclosed volume 625. Figure 7 and 8 As shown, the first body 610 and the second body 620 are coupled together using mating elements, such as at the coupling portion 630, such as a snap fit, a form fit, or other methods known in the art. Fig.10 and 11A top perspective view and a bottom perspective view of the cell culture chamber 601 are depicted, respectively, wherein the first body 610 and the second body 620 are coupled together. Additionally, it is contemplated that the first body 610 and the second body 620 may be formed as a unified body without coupling the first body 610 and the second body 620 together.

[0081] Fig. 9 Exemplary dimensions of the cell culture chamber 601 are provided in a top view. For example, the length of the cell culture chamber 601 (i.e., the length of the first body 610 and / or the second body 620) can be approximately 360 mm. In an embodiment, the length of the cell culture chamber can be between 200-600 mm. In an embodiment, the length of the cell culture chamber can be between 300-400 mm. The width of the cell culture chamber 601 (i.e., the width of the first body 610 and / or the second body 620) can be approximately 90 mm. In an embodiment, the width of the cell culture chamber 601 can be between 50-150 mm. In an embodiment, the width of the cell culture chamber can be between 75-100 mm.

[0082] In order to allow gas exchange between the enclosed volume 625 and the external environment, the first body 610 and / or the second body 620 may include a plurality of openings 655 configured to allow gas to be exposed to the external environment through the outer surface of the first non-porous gas-permeable material 640 and / or the second non-porous gas-permeable material 645 disposed in the cell culture chamber 601, such as Figure 7-11 For example, Figure 7-10 A plurality of openings 655 are shown formed in the first body 610 or disposed through the first body or through the first body in a staggered arrangement. Fig.11A plurality of openings 655 are shown formed in the second body 620 or disposed through the second body or in a staggered arrangement. The plurality of openings 655 can take a variety of shapes, including geometric shapes (e.g., hexagons, heptagons, octagons, etc.), symmetrical shapes, honeycombs, circles, rings, rectangles, ellipses, squares, etc. Additionally, the design of the plurality of openings 655 can be used to provide additional structural support and strength for the first body 610 and the second body 620. For example, the use of a honeycomb structure allows a high ratio of open space to structure, allowing a higher volume of gas to be exchanged while reducing the amount of structure required to support the first body 610 and / or the second body 620. In an embodiment, the plurality of openings 655 can be formed in the first body 610 and / or the second body 620 or through the first body and / or the second body / arranged on the first body and / or the second body in a sequential arrangement. In an embodiment, the first body 610 and / or the second body 620 can alternatively be formed of a solid thermoplastic in which the plurality of openings 655 are not formed. At least about 30% of the first body 610 and / or the second body 620 may be covered by the plurality of openings 655, more suitably at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or at least about 100% (i.e., all).

[0083] The inner surface 611 of the first body 610 of the cell culture chamber 601, also referred to as the culture medium contact surface, and the inner surface 621 or culture medium contact surface of the second body 620 are suitably lined or covered with a non-porous gas-permeable material to prevent water (i.e., in liquid form) or other liquids (e.g., cell culture medium) from evaporating and flowing out of the cell culture chamber 601, while allowing gases such as oxygen, nitrogen, carbon dioxide, etc. to pass through. As used herein, "non-porous gas-permeable material" means any composition, coating, film or material for a gas-permeable cell culture device that allows gas to pass through and enter the cell culture chamber 601, but does not contain holes or holes that allow liquid (i.e., cell culture medium) to pass through or leak. Exemplary non-porous gas-permeable materials include, but are not limited to, silicone, fluoroethylene polypropylene (FEP), polyolefins, ethyl vinyl olefin (EVO) and ethylene vinyl acetate copolymers. As described herein, the non-porous gas-permeable material is suitably helpful in delivering one or more gases including oxygen, nitrogen, CO2, etc. to the cells in the cell culture chamber 601. Additionally, the thickness of the non-porous gas-permeable material can be reduced or increased to increase or decrease gas exchange through the material.

[0084] like Figure 7As shown, in an embodiment, a first non-porous gas-permeable material 640 is positioned at the inner surface 611 of the first body 610, and a second non-porous gas-permeable material 645 is positioned at the inner surface 621 of the second body 620. During the formation of the cell culture chamber 601, for example, by using molding, the first and second non-porous gas-permeable materials 640 / 645 can be simultaneously produced at the inner surface 611 / 621 by a double shot molding technique. In one embodiment, this molding technique includes an injection molding technique. For example, the first body 610 and the second body 620 of the cell culture chamber 601 are formed by the first injection of the double shot molding technique. After a short delay allowing the thermoplastic to harden, the core portion of the mold is retracted with a small volume to create space for the second shot of silicone (or any other suitable gas-permeable material) to form gas-permeable materials 640 / 645 respectively arranged on the first body 610 and the second body 620. The measured thickness of the gas-permeable material 640 / 645 depends on the amount of mold retracted from the first injection, and can therefore be adjusted according to the needs of the end user. Additionally, the non-porous breathable material can be cast onto the cell culture chamber 601 or therein to provide the breathable properties described herein. The casting of the non-porous breathable material can be carried out using various methods known in the art. By casting the non-porous breathable material onto the cell culture chamber 601 or therein, the structural strength and integrity of the breathable membrane can be increased while maintaining the breathable properties. The non-porous breathable material can also be coated, sprayed, painted, layered or otherwise arranged on the inner surface 611 / 621 of the first 610 body and the second 620 body. The first and second non-porous breathable layers 640 / 645 can be made of the same or different materials, and in an embodiment, each layer 640 / 645 can include a variety of different materials.

[0085] In an exemplary embodiment, "a portion" of the inner surface, such as 611, comprises a non-porous, gas-permeable material. As used herein, "a portion" refers to at least about 20% of the cell contact surface or non-cell contact surface being composed of a non-porous, gas-permeable material. In an embodiment, both the culture medium contact surfaces 611 and 621 comprise a non-porous, gas-permeable material. In such embodiments in which less than 100% of the culture medium contact surface is made of a non-porous, gas-permeable material, the remainder of the culture medium contact surface 611 may comprise other suitable materials, including various plastics (e.g., polypropylene, polystyrol, polystyrene, etc.) that promote adhesion and promote cell adhesion and growth. In embodiments in which less than 100% of the culture medium contact surface is made of a non-porous, gas-permeable material, the remainder of the culture medium contact surface 621 may include other suitable materials, including various plastics (e.g., polypropylene, polystyrol, polystyrene, etc.) that provide structural support. In an embodiment, the inner surface 611 of the first body 610 may lack a non-porous, gas-permeable material so that no (i.e., 0%) inner surface 611 is covered. In an embodiment, the inner surface 621 of the second body 620 may lack the non-porous breathable material such that none (ie, 0%) of the inner surface 621 is covered.

[0086] In an embodiment, at least about 30% of the culture medium contact surface is made of a non-porous, breathable material, more suitably at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or at least about 100% of the culture medium contact surface (i.e., the entire culture medium contact surface) comprises a non-porous, breathable material.

[0087] In an embodiment, the inner surface of the first body 610 and the second body 620 may further include a surface coating, such as a surface coating that activates cells; a surface coating that regulates biological pathways in cells; a surface coating that enhances the growth of cells; a surface coating that increases the transduction efficiency of cells; a surface coating that improves the selection of a certain type of cells; a surface coating that improves the adhesion of cells; a surface coating that inhibits cells; a surface coating with controlled solubility. Therefore, in an embodiment, a non-porous, gas-permeable material that suitably constitutes part or all of the culture medium contact surface may further include a surface coating as described herein. In other embodiments, the remainder of the culture medium contact surface that does not include a non-porous, gas-permeable material may also include a surface coating as described herein. In other embodiments, the entire culture medium contact surface does not include a non-porous, gas-permeable material, but still includes a surface coating as described herein. The adhesion of cells can also provide an opportunity for cell selection to promote the growth of a desired population while not allowing adhesion of an undesirable population.

[0088] In an embodiment, the cell culture chamber 601 is a non-flexible contained chamber. The use of the non-flexible chamber allows the cells to maintain a substantially undisturbed state. As described above, the orientation of the cell culture chamber 601 can allow the cell culture to spread throughout the entire cell culture chamber 601. In an embodiment, the total thickness of the cell culture chamber 601 is on the order of about 0.5cm to about 5cm. Suitably, the volume of the cell culture chamber can be between about 0.50ml and about 1L, more suitably about 0.5 to about 900ml, or the volume of the cell culture chamber is about 825ml. Suitably, the volume of the cell culture chamber can be between about 0.5ml to about 500ml, between about 10ml to about 300ml, or more suitably about 50ml to about 200ml, or the volume of the cell culture chamber is about 180ml. Use a lower chamber height (less than 5cm, suitably less than 4cm, less than 3cm or less than 2cm) to allow effective culture medium and gas exchange near the cell. The ports are configured to allow mixing by recirculation of fluid without disturbing the cells. Static vessels of greater height can create concentration gradients so that oxygen and fresh nutrients are limited in the vicinity of the cells. Through controlled flow dynamics, medium exchange can be performed without cell disturbance. Medium can be removed from an additional chamber (where cells are not present) without risk of cell loss.

[0089] In some embodiments, the cell culture chamber 601 may include an input / output port 636 or a coupling, which may be configured to be connected to an automated cell engineering system. Such components allow external machinery such as pumps, pipelines, connectors, or other machinery to interact with the culture medium and cells in the closed volume 625 and the components of the automated cell engineering system and provide culture medium and cells or additional support. In an embodiment, the input / output port may be configured to receive a valve or a coupling. For example, a valve may be a trumpet valve, i.e., a valve with two positions (open "0" or close "1"), or a valve that can compress or clamp a flow path. The inlet / outlet port 636 may also include a coupling (not shown), allowing various syringes, sampling devices, etc. to be connected.

[0090] In an embodiment, the top surface 613 of the first body 610 includes or is integrally formed with at least one bubble trap 700. The at least one bubble trap 700 is configured to collect any bubbles that may form within the enclosed volume 625 of the cell culture chamber 601 and rise to the top of the culture medium. Fig. 9As shown in the figure, at least two bubble catchers 700A, 700B are respectively arranged on each end of the top surface 613 of the first body 610 (one catcher 700A, 700B at each end), wherein the bubble catcher track 710 is included together with the first body 610 or formed integrally therewith, and the bubble catcher track 710 further extends between each bubble catcher 700A, 700B and fluidically connects each of the bubble catchers. Figures 13A-13B Various embodiments are described in which at least two bubble traps 700A, 700B and a bubble trap track 710 may be formed. The width 722 of the bubble trap track 710 may vary along its length such that the bubble trap track 710 widens toward its ends 712 (i.e., closer to the bubble trap 700) and narrows toward the middle of its length (i.e., at or near the center of the top surface 613). Thus, the bubble trap track 710 may taper outwardly from the middle of the bubble trap track 710 to the ends of the bubble trap track 710 adjacent to the at least two bubble traps 700A, 700B. For example, the width 722 of the bubble trap track 710 can be such that there is almost no width / space at the center of the bubble trap track 710 (i.e., near the center of the plane of the inner surface 611), and the width 722 gradually increases (becomes smaller in width) along the length of the bubble trap track 710 (i.e., toward the at least one bubble trap 700), increasing by up to 1 mm or more toward the end 712 of the bubble trap track 710 where it meets / connects with the at least one bubble trap 700, thereby forming a portion of the additional volume 711, for example, as Fig.13A . In an embodiment, the width 722 of the bubble trap track 710 can increase from any value between 0.01 mm and 10 mm, starting from the center of the bubble trap track 710 and ending toward the end 712 of the bubble trap track 710 connected to at least one bubble trap 700, respectively. In an embodiment, the bubble trap track 710 can taper inward from the middle of the bubble trap track 710 to the end of the bubble trap track 710 adjacent to at least two bubble traps 700A, 700B (not shown). In an embodiment, the at least two bubble trap tracks 710A, 710B can be fluidly connected to each bubble trap of the at least two bubble traps 700A, 700B, such that each bubble trap 700A, 700B is connected to one bubble trap track 710A, 710B, and each bubble trap track 710A, 710B is disconnected from each other (i.e., there is space between the bubble trap track sections), for example, as Fig. 13C The bubble trap tracks 710A, 710B may taper outward from the end of the bubble trap track farthest from the bubble trap 700A, 700B, for example. Fig.13A In an embodiment, each bubble trap track 710A, 710B may taper inwardly from the end of the bubble trap track farthest from the bubble trap 700A, 700B (not shown). In an embodiment, the bubble trap track 710A, 710B may have a uniform width throughout its length, from the end of the bubble trap track farthest from the bubble trap 700A, 700B to the end 712 of the bubble trap track 710 closest to or fluidly connected to the bubble trap 700A, 700B, for example, as shown in FIG. Fig. 13B As shown in .

[0091] In an embodiment, at least one bubble trap 700 is fluidly connected to a bubble trap track 710. In an embodiment, at least two bubble traps 700A, 700B may each be fluidly connected to at least two bubble trap tracks 710A and 710B ( Fig. 13C ), forming at least two additional volumes 711A, 711B. In an embodiment, a plurality of bubble traps 700 may be fluidly connected to a plurality of bubble trap tracks 710. In an embodiment, a plurality of bubble traps 700 may be fluidly connected to a bubble trap track 710. In an embodiment, a plurality of bubble traps 700 may be fluidly connected to a bubble trap track 710. In an embodiment, a plurality of bubble traps 700 may be fluidly connected to a single bubble trap track 710. For example, the first body 610 may include or be integrally formed with a plurality of bubble traps 700, and a bubble trap track 710 fluidly connected to each of the plurality of bubble traps 700. In an embodiment, a plurality of bubble traps 700 may be fluidly connected to respective bubble trap tracks 710 such that the bubble trap tracks 710 are disconnected from one another.

[0092] The bubble trap rail 710 / 710A / 710B may also be configured to have a gradually increasing height. For example, Figure 8 and 12 The bubble trap track 710 is shown forming a slight gradient 720 on the inner surface 611 of the first body 610 so as to direct bubbles collected near the center of the bubble trap track 710 (i.e., near the center of the plane of the inner surface 611 of the first body 610) outwardly toward at least one bubble trap 700. Fig.12, the gradient 720 of the bubble trap track 710 can be such that there is almost no height / space starting at the center of the bubble trap track 710 (i.e., near the center of the plane of the inner surface 611), and the height gradually increases along the length of the bubble trap track 710 (i.e., the height gradually decreases toward the at least one bubble trap 700), increasing by up to 1 mm or more toward the end 712 of the bubble trap track 710 that meets / connects to the at least one bubble trap 700, thereby forming a portion of the additional volume 711. Therefore, the bubble trap track 710 can gradually decrease from the middle of the bubble trap track 710 to the end of the bubble trap track 710 adjacent to the at least two bubble traps 700A, 700B. In an embodiment, the gradient 720 of the bubble trap track 710 can increase from any value between 0.01 mm and 10 mm in height on the inner surface 611, starting from the center of the bubble trap track 710, and ending toward the end 712 of the bubble trap track 710 connected to the at least one bubble trap 700, respectively. The at least one bubble trap 700 and the bubble trap track 710 may further form an additional volume 711 within the enclosed volume 625 for collecting / holding bubbles as described herein. This additional volume 711 is structured solely for collecting bubbles formed within the culture medium disposed within the enclosed volume 625, such that the fluid height of the culture medium within the enclosed volume 625 is maintained below 20 mm (i.e., the culture medium does not enter the bubble trap track 710 nor the at least one bubble trap 700). The curved long profile (i.e., gradient 720) of the bubble trap track 710 helps guide bubbles along the length of the bubble trap track 710 into the at least one bubble trap 700 located at the end of the bubble trap track 710.

[0093] In an embodiment, at least one bubble trap 700 described herein may include or be integrally formed with the bottom surface 623 of the second body 620 (not shown). A plurality of bubble traps 700 may be disposed on each end of the bottom surface 623 of the second body, wherein a bubble trap track 710 has a gradient 720 running between and connected to each bubble trap 700. The width of the bubble trap track 710 may vary along its length, and its gradient 720 may vary, as previously described herein. This configuration will allow at least one bubble trap 700 and bubble trap track 710 to collect bubbles formed within the culture medium in the event that the cell culture chamber 601 is flipped (i.e., axially rotated) so that the second body 620 faces upward. The bubble trap 700 / 700A / 700B and the bubble trap track 710 / 710A / 710B may include or be integrally formed with the second body 620 in a similar manner as previously described with respect to the first body 610.

[0094] Although various embodiments according to the present invention have been described above, it should be understood that the embodiments are presented only by illustration and example and in a non-restrictive manner. It is obvious to those skilled in the relevant art that various changes can be made to form and details therein without departing from the spirit and scope of the present invention. Therefore, the breadth and scope of the present invention should not be limited to any one of each embodiment discussed herein, and each reference cited herein can be used in combination with the features of any other embodiment. All patents and public documents discussed herein are incorporated herein by reference as a whole.

[0095] Embodiments of the present disclosure include the following examples.

[0096] Example 1. A cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: a first body, the first body forming a first part of the cell culture chamber; a second body, the second body forming a second part of the cell culture chamber, the first body and the second body being configured to be connected together to form a closed volume, wherein a first non-porous gas-permeable material is disposed on the first body; and wherein the first non-porous gas-permeable material and the first body are formed together.

[0097] Example 2. The cell culture chamber according to Example 1, further comprising a second non-porous gas-permeable material, which is disposed on the second body; and wherein the second non-porous gas-permeable material and the second body are formed together.

[0098] Example 3. The cell culture chamber according to Example 1, further comprising at least one bubble trap, wherein the at least one bubble trap is integrally formed with the cell culture chamber.

[0099] Example 4. The cell culture chamber according to Example 1, further comprising one or more structural islands.

[0100] Example 5. A cell culture chamber according to Example 2, wherein the first body and the second body are connected together using mating elements.

[0101] Example 6. A cell culture chamber according to Example 2, wherein the first body comprises a plurality of openings configured to allow gas to be exposed to the external environment through the outer surface of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

[0102] Example 7. A cell culture chamber according to Example 6, wherein the plurality of openings comprises a geometric shape, a hexagonal shape, a heptagonal shape, an octagonal shape, a symmetrical shape, a honeycomb shape, a circular shape, a ring shape, a rectangular shape, an elliptical shape or a square shape.

[0103] Example 8. The cell culture chamber according to Example 6, wherein the first non-porous gas-permeable material is positioned on the inner surface of the first body, and the first non-porous gas-permeable material is configured to cover the plurality of openings of the first body.

[0104] Example 9. A cell culture chamber according to Example 2, wherein the structural section of the second body includes a plurality of openings, and the plurality of openings are configured to allow gas to be exposed to the external environment through the outer surface of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

[0105] Example 10. A cell culture chamber according to Example 9, wherein the plurality of openings comprises a geometric shape, a hexagonal shape, a heptagonal shape, an octagonal shape, a symmetrical shape, a honeycomb shape, a circular shape, a ring shape, a rectangular shape, an elliptical shape or a square shape.

[0106] Example 11. The cell culture chamber according to Example 10, wherein the second non-porous gas-permeable material is positioned on the inner surface of the first body, and the second non-porous gas-permeable material is configured to cover the plurality of openings of the second body.

[0107] Example 12. A cell culture chamber according to Example 3, wherein the at least one bubble trap is integrally formed with the first body of the cell culture chamber, and the cell culture chamber further comprises a bubble trap track, which is integrally formed with the first body and fluidically connected to the at least one bubble trap.

[0108] Example 13. The cell culture chamber of Example 12, wherein at least two bubble traps are integrally formed with the first body of the cell culture chamber, and the bubble trap track is fluidly connected to the at least two bubble traps.

[0109] Example 14. A cell culture chamber according to Example 13, wherein the width of the bubble trap track gradually decreases.

[0110] Example 15. A cell culture chamber according to Example 13, wherein the height of the bubble trap track gradually decreases.

[0111] Example 16. A cell culture chamber according to Example 12, wherein at least two bubble traps are integrally formed with the first body of the cell culture chamber, and at least two corresponding bubble trap tracks are fluidly connected to each of the at least two bubble traps, so that the at least two corresponding bubble trap tracks are disconnected from each other.

[0112] Example 17. An automated cell engineering system, the automated cell engineering system comprising: a box, the box including: a cell culture chamber, the cell culture chamber having at least one bubble catcher disposed thereon and further comprising: a first body, the first body forming a first part of the cell culture chamber; a second body, the second body forming a second part of the cell culture chamber, the first body and the second body being configured to be connected together to thereby form a closed volume; wherein a first non-porous gas-permeable material is disposed on the first body and / or a second non-porous gas-permeable material is disposed on the second body; and wherein the first non-porous gas-permeable material and the first body are formed together, and the second non-porous gas-permeable material and the second body are formed together.

[0113] Example 18. According to the automated cell engineering system of Example 17, the cell culture chamber is in a horizontal orientation.

[0114] Example 19. An automated cell engineering system according to Example 17, wherein the cell culture chamber comprises a plurality of bubble traps disposed thereon.

[0115] Example 20. An automated cell engineering system according to Example 17, wherein the first body comprises a plurality of openings and the second body comprises a plurality of openings.

[0116] Example 21. An automated cell engineering system according to Example 20, wherein the first non-porous, gas-permeable material is positioned on the inner surface of the first body, and the first non-porous, gas-permeable material is configured to cover the multiple openings of the first body.

[0117] Example 22. An automated cell engineering system according to Example 20, wherein the second non-porous, gas-permeable material is positioned on the inner surface of the second body, and the second non-porous, gas-permeable material is configured to cover the plurality of openings of the second body.

[0118] Example 23. An automated cell engineering system, the automated cell engineering system comprising: a cell culture chamber, the cell culture chamber comprising: a first body, the first body forming the first part of the cell culture chamber and having a plurality of bubble catchers disposed thereon, and a bubble catcher track, the bubble catcher track being disposed between the plurality of bubble catchers and connected to the plurality of bubble catchers; a second body, the second body forming the second part of the cell culture chamber, the first body and the second body being configured to be connected together to thereby form a closed volume; wherein a first non-porous gas-permeable material is disposed on the first body and a second non-porous gas-permeable material is disposed on the second body; and wherein the first non-porous gas-permeable material and the first body are formed together, and the second non-porous gas-permeable material and the second body are formed together.

[0119] Example 24. An automated cell engineering system according to Example 23, wherein the cell culture chamber is in a horizontal orientation.

[0120] Example 25. An automated cell engineering system according to Example 24, wherein the height and / or width of the bubble trap track gradually decreases.

[0121] Example 26. An automated cell engineering system according to Example 25, wherein each of the plurality of bubble traps is formed at opposite ends of the first body, respectively.

[0122] Example 27. An automated cell engineering system according to Example 26, wherein the first non-porous, gas-permeable material is positioned on the inner surface of the first body, and the first non-porous, gas-permeable material is configured to cover the plurality of openings of the first body.

[0123] Example 28. An automated cell engineering system according to Example 27, wherein the second non-porous, gas-permeable material is positioned on the inner surface of the second body, and the second non-porous, gas-permeable material is configured to cover the plurality of openings of the second body.

[0124] Example 29. A cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: at least one body, the at least one body forming a closed volume; and a non-porous, gas-permeable material, the non-porous, gas-permeable material being disposed on the at least one body such that the non-porous, gas-permeable material and the at least one body are formed together; wherein no structure is disposed within the closed volume.

Claims

1. A cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: a first body, the first body forming a first portion of the cell culture chamber; a second body, the second body forming a second portion of the cell culture chamber, the first body and the second body being configured to be coupled together to thereby form an enclosed volume; wherein the first non-porous breathable material is placed on the first body; and The first non-porous breathable material and the first main body are formed together.

2. The cell culture chamber according to claim 1, further comprising a second non-porous gas-permeable material, wherein the second non-porous gas-permeable material is disposed on the second body; and The second non-porous breathable material and the second main body are formed together. 3 . The cell culture chamber according to claim 1 , further comprising at least one bubble trap, wherein the at least one bubble trap is integrally formed with the cell culture chamber. The cell culture chamber of claim 1 , further comprising one or more structural islands.

5. The cell culture chamber of claim 2, wherein the first body and the second body are coupled together using mating elements.

6. The cell culture chamber according to claim 2, wherein the first body comprises a plurality of openings configured to allow gas to be exposed to the external environment through an outer surface of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

7. The cell culture chamber of claim 6, wherein the plurality of openings comprises a geometric shape, a hexagonal shape, a heptagonal shape, an octagonal shape, a symmetrical shape, a honeycomb shape, a circular shape, a ring shape, a rectangular shape, an elliptical shape, or a square shape. 8 . The cell culture chamber according to claim 6 , wherein the first non-porous gas-permeable material is positioned on an inner surface of the first body, the first non-porous gas-permeable material being configured to cover the plurality of openings of the first body.

9. The cell culture chamber according to claim 2, wherein the structural section of the second body comprises a plurality of openings, and the plurality of openings are configured to allow gas to be exposed to the external environment through the outer surface of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

10. The cell culture chamber of claim 9, wherein the plurality of openings comprises a geometric shape, a hexagonal shape, a heptagonal shape, an octagonal shape, a symmetrical shape, a honeycomb shape, a circular shape, a ring shape, a rectangular shape, an elliptical shape, or a square shape. 11 . The cell culture chamber according to claim 10 , wherein the second non-porous gas-permeable material is positioned on an inner surface of the first body, the second non-porous gas-permeable material being configured to cover the plurality of openings of the second body.

12. A cell culture chamber according to claim 3, wherein the at least one bubble trap is integrally formed with the first body of the cell culture chamber, and the cell culture chamber further comprises a bubble trap track, which is integrally formed with the first body and fluidly connected to the at least one bubble trap.

13. The cell culture chamber of claim 12, wherein at least two bubble traps are integrally formed with the first body of the cell culture chamber, and the bubble trap track is fluidly connected to the at least two bubble traps.

14. The cell culture chamber of claim 13, wherein the bubble trap track tapers in width.

15. The cell culture chamber of claim 13, wherein the bubble trap track tapers in height.

16. A cell culture chamber according to claim 12, wherein at least two bubble traps are integrally formed with the first body of the cell culture chamber, and at least two corresponding bubble trap tracks are fluidly connected to each of the at least two bubble traps, so that the at least two corresponding bubble trap tracks are disconnected from each other.

17. An automated cell engineering system, comprising: A box, comprising: A cell culture chamber having at least one bubble trap disposed thereon and further comprising: a first body, the first body forming a first portion of the cell culture chamber; a second body, the second body forming a second portion of the cell culture chamber, the first body and the second body being configured to be coupled together to thereby form an enclosed volume; wherein a first non-porous breathable material is disposed on the first body and / or a second non-porous breathable material is disposed on the second body; and The first non-porous air-permeable material is formed together with the first main body, and the second non-porous air-permeable material is formed together with the second main body.

18. The automated cell engineering system of claim 17, wherein the cell culture chamber is in a horizontal orientation.

19. The automated cell engineering system of claim 17, wherein the cell culture chamber comprises a plurality of bubble traps disposed thereon.

20. The automated cell engineering system of claim 17, wherein the first body comprises a plurality of openings, and the second body comprises a plurality of openings.

21. The automated cell engineering system of claim 20, wherein the first non-porous gas-permeable material is positioned on an inner surface of the first body, the first non-porous gas-permeable material being configured to cover the plurality of openings of the first body.

22. The automated cell engineering system of claim 20, wherein the second non-porous gas-permeable material is positioned on an inner surface of the second body, the second non-porous gas-permeable material being configured to cover the plurality of openings of the second body.

23. An automated cell engineering system, comprising: A cell culture chamber, comprising: a first body forming a first portion of the cell culture chamber and having a plurality of bubble traps disposed thereon, and a bubble trap track disposed between and connected to the plurality of bubble traps; a second body, the second body forming a second portion of the cell culture chamber, the first body and the second body being configured to be coupled together to thereby form an enclosed volume; wherein a first non-porous breathable material is disposed on the first body and a second non-porous breathable material is disposed on the second body; and The first non-porous air-permeable material is formed together with the first main body, and the second non-porous air-permeable material is formed together with the second main body.

24. The automated cell engineering system of claim 23, wherein the cell culture chamber is in a horizontal orientation.

25. The automated cell engineering system according to claim 24, wherein the height and / or width of the bubble trap track gradually decreases. 26 . The automated cell engineering system according to claim 25 , wherein each of the plurality of bubble traps is formed at opposite ends of the first body, respectively.

27. The automated cell engineering system of claim 26, wherein the first non-porous gas-permeable material is positioned on an inner surface of the first body, the first non-porous gas-permeable material being configured to cover a plurality of openings of the first body.

28. The automated cell engineering system of claim 27, wherein the second non-porous gas-permeable material is positioned on an inner surface of the second body, the second non-porous gas-permeable material being configured to cover the plurality of openings of the second body.

29. A cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: at least one body forming an enclosed volume; and a non-porous, breathable material disposed on the at least one body such that the non-porous, breathable material and the at least one body are formed together; Wherein no structure is placed in the closed volume.

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