Cell culture chamber for increasing cell yield

Abstract

The present disclosure provides a cell culture chamber for use in an automated cell engineering system, particularly a cell culture chamber that results in an increase in cell yield. The cell culture chamber can include a first body that forms a first portion of the cell culture chamber. The cell culture chamber can include a second body that forms a second portion of the cell culture chamber, and the first body and the second body are configured to be coupled to each other to form a sealed volume, and a first non-porous gas-permeable material is disposed on the first body. The first non-porous gas-permeable material and the first body are formed together. A second non-porous gas-permeable material can be disposed on the second body, and the second non-porous gas-permeable material and the second body can 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 a cell culture chamber for use in an automated cell engineering system, particularly a cell culture chamber that results in increased cell yield.

Background Art

[0002] As the clinical introduction of advanced cell therapies is expected to accelerate, much attention is being focused on the basic manufacturing strategies that will enable these therapies to benefit patients worldwide. Although cell therapies hold great clinical promise, their high manufacturing costs relative to reimbursement rates pose a significant barrier to commercialization. Therefore, due to the need for cost-effectiveness, process efficiency, and product consistency, efforts are being made to automate many areas of cell therapy, particularly in T cell immunotherapy.

[0003] Integrating cell activation, transduction, and expansion into commercial manufacturing platforms is important for the translation of these important immunotherapies to a wide patient population. To apply these life-saving treatments to patients worldwide, it is necessary to implement a transformation of manufacturing technology to support personalized medicine. The advantages of automation include savings in labor hours associated with the use of automation, as well as improvements in product consistency, reduced room classification, reduced cleanroom footprint, reduced training complexity, and improved scale-up and tracking logistics. Additionally, the documentation process can be rationalized by using software and providing a history of all processing equipment, reagents, patient identification, operator identification, in-process sensor data, etc. using automatically generated electronic batch records.

[0004] What is needed to advance these therapies and automation systems are components of cell expansion systems, such as cell culture chambers, that increase cell production or provide desirable cell characteristics. The present application meets these needs.

Summary of the Invention

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

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

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

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

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

[0010] In some aspects, the technology described herein relates to a cell culture chamber in which the first body includes a plurality of openings configured such that gas can be exposed to the external environment through the outer surfaces 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 in which the plurality of openings include a geometric, hexagonal, heptagonal, octagonal, symmetric, honeycomb, round, circular, oval, elliptical, or square shape.

[0012] In some embodiments, the techniques described herein relate to a cell culture chamber in which a first non-porous gas-permeable material is disposed on an inner surface of a first body, and the first non-porous gas-permeable material is configured to cover a plurality of openings of the first body.

[0013] In some embodiments, the techniques described herein relate to a cell culture chamber in which a structural section of a second body includes a plurality of openings configured such that gas can be exposed to an external environment through an outer surface of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

[0014] In some embodiments, the techniques described herein relate to a cell culture chamber in which the plurality of openings include a geometric, hexagonal, heptagonal, octagonal, symmetric, honeycomb, round, circular, oval, elliptical, or square shape.

[0015] In some embodiments, the techniques described herein relate to a cell culture chamber in which a second non-porous gas-permeable material is disposed on an inner surface of the first body, and the second non-porous gas-permeable material is configured to cover a plurality of openings of the second body.

[0016] In some embodiments, the techniques described herein relate to a cell culture chamber in which at least one bubble trap is integrally formed with a first body of the cell culture chamber, and further includes a bubble trap track integrally formed with the first body and fluidly connected to the at least one bubble trap.

[0017] In some embodiments, the techniques described herein relate to a cell culture chamber in which at least two bubble traps are integrally formed with a first body of the cell culture chamber, and a bubble trap track is fluidly connected to the at least two bubble traps.

[0018] In some embodiments, the techniques described herein relate to a cell culture chamber in which the width of a bubble trap track gradually decreases.

[0019] In some embodiments, the techniques described herein relate to a cell culture chamber in which the height of a bubble trap track gradually decreases.

[0020] In some embodiments, the techniques described herein relate to a cell culture chamber in which at least two bubble traps are integrally formed with a 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, whereby the at least two corresponding bubble trap tracks are separated from each other.

[0021] In some embodiments, the techniques described herein include a cassette, the cassette being a cell culture chamber in which at least one bubble trap is disposed on top, the cassette further including a first body forming a first portion of the cell culture chamber and a second body forming a second portion of the cell culture chamber, the first body and the second body being coupled to each other so as to form a sealed volume, and a first non-porous gas-permeable material being disposed on the first body and / or a second non-porous gas-permeable material being disposed on the second body, the first non-porous gas-permeable material and the first body being formed together, and the second non-porous gas-permeable material and the second body being formed together, relating to an automated cell engineering system.

[0022] In some embodiments, the techniques described herein relate to an automated cell engineering system in which the cell culture chamber is in a horizontal orientation.

[0023] In some embodiments, the techniques described herein relate to an automated cell engineering system that includes a plurality of bubble traps disposed on top of the cell culture chamber.

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

[0025] In some aspects, the technology described herein relates to an automated cell engineering system in which a first non-porous gas-permeable material is disposed on an 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.

[0026] In some aspects, the technology described herein relates to an automated cell engineering system in which a second non-porous gas-permeable material is disposed on an 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.

[0027] In some aspects, the technology described herein includes a first body that forms a first portion of a cell culture chamber and has a plurality of bubble traps disposed on top and a bubble trap track disposed between the plurality of bubble traps and connected to the plurality of bubble traps, and a second body that forms a second portion of the cell culture chamber, wherein the first body and the second body are configured to be coupled to each other to thereby form a sealed volume, and 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 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, and relates to an automated cell engineering system.

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

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

[0030] In some aspects, the technology described herein relates to an automated cell engineering system in which a plurality of bubble traps are formed at both ends of a first body.

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

[0033] In some aspects, the technology described herein is a cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising at least one body forming a sealed volume, and a non-porous gas-permeable material disposed on the at least one body such that the non-porous gas-permeable material and the at least one body are formed together, and in which no structure is disposed within the sealed volume.

Brief Description of the Drawings

[0034]

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DETAILED DESCRIPTION OF THE INVENTION

[0035] It should be understood that the various aspects disclosed herein can be combined in combinations different from those specifically shown in the description and the accompanying drawings. Also, by way of example, it should be understood that any particular operation or event of any of the processes or methods described herein can be performed in a different order, added, combined, or completely omitted (e.g., not all of the operations or events described may be necessary to practice this technique).

[0036] Any published patents, patent applications, websites, company names, and scientific literature referred to in this specification are hereby incorporated by reference in their entirety to the same extent as if each was specifically and individually indicated to be incorporated by reference. In the event of any conflict between any reference cited in this specification and the specific teachings of this specification, the latter shall prevail. Similarly, in the event of a conflict between the definition of a word or phrase as understood in the relevant art and the definition of a word or phrase specifically taught in this specification, the latter shall prevail.

[0037] As used herein, the singular forms "a", "an", and "the" specifically include the plural forms of the terms to which they refer unless the context clearly dictates otherwise. The term "about" as used herein means approximately, within the region of, roughly, or around. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the recited numerical values. In general, the term "about" as used herein is used to modify a numerical value by plus or minus 20% of the recited value.

[0038] The automated cell engineering system 102 described herein is a fully enclosed automated system for performing the steps of activation, transduction, proliferation, concentration, and / or recovery of cell cultures. The automated cell engineering system (also referred to herein as an overall automated biological processing unit) provides for the automated production of cell cultures. As used herein, "cell culture" refers to any suitable cell type, including individual cells, as well as cells that can form multiple cells or tissue structures. Exemplary cell cultures include blood cells, skin cells, muscle cells, bone cells, cells from various tissues and organs, and the like. In embodiments, genetically modified immune cells, including the CAR T cells described herein, can be produced. Exemplary automated cell engineering systems are also referred to herein as COCOON (registered trademark) or COCOON (registered trademark) systems throughout (see, e.g., U.S. Patent Application Publication No. 2019 / 0169572, the disclosure of which is hereby incorporated by reference in its entirety).

[0039] A cell culture chamber 101 for use in an automated cell engineering system includes a first body 110 and a second body 120 configured to be coupled to each other, as shown in FIGS. 1 and 4, for example. The first body 110 forms a first portion 105 of the cell culture chamber 101, and the second body 120 forms a second portion 115 of the cell culture chamber 101. When coupled to each other, the first body 110 and the second body 120 form a sealed volume 325. At least one structural member 130 extends from the first body 110 through the sealed volume 325 to the second body 120 (see FIGS. 1 and 2-3). The structural member 130 includes a series of lumens 135 that extend through the sealed volume 325 from the first body 110 to the second body 120. A first non-porous gas-permeable material 140 is disposed on the first body 110, and a second non-porous gas-permeable material 145 is disposed on the second body 120 (see FIG. 4). In an embodiment, the first non-porous gas-permeable material 140 can be formed together with the first body 110, and the second non-porous gas-permeable material 145 can be formed together with the second body 120, and the forming procedures will be described in more detail below.

[0040] In a further embodiment, the cell culture chamber includes at least one body 110' that forms a sealed volume 325'. The at least one body can be formed as a single solid element that forms the sealed volume 325', and the sealed volume further includes a gas-permeable material 140' disposed on top. At least one structural member 130' extends from the at least one body 110' through the sealed volume 325'. A series of lumens 135' further extend within the sealed volume of the at least one body. As described herein, the sealed volume 325' and the gas-permeable material 140' are appropriately prepared using dual-shot molding techniques.

[0041] As described above, the sealed volume 325 is formed in the region between the first body 110 and the second body 120. The sealed volume 325 is configured to contain a cell culture and is configured to be fluidly sealed (as described herein, it is not allowed for liquid to flow into or out of the sealed volume, but gas flow is allowed). As shown in FIG. 4, the first body 110 and the second body 120 are coupled to each other using mating elements such as snap fitting, shape fitting, or other methods known in the art, for example at the coupling portion 321. Further, it is envisioned that the first body 110 and the second body 120 can be formed as an integral body, eliminating the need to couple the first body 110 and the second body 120.

[0042] The structural member 130 extends from the first body 110 through the sealed volume 325 to the second body 120 and includes a first side surface 132 and a second side surface 134. The first side surface 132 is disposed on and formed as part of the first body 110, and the second side surface 134 is disposed on and formed as part of the second body 120. The structural member 130 is configured to maintain the integrity of the sealed volume 325 while defining a lumen 135 therethrough. In some embodiments, the lumen 135 is configured to receive a valve 136 or a coupler that can be configured to be connected to an automated cell engineering system. Such components enable external machinery such as pumps, piping, or other machines to interact with the media and cells within the sealed volume 325 and the components of the automated cell engineering system and provide additional support thereto. A series of lumens 135 is envisioned to be configured to receive the valve or coupler. The valve 136 can be, for example, a trumpet valve, i.e., 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) that enables connection of various syringes, sampling devices, etc.

[0043] As shown in FIGS. 2 and 3, in some embodiments, the cell culture chamber 101 includes at least one or more structural islands 150 that extend from a first body 110 through a sealed volume 325 to a second body 120. In an embodiment, each of the first body 110 and the second body 120 can include a structural island 150 that extends into the sealed volume 325. Each structural island 150 provides additional structural support to the cell culture chamber 101 and allows a larger volume to be placed within the cell culture chamber 101 without compromising structural integrity. Further, the structural islands 150 provide a larger surface area for cells to rest within the cell culture chamber, allowing for increased growth. The structural islands 150 can also result in an increase in turbulent flow (i.e., non-laminar flow) during filling and movement of fluid within the cell culture chamber, assisting in cell mixing and / or removal from the adhesion surface. In an embodiment, a lumen can be disposed within the structural island 150 to allow air to flow around and through the cell culture chamber and cassette, enhancing the potential for cooling and temperature management of the cassette.

[0044] To allow gas exchange between the sealed volume 325 and the external environment, the first body 110 and / or the second body 120 may include a plurality of openings 155 configured such that gas can be exposed to the external environment through the outer surfaces of a first non-porous gas permeable material 140 and / or a second non-porous gas permeable material 145 disposed within the cell culture chamber 101. The plurality of openings 155 can take on various shapes including geometric, symmetric, honeycomb, round, circular, oval, elliptical, square, and the like. Further, the design of the plurality of openings 155 can be used to provide additional structural support and strength to the first body 110 and the second body 120. For example, using a honeycomb structure allows for a high ratio of open space to structure, allows for a greater amount of gas exchange, and at the same time reduces the amount of structure required to support the first body 110 and / or the second body 120.

[0045] The inner surface 311 (otherwise known as the culture medium contact surface) of the first main body 110 of the cell culture chamber 101 and the inner surface 316 (or culture medium contact surface) of the second main body 120 are appropriately lined or covered with a non-porous gas-permeable material to prevent water and other liquids from evaporating from within the cell culture chamber 101. As used herein, "non-porous gas-permeable material" means any compositional film or material used in a gas-permeable cell culture device that allows gas to pass through and enter the cell culture chamber 101, but does not contain pores or holes through which liquid (i.e., cell culture medium) can pass or leak. Exemplary non-porous gas-permeable materials include, but are not limited to, silicone, fluorinated ethylene propylene (FEP), polyolefin, ethyl vinyl olefin (EVO), and ethylene vinyl acetate copolymer. The non-porous gas-permeable materials described herein are suitably useful for delivering one or more gases, including oxygen, nitrogen, CO2, etc., to the cells within the cell culture chamber 101. Further, the width of the non-porous gas-permeable material can be decreased or increased to increase or decrease gas exchange through the material.

[0046] As shown in FIG. 4, in an embodiment, the first non-porous gas permeable layer or material 140 is disposed on the inner surface 311 of the first body 110, and the second non-porous gas permeable material 145 is disposed on the inner surface 316 of the second body 120. During the formation of the cell culture chamber 101, for example, through the use of molding, the first non-porous gas permeable layer 140 and the second non-porous gas permeable layer 145 can be simultaneously created on the inner surfaces 311 / 316 by a dual-shot molding technique. In one embodiment, this 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 shot of a dual-shot molding technique. After a short delay until the thermoplastic hardens, the core portion of the mold retracts in small amounts to create a space for the second shot of silicone (or other suitable gas permeable material), and form the gas permeable materials 140 / 145 within the first body 110 and the second body 120, respectively. Since the measured thickness of the gas permeable materials 140 / 145 is determined by the amount of the mold that retracts from the first shot, it can be adjusted according to the needs of the end user. Further, a non-porous gas permeable material can be cast onto or within the cell culture chamber 101 to provide the gas permeable characteristics described herein. The casting of the non-porous gas permeable material can be performed using various methods known in the art. Through the process of casting a non-porous gas permeable material onto or within the cell culture chamber 101, the structural strength and integrity of the gas permeable membrane can be enhanced while maintaining the gas permeable characteristics. The non-porous gas permeable material may also be disposed on the inner surfaces 311 / 316 of the first body 110 and the second body 120 by coating, spraying, painting, laminating, or other methods. The first non-porous gas permeable layer 140 and the second non-porous gas permeable layer 145 can be composed of the same or different materials, and in an embodiment, each layer 140 / 145 can include a plurality of different materials.

[0047] In an exemplary embodiment, "a portion" of the inner surface, such as 311, includes a non-porous gas permeable material. As used herein, "a portion" refers to at least about 20% of the surface composed of the non-porous gas permeable material. In embodiments, both the media contact surfaces 311 and 316 include a non-porous gas permeable material. In such embodiments where less than 100% of the media contact surface is made of the non-porous gas permeable material, the remaining portion of the media contact surface 311 can include other suitable materials including various plastics (e.g., polypropylene, polystyrene, etc.) that promote cell adhesion and growth. In embodiments where less than 100% of the media contact surface is made of the non-porous gas permeable material, the remaining portion of the media contact surface 316 can include other suitable materials including various plastics (e.g., polypropylene, polystyrene, etc.) that provide structural support. In embodiments, the media 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 encloses the chamber volume without a gas permeable material. The first body 110 of the cell culture chamber 101 can be suitably made of a transparent material that enables visual inspection or imaging of the cell culture within the cell culture chamber 101.

[0048] In embodiments, at least about 30% of the media contact surface is made of the non-porous gas permeable material, and 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., the whole) includes the non-porous gas permeable material.

[0049] In an embodiment, the inner surfaces of the first main body 110 and the second main body 120 can further include surface coatings such as a surface coating that activates cells, a surface coating that regulates biological pathways within cells, a surface coating that promotes cell growth, a surface coating that enhances the transfection efficiency of cells, a surface coating that improves the selection of specific types of cells, a surface coating that improves cell adhesion, a surface coating that inhibits cells, and a surface coating with controlled solubility. Therefore, in an embodiment, the non-porous gas-permeable material that appropriately constitutes part or all of the culture medium contact surface can further include a surface coating as described herein. In other embodiments, the remaining portion of the culture medium contact surface that does not include the non-porous gas-permeable material can also include a surface coating as described herein. In other embodiments, the entire culture medium contact surface does not include the non-porous gas-permeable material but still includes a surface coating as described herein. Cell adhesion can also provide an opportunity for cell selection that promotes the growth of a desired population without permitting the adhesion of an undesired population.

[0050] As used herein, "a surface coating that activates cells" refers to a material, substrate, or component that causes cell growth and / or differentiation.

[0051] As used herein, "a surface coating that regulates biological pathways within cells" refers to a material, substrate, or component that causes one or more actions among molecules within a cell, resulting in a specific product or change within the cell. For example, such a surface coating may cause the assembly of new molecules such as fats or proteins, switch genes on and off, or cause cell movement.

[0052] As used herein, "a surface coating that promotes cell growth" refers to a material, substrate, or component that causes cells to grow faster or more than in the absence of the material.

[0053] As used herein, a "surface coating that improves cell adhesion" refers to a material, substrate, or component that enables cells to interact better with the surface, adhere to the surface, and interact with other cells when the cells adhere to the surface.

[0054] As used herein, a "surface coating that inhibits cells" refers to a material, substrate, or component that does not allow cells to proliferate and / or does not allow them to adhere to the medium contact surface.

[0055] As used herein, a "surface coating that responds to the state of the medium" refers to a material, substrate, or component that changes when the state of the medium changes. Exemplary changes include changes in temperature, pH, oxygen level or concentration, level of toxic gas, presence of toxic substances, and also include, for example, a change in color as an aid for monitoring.

[0056] As used herein, a "surface coating with controlled solubility" refers to a material, substrate, or component that is released from the surface at a specific time or in response to a specific temperature or pH, for example, to achieve controlled release of the coating content.

[0057] Surface treatments (such as corona, plasma, etching, etc.) can be used to promote the bonding (or rejection) of the coating. This enables selective surface modification, allowing two different types of cells to be grown in the same cell culture chamber or enabling selective unit operations to be performed within the same chamber.

[0058] As shown in FIG. 1, in some embodiments, the cell culture chamber 101 is configured to be arranged as part of a cassette 100 for use in an automated cell engineering system 102 (see FIG. 5). As used herein, a "cassette" refers to a largely self - contained, removable, and replaceable element of the automated 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, vectors, etc.

[0059] Cassette 100 includes a low - temperature chamber suitable for storing cell - culture media, and a high - temperature chamber suitable for performing activation, transduction, and / or proliferation of cell cultures, including immune - cell cultures. The high - temperature chamber may be separated from the low - temperature chamber by thermal shielding. As used herein, a “low - temperature chamber” refers to a chamber that is maintained, suitably below room temperature and more suitably at about 4°C to about 8°C, for maintaining cell media, etc. at refrigeration temperatures. The low - temperature chamber can include a bag or other holder for media containing about 1L, about 2L, about 3L, about 4L, or about 5L of fluid. Additional media bags or other fluid sources can be connected outside the cassette and connected to the cassette via access ports. The cassette can further include one or more fluid pathways connected to the cell - culture chamber, and the fluid pathways provide for recirculation to the cell - culture chamber, waste removal, and homogeneous gas exchange and nutrient distribution without disturbing the cells within the cell - culture chamber. Cassette 100 also includes a pump system that includes one or more pumps for driving fluid through the cassette, such as a peristaltic pump, and one or more valves for controlling the flow through various fluid pathways, as described herein.

[0060] In an embodiment, the cell culture chamber 101 is a sealed non-flexible container. The use of a non-flexible chamber enables the cells to be maintained in a substantially undisturbed state. As described, the cell culture chamber 101 can be oriented to allow the immune cell culture to spread throughout the cell culture chamber. As shown in FIG. 5, the cell culture chamber 101 is preferably maintained in a horizontal position parallel to the floor or table, the cell culture is maintained in an undisturbed state, and the cell culture can diffuse throughout a large area of the cell culture chamber. In an embodiment, the overall thickness of the cell culture chamber 101 is on the order of about 0.5 cm to about 5 cm. Suitably, the cell culture chamber can have a volume between about 0.50 ml and about 1 L, more suitably between about 0.5 and about 900 ml, or the cell culture chamber can have a volume of about 825 ml. Suitably, the cell culture chamber can have a volume between about 0.5 ml and about 500 ml, between about 10 ml and about 300 ml, more suitably between about 50 ml and about 200 ml, or the cell culture chamber can have a volume of about 180 ml. By using a low chamber height (less than 5 cm, suitably less than 4 cm, less than 3 cm, or less than 2 cm), effective medium and gas exchange close to the cells is possible. The ports are configured to allow mixing through fluid recirculation without disturbing the cells. A static container with a greater height can create a concentration gradient, which may limit oxygen and fresh nutrients in the area near the cells. By controlling the fluid dynamics, medium exchange can be performed without disturbing the cells. The medium can be removed from an additional chamber (where there are no cells) without risk of cell loss.

[0061] As described herein, in an exemplary embodiment, the cassette is pre-filled with one or more of a cell culture, a culture medium, an activation reagent, and / or a vector containing any combination thereof. In further embodiments, these various elements can be added later via a suitable injection port or the like.

[0062] The cell engineering system 102 may include components such as a gas control seal, a heating zone, an actuator, a pivot for rocking or tilting the cell engineering system as desired, and a low-temperature zone for holding a low-temperature chamber. The cell engineering system 102 may also include a user interface that can include a barcode reader, and a function for receiving input by a touchpad or other similar device.

[0063] As shown in FIG. 1, the cassette 100 may further include an additional chamber volume, or satellite volume 165, for increasing the working volume of the cell culture chamber by providing an additional volume for the culture medium and other working fluids, or may be used to contain the cell culture or a portion of the cell culture during the steps of activation, transduction, proliferation, concentration, and / or recovery of the cell culture. Suitably, the satellite volume 165 is oriented substantially vertically and fluidly connected to the cell culture chamber 101 such that the culture medium is exchanged between the culture chamber without disturbing the cell culture. In an exemplary embodiment, the satellite volume is a bag, and in other embodiments, the satellite volume is a non-yield chamber as shown in FIG. 1. In embodiments, the satellite volume is from about 0.50 ml to about 1 L, or from about 10 ml to 800 ml, or from about 100 ml to about 500 ml, or from about 100 ml to about 300 ml, more suitably from about 150 ml to about 200 ml. In embodiments, the satellite volume is further configured such that the culture medium can be removed without loss of cells from the immune cell culture. That is, the exchange of the culture medium between the satellite volume and the cell culture chamber is performed in such a way that the cells are not disturbed and are not removed from the cell culture chamber. In some embodiments, the satellite volume may further include a port, inlet, cutout, lumen, or pass-through that allows air flow to be transferred from the outside of the cassette to the inside of the cassette.

[0064] As shown in FIG. 1, the satellite volume 165 suitably includes a satellite volume lumen 170, which is a passage through the satellite volume 165 that allows gas to flow through the satellite volume 165 while maintaining the fluid integrity of the satellite volume 165. FIG. 5 shows the orientation of the cell culture chamber 101 within the cell engineering system 102, where the cell culture chamber is oriented substantially horizontally and the satellite volume 165 is oriented substantially vertically with the shell 500 of the cell engineering system, which is a movable cover surrounding the system. As shown, the satellite volume lumen 170 is arranged such that air 502 can move through the lumen to the center of the cell engineering system 102 and the cassette 100 to provide an increased air flow as well as temperature control and regulation.

[0065] Positioning the cell culture chamber 101 in a horizontal position within the cell engineering system 102 is accomplished by rotating the entire cell engineering system 102 by approximately 90 degrees, whereby the cell culture chamber 101, which was originally oriented vertically (see FIG. 1 for the position on the cassette 100), is changed to a horizontal position as shown in FIG. 5. This allows for the utilization of a larger volume of the cell culture chamber 101 since the surface of the cassette 100 on which the cell culture chamber 101 is placed is larger than the upper section of the cassette.

[0066] FIG. 6 shows a temperature gradient diagram of the satellite volume 165 showing the satellite volume lumen 170.

[0067] 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 configured to be coupled to each other, as shown, for example, in FIGS. 7-12. The 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 to each other, the first body 610 and the second body 620 form a sealed volume 625, as shown in the cross-sectional views of FIGS. 7 and 8. The cell culture chamber 601 can be formed without any structure disposed within the sealed volume 625 so as to increase the amount of medium disposed within the sealed volume 625, as shown in FIGS. 7 and 8. A first non-porous gas-permeable material 640 can be disposed on the first body 610, and a second non-porous gas-permeable material 645 can be disposed on the second body 620 (see FIG. 7). In an embodiment, using the formation procedure described above herein, the first non-porous gas-permeable material 640 can be formed together with the first body 610, and the second non-porous gas-permeable material 645 can be formed together with the second body 620. In an embodiment, the first body 610 may not include the first non-porous gas-permeable material 640, but 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, but 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, an advantageous maximization of the oxygen level within the cell culture chamber 601 can be achieved. In another embodiment, the first body 610 can include the first non-porous gas-permeable material 640, and the second body 620 can also include the second non-porous gas-permeable material 645.

[0068] As described above, the sealed volume 625 is formed in the region between the first body 610 and the second body 620. The sealed volume 625 can be configured to contain a cell culture and can be configured to seal the liquid, except when intended through a designated valve, port, etc. (as described herein, it is not allowed for liquid to flow into or out of the sealed volume, but gas flow is allowed). For example, the inlet / outlet port 636 can be connected to the cassette 100 or other elements of the cell engineering system 102 to provide for the exchange of medium or other reagents with the sealed volume 625 and to enable the recovery of the cell culture within the sealed volume 625. As shown in FIGS. 7 and 8, the first body 610 and the second body 620 are coupled to each other using mating elements such as snap fits, shape fits, or other methods known in the art, for example, at the joint 630. FIGS. 10 and 11 show, respectively, a top perspective view and a bottom perspective view of the cell culture chamber 601 with the first body 610 and the second body 620 coupled. Further, it is envisioned that the first body 610 and the second body 620 can be formed as an integral body, eliminating the need to couple the first body 610 and the second body 620.

[0069] FIG. 9 shows a top view of exemplary dimensions of the cell culture chamber 601. 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) may be about 360 mm. In an embodiment, the length of the cell culture chamber may be 200 - 600 mm. In an embodiment, the length of the cell culture chamber may be 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 about 90 mm. In an embodiment, the width of the cell culture chamber 601 may be 50 - 150 mm. In an embodiment, the width of the cell culture chamber may be 75 - 100 mm.

[0070] To enable gas exchange between the sealed volume 625 and the external environment, the first body 610 and / or the second body 620 may include a plurality of openings 655 configured such that gas can be exposed to the external environment through the outer surfaces of the first non-porous gas permeable material 640 and / or the second non-porous gas permeable material 645 disposed within the cell culture chamber 601, as shown in FIGS. 7-11. For example, FIGS. 7-10 show a plurality of openings 655 formed in or passing through the first body 610 in a staggered arrangement, or disposed through the first body 610, while FIG. 11 shows a plurality of openings 655 formed in or passing through the second body 620 in a staggered arrangement, or disposed through the second body 620. The plurality of openings 655 can take on various shapes including geometric shapes (e.g., hexagon, heptagon, octagon, etc.), symmetric shapes, honeycomb shapes, round, circular, rectangular, elliptical, square, etc. Further, the design of the plurality of openings 655 can be used to provide additional structural support and strength to the first body 610 and the second body 620. For example, using a honeycomb structure increases the ratio of open space to the structure, allowing for more gas exchange while reducing the amount of structure necessary to support the first body 610 and / or the second body 620. In an embodiment, the plurality of openings 655 can be disposed within, formed through, or in a continuous arrangement on the first body 610 and / or the second body 620. In an embodiment, the first body 610 and / or the second body 620 may instead be formed from a solid thermoplastic plastic in which no plurality of openings 655 are formed. With the plurality of openings 655, at least about 30% of the first body 610 and / or the second body 620 may be covered, more preferably 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., the whole) may be covered.

[0071] The inner surface 611 of the first main body 610 of the cell culture chamber 601 (otherwise known as the culture medium contact surface), and the inner surface 621 (or culture medium contact surface) of the second main body 620 are appropriately lined or covered with a non-porous gas permeable material(s) such that water (i.e., liquid) or other liquids (e.g., cell culture medium) are prevented from evaporating out of the cell culture chamber 601 while at the same time allowing the passage of gases such as oxygen, nitrogen, carbon dioxide, etc. As used herein, "non-porous gas permeable material" means any composition, coating, film, or material used in a gas permeable cell culture device that allows gases to pass into the cell culture chamber 601 but does not contain pores or holes through which a liquid (i.e., cell medium) could pass or leak. Exemplary non-porous gas permeable materials include, but are not limited to, silicone, fluorinated ethylene propylene (FEP), polyolefin, ethyl vinyl olefin (EVO), and ethylene vinyl acetate copolymer. The non-porous gas permeable materials described herein are suitably useful for delivering one or more gases including oxygen, nitrogen, CO2, etc. to the cells within the cell culture chamber 601. Further, the thickness of the non-porous gas permeable material can be decreased or increased to increase or decrease gas exchange through the material.

[0072] As shown in FIG. 7, in the embodiment, the first non-porous gas permeable material 640 is disposed on the inner surface 611 of the first body 610, and the second non-porous gas permeable material 645 is disposed on the inner surface 621 of the second body 620. During the formation of the cell culture chamber 601, for example, through the use of molding, the first non-porous gas permeable material 640 and the second non-porous gas permeable material 645 can be simultaneously created on the inner surfaces 611 / 621 by dual-shot molding techniques. In one embodiment, this molding technique includes injection molding techniques. For example, the first body 610 and the second body 620 of the cell culture chamber 601 are formed by the first shot of the dual-shot molding technique. After a short delay until the thermoplastic hardens, the core part of the mold retracts little by little to create a space for the second shot of silicone (or other suitable gas permeable material), and the gas permeable materials 640 / 645 disposed on the first body 610 and the second body 620 respectively are formed. Since the measured thickness of the gas permeable materials 640 / 645 is determined by the amount of the mold that retracts from the first shot, it can be adjusted according to the needs of the end user. Further, the non-porous gas permeable material can be cast on or within the cell culture chamber 601 to provide the gas permeable characteristics described herein. Casting of the non-porous gas permeable material can be performed using various methods known in the art. Through the process of casting the non-porous gas permeable material on or within the cell culture chamber 601, the structural strength and integrity of the gas permeable membrane can be enhanced while maintaining the gas permeable characteristics. The non-porous gas permeable material may also be disposed on the inner surfaces 611 / 621 of the first body 610 and the second body 620 by coating, spraying, painting, laminating, or other methods. The first and second non-porous gas permeable layers 640 / 645 can be composed of the same or different materials, and in the embodiment, each layer 640 / 645 can include a plurality of different materials.

[0073] In an exemplary embodiment, "a portion" of the inner surface, such as 611, includes a non-porous gas permeable material. As used herein, "a portion" refers to at least about 20% of the surface made of the non-porous gas permeable material. In embodiments, both of the media contact surfaces 611 and 621 include a non-porous gas permeable material. In embodiments where less than 100% of the media contact surface is made of the non-porous gas permeable material, the remaining portion of the media contact surface 611 can include other suitable materials, such as various plastics (e.g., polypropylene, polystyrene, etc.) that promote cell adhesion and growth. In embodiments where less than 100% of the media contact surface is made of the non-porous gas permeable material, the remaining portion of the media contact surface 621 can include other suitable materials, such as various plastics (e.g., polypropylene, polystyrene, etc.) that provide structural support. In embodiments, the inner surface 611 of the first body 610 may lack the non-porous gas permeable material such that the inner surface 611 is not completely (i.e., 0%) covered. In embodiments, the inner surface 621 of the second body 620 may lack the non-porous gas permeable material such that the inner surface 621 is not completely (i.e., 0%) covered.

[0074] In embodiments, at least about 30% of the media contact surface is made of the non-porous gas permeable material, more preferably 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., the whole) includes the non-porous gas permeable material.

[0075] In an embodiment, the inner surfaces of the first main body 610 and the second main body 620 can further include surface coatings such as a surface coating that activates cells, a surface coating that regulates biological pathways within cells, a surface coating that promotes cell growth, a surface coating that enhances the transduction efficiency of cells, a surface coating that improves the selection of specific types of cells, a surface coating that improves cell adhesion, a surface coating that inhibits cells, and a surface coating with controlled solubility. Therefore, in an embodiment, the non-porous gas-permeable material that appropriately constitutes part or all of the culture medium contact surface can further include a surface coating as described herein. In other embodiments, the remaining portion of the culture medium contact surface that does not include the non-porous gas-permeable material can also include a surface coating as described herein. In other embodiments, the entire culture medium contact surface does not include the non-porous gas-permeable material but still includes a surface coating as described herein. Cell adhesion can also provide an opportunity for cell selection that promotes the growth of a desired population without allowing the adhesion of an undesired population.

[0076] In an embodiment, the cell culture chamber 601 is a sealed non-flexible chamber. The use of a non-flexible chamber enables the cells to be maintained in a substantially undisturbed state. As described, the cell culture chamber 601 can be oriented to allow the cell culture to spread throughout the cell culture chamber 601. In an embodiment, the overall thickness of the cell culture chamber 601 is on the order of about 0.5 cm to about 5 cm. Suitably, the cell culture chamber can have a volume between about 0.50 ml and about 1 L, more suitably between about 0.5 and about 900 ml, or the cell culture chamber can have a volume of about 825 ml. Suitably, the cell culture chamber can have a volume between about 0.5 ml and about 500 ml, between about 10 ml and about 300 ml, more suitably between about 50 ml and about 200 ml, or the cell culture chamber can have a volume of about 180 ml. By using a low chamber height (less than 5 cm, suitably less than 4 cm, less than 3 cm, or less than 2 cm), effective medium and gas exchange close to the cells is possible. The ports are configured to allow mixing via fluid recirculation without disturbing the cells. Larger height static vessels can create concentration gradients, which can limit oxygen and fresh nutrients in the regions near the cells. By controlling the fluid dynamics, medium exchange can be performed without disturbing the cells. The medium can be removed from an additional chamber (where there are no cells) without risk of cell loss.

[0077] In some embodiments, the cell culture chamber 601 may include an inlet / outlet port 636 or a coupler configured to be connected to an automated cell engineering system. Such components enable external machinery, such as pumps, tubing, connectors, or other machines, to interact with the media and cells within the sealed volume 625 to provide the media and cells, provide additional support to the media and cells, or provide components of the automated cell engineering system. In an embodiment, the inlet / outlet port may be configured to receive a valve or a coupler. The valve can be, for example, a trumpet valve, i.e., a valve having two positions (open "0" or closed "1"), or a pinch valve that can compress or pinch the flow path. The inlet / outlet port 636 can also include a coupler (not shown) that allows connection of various syringes, sampling devices, and the like.

[0078] In an embodiment, the upper surface 613 of the first main body 610 includes or is integrally formed with at least one bubble trap 700. The at least one bubble trap 700 is formed within the sealed volume 625 of the cell culture chamber 601 and is configured to collect any bubbles that may rise to the upper portion of the culture medium. As shown in FIG. 9, at least two bubble traps 700A, 700B are disposed at each end of the upper surface 613 of the first main body 610 (one trap 700A, 700B at each end), and a bubble trap track 710 is included or integrally formed within the first main body 610. The bubble trap track 710 further extends between each bubble trap 700A, 700B and fluidly connects each bubble trap 700A, 700B. FIGS. 13A-13B show various embodiments in which at least two bubble traps 700A, 700B and a bubble trap track 710 can be formed. Since the width 722 of the bubble trap track 710 can vary along its length, the bubble trap track 710 widens towards its ends 712 (i.e., closer to the bubble traps 700) and narrows towards the center of its length (i.e., the center of the upper surface 613 or near it). Thus, the bubble trap track 710 can taper outwardly from the center of the bubble trap track 710 to the ends 712 of the bubble trap track 710 adjacent to at least two bubble traps 700A, 700B. For example, the width 722 of the bubble trap track 710 starts with little width / space near the center of the bubble trap track 710 (i.e., near the planar center of the inner surface 611), and the width 722 gradually increases (the width becomes finer) along the length of the bubble trap track 710 (i.e., towards at least one bubble trap 700), increasing by up to 1 mm or more towards the ends 712 of the bubble trap track 710 where it meets / connects to at least one bubble trap 700, thereby forming a part of an additional volume 711 as shown, for example, in FIG. 13A. In an embodiment, the width 722 of the bubble trap track 710 may increase from any value between 0.01 mm and 10 mm in width from the center of the bubble trap track 710 towards the ends 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 inwardly from the center of the bubble trap track 710 to the ends of the bubble trap track 710 adjacent to at least two bubble traps 700A, 700B (not shown). In an embodiment, for example, as shown in FIG. 13C, at least two bubble trap tracks 710A, 710B are fluidly connected to each of 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 separated from each other (i.e., there is a space between the bubble trap track sections). The bubble trap tracks 710A, 710B can taper outwardly from the ends of the bubble trap track farthest from the bubble traps 700A, 700B, as shown, for example, in FIG. 13A. In an embodiment, each bubble trap track 710A, 710B can taper inwardly starting from the end of the bubble trap track farthest from the bubble traps 700A, 700B (not shown). In an embodiment, for example, as shown in FIG. 13B, the bubble trap tracks 710A, 710B can have a uniform width over their entire length from the end of the bubble trap track farthest from the bubble traps 700A, 700B to the end 712 of the bubble trap track 710 closest to or fluidly connected to the bubble traps 700A, 700B.

[0079] 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 can each be fluidly connected to at least two bubble trap tracks 710A and 710B (FIG. 13C) to form at least two additional volumes 711A, 711B. In an embodiment, a plurality of bubble traps 700 can be fluidly connected to a plurality of bubble trap tracks 710. In an embodiment, a plurality of bubble traps 700 can be fluidly connected to a bubble trap track 710. In an embodiment, a plurality of bubble traps 700 can be fluidly connected to a single bubble trap track 710. For example, the first body 610 can 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, the plurality of bubble traps 700 can be fluidly connected to respective bubble trap tracks 710 such that the bubble trap tracks 710 are separated from each other.

[0080] The bubble trap track 710 / 710A / 710B can also be configured such that its height gradually increases. For example, FIGS. 8 and 12 show that a slight gradient 720 is formed on the inner surface 611 of the first body 610 for the bubble trap track 710, whereby bubbles collected near the center of the bubble trap track 710 (i.e., near the planar center of the inner surface 611 of the first body 610) are guided outward toward at least one bubble trap 700. In FIG. 12, the gradient 720 of the bubble trap track 710 starts from a state where the center of the bubble trap track 710 (i.e., near the planar center of the inner surface 611) has little height / space, and the height gradually increases along the length of the bubble trap track 710 (i.e., the height gradually decreases toward 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 with at least one bubble trap 700, thereby forming part of an additional volume 711. Thus, the bubble trap track 710 can taper from the center of the bubble trap track 710 toward the ends of the bubble trap track 710 adjacent to at least two bubble traps 700A, 700B. In an embodiment, the gradient 720 of the bubble trap track 710 starts from the center of the bubble trap track 710 and may increase from any value between 0.01 mm and 10 mm in height on the inner surface 611 toward the end 712 of the bubble trap track 710 connected to at least one bubble trap 700 respectively. The at least one bubble trap 700 and the bubble trap track 710 can further form an additional volume 711 for collecting / holding the bubbles described herein within the sealed volume 625. This additional volume 711 is structured only for collecting bubbles formed in the culture medium disposed within the sealed volume 625 such that the fluid height of the culture medium within the sealed volume 625 remains less than 20 mm (i.e., the culture medium does not enter either the bubble trap track 710 or the at least one bubble trap 700).The curved, long profile of the bubble trap track 710 (i.e., the gradient 720) serves to guide bubbles along the length of the bubble trap track 710 into at least one bubble trap 700 located at the end of the bubble trap track 710.

[0081] In an embodiment, at least one bubble trap 700 described herein can be included in or integrally formed with the bottom surface 623 of the second body 620 (not shown). A plurality of bubble traps 700 can be disposed at each end of the bottom surface 623 of the second body, and a bubble trap track 710 having a gradient 720 runs between each bubble trap 700 and is connected to each bubble trap 700. As previously described herein, the bubble trap track 710 may vary in width along its length and its gradient 720 may also vary. With this configuration, when the cell culture chamber is inverted 601 (i.e., axially rotated) so that the second body 620 faces upward, at least one bubble trap 700 and the bubble trap track 710 can collect the bubbles formed in the culture medium. The bubble traps 700 / 700A / 700B and the bubble trap tracks 710 / 710A / 710B can be included in or integrally formed with the second body 620 in the same manner as described above with respect to the first body 610.

[0082] Although various embodiments according to the present invention have been described above, it should be understood that they are not limiting and have been presented by way of example only. It will be apparent to those skilled in the art that various forms and details can be changed therein without departing from the spirit and scope of the present invention. Therefore, the width and scope of the present invention should not be limited by any of the embodiments discussed herein, and each reference cited herein can be used in combination with the features of other embodiments. All patents and publications discussed herein are hereby incorporated by reference in their entirety.

[0083] Embodiments of the present disclosure include the following examples.

[0084] Example 1. A cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising a first body forming a first portion of the cell culture chamber and a second body forming a second portion of the cell culture chamber, the first body and the second body being coupled to each other so as to form a sealed volume, and a second body, wherein a first non-porous gas-permeable material is disposed on the first body, and the first non-porous gas-permeable material and the first body are formed together, the cell culture chamber.

[0085] Example 2. The cell culture chamber according to Example 1, further comprising a second non-porous gas-permeable material disposed on the second body, and the second non-porous gas-permeable material and the second body are formed together.

[0086] Example 3. The cell culture chamber according to Example 1, further comprising at least one air bubble trap integrally formed with the cell culture chamber.

[0087] Example 4. The cell culture chamber according to Example 1, further comprising one or more structural islands.

[0088] Example 5. The cell culture chamber according to Example 2, wherein the first body and the second body are coupled to each other using a fitting element.

[0089] Example 6. The cell culture chamber according to Example 2, wherein the first body includes a plurality of openings configured such that gas can be exposed to the external environment through the outer surfaces of the first non-porous gas-permeable material and / or the second non-porous gas-permeable material.

[0090] Example 7. The cell culture chamber according to Example 6, wherein the plurality of openings include a geometric, hexagonal, heptagonal, octagonal, symmetric, honeycomb, round, circular, rectangular, elliptical, or square shape.

[0091] Example 8. The cell culture chamber according to Example 6, wherein the first non-porous gas permeable material is disposed on the inner surface of the first body, and the first non-porous gas permeable material is configured to cover a plurality of openings of the first body.

[0092] Example 9. The cell culture chamber according to Example 2, wherein the structural section of the second body includes a plurality of openings configured such that gas can 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.

[0093] Example 10. The cell culture chamber according to Example 9, wherein the plurality of openings include geometric, hexagonal, heptagonal, octagonal, symmetric, honeycomb, round, circular, rectangular, elliptical, or square shapes.

[0094] Example 11. The cell culture chamber according to Example 10, wherein the second non-porous gas permeable material is disposed on the inner surface of the first body, and the second non-porous gas permeable material is configured to cover a plurality of openings of the second body.

[0095] Example 12. The cell culture chamber according to Example 3, wherein at least one bubble trap is integrally formed with the first body of the cell culture chamber, and further includes a bubble trap track integrally formed with the first body and fluidly connected to the at least one bubble trap.

[0096] Example 13. The 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 the bubble trap track is fluidly connected to the at least two bubble traps.

[0097] Example 14. The cell culture chamber according to Example 13, wherein the width of the bubble trap track is gradually decreasing.

[0098] Example 15. The cell culture chamber according to Example 13, wherein the height of the bubble trap track is gradually decreasing.

[0099] Example 16. The 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, whereby the at least two corresponding bubble trap tracks are separated from each other.

[0100] Example 17. A cell culture chamber comprising a cassette, the cassette being a cell culture chamber in which at least one bubble trap is disposed at the upper part, the cell culture chamber comprising a first body forming a first part of the cell culture chamber, and a second body forming a second part of the cell culture chamber, the first body and the second body being coupled to each other so as to form a sealed volume, the second body, a first non-porous gas-permeable material being disposed on the first body, and / or a second non-porous gas-permeable material being disposed on the second body, the first non-porous gas-permeable material and the first body being formed together, and the second non-porous gas-permeable material and the second body being formed together.

[0101] Example 18. The automated cell engineering system according to Example 17, wherein the cell culture chamber is in a horizontal orientation.

[0102] Example 19. The automated cell engineering system according to Example 17, wherein the cell culture chamber includes a plurality of bubble traps disposed at the upper part.

[0103] Example 20. The automated cell engineering system according to Example 17, wherein the first body includes a plurality of openings and the second body includes a plurality of openings.

[0104] Example 21. The first non-porous gas permeable material is disposed on the inner surface of the first body, and the first non-porous gas permeable material is configured to cover a plurality of openings of the first body, the automated cell engineering system according to Example 20.

[0105] Example 22. The second non-porous gas permeable material is disposed on the inner surface of the second body, and the second non-porous gas permeable material is configured to cover a plurality of openings of the second body, the automated cell engineering system according to Example 20.

[0106] Example 23. An automated cell engineering system, comprising a cell culture chamber, the cell culture chamber forming a first portion of the cell culture chamber, a plurality of bubble traps being disposed at the top, a bubble trap track being disposed between the plurality of bubble traps and connected to the plurality of bubble traps, a first body, a second body forming a second portion of the cell culture chamber, the first body and the second body being coupled to each other so as to form a sealed volume, the second body, the first non-porous gas permeable material being disposed on the first body, the second non-porous gas permeable material being disposed on the second body, the first non-porous gas permeable material and the first body being formed together, the second non-porous gas permeable material and the second body being formed together, the automated cell engineering system.

[0107] Example 24. The automated cell engineering system according to Example 23, wherein the cell culture chamber is in a horizontal orientation.

[0108] Example 25. The automated cell engineering system according to Example 24, wherein the bubble trap track has a gradually decreasing height and / or a gradually decreasing width.

[0109] Example 26. The automated cell engineering system according to Example 25, wherein each of the plurality of bubble traps is formed at both ends of the first body.

[0110] Example 27. The automatic cell engineering system according to Example 26, wherein a first non-porous gas permeable material is disposed on the inner surface of the first main body, and the first non-porous gas permeable material is configured to cover a plurality of openings of the first main body.

[0111] Example 28. The automatic cell engineering system according to Example 27, wherein a second non-porous gas permeable material is disposed on the inner surface of the second main body, and the second non-porous gas permeable material is configured to cover a plurality of openings of the second main body.

[0112] Example 29. A cell culture chamber for use in an automatic cell engineering system, the cell culture chamber including at least one main body that forms a sealed volume, and a non-porous gas permeable material disposed on the at least one main body such that the non-porous gas permeable material and the at least one main body are formed together, and no structure is disposed within the sealed volume.

Claims

1. A cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: a first body forming a first portion of the cell culture chamber; a second body forming a second portion of the cell culture chamber, the first body and the second body being configured to be coupled to each other to thereby form a sealed volume, the second body; a first non-porous gas permeable material disposed on the first body; A cell culture chamber, wherein the first non-porous gas permeable material and the first body are integrally formed.

2. Further comprising a second non-porous gas permeable material disposed on the second body; The cell culture chamber according to claim 1, wherein the second non-porous gas permeable material and the second body are integrally formed.

3. The cell culture chamber according to claim 1, further comprising at least one bubble trap integrally formed with the cell culture chamber.

4. The cell culture chamber according to claim 1, further comprising one or more structural islands.

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

6. The cell culture chamber according to claim 2, wherein the first body includes a plurality of openings configured such that gas can 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 according to claim 6, wherein the plurality of openings include geometric, hexagonal, heptagonal, octagonal, symmetric, honeycomb, round, circular, rectangular, elliptical, or square shapes.

8. The cell culture chamber according to claim 6, wherein the first non-porous gas permeable material is disposed on an 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.

9. The cell culture chamber according to claim 2, wherein a structural section of the second body includes a plurality of openings configured such that gas can 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.

10. The cell culture chamber according to claim 9, wherein the plurality of openings include geometric, hexagonal, heptagonal, octagonal, symmetric, honeycomb, round, circular, rectangular, elliptical, or square shapes.

11. The second non-porous gas permeable material is disposed 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. The cell culture chamber according to claim 10.

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

13. 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. The cell culture chamber according to claim 12.

14. The width of the bubble trap track is gradually decreasing. The cell culture chamber according to claim 13.

15. The height of the bubble trap track is gradually decreasing. The cell culture chamber according to claim 13.

16. 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, whereby the at least two corresponding bubble trap tracks are separated from each other. The cell culture chamber according to claim 12.

17. An automated cell engineering system, Comprising a cassette, the cassette being, A cell culture chamber having at least one bubble trap disposed thereon, A first body forming a first portion of the cell culture chamber, and A second body forming a second portion of the cell culture chamber, the first body and the second body being coupled to each other and configured to form a sealed volume. A second body, further comprising, A cell culture chamber in which 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, An automated cell engineering system in which 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.

18. The automatic cell engineering system according to claim 17, wherein the cell culture chamber is in a horizontal orientation.

19. The automatic cell engineering system according to claim 17, wherein the cell culture chamber includes a plurality of bubble traps disposed at the upper part.

20. The automatic cell engineering system according to claim 17, wherein the first main body includes a plurality of openings, and the second main body includes a plurality of openings.

21. The automatic cell engineering system according to claim 20, wherein the first non-porous gas-permeable material is disposed on the inner surface of the first main body, and the first non-porous gas-permeable material is configured to cover the plurality of openings of the first main body.

22. The automatic cell engineering system according to claim 20, wherein the second non-porous gas-permeable material is disposed on the inner surface of the second main body, and the second non-porous gas-permeable material is configured to cover the plurality of openings of the second main body.

23. An automatic cell engineering system, comprising a cell culture chamber, a first main body forming a first part of the cell culture chamber, having a plurality of bubble traps disposed thereon, and a bubble trap track disposed between the plurality of bubble traps and connected to the plurality of bubble traps, a second main body forming a second part of the cell culture chamber, wherein the first main body and the second main body are coupled to each other and configured to form a sealed volume, a first non-porous gas-permeable material is disposed on the first main body, and a second non-porous gas-permeable material is disposed on the second main body, The automatic cell engineering system, wherein the first non-porous gas-permeable material and the first main body are integrally formed, and the second non-porous gas-permeable material and the second main body are integrally formed.

24. The automatic cell engineering system according to claim 23, wherein the cell culture chamber is in a horizontal orientation.

25. The automatic cell engineering system according to claim 24, wherein the bubble trap track has a gradually decreasing height and / or a gradually decreasing width.

26. The automatic cell engineering system according to claim 25, wherein each of the plurality of bubble traps is formed at both ends of the first main body.

27. The first non-porous gas-permeable material is disposed on the inner surface of the first main body, and the first non-porous gas-permeable material is configured to cover a plurality of openings of the first main body. The automated cell engineering system according to claim 26.

28. The second non-porous gas-permeable material is disposed on the inner surface of the second main body, and the second non-porous gas-permeable material is configured to cover the plurality of openings of the second main body. The automated cell engineering system according to claim 27.

29. A cell culture chamber for use in an automated cell engineering system, the cell culture chamber comprising: At least one main body that forms a sealed volume; A non-porous gas-permeable material disposed on the at least one main body such that the non-porous gas-permeable material and the at least one main body are formed together; and A cell culture chamber in which no structure is disposed within the sealed volume.

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