Systems, devices, and methods for fluid control within cell processing systems

Abstract

This disclosure relates to systems, devices, and methods for automatic fluid control in cell processing systems. In embodiments, this disclosure relates to a fluid manifold comprising a cartridge, a first end panel, a second end panel, and a central panel connecting the first and second end panels, wherein each of the first and second end panels comprises a plurality of fluid paths formed therein and a plurality of valves for controlling the flow of fluid through the plurality of fluid paths.

Description

Technical Field

[0001] (Cross - reference to Related Applications) This application claims the priority of U.S. Provisional Patent Application No. 63 / 520,859, filed on August 21, 2023, the entire content of which is incorporated herein by reference for all purposes.

[0002] (Field of the Invention) The present disclosure relates to systems, devices, and methods for fluid control in a bioprocessing system, and more particularly, to a complex automated cell processing system having multiple cell processing modules within a single cartridge.

Background Art

[0003] Cell therapy involves collecting cells from an individual, processing the cells, and using the processed cells to achieve a clinical response in the same or a different individual. Cell processing is a complex workflow involving multiple steps, typically requiring separate cell processing devices and / or systems to accomplish a specific step. Fluids, such as cell material, may need to be transferred between different cell processing devices to achieve the final cell output. Improvements have been made to cell processing systems, replacing multiple cell processing devices with multiple modules within a single cartridge. However, even in these improved systems, fluids and cell material still need to be transferred between modules to perform separate cell processing steps. Therefore, each module is typically fluidically connected to a fluid source by fluid conduits. As more cell processing steps are included in a single cartridge or workflow, or as the desired throughput of cell material increases, the number of modules typically increases proportionally. As a result, the number of fluid conduits connecting the required modules often becomes extremely complex. As the complexity of the fluid pathways increases, entanglement of fluid conduits during normal use and / or during repair work in the event of breakage or damage becomes a significant problem. Furthermore, the overall throughput is generally limited by complexity, as fluid conduits may be routed inefficiently, potentially increasing the time required to transfer necessary fluids to or from the relevant modules. Complexity can also limit the number of modules that can be included in a cell processing system, because the spatial requirements of modules and / or fluid conduits may be critical. Therefore, additional systems and methods for routing fluids in cell processing systems are desirable. [Overview of the project]

[0004] This disclosure relates, in general, to systems, devices, and methods for routing fluid flows within bioprocessing systems, such as automated cell processing systems. Generally, a cartridge for cell processing may include a fluid manifold comprising a first end panel, a second end panel, and a central panel connecting the first and second end panels. Each of the first and second end panels may include a plurality of fluid paths formed therein and a plurality of valves for controlling the flow of fluid through the plurality of fluid paths. In some modifications, at least one of the plurality of valves may be a pinch valve.

[0005] The first end panel may further comprise at least one window configured for optical detection. In some modifications, at least one window may be a bubble-sensing window. The first end panel may further comprise a bubble trap. The second end panel may further comprise at least one fluid extraction port. In some modifications, at least one fluid extraction port may be a needle-free injection port. The first end panel may be fluidically connected to a first bioreactor module, and the second end panel may be fluidically connected to a second bioreactor module.

[0006] The central panel may have multiple fluid pathways. These fluid pathways in the central panel may be fluidically connected to multiple fluid pathways in the first and second end panels, respectively. The fluid pathways in the central panel may be fluidically connected to one or more pumps. In some modifications, the central panel may further include at least one pressure sensor configured to monitor the fluid flow through the fluid pathways in the central panel. The central panel may be connected to a ventilation manifold configured to provide sterile air to the fluid pathways in the central panel. The central panel may also be connected to a degassing module, which may include an air-permeable membrane. The central panel may be connected to the first and second end panels via first and second bridges.

[0007] In some variations, the fluid manifold may be fluidically connected to one or more modules of the cartridge. One or more modules of the cartridge may be selected from the group consisting of elutriation modules, electroporation modules, spinoculation modules, and cell sorting modules.

[0008] Further embodiments, features, and advantages of the present invention will become apparent from the following detailed description and through the implementation of the present invention. [Brief explanation of the drawing]

[0009] [Figure 1A] This is a block diagram of an exemplary modified example of a cell processing system. [Figure 1B] Figure 1A is a block diagram of a cartridge that may be provided to the cell processing system. [Figure 1C] Figure 1B is a block diagram of the fluid manifold for the cartridge. [Figure 1D] Figure 1C is a block diagram of the first end panel of the fluid manifold shown. [Figure 1E] Figure 1C is a block diagram of the second end panel of the fluid manifold shown. [Figure 1F] Figure 1C is a block diagram of the central panel of the fluid manifold shown. [Figure 1G] Figure 1C is a block diagram of the vent manifold of the fluid manifold shown. [Figure 1H] Figure 1C is a block diagram of the degassing module of the fluid manifold shown. [Figure 2A] This is a front perspective view of an exemplary and modified example of a cartridge that may be provided to a cell processing system. [Figure 2B] Figure 2A is a rear perspective view of the cartridge shown. [Figure 2C] This is a front perspective view of another exemplary variation of the cartridge. [Figure 3A]It is a front perspective view showing the upper side of an exemplary modification of a fluid manifold of a cartridge. [Figure 3B] It is a rear perspective view showing the lower side of the fluid manifold illustrated in FIG. 3A. [Figure 3C] It shows an exploded view of the fluid manifold illustrated in FIG. 3A. [Figure 4A] It shows a front view of an exemplary modification of a first end panel of a fluid manifold. [Figure 4B] It shows a rear view of the first end panel illustrated in FIG. 4A. [Figure 4C] It shows a perspective view of the first end panel illustrated in FIG. 4A. [Figure 5A] It shows a front view of an exemplary modification of a second end panel of a fluid manifold. [Figure 5B] It shows a rear view of the first end panel illustrated in FIG. 5A. [Figure 5C] It shows a perspective view of the first end panel illustrated in FIG. 5A. [Figure 6A] It shows a front perspective view of an exemplary modification of a central panel of a fluid manifold. [Figure 6B] It shows a rear perspective view of the central panel illustrated in FIG. 6A. [Figure 6C] It shows a front view of an exemplary modification of an intermediate body of the central panel illustrated in FIG. 6A. [Figure 7A] It shows a front perspective view of an exemplary modification of a degassing module of a fluid manifold. [Figure 7B] It shows a front view of an intermediate body of the degassing module illustrated in FIG. 7A. [Figure 8A] [[ID=?]]It shows a front perspective view of an exemplary modification of a vent manifold of a fluid manifold?. [Figure 8B] It shows a front view of an intermediate body of the vent manifold illustrated in FIG. 8A. [Figure 9A] It shows a front perspective view of an exemplary modification of a first bridge of a fluid manifold. [Figure 9B] It seems there is a formatting or tagging issue in the original text for line ID 42. I've translated it as best as possible based on the context, but it might need further clarification in the original source.Shows a rear perspective view of the first bridge illustrated in FIG. 9A. [Figure 9C] Shows a side view of the first bridge illustrated in FIG. 9A. [Figure 9D] Shows an exemplary modification of the fluid path of the first bridge illustrated in FIG. 9A. [Figure 9E] Shows an exemplary modification of the fluid path of the first bridge illustrated in FIG. 9A. [Figure 10A] Shows a front perspective view of an exemplary modification of the second bridge of the fluid manifold. [Figure 10B] Shows a rear perspective view of the second bridge illustrated in FIG. 10A. [Figure 10C] Shows a side view of the second bridge illustrated in FIG. 10A. [Figure 10D] Shows an exemplary modification of the fluid path of the second bridge illustrated in FIG. 10A. [Figure 10E] Shows an exemplary modification of the fluid path of the second bridge illustrated in FIG. 10A. [Figure 11] Is a flowchart of an exemplary modification of the control of the flow of fluid using a fluid manifold.

Best Mode for Carrying Out the Invention

[0010] This specification discloses devices, systems, and methods for controlling the flow of fluids through and between cell processing modules of one or more cell processing cartridges in order to facilitate cell processing. Multiple cell processes or cell processing steps may be performed on cells in a cell processing system (e.g., a working cell). Each cell processing step may require one or more fluids (e.g., cell suspensions, culture media, buffers, reagents). One or more fluids may be supplied to one or more modules of the working cell cartridge according to a predetermined workflow. Thus, one or more fluids may flow through a fluid manifold of the cartridge, and as a result, the fluid manifold can control the type, amount, flow rate, timing, and / or destination of any fluid flowing through it. That is, the fluid manifold may be connected to one or more modules of the cartridge by one or more fluid conduits (e.g., tubes or channels). For example, the fluid manifold may be fluidly connected to one or more of the following: elutriation modules, electroporation modules, spinoculation modules, and cell sorting modules. A fluid manifold may comprise one or more valves configured to control the flow of fluid through one or more fluid conduits. A fluid manifold may also comprise one or more fluid paths. A fluid manifold described herein may be configured to automatically control the flow of fluid through a cartridge.

[0011] A fluid manifold can be optimized to reduce the number and / or length of fluid conduits connected to it. Optimizing the fluid manifold can avoid entanglement of fluid conduits. Otherwise, entanglement can cause problems, including delays in troubleshooting due to the difficulty in identifying specific fluid conduits and / or fluid paths. Furthermore, if the length and / or routing is not optimized, entanglement can cause significant pressure drops in the fluid flowing through one or more fluid conduits, which may necessitate additional pumping capacity to deliver the fluid to its destination at the desired flow rate. Therefore, the optimization of the fluid manifold described herein may allow for the fluidic connection of an adjustable number of modules. These adjustable modules can then facilitate flexible workflows, which can be modified to increase the total throughput of cell byproducts for use in cell therapy. Thus, the fluid control system can be configured to automatically perform high-throughput cell processing in an automated cell processing system.

[0012] I. Cell Processing Systems The cell processing systems described herein may be configured to perform one or more cell processing steps within a working cell. The working cell may comprise a closed, automated environment that can be configured to maintain a sterile environment. The working cell may receive a cartridge and perform one or more cell processing steps on the cells in the cell solution (e.g., cell suspension) contained within the cartridge. For example, a cell processing system may comprise a working cell comprising a plurality of instruments, each configured to independently perform one or more cell processing steps on cells and / or cell solutions, and a robot capable of moving cartridges within the working cell (e.g., between one or more bays). The robot and / or instruments may be configured to operate automatically so as not to require operator assistance at any point in the workflow. For example, the robot may receive a cartridge and move it between locations within the working cell (e.g., instruments, bays, storage, feedthroughs) according to a pre-programmed workflow, each location being associated with one or more cell processing steps. After performing one or more cell processing steps of the pre-programmed workflow, the working cell may be configured to transfer the cartridge out of the working cell (e.g., via the robot). Additionally or alternatively, at least a portion of the cell solution may be transferred to a second cartridge (for example, via a fluid device or fluid manifold).

[0013] The cell solutions (e.g., cell suspensions) described herein may contain cells that can subsequently be processed for use in cell therapy. The cell solutions may contain cells (e.g., allogeneic cells) in a fluid such as a culture medium (e.g., cell culture medium). The cell solutions may contain cells from the same or different donors. Cells from the same donor may be divided between one or more cartridges, so that separate cell processing steps are performed in each cartridge, thereby increasing the overall throughput of the cell processing system described herein. The cell solutions may be transferred to the cartridges, for example, by an operator, before loading the cartridges into the working cell. In some modifications, the cartridges may be empty when loaded into the working cell so that the working cell can transfer the cell solutions into the cartridges. In some modifications, cells from two or more cartridges may be combined according to a predetermined ratio that can correspond to an intended therapeutic procedure for a patient.

[0014] An exemplary cell processing system for use in automated devices, systems, and methods is illustrated in Figure 1A. A block diagram of a cell processing system 100 comprising a working cell 110 and a controller 120 is shown. The working cell 110 may comprise one or more of the following: instruments 112, a robot 116 (e.g., a robotic arm), a reagent storage unit 118, a sterile liquid transfer port 132, a sterilizer source 129, a fluid source 136, a pump 138, and a sensor 151. Cartridges 114 and fluid devices 142, which may be located outside the working cell 110 and can be used inside the working cell 110, are shown by dashed lines. In some modifications, the fluid device 142 may be a sterile liquid transfer device (SLTD). However, it should be understood that the fluid device 142 may be configured to transfer any fluid (including liquids), whether sterile or not. The controller 120 may include one or more of the following: a processor 122, memory 124, communication device 126, input device 128, and display 130.

[0015] The working cell 110 may comprise a housing that is fully or at least partially sealed, within which one or more cell processing steps may be performed in a fully or at least partially automated process. As described in detail herein, the cartridge 114 may be moved using a robot 116 to reduce manual labor in the cell processing steps, and the transfer of fluids into and out of the cartridge 114 may also be performed in a fully or partially automated process. For example, one or more fluids may be stored in a fluid device 142 so that one or more fluids can be transferred to and / or taken out of the cartridge 114 via the fluid device 142. In some modifications, the fluid device 114 may be moved within the system 100 by a robot 116. Thus, the working cell 110 described herein advantageously allows for the transfer of fluids in an automated and metered manner to automate the production of cell therapy.

[0016] The working cell 110 can facilitate fluid transfer and / or cartridge transfer. For example, in some modifications, the robot 116 may be configured to move two or more cartridges 114 between different bays to perform a predetermined sequence of cell processing steps (e.g., a workflow). In this way, multiple cartridges 114 can be processed in parallel, as different steps of the cell processing workflow can be performed simultaneously on different cartridges. In another example, a sterile fluid transfer port 132 may be connected between two or more cartridges 114 to transfer cell products and / or other fluids between the cartridges 114. Furthermore, the sterile fluid transfer port 132 may be connected between any set of fluid transport components of the system 100 (e.g., cartridges 114, reagent storage unit 118, fluid source 136, fluid device 142, etc.). For example, a first sterile fluid transfer port may be connected between a first cartridge and the corresponding sterile fluid transfer port of a fluid device.

[0017] Other preferred cell processing systems and embodiments thereof are provided, for example, in U.S. Patent Application No. 17 / 198,134, U.S. Patent Application No. 18 / 731,095, U.S. Patent Application No. 18 / 759,602, and U.S. Patent Application No. 18 / 807,699, published as U.S. Patent Application Publication No. 2021 / 0283565, the entire contents of each of these are incorporated herein by reference.

[0018] A. Cartridge The cell processing systems described herein may comprise one or more cartridges having one or more modules configured to interface with or detachably connect to one or more instruments in a working cell. Some or all of the modules may, but are not required, be integrated in a fixed configuration within the cartridge. In fact, one or more of the modules are configurable or movable within the cartridge, so that various forms of cartridges can be assembled. For example, the cartridge may be a single closed unit with fixed components for each module, or the cartridge may house configurable modules connected by configurable fluidic, mechanical, optical, and electrical connections. In some modifications, one or more subcartridges, each housing a set of modules, may be used to perform various cell processing workflows. The modules may each be housed in a separate housing, or they may be incorporated together in a cartridge or subcartridge with other modules. While this disclosure generally presents modules as separate groups of components for simplicity, it should be noted that these modules may be arranged in any preferred configuration. For example, components for different modules may be scattered amongst themselves, such that each module may be defined by a set of connected components that collectively perform a predetermined function. However, the components of each module may or may not be physically grouped within the cartridge. In some embodiments, multiple cartridges may be used to process a single cell product by transferring the cell product from one cartridge to another cartridge of the same or different type, and / or by dividing the cell product into more cartridges, and / or by pooling multiple cell products into fewer cartridges.

[0019] Generally, each piece of equipment within a working cell is interfaced to or detachably connected to its respective module in a cartridge to perform a specific cell processing step. For example, if a cartridge has an electroporation module, a robot may move the electroporation equipment within the working cell to a bay in the working cell, and electroporation may be performed on the cells in the cartridge. One advantage of such a segmented module / equipment design is that expensive components (e.g., motors, sensors, heaters, lasers, etc.) can be kept within the equipment of the system, while less expensive components reside within the cartridge.

[0020] As illustrated in Figure 1B, the cartridge 114 may be configured to contain (e.g., store) a cell solution (e.g., a cell suspension) for cell processing. Any number of cell processing steps may be performed on the cells in the cartridge. Thus, the cartridge 114 may comprise one or more of the following: a bioreactor module 150, an electroporation module 160, an elutriation module 162, a spinoculation module 164, a cell sorting module 166, a fluid manifold 168, and a pump module 169. The fluid manifold 168 may be configured to transfer one or more fluids between one or more modules of the cartridge 114. For example, the fluid manifold 168 may transfer a fluid from the pump module 169 to the bioreactor module 150. In another example, the fluid manifold 168 may transfer a fluid (e.g., a cell solution) from the bioreactor module 150 to the cell sorting module 166. The cell solution may contain cell material including target cells linked to magnetic particles. In another example, the fluid manifold 168 may transfer fluid from, for example, the cell sorting module 166 to any other module 114 after a cell sorting process may have been performed. The fluid manifold 168 may be configured to transfer sorted cells (e.g., magnetically tagged cells) to one module and non-target cell material to a different module.

[0021] The bioreactor module 150 may be configured to contain a cell solution. The bioreactor module 150 may further include a mixing chamber in which the cell solution may be mixed with one or more reagents. One or more reagents may comprise magnetic particles configured to bind to a specific type of cell (e.g., target cells). The elutriation module 162 may be configured to perform an elutriation process that can separate cell material according to size, shape, and / or density. The spinoculation module 164 may be configured to perform a spinoculation process that can bind different types of cells together.

[0022] Other suitable cartridges and cell processing modules that may be used in the automated cell processing work cell described herein are provided, for example, in U.S. Patent Application No. 18 / 652,602, U.S. Patent Application No. 18 / 532,621, U.S. Patent Application No. 18 / 620,826, and U.S. Patent Application No. 18 / 611,632, the entire contents of each of these are incorporated herein by reference.

[0023] Referring to Figures 2A and 2B, exemplary variations of the cartridge 200 are illustrated. The cartridge 200 may comprise a water elutriation module 210, a fluid manifold 222, a first cell sorting module 224a, a second cell sorting module 224b, an auxiliary module 226, a fluid device tray 228, a liquid container 230, and a pump module 232. Although these figures illustrate the presence of two cell sorting modules, it should be understood that any number of cell sorting modules may be used as desired. For example, the cartridge may contain one, two, three, four, or more cell sorting modules, depending on the size of the cartridge, the presence of other cell processing modules within the cartridge, etc. Cell sorting modules 224a and 224b may perform a magnetic cell sorting process. The electroporation module 220 may be configured to facilitate intracellular delivery of macromolecules (i.e., transfection by electroporation). Discharge from one or more capacitors or current sources can generate a sufficient current within the chamber to facilitate the transfer of polynucleotides, proteins, nucleoprotein complexes, or other macromolecules into cells within the cell products. The fluid manifold 222 may have at least one fluid conduit. At least one fluid conduit of the fluid manifold 222 may be configured to allow a fluid to pass through it. For example, at least one fluid may be a liquid or a gas. In some modifications, at least one fluid may contain a solution of cells of varying sizes and densities. The fluid manifold 222 may have at least one fluid inlet and at least one fluid outlet, and may have at least one valve. The fluid manifold 222 may be fluidically connected to at least one module in the cartridge 200. For example, the fluid manifold 222 may be configured to transfer at least one fluid to first and / or second cell sorting modules 224a, 224b. The fluid manifold 222 may communicate with a controller, such as the controller 120 described with reference to Figure 1A.For example, at least one valve of the fluid manifold 222 may open and / or close in response to a command transmitted by the controller 120 to transfer fluid between various modules of the cartridge according to a predetermined workflow.

[0024] The fluid transfer port tray 228 may have one or more ports configured to transfer fluid to or from one or more fluid devices. That is, each port of the fluid transfer port tray 228 may be configured to facilitate the transfer of sterile fluid. In some modifications, each port may be fluidically connected to a fluid conduit configured to fluidically connect to at least one module of the cartridge 114. For example, each port of the fluid transfer port tray 228 may be fluidically connected to a fluid manifold 222. In this way, fluid can flow from a fluid device connected to a port of the fluid transfer port tray 228 to the fluid manifold 222, or vice versa. In some modifications, each port of the fluid transfer port tray 228 may be fluidically connected to a liquid storage container 230. The liquid storage container 230 may be configured to contain fluid. In some modifications, the fluid may be a liquid or a gas. In some modifications, the liquid storage container 230 comprises multiple liquid containers. For example, the liquid storage container 230 may comprise one container, two containers, or three containers. The liquid storage container 230 can be fluidically connected to at least one module of the cartridge 200. In some variations, the liquid container 230 can be fluidically connected to a fluid manifold 222. Thus, the fluid can flow between the ports of the fluid transfer port tray 228, the fluid manifold 222, and the liquid storage container 230.

[0025] The cartridge may further comprise a pump module 232 having a pump configured to pump fluid in one or more directions along at least one fluid path. For example, the pump module 232 may be configured to pump fluid to or from one or more of the following modules in the cartridge: the elutriation module 210, the fluid manifold 222, the cell sorting modules 224a, 224b, the auxiliary module 226, the fluid device tray 228, the liquid container 230, and any other modules in the cartridge. The auxiliary module 226 may be configured to engage with at least one device and / or module. The auxiliary module 226 may comprise at least one electrical connector and / or at least one fluid connector. In some modifications, the auxiliary module 226 may be removed and replaced by any other module.

[0026] Figure 2C shows an exemplary modification of the cartridge 200 with the fluid device tray 228 removed. As shown, the fluid manifold 222 may comprise a first end panel 252, a central panel 254, and a second end panel 256. The first end panel 252 and the second end panel 256 may each define the outer surface of the cartridge 200. The central panel 254 may be positioned within the cartridge 200, as a result of which the central panel 254 cannot define the outer surface of the cartridge 200. The central panel 254 may extend between the first end panel 252 and the second end panel 256. That is, the central panel 254 may be connected to each of the first and second end panels 252 and 256. Thus, one or more fluid paths extend between the first end panel 252 and the second end panel 256 via the central panel 254. One or more of the first end panel, the second end panel, and the central panel may be fluidically connected to one or more of the other modules of the cartridge 200, including one or more of the elutriation module 210, cell sorting modules 224a, 224b, auxiliary module 226, fluid device tray 228 (see Figures 2A and 2B), and liquid container 230.

[0027] Various materials, including metal, plastic, rubber, and / or glass, or combinations thereof, may be used to construct the cartridge (including its modules) and the cartridge housing. The cartridge, its components, and its housing may be molded, machined, extruded, 3D printed, or any combination thereof. The cartridge may house commercially available components (e.g., tubes, valves, fittings). Commercially available components may be attached to or integrated with custom components or devices. The cartridge housing may constitute an additional sealing layer to further protect the sterility of the cell products.

[0028] i. Fluid manifold To allow fluid to move between various modules of the cartridge, the cartridge described herein comprises a novel fluid manifold. The fluid manifold may be configured to deliver fluid (e.g., cell products) to one or more modules according to a predefined workflow that can be preprogrammed into a controller of the working cell, as described throughout this specification. Advantageously, the fluid manifold can replace some or all of the tubing in the cartridge. The fluid manifold may be controlled to deliver fluid to the cartridge modules in a predefined order according to the workflow, or one or more modules may be bypassed collectively using one or more valves. The fluid manifold may be fluidically connected to a plurality of fluid devices used to provide solutions or reagents, to store cell products, or to collect waste liquid or reagents. The fluid may be a liquid or a gas. In some modifications, the fluid may be a solution (e.g., cell solution, cell suspension). For example, the solution may contain one or more of cells, culture media, buffers, and reagents.

[0029] The cartridge may include one or more pumps (e.g., pump modules) fluidically connected to a fluid manifold. The pumps may be direct lift pumps, positive displacement pumps, gravity pumps, reciprocating pumps, rotary pumps, or peristaltic pumps. In some embodiments, one or more of the system's components may have one or more integrated pump actuators. This allows the system to transport fluid between modules, fluid containers, or other components, while simultaneously allowing the cartridge to interface with its modules. The system (e.g., a working cell) may also include dedicated pump equipment configured to interface with a pump module containing pumps.

[0030] A fluid manifold may be used to facilitate cell processing steps. For example, in some variations, the concentration step may include concentrating a selected cell population in a solution by transporting the solution to a cartridge elutriation module via a fluid manifold; operating a robot to move the cartridge to the elutriation device so that the elutriation device can interface with the elutriation module; and operating the elutriation device to concentrate the selected cell population in the elutriation module.

[0031] In some variations, the washing step may include washing a selected cell population in a solution by transporting the solution to a cartridge elutriation module via a fluid manifold; operating a robot to move the cartridge to the elutriation instrument so that the elutriation module can interface with the instrument; and operating the elutriation instrument to cause the elutriation module to remove culture medium from the solution, introduce culture medium into the solution, and / or replace the culture medium in the solution. The removal and / or introduction of culture medium may be performed by a fluid manifold. During the concentration step, target cells can be concentrated. Different concentration steps may include, but are not limited to, platelet depletion, cytokine depletion, erythrocyte depletion, and volume concentration.

[0032] In some variations, the selection step may include selecting a selected population of cells in a solution (e.g., by cell surface proteins) by transporting the solution to a selection module in a cartridge via a fluid manifold; operating a robot to move the cartridge to a selection device so that the selection module interfaces with the selection device; and operating the selection device to cause the selection module to select the selected population of cells.

[0033] In some variations, the sorting step may include sorting a population of cells in a solution by transporting the solution to a sorting module in a cartridge via a fluid manifold; operating a robot to move the cartridge to a sorting device so that the sorting module can interface with the sorting device; and operating the sorting device to allow the sorting module to sort the population of cells.

[0034] In some variations, the static step may be configured to maintain the fluid in an unagitated state. For example, an unagitated state may relate to maintaining the fluid within a fluid device without agitating it using an impeller or the like. In contrast, an agitated state may relate to agitating the fluid via an impeller. In another example, the static step may include transporting the solution from the cartridge to the bioreactor module via a fluid manifold, operating a robot to move the cartridge to the bioreactor instrument so that the bioreactor module can interface with the bioreactor instrument, and operating the bioreactor instrument to maintain the cells in the bioreactor module.

[0035] In some variations, the proliferation step may include growing cells in solution by transporting the solution to a bioreactor module in a cartridge via a fluid manifold; operating a robot to move the cartridge to the bioreactor device so that the bioreactor device interfaces with the bioreactor module; and operating the bioreactor device to grow cells in the bioreactor module by cell replication. The bioreactor device may provide closed-loop control of one or more of the bioreactor temperature, dissolved oxygen concentration, acidity (pH), and mixing intensity. A single bioreactor device may interface with one or more bioreactors (e.g., multiple bioreactors of the same or different sizes), or the system may comprise multiple bioreactor devices. The bioreactor device may be designed to interface with several cartridges simultaneously.

[0036] In some variations, the tissue digestion step includes transporting the enzyme reagent via a fluid manifold to a module containing a tissue-containing solution, so that the enzyme reagent can cause tissue digestion and release a selected population of cells into the solution.

[0037] In some variations, an activation step, such as a T cell activation step or an NK cell activation step, may include activating a selected cell population in a solution by transporting an activation reagent via a fluid manifold to a module containing a solution of cells.

[0038] In some variations, the transduction step includes transducing a selected cell population in a solution by transporting an effective amount of vector to a module containing a solution of cells via a fluid manifold. Multiple vectors can be used in a single transduction step. The vector may be delivered with one or more proteins (e.g., proteins delivered in liposomes or lipid nanoparticles) or using cell-permeable peptides. The transduction step may include modifying the cells by inserting, deleting, or mutating one or more polynucleotides in the cells (e.g., the cell's genome, or any other polynucleotides in the cells).

[0039] In some variations, the fluid manifold may be used more than once in the cell processing method. In an exemplary method, the method may include: culturing a cell product in a first bioreactor module; transferring the cell product to an elutriation module via a fluid manifold to concentrate a desired cell type; transferring the cell product to a second bioreactor module via a fluid manifold for a second culture step; washing the elutriation module with a washing solution using the fluid transferred by the fluid manifold; and transferring the cell product to the elutriation module via a fluid manifold for a second concentration step.

[0040] Referring to Figure 1C, a block diagram of an exemplary modification of the fluid manifold 168 is illustrated. The fluid manifold 168 may comprise a first end panel 172, a second end panel 174, a first bridge 180, a second bridge 182, a central panel 176, a ventilation manifold 177, and a degassing module 178. The first and second end panels 172, 174 may be connected to the central panel by the first and second bridges 180, 182. That is, the first end panel 172 may be connected to the first bridge 180, which may also be connected to the central panel 176. Similarly, the second end panel 174 may be connected to the second bridge 182, which may also be connected to the central panel 176. The end panels 172 and 174 can be connected to the respective bridges 180 and 182 by mechanical fasteners (e.g., screws, nails, bolts), adhesives (e.g., glue), friction fittings (e.g., projections of one component that are received in corresponding openings of one component, such as a fluid conduit), or a combination thereof.

[0041] One or more components of a fluid manifold can be fluidically connected. For example, a first end panel 172 can be fluidically connected to a central panel 176 via a first bridge 180. That is, the first end panel 172 may have a fluid path that is fluidically connected to the fluid path of the first bridge 180, which can then be fluidically connected to the fluid path of the central panel 176. Similarly, a second end panel 174 can be fluidically connected to a central panel 176 via a second bridge 182. That is, the second end panel 174 may have a fluid path that is fluidically connected to the fluid path of the second bridge 182, which can then be fluidically connected to the fluid path of the central panel 176. In some modifications, the first end panel 172 may be directly fluidically connected to the central panel 176, and as a result, the fluid connection between them can bypass the first bridge 180. Additionally or alternatively, the second end panel 174 may be directly fluidically connected to the central panel 176, thereby bypassing the second bridge 182 for the fluid connection between them. The central panel 176 may be fluidically connected to one or more of the vent manifold 177 and the degassing module 178. For example, the fluid path of the vent manifold 177 may be fluidically connected to the fluid path of the central panel 176. Additionally or alternatively, the fluid path of the degassing module 178 may be fluidically connected to the fluid path of the central panel 176. In some modifications, the fluid paths of the first and / or second end panels 172, 174 may be fluidly connected to the vent manifold 177 and / or the degassing module 178.

[0042] Figure 1D shows a block diagram of an exemplary modification of the first end panel 172. The first end panel 172 may comprise a fluid path 1010, a window 1012, a bubble trap 1014, an outlet port 1015, and a valve 1016. The fluid path 1010 may be configured for fluid flow. In some modifications, the first end panel 172 may comprise multiple fluid paths. The fluid path 1010 may be defined by grooves, recesses, or channels. In some modifications, there may be 1 to 70 fluid paths, 5 to 65 fluid paths, 10 to 60 fluid paths, or 15 to 55 fluid paths (including any value or sub-range thereof). For example, in some modifications, there may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 fluid paths.

[0043] The fluid path 1010 may be configured to facilitate one or more measurements. For example, the fluid path 1010 may be covered by a film. The film may be transparent, and as a result, one or more measurements of the fluid flowing through the fluid path 1010 may be generated. The film may be removable, and as a result, the user may remove the film to clean and / or repair the fluid path 1010. In some modifications, the film may be transparent so that the fluid in the fluid path 1010 can be observed while maintaining a fluid seal between the fluid in the fluid path 1010 and the external environment. In some modifications, the film may be formed integrally with the fluid path, and as a result, the film is not removable. Thus, the first end panel 172 may be manufactured from any biocompatible material that can facilitate the fluid path described herein. For example, the first end panel 172 containing the fluid path 1010 may be formed by compression, rotation, blowing, and / or injection molding, etc. In some modifications, the fluid path 1010 may be machined, extruded, or 3D printed. The fluid path 1010 may be formed integrally with the first end panel, or, in some modifications, may be connected to it. In some modifications, the first end panel 172 may be made from a metal (e.g., aluminum), a polymer (e.g., polyethylene terephthalate glycol, polymethyl methacrylate), or a combination thereof. The film may be made from a polymer (e.g., polyethylene terephthalate glycol, polymethyl methacrylate). In further modifications, the film and the first end panel 172 may be formed integrally using the same material.

[0044] The fluid path 1010 may be fluidically connected to one or more fluid control mechanisms. For example, the fluid path 1010 may be fluidly connected to a valve 1016, an outlet port 1015, and / or a bubble trap 1014. The valve 1016 may be configured to control the flow of fluid through the fluid path 1010. For example, the valve 1016 may be configured to transfer fluid from the module to the central panel 176. The valve 1016 may be actuated, thereby allowing it to transition between an open configuration and a closed configuration. The valve 1016 may transition between configurations in response to manual input, such as a user manually opening and closing the valve 1016 (e.g., by applying a translational force). In further modifications, the valve 1016 may transition between configurations in response to electrical signals, such as signals transmitted by a controller, such as the controller 120 in Figure 1A. In some variations, there may be 1 to 60 valves, 1 to 50 valves, 10 to 50 valves, 20 to 50 valves, or 40 to 50 valves (including any value or subrange thereof). For example, in some variations, there may be 1, 10, 20, 30, 35, 40, 43, 45, or 50 valves. In variations with multiple valves, each valve may be individually operable. For example, in variations with two or more valves, one or more valves may remain open while the remaining valves remain closed. Similarly, one or more valves may remain closed while the remaining valves remain open. Valve 1016 may include pinch valves, check valves, ball valves, diaphragm valves, or gate valves. For example, valve 1016 may include pinch valves. A pinch valve may comprise a button, a fluid conduit, a spring, and a rod, and as a result, the pinch valve can be switched from a closed configuration to an open configuration, or vice versa, by applying force to the button (e.g., by pressing the button with a translational force). In some modifications, the open configuration may correspond to a spring in an extended (e.g., unpressed) configuration, as a result, fluid can flow through the fluid conduit. The closed configuration may correspond to a spring in a retracted (e.g., pressed) configuration, as a result, the rod blocks the fluid conduit, preventing fluid from flowing through it.In further modifications, the configurations may be reversed, with the open configuration corresponding to the retraction spring configuration and the closed configuration corresponding to the extension configuration. In yet another modification, the pinch valve may have two or more fluid conduits. In such modifications, the first fluid conduit may be open and the second fluid conduit may be closed. Applying force to a button allows the configuration of each fluid conduit to be switched.

[0045] The bubble trap 1014 may be configured to perform a bubble trapping process. A fluid, such as a fluid having a mixture of liquid and gas (e.g., bubbles), may flow through the fluid path 1010 and through the bubble trap 1014. The bubble trap 1014 may be configured to hold gas while allowing liquid to continue to flow out of the bubble trap 1014 through the fluid path 1010. For example, the bubble trap 1014 may contain a certain amount of fluid (e.g., liquid and gas), so that any additional fluid entering the bubble trap can interact with the fluid already in the bubble trap 1014. That is, any gas in the additional fluid can rise to the surface of the liquid already in the bubble 1014, and any liquid in the additional fluid can mix with the liquid already in the bubble 1014. The gas may remain in the bubble trap 1014, and the liquid may flow out of the bubble trap 1014. In some modifications, the bubble trap 1014 may comprise a gas-permeable membrane (e.g., unidirectional gas-permeable). That is, the fluid may flow into the bubble trap 1014 such that the fluid comes into contact with (e.g., collides with) the membrane, so that any gas in the fluid can permeate the membrane, but any liquid in the fluid cannot. The liquid may continue to flow out of the bubble trap 1014, but the gas may remain trapped by the membrane. Thus, the bubble trap 1014 may be configured to provide the rest of the fluid manifold 168 with a liquid that is substantially free of bubbles. The bubble trap 1014 may have a shape suitable for performing the bubble trapping process. For example, the bubble trap may be shaped as a triangle, rectangle, trapezoid, circle, or a combination thereof. Any number of bubble traps may be used as desired. In some modifications, there may be 1 to 5 bubble traps, 1 to 4 bubble traps, or 1 to 3 bubble traps, including 1, 2, 3, 4, or 5 bubble traps.

[0046] The outlet port 1015 may be configured to transfer fluid to and / or from another component of the fluid manifold. For example, the outlet port 1015 may be configured to transfer fluid from the first end panel 172 to the central panel 176. That is, the fluid can flow from the valve 1016 through the outlet port 1015. In another example, the outlet port 1015 may be configured to receive fluid from the central panel 176. That is, the fluid can flow through the outlet port 1015 to the valve 1016. The outlet port 1015 may be fluidically connected to a bridge connected to the central panel, such as the first bridge 180. That is, sterile fluid transfer may occur between the outlet port 1015 and the corresponding fluid transfer port of the first bridge 180. In some modifications, the first end panel may have multiple outlet ports. For example, in some variations, there may be 1 to 60 outlet ports, 10 to 60 outlet ports, 20 to 60 outlet ports, or 40 to 60 outlet ports, or any value or subrange thereof. For example, in some variations, there may be 1, 5, 10, 20, 30, 40, 45, 50, 55, or 60 outlet ports. The number of outlet ports may correspond to the number of fluid transfer ports on the corresponding bridge (e.g., end panel transfer ports).

[0047] The window section 1012 may be configured to facilitate fluid measurement. For example, the measurement may include a bubble count value. The bubble count value may represent the amount of bubbles in the fluid. The bubble count value may be compared to a predetermined condition (e.g., a threshold). The comparison may be performed by the controller 120, which in some modifications may determine the response by the fluid manifold. For example, the response may include routing the fluid through the fluid path 1010 and through the bubble trap 1014 if the comparison indicates that the bubble count value meets or exceeds a predetermined condition. Therefore, the window section 1012 may be transparent. In some modifications, the window section 1012 may be manufactured using a suitable translucent material such as a polymer (e.g., polyethylene terephthalate glycol, polymethyl methacrylate). Any number of windows may be used as desired. In some modifications, there may be 1 to 60 windows, 10 to 60 windows, 20 to 60 windows, or 40 to 60 windows, or any value or subrange thereof. For example, in some variants, there may be 1, 5, 10, 20, 30, 40, 45, 50, 55, 57, or 60 window sections.

[0048] Figure 1E shows a block diagram of an exemplary modification of the second end panel 174. The second end panel 174 may comprise a fluid path 1020, a window 1022, a fluid extraction port 1024, an outlet port 1025, and a valve 1026. The fluid path 1020, the window 1022, the outlet port 1025, and the valve 1026 may correspond to the descriptions provided for the fluid path 1010, the window 1012, the outlet port 1015, and the valve 1016 with reference to Figure 1D. The fluid extraction port 1024 may be configured to transfer fluid from the second end panel. That is, fluid from any position in the fluid manifold can be flowed to the fluid extraction port of the second end panel 174. Thus, the fluid extraction port 1024 can be used to extract at least a portion of the fluid in the fluid manifold 168. The fluid extraction port 1024 may, advantageously, provide an extraction path that can be used to quickly remove fluid from the fluid manifold 168 in emergencies, etc. Emergencies may include power loss, leakage elsewhere in the fluid manifold, fluid blockage at one or more points in the fluid manifold and / or cartridge, or interruption of another workflow. In some modifications, the fluid extraction port 1024 may be equipped with a needleless injection port. Thus, the fluid extraction port 1024 may form a liquid and / or gas impermeable barrier that can be perforated by the user (e.g., using a syringe) during the fluid extraction process and subsequently reformed at the end of the fluid extraction process. The fluid extraction process may include connecting to the fluid extraction port 1024 using a fluid conduit, receiving fluid from a fluid path associated with the fluid extraction port 1024, and then separating it from the fluid extraction port 1024. Any number of fluid extraction ports may be used as desired. In some variations, there may be 1 to 20 fluid extraction ports, 1 to 15 fluid extraction ports, or 5 to 15 fluid extraction ports, or any value or subrange thereof. For example, in some variations, there may be 1, 5, 8, 10, 11, 12, 15, or 20 fluid extraction ports.

[0049] Figure 1F shows a block diagram of an exemplary modification of the central panel 176. The central panel 176 may comprise a fluid path 1030, a fluid transfer port 1035, a bridge transfer port 1034, and a sensor 1038. The fluid path 1030 may define the flow path of fluid through the central panel 176. In some modifications, the central panel 176 may comprise multiple fluid paths. The fluid path 1030 may be defined by grooves, depressions, channels, etc. The fluid path 1030 may have a cross-sectional shape suitable for transporting fluid, such as circular, square, triangular, trapezoidal, or a combination thereof. In some modifications, the fluid path 1030 may have side walls (e.g., circular cross-section) or two or more side walls (e.g., rectangular cross-section). The fluid path 1030 may be fluidically connected to one or more of the fluid transfer port 1035, the bridge transfer port 1034, and the sensor 1038. Any number of fluid paths may be used as desired. In some modifications, there may be 1 to 50 fluid pathways, 10 to 50 fluid pathways, 20 to 50 fluid pathways, or 30 to 50 fluid pathways, or any value or subrange thereof. For example, in some modifications, there may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 fluid pathways. The central panel 176 may be manufactured from any biocompatible material capable of facilitating the fluid pathways described herein. For example, the central panel 176 including the fluid pathway 1030 may be formed by compression, rotation, blowing, and / or injection molding, etc. In some modifications, the central panel 176 including the fluid pathway 1030 may be machined, extruded, or 3D printed. The fluid pathway 1030 may be formed integrally with the central panel 176, or, in some modifications, may be connected to it. In some variations, the central panel 176 may be made from a metal (e.g., aluminum), a polymer (e.g., polyethylene terephthalate glycol, polymethyl methacrylate), or a combination thereof.

[0050] The fluid transfer port 1035 may be configured to receive fluid from one or more modules of the cartridge. That is, a fluid conduit (e.g., a tube) may fluidically connect one or more modules to the fluid transfer port 1035. The fluid transfer port 1035 may be equipped with a seal, thereby allowing sterile fluid to be transferred through it. Thus, fluid can flow from one or more modules into the fluid path 1030, thereby allowing the fluid to be transferred to other components of the fluid manifold 168. The bridge transfer port 1034 may be configured to receive fluid from a bridge, such as a first bridge 180 or a second bridge 182. For example, the fluid path of the first bridge 180 may be fluidically connected to the bridge transfer port 1034. In some modifications, the fluid path of the second bridge 182 may be fluidically connected to the bridge transfer port 1034. The bridge transfer port 1036 may be equipped with a seal, thereby allowing sterile fluid to be transferred through it. Therefore, the fluid can flow from one or more bridges (and in some variations, one or more end panels) into the fluid path 1030, thereby enabling the fluid to be transferred to other components of the fluid manifold 168.

[0051] Sensor 1038 may be configured to measure one or more parameters of the fluid flowing through the central panel 176 (e.g., via the fluid path 1030). In some variations, sensor 1038 may include a pressure sensor (e.g., a piezoelectric sensor, a strain gauge sensor), an optical sensor (e.g., a camera), a temperature sensor, or a humidity sensor. One or more parameters may include a pressure value, a dissolved oxygen value, a pH value, or a cell count value. For example, sensor 1038 may include a pressure sensor configured to measure the pressure value of the fluid. The pressure value may be approximately 1 psi to approximately 25 psi, approximately 1 psi to approximately 20 psi, or approximately 1 psi to approximately 15 psi, including approximately 1 psi, approximately 5 psi, approximately 10 psi, approximately 15 psi, or approximately 25 psi. The pressure value may be useful in indicating the state of the fluid in the relevant fluid channel. For example, a non-zero pressure value may indicate that fluid is likely flowing through the relevant fluid channel. In another example, the pressure value can be compared to an expected value to determine whether an expected volume of fluid is likely flowing through the relevant fluid channel. Pressure values ​​that do not match the expected values ​​may indicate a problem somewhere in the system (e.g., blockage, leak, air bubbles), which may result in a decrease in flow velocity. In some variations, the central panel may be equipped with multiple sensors. For example, in some variations, there may be 1 to 20 sensors, 1 to 16 sensors, 6 to 14 sensors, or 8 to 12 sensors, including 1, 2, 4, 6, 8, 10, or 12 sensors. In some variations, multiple sensors may define an analytical instrument, which in turn allows for the parallel generation and / or scrutiny of multiple measured parameters.

[0052] In some modifications, the sensor 1038 may correspond to one or more pumps of a pump module 169 that can be fluidically connected to a central panel 176. In some modifications, the central panel 176 may be fluidically connected to one, two, three, four, five, six, seven, eight, nine, or ten pumps of the pump module 169. Thus, the sensor 1038 may be configured to monitor the fluid flow to and from the central panel 176 via one or more pumps. A flex circuit may be wired to the sensor 1038. The flex circuit may be connected to the central panel 176. In some modifications, the flex circuit may be located elsewhere in the fluid manifold 168, or in further modifications, at any other suitable location in the cartridge 114. The flex circuit may be wired to communicate with a controller 120, as a result allowing a user to remotely monitor the measurements generated by the sensor 1038. In some variations, one or more of the sensors 1038 and flex circuits can communicate wirelessly with the controller 120, thereby reducing or eliminating the amount of wiring in the cartridge 114.

[0053] Figure 1G shows a block diagram of an exemplary modification of the vent manifold 177. The vent manifold 177 may comprise a fluid path 1040, an inlet port 1042, an outlet port 1044, a gas port 1045, a fluid channel 1046 (e.g., a fluid cavity), and a filter 1048. The inlet port 1042 and / or the outlet port 1044 may be fluidically connected to the fluid path 1040. The inlet port 1042 and / or the outlet port 1044 may be configured to receive fluid. In some modifications, the inlet port 1042, the outlet port 1044, and / or the gas port 1045 may be fluidically connected to the central panel 176 via a fluid conduit (e.g., a tube). For example, the first end of a first fluid conduit may be connected to the inlet port 1042, and the second end of the fluid conduit may be connected to a fluid transfer port 1035, such as a first fluid transfer port. In another example, the first end of a second fluid conduit may be connected to an outlet port 1044, and the second end of the fluid conduit may be connected to a fluid transfer port 1035, such as a second fluid transfer port. In yet another example, the first end of a third fluid conduit may be connected to a gas port 1045, and the second end of the fluid conduit may be connected to a fluid transfer port 1035, such as a third fluid transfer port. Thus, fluid can flow into the vent manifold 177 through the inlet port 1042 and out of the vent manifold 177 through the outlet port 1044. Gas can flow through the gas port 1045 before, during, and / or after the fluid can flow through the inlet port 1042 and / or outlet port 1044. Any number of inlets, gases, and / or outlet ports may be used as desired, and there may be more inlet ports than outlet ports, or vice versa. In some variations, there may be 1 to 10 inlet ports, 1 to 8 inlet ports, or 1 to 3 inlet ports, including 1, 2, 3, 4, or 5 inlet ports. In some variations, there may be 1 to 10 outlet ports, 1 to 8 outlet ports, or 1 to 3 outlet ports, including 1, 2, 3, 4, or 5 outlet ports.In some variations, there may be 1 to 10 gas ports, 1 to 8 gas ports, or 1 to 3 gas ports, including 1, 2, 3, 4, or 5 gas ports.

[0054] The fluid path 1040 can define the flow path of fluid through the vent manifold 177. In some modifications, the vent manifold 177 may have multiple fluid paths. The fluid path 1040 may be defined by grooves, depressions, channels, etc. The fluid path 1040 may have a cross-sectional shape suitable for transporting fluid, such as circular, square, triangular, trapezoidal, or a combination thereof. In some modifications, the fluid path 1040 may have side walls (e.g., circular cross-section) or two or more side walls (e.g., rectangular cross-section). In some modifications, there may be 1 to 10 fluid paths, 1 to 8 fluid paths, or 1 to 5 fluid paths, including 1, 2, 3, 4, or 5 fluid paths. The fluid path 1040 may be fluidically connected to one or more of the inlet port 1042, outlet port 1044, gas port 1045, fluid channel 1046, and filter 1048. For example, a fluid may flow through the inlet port 1042, along the fluid path 1040, through the filter 1048, and through the outlet port 1044. Thus, the filter 1048 may be configured to remove one or more particles (e.g., solid particles) from the fluid flowing along the fluid path 1040. Additionally or alternatively, the fluid may flow through the gas port 1045, along the fluid path 1040, into the fluid channel 1046. The vent manifold 177, including the fluid path 1040 and / or any other components thereof, may be formed by compression, rotation, blowing, and / or injection molding, etc. The fluid path 1040 may be formed integrally with the vent manifold 177, or, in some modifications, may be connected thereto. In some modifications, the vent manifold 177 may be manufactured from a metal (e.g., aluminum), a polymer (e.g., polyethylene terephthalate glycol, polymethyl methacrylate), or a combination thereof.

[0055] The vent manifold 177 may be configured to perform a venting process. The venting process may be configured to mitigate the effects associated with clogging and / or condensation. For example, a filter 1048 may remove a certain number of particles from the fluid flowing along the fluid path 1040, thereby restricting the flow through the filter 1048. The restricted flow may correspond to a decrease in flow velocity, an increase in the pressure drop due to the fluid, and / or an increase in the fluid pressure. In another example, the fluid may condense on one or more internal surfaces of the vent manifold 177 or the central panel, such as the fluid paths 1030, 1040. If not adequately mitigated, condensation may obstruct the flow of fluid through the vent manifold 177. Therefore, the venting process can mitigate the effects of clogging and / or condensation by transferring gas from the external environment into the fluid path 1040 of the vent manifold 177 and / or transferring gas from the fluid path 1040 to the external environment. For example, the gas may be contained within the fluid channel 1046, and as a result, the gas can be selectively released into the fluid path 1040 to release any particles that may have clogged the filter 1048 and / or remove any condensed fluid in the fluid path 1040. The fluid channel 1046 may have a volume corresponding to the volume of gas required to perform the aeration process. For example, in some modifications, the volume of the fluid channel 1046 may be approximately 0.25 mL to approximately 5 mL, approximately 0.5 mL to approximately 2.5 mL, approximately 0.5 mL to approximately 1.5 mL, or approximately 0.75 mL to approximately 1.25 mL, including approximately 0.25 mL, approximately 0.5 mL, approximately 0.75 mL, approximately 1 mL, approximately 2 mL, or approximately 3 mL. In some modifications, there may be multiple fluid channels, such as 1 to 10 fluid channels, including 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 fluid channels. The fluid channels may be fluidically connected to one another, or, in some variations, may not be. Each fluid channel may be configured to perform the ventilation process described herein.

[0056] Figure 1H shows a block diagram of an exemplary modification of the degassing module 178. The degassing module 178 may comprise a fluid path 1050, an inlet port 1052, an outlet port 1054, a gas port 1058, and a membrane 1056. The inlet port 1052 and / or the outlet port 1054 may be fluidically connected to the fluid path 1040. The inlet port 1052 and / or the outlet port 1054 may be configured to receive fluid. In some modifications, the inlet port 1052 and / or the outlet port 1054 may be fluidically connected to the central panel 176 via a fluid conduit (e.g., a tube). For example, the first end of a first fluid conduit may be connected to the inlet port 1052, and the second end of the fluid conduit may be connected to a fluid transfer port 1035, such as a first fluid transfer port. In another example, the first end of the second fluid conduit may be connected to the outlet port 1054, and the second end of the fluid conduit may be connected to a fluid transfer port 1035, such as a second fluid transfer port. Thus, fluid can flow into the degassing module 178 via the inlet port 1052 and out of the vent manifold 177 via the outlet port 1054. In some modifications, there may be 1 to 10 inlet ports, 1 to 8 inlet ports, or 1 to 3 inlet ports, including 1, 2, 3, 4, or 5 inlet ports. In some modifications, there may be 1 to 10 outlet ports, 1 to 8 outlet ports, or 1 to 3 outlet ports, including 1, 2, 3, 4, or 5 outlet ports.

[0057] The fluid path 1050 can define the flow path of the fluid through the degassing module 178. In some modifications, the degassing module 178 may have multiple fluid paths. For example, in some modifications, there may be 1 to 10 fluid paths, 1 to 5 fluid paths, or 1 to 3 fluid paths, including 1, 2, 3, 4, 5, or 10 fluid paths. The fluid path 1050 can be defined by grooves, recesses, or channels. The fluid path 1050 may have a cross-sectional shape suitable for transporting fluid, such as circular, square, triangular, trapezoidal, or a combination thereof. In some modifications, the fluid path 1050 may have side walls (e.g., circular cross-section) or two or more side walls (e.g., rectangular cross-section). The membrane 1056 can cover at least a portion of the fluid path 1050, resulting in a contact surface area between the membrane and the fluid flowing through the fluid path. The gas port 1058 may be positioned on the opposite side of the membrane 1056 from the fluid path 1050. The membrane 1056 may be a gas-permeable membrane. For example, the membrane 1056 may be configured to allow a gas (e.g., air) to permeate from the fluid path 1050 to the gas port 1058 while retaining a liquid in the fluid path 1050. In some modifications, the membrane may be made from superhydrophobic polyvinylidene fluoride (PVDF), polyethersulfone (PES), polydimethylsiloxane (PDMS), or polytetrafluoroethylene (PTFE). The degassing module 178, including the fluid path 1050 and / or any other components thereof, may be formed by compression, rotation, blowing, and / or injection molding, etc. The fluid path 1050 may be formed integrally with the degassing module 178, or, in some modifications, may be connected to it. In some variations, the degassing module 178 may be made from a metal (e.g., aluminum), a polymer (e.g., polyethylene terephthalate glycol, polymethyl methacrylate), or a combination thereof.

[0058] The degassing module 178 may be configured to perform a degassing process. The degassing process may be configured to remove gas (e.g., dissolved gas) from a liquid. For example, during the degassing process, suction (e.g., vacuum) may be applied to the gas port 1058, thereby allowing gas (e.g., air) to be drawn from the fluid flowing along the fluid path 1050 of the degassing module 178. In some modifications, the fluid path 1050 may define a meandering fluid path configured to maximize the ratio of contact surface area to fluid volume. The meandering path may have multiple bends within the fluid path 1050. Maximizing the ratio of contact surface area to fluid volume can increase the effectiveness of the degassing process by increasing the duration of the fluid's exposure to suction as the fluid flows along the fluid path 1050. The degassing module 178 may be used alone or in combination with the bubble trap 1014 as described with reference to Figure 1D.

[0059] The central panel 176 may be configured to selectively route fluid through one or more of the aeration manifold 177 and the degassing module 178. That is, in some modifications, the central panel 176 can route substantially all fluid through one or more of the aeration manifold 177 and the degassing module 178. In further modifications, the central panel 176 may not be able to route at least a portion of the fluid through either the aeration manifold 177 or the degassing module 178. The decision to route fluid through the aeration manifold 177 and / or the degassing module 178 may correspond to a given workflow and / or type of fluid. For example, a fluid containing cells may not be routed through either the aeration manifold 177 or the degassing module 178 to avoid clogging of components, while a fluid containing culture medium, buffer, and / or reagents may be routed through one or more of the aeration manifold 177 and the degassing module 178.

[0060] Figures 3A to 3C show exemplary variations of the fluid manifold 300. The fluid manifold 300 may comprise a first end panel 310, a second end panel 314, a central panel 312, a first bridge 320, a second bridge 322, a degassing module 340, and a vent manifold 342. The first end panel 310 may be connected to the first bridge 320. The first bridge 320 may be connected to the first end panel 310 parallel to its lateral dimension. The first bridge 320 may be connected to the first end panel 310 at a predetermined position along its longitudinal dimension. For example, in some variations, the predetermined position may be about 1 / 4, 1 / 3, 1 / 2, or 3 / 4 of the longitudinal dimension. The first bridge 320 may be connected to the central panel 312 at its first end. The first end panel 310 may be perpendicular to the central panel 312, so that the longitudinal dimension of the first end panel 310 may be perpendicular to the longitudinal dimension of the central panel 312. Thus, a given position of the first bridge 320 relative to the first end panel 310 may provide volume on one or more sides of the central panel 312, thereby allowing the user to access one or more fluid transfer ports of the central panel 312.

[0061] The second end panel 314 may be connected to the second bridge 320. The second bridge 322 may be connected to the central panel 312 at its second end. The first end of the central panel 312 may be opposite the second end of the central panel 312. The second bridge 322 may be connected to the second end panel 320 in the same manner as described for the first bridge 320 and the first end panel 310. Thus, in some modifications, at least portions of the first and second bridges 320, 322 may be coplanar.

[0062] The central panel 312 may be further connected to a degassing module 340 and a ventilation manifold 342. For example, the degassing module 340 and the ventilation manifold 342 may be connected to the front of the central panel 312. The front of the central panel 312 may be opposite the rear of the central panel 312. A sensor 344 may be connected to the rear of the central panel 312. In some modifications, there may be multiple sensors connected to the rear of the central panel 312. The sensor 344 may be configured to measure one or more parameters of the fluid flowing through the central panel 312. The connections described herein may be facilitated by mechanical fasteners (e.g., screws, nails, bolts), adhesives (e.g., glue), friction fits (e.g., protrusions of one component received in a corresponding opening of another component), or a combination thereof.

[0063] a. End panel The fluid manifold described herein may comprise one or more end panels. An end panel may comprise one or more fluid pathways. For example, an end panel may be configured to transport fluid to and / or from one or more modules of a cartridge via the fluid pathways. An end panel may be connected to a central panel. The fluid pathways of an end panel may be fluidically connected to one or more fluid pathways of the central panel. In some modifications, multiple end panels may be connected to the central panel. For example, an end panel may be connected to a separate end of the central panel. Thus, the number, size, and / or shape of the end panels may correspond to the size and / or shape of the central panel. In some modifications, the end panels may have cross-sectional shapes such as rectangle, triangle, square, trapezoid, circle, or a combination thereof. In some modifications, there may be 1 to 10 end panels, 1 to 6 end panels, or 1 to 4 end panels, including 1, 2, 3, 4, 5, or 6 end panels.

[0064] Figures 4A to 4C show the first end panel 400. The first end panel 400 may comprise a plurality of fluid paths 410a to 410c, a plurality of valves 420a to 420d, a bubble trap 430, a plurality of windows 440a to 440c, a plurality of outlet ports 450a to 450d, and fasteners 460. Figure 4A shows the first body 402 of the first end panel 400. The first body 402 of the first end panel 400 may define one or more fluid paths, such as a first fluid path 410a, a second fluid path 410b, a third fluid path 410c, etc. The fluid paths of the first body 402 of the first end panel 400 may be covered by a film (not shown). The film may be configured to maintain a fluid seal between the fluid flowing through the fluid paths and the external environment. In some modifications, the film may also cover the bubble trap 430. The windows 440a to 440c may be configured to facilitate the measurement of one or more fluids. That is, the windows 440a to 440c may correspond to one or more fluid paths, and as a result, sensors operably connected to the windows 440a to 440c may generate measurements of the fluid in the fluid paths. For example, the windows 440a to 440c may be transparent, and as a result, optical measurements may be generated. The optical measurements may correspond to, for example, bubble count values.

[0065] The fluid can flow through the fluid pathways, valves, and windows of the first end panel 400 in various paths. That is, multiple valves 420a to 420d can be used to guide (e.g., control) the fluid according to a desired path. For example, as illustrated in Figure 4A, valve 420a can be fluidically connected to fluid pathway 410a. Thus, when valve 420a is in the open position, the fluid can flow through valve 420a to fluid pathway 410a. In some modifications, valve 420a can be driven to the closed position (e.g., via a controller), as a result preventing the fluid from flowing into fluid pathway 410a. The fluid can then flow along fluid pathway 410a to valve 420b. Similarly, the position of valve 420b (e.g., open or closed) can be used to determine whether the fluid can flow through it. Valve 420b can be fluidically connected to window 440b. That is, the fluid flowing through valve 420b can flow close to window 440b. Window 440b can be fluidically connected to valve 420c so that the fluid can flow through valve 420c when it is open, or not through it when valve 420c is closed. When valve 420c is open, the fluid can flow into fluid path 410c. Fluid path 410c can be fluidically connected to bubble trap 430. Therefore, any bubbles in the fluid flowing along fluid path 410c can be removed as the fluid flows through bubble trap 430. The fluid can then exit bubble trap 430 and continue flowing along fluid path 410c. Fluid path 410c can be fluidically connected to window 440c, as a result allowing observation of any remaining bubbles in the fluid. Next, the window 440c may be fluidically connected to the valve 420d, and as a result, the position of the valve 420d can be used to determine whether fluid can flow through it. The valve 420d may be fluidically connected to an outlet port, such as the outlet port 450b. Thus, fluid can flow from the valve 420a to the outlet port 450b. Alternative paths can be realized by the position of any of the valves on the first end panel 400, according to one or more valve actuation inputs provided by the controller.

[0066] Figure 4B shows the second body 404 of the first end panel 400. As shown, the multiple outlet ports 450a to 450d may extend along the lateral dimension of the second body 404 of the first end panel 400. That is, the multiple outlet ports 450a to 450d are collinear along a line extending from the first end of the first end panel 400 to the second end of the first end panel 400. In some modifications, the multiple outlet ports 450a to 450d are arranged in one or more rows. For example, outlet ports 450a and 450c may form a first row, and outlet ports 450b and 450d may form a second row. The multiple outlet ports 450a to 450d may be fluidically connected to one or more fluid paths of the first end panel 400. Multiple outlet ports 450a to 450d may be configured to transfer fluid from one or more fluid paths to other components of the fluid manifold. For example, the multiple outlet ports 450a to 450d may have a size, shape, and / or position that can correspond to multiple ports of a first bridge (not shown). In some modifications, the second body 404 may have one or more fluid paths that can correspond to the fluid paths of the first body 402 as described.

[0067] The two bodies 402 and 404 of the first end panel 400 can be connected to one another. For example, the first body 402 shown in Figures 4A and 4C and the second body 404 shown in Figures 4B and 4C can together form the first end panel 400. In some variations, the first and second bodies 402 and 404 can be connected to one another by mechanical fasteners (e.g., screws, nails, bolts), adhesives (e.g., glue), friction fittings (e.g., protrusions of one component receiving into a corresponding opening of another component), or a combination thereof. For example, the first and second bodies 402 and 404 can be connected to one another by a fastener 460, that is, the fastener 460 may extend through each of the first and second bodies 402 and 404, thereby securely connecting the first and second bodies 402 and 404. In some modifications, the first and second bodies 402, 404 may be connected to each other by a plurality of fasteners. In such modifications, the plurality of fasteners may be arranged in any manner suitable for securely connecting the first and second bodies 402, 404 together. For example, the plurality of fasteners may be arranged collinearly, for example, along the longitudinal and / or transverse dimensions. The plurality of fasteners may be arranged such that each fastener 460 avoids any fluid pathways, valves, bubble traps, or other components of the first end panel 400. One or more seals may be positioned between the two bodies 402, 404, thereby forming a fluid-sealing seal between the two bodies 402, 404. Thus, the first end panel 400 may have an adhesive fluid-sealing body. In some modifications, the first end panel 400 may be integrally formed so that fasteners 460 are not required.

[0068] Figures 5A to 5C show the second end panel 500. The second end panel 500 may comprise a plurality of fluid paths 510a to 510c, a plurality of valves 520a to 520f, a plurality of fluid extraction ports 522a to 522c, a plurality of windows 540a to 540c, a plurality of outlet ports 550a to 550d, and fasteners 560. Figure 5A shows the first body 502 of the second end panel 500. The first body 502 of the first end panel 500 may define one or more fluid paths, such as a first fluid path 510a, a second fluid path 510b, a third fluid path 510c, etc. The fluid paths of the first body 502 of the first end panel 500 may be covered by a film (not shown). The film may be as described with reference to Figures 4A to 4C. Similarly, the descriptions of the fluid paths 510a to 510c, the multiple valves 520a to 520f, and the multiple window sections 540a to 540c may correspond to the descriptions of the fluid paths 410a to 410c, the valves 420a to 420f, and the window sections 440a to 440c provided with reference to Figures 4A to 4C.

[0069] The first body 502 may further comprise a plurality of fluid extraction ports 522a to 522c. The fluid extraction ports 522a to 522c may be configured to extract fluid from fluid paths 510a to 510c. For example, each of the fluid extraction ports 522a to 522c may comprise a needle-free injection port. The fluid extraction ports 522a to 522c may form a fluid seal that is impermeable to liquids and / or gases. The fluid seal may be temporarily destroyed (e.g., punctured) by a conduit and / or a needle such as a syringe. The fluid extraction ports 522a to 522c may be configured to reform the fluid seal after the fluid extraction process has been performed. The fluid extraction ports 522a to 522c may be collinear, so that they can be positioned along a line extending from the first side of the first body 502 to the second side of the first body 502. In some variations, there may be 11 fluid extraction ports that are arranged on the same line.

[0070] The fluid can flow through the fluid paths, valves, and windows of the second end panel 500 in various paths. That is, multiple valves 520a to 520f can be used to guide (e.g., control) the fluid according to a desired path. For example, as illustrated in Figure 5A, valve 520a may be fluidically connected to window 540a. Window 540a may be fluidically connected to fluid path 510a. Thus, when valve 520a is in the open position, the fluid can pass through window 540a and flow through valve 520a into fluid path 510a. In some modifications, valve 520a may be driven to the closed position (e.g., via a controller), as a result preventing the fluid from flowing into fluid path 510a. The fluid can then flow along fluid path 510a to valve 520d. Similarly, the position of valve 520d (e.g., open or closed) can be used to determine whether the fluid can flow through it. Valve 520d may be fluidically connected to an outlet port such as outlet port 550c. Thus, fluid may flow from valve 520a to outlet port 550c. An alternative path may be realized by the position of any of the valves on the second end panel 500, according to one or more valve actuation inputs provided by the controller.

[0071] Figure 5B shows the second body 504 of the second end panel 500. The second body 504 may have a plurality of outlet ports 550a to 550d. A description of the plurality of outlet ports 550a to 550d may correspond to the description of outlet ports 450a to 450d provided with reference to Figures 4A to 4C. In some variations, the second body 504 may have one or more fluid paths, which may correspond to the description provided for the fluid paths of the first body 502. The two bodies 502 and 504 of the second end panel 500 may be connected to each other. For example, the first and second bodies 502 and 504 may be connected to each other by a fastener 560. A description of connecting the two bodies 502 and 504 to the fastener 560 may correspond to the description of connecting the two bodies 402 and 404 to the fastener 460 provided with reference to Figures 4A to 4C.

[0072] b. Central panel The fluid manifold described herein may comprise one or more central panels. The central panels may comprise one or more fluid pathways. For example, the central panel may be configured to transport fluid to and from the end panels and / or one or more modules of the cartridge via the fluid pathways. The central panel may be connected to one or more end panels. The fluid pathways of the central panel may be fluidically connected to one or more fluid pathways of the end panels. In some modifications, the central panel may have a cross-sectional shape such as rectangular, triangular, square, trapezoidal, circular, or a combination thereof. In some modifications, there may be 1 to 10 central panels, 1 to 6 central panels, or 1 to 4 central panels, including 1, 2, 3, or 4 central panels.

[0073] Figures 6A and 6B show exemplary variations of the central panel 600. The central panel 600 may comprise a first body 602, a second body 604, and an intermediate body 606. The intermediate body 606 may form a layer between the first body 602 and the second body 604. The first and second bodies 602 and 604 may define one or more fluid ports configured to transfer fluid. For example, the first body 602 may comprise a plurality of fluid transfer ports, such as fluid transfer ports 610a to 610c, so that it can be configured to receive fluid from and / or transfer fluid to another module. Thus, the fluid transfer ports 610a to 610c may be configured for bidirectional fluid flow. As illustrated in Figure 6A, the fluid transfer ports 610a to 610c may be located on the first body 602 of the central panel 600. Additionally or alternatively, the second body 604 may define one or more fluid ports configured for fluid transfer. For example, the second body 604 may have a plurality of bridge transfer ports, such as bridge transfer ports 620a to 620d as illustrated in Figure 6B. The bridge transfer ports 620a to 620d may be configured to receive fluid from and / or transfer fluid to the bridge.

[0074] Figure 6C shows an exemplary modification of the intermediate body 606. The intermediate body 606 may define a plurality of fluid paths and a plurality of fluid openings. For example, the intermediate body 606 may have a first fluid path 650a, a second fluid path 650b, and so on. The multiple openings may correspond to one or more transfer ports. For example, the first fluid opening 612a may correspond to a fluid transfer port, and the second fluid opening 612b may correspond to a bridge transfer port. Thus, the fluid paths of the intermediate body 606 may be fluidically connected to one or more modules and / or one or more bridges. In some modifications, the fluid paths of the intermediate body 606 may be fluidically connected to both fluid transfer ports and bridge transfer ports. For example, fluid path 650a may be fluidly connected to fluid openings 612a, 612b, and 612c. Fluid openings 612b and 612c may correspond to bridge transfer ports, while fluid opening 612a may correspond to a fluid transfer port. Therefore, as illustrated in Figure 6C, the fluid may flow from fluid opening 612b (which may be fluidically connected to a bridge transfer port of the first bridge) along fluid path 650a to fluid opening 612c (which may be connected to a bridge transfer port of the second bridge). Fluid path 650a may also be fluidically connected to fluid opening 612a, which may be fluidically connected to a fluid transfer port. Thus, the fluid may be transported to and from fluid path 650a through any of the fluid openings 612a-c. In another example, fluid path 650b may be fluidically connected to fluid openings 612d (which may be fluidically connected to another bridge transfer port of the first bridge) and 612e (which may be fluidically connected to a fluid transfer port). Thus, the fluid may be transported to and from fluid path 650b through any of the fluid openings 612a and 612b. In this way, the fluid may move between bridges and / or one or more modules via the fluid paths of the intermediate body 606.

[0075] In some variations, the first body 602, the second body 604, and the intermediate body 606 may be connected to one another by mechanical fasteners (e.g., screws, nails, bolts), adhesives (e.g., glue), friction fittings (e.g., projections of one component received within a corresponding opening of another component), or a combination thereof. For example, the bodies 602, 604, and 606 may be connected to one another by fasteners 630a to 630c. That is, the fasteners 630a to 630c may extend through each of the bodies 602, 604, and 606 so that the first and second bodies 602 and 604 can be securely connected to both sides of the intermediate body 606. The fasteners 630a to 630c may be arranged in any manner suitable for securely connecting the bodies 602, 604, and 606 to one another. For example, fasteners 630a to 630c may be positioned near the outer edges of the first and second bodies 602 and 604, respectively. In another example, multiple fasteners may be arranged such that each fastener avoids any fluid path in the central panel 600.

[0076] In some variations, the degassing module and / or ventilation manifold may be connected to the central panel. For example, the degassing module and / or ventilation manifold may be mechanically fastened to the central panel and / or fluidically connected to the central panel. Thus, fluid may flow through the central panel, through one or more degassing modules and / or ventilation manifolds, and then flow back to the central panel.

[0077] Figures 7A and 7B show the degassing module 700. The degassing module 700 may comprise a first body 702, a second body 704, and an intermediate body 706. The first body 702 may comprise a plurality of fluid transfer ports. For example, the first body 702 may comprise an inlet port 710, an outlet port 712, and a gas port 714. The inlet port 710 may be configured to receive fluid from a fluid conduit connected to a fluid transfer port on the central panel. The outlet port 712 may be configured to transfer fluid (e.g., liquid) from the degassing module 700 to a fluid conduit connected to a fluid transfer port on the central panel. The gas port 714 may be configured to supply another fluid (e.g., gas) to the fluid transfer port on the central panel.

[0078] A port can be fluidically connected to one or more openings. For example, the intermediate body 706 may have an inlet opening 711, an outlet opening 713, and a gas opening 715. The inlet opening 711 can be fluidically connected to the inlet port 710. Thus, fluid can flow from the inlet port 710 into the fluid opening 711 and into the fluid path 750. The outlet opening 713 can be fluidically connected to the outlet port 712. Thus, fluid can flow from the fluid path 750 into the outlet opening 713 and through the outlet port 712. The fluid path 750 can define a meandering path. For example, as illustrated in Figure 7B, the fluid path 750 may have multiple bends.

[0079] While the fluid can flow along the fluid path 750, the gas can be extracted from the fluid. For example, a gas opening 715 may be fluidically connected to the fluid path 750. The gas opening 715 may be separated from the fluid path 750 by a membrane, such as a gas-permeable membrane (not shown). Thus, the gas can be transferred through the gas opening 715 to a gas port 714. The gas port 714 may be connected to a fluid conduit (e.g., a tube) configured to apply suction. That is, the gas port 714 can facilitate degassing by providing a channel for the gas removed from the fluid flowing along the fluid path 750.

[0080] The bodies 702, 704, and 706 can be connected to each other to form a fluid-sealing seal. For example, the bodies 702, 704, and 706 can be connected to each other by fasteners 730a to 730c. That is, the fasteners 730a to 730c can extend through each of the bodies 702, 704, and 706 so that the first and second bodies 702 and 704 can be securely connected to both sides of the intermediate body 706. The fasteners 730a to 730c can be arranged in any manner suitable for securely connecting the bodies 702, 704, and 706 to each other. For example, the fasteners 730a to 730c can be arranged so that each fastener avoids any fluid path in the intermediate body 706.

[0081] Figures 8A and 8B show a ventilation manifold 800. The ventilation manifold 800 may comprise a first body 802 and a second body 804. The first body 802 may comprise a plurality of fluid transfer ports, such as a first fluid transfer port 810a and a second fluid transfer port 810b. The second body 804 may comprise a plurality of fluid channels (e.g., fluid cavities), such as a first fluid channel 840a, a second fluid channel 840b, and a third fluid channel 840c. The second body 804 may further comprise a plurality of fluid paths and a plurality of fluid openings. For example, the second body 804 may comprise a first fluid path 850a, a second fluid path 850b, a third fluid path 850b, and so on. Similarly, the second body 804 may comprise a first fluid opening 820a, a second fluid opening 820b, a third fluid opening 820c, and so on. In some modifications, a filter may be positioned between the first body 802 and the second body 804. That is, the filter may be positioned within the flow path between the fluid channel and the fluid transfer port. The filter may be configured to remove particles from the fluid flowing through the vent manifold. The first and second bodies 802 and 804 may be connected by a plurality of fasteners, such as fasteners 830a to 830c. Fasteners 830a to 830c may be positioned to avoid the fluid paths, ports, and channels described herein. In some modifications, a seal may be present between the first body 802 and the second body 804 so that the connection is fluid-sealed.

[0082] Fluid transfer ports, fluid openings, fluid paths, and fluid channels can be fluidically connected. For example, a first fluid channel 840a can be fluidically connected to a first fluid transfer port 810a. The first fluid transfer port 810 can be connected to a fluid conduit which can be connected to another fluid transfer port on the cartridge module or the central panel. The first fluid transfer port 810a can also be fluidically connected to a fluid transfer opening 820a, which can then be fluidically connected to a fluid path 850a. Thus, fluid can flow to and from another fluid path on the central panel via the fluid conduit connected to the first fluid transfer port 810a. In this way, the vent manifold can perform a venting process by transferring a gas (e.g., sterile air) to one or more fluid paths on the central panel. The gas can release any particles that may have clogged the filters of the vent manifold and / or remove any condensed fluid in the fluid paths on the central panel.

[0083] c. Bridge One or more components of a fluid manifold may be connected via bridges. A bridge may have one or more fluid paths, thereby enabling fluidic connection of the connected components. For example, an end panel may be fluidically connected to a central panel via a first bridge. The central panel may be connected to two or more bridges, thereby enabling fluidic connection of two or more end panels to the central panel. The size and / or number of bridges may correspond to the central panel. For example, a bridge may have a lateral dimension (e.g., width) equal to the lateral dimension of the central panel. In some modifications, the lateral dimension of a bridge may correspond to only a portion of the lateral dimension of the central panel. The number of bridges may correspond to the shape of the central panel, in that each end of the central panel can be configured to receive a bridge. For example, a central panel with four ends may be configured to receive up to four bridges. In some further modifications, two or more bridges may be connected to a single end of the central panel. For example, in some variations, there may be 1 to 10 bridges, 1 to 6 bridges, or 1 to 4 bridges, including 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bridges.

[0084] Figures 9A–9E show exemplary variations of the first bridge 900. The first bridge 900 may have a plurality of fluid transfer ports, which may allow the first bridge 900 to be fluidically connected to one or more of the central panel and end panels. For example, as shown, the first bridge 900 may have a plurality of central panel transfer ports, such as a first central panel transfer port 910a, a second central panel transfer port 910b, and so on. The first bridge may also have a plurality of end panel transfer ports, such as a first end panel transfer port 920a, a second end panel transfer port 920a, and so on. The central panel transfer ports 910a, 910b may be configured to be fluidically connected to bridge transfer ports on the central panel, such as bridge transfer ports 620a–6260d, as described with reference to Figure 6B. For example, transfer port 910a may be fluidly connected to a bridge transfer port. Seals may be present between the central panel transfer ports 910a and 910b and their respective bridge transfer ports, thereby enabling the transfer of sterile fluid between them. The central panel transfer ports 910a and 910b may be configured for bidirectional fluid flow, thereby enabling fluid to flow from the central panel to the bridge 900 and vice versa. The end panel transfer ports 920a and 920b may be configured to fluidically connect to the outlet ports of the end panels, such as the outlet ports 450a to 450d as described with reference to Figure 4B or the outlet ports 550a to 550d as described with reference to Figure 5B. For example, the end panel transfer ports 920a and 920b may be fluidically connected to the outlet ports. Seals may be present between the end panel transfer ports 920a and 920b and their respective outlet ports, thereby enabling the transfer of sterile fluid between them. The end panel transfer ports 920a and 920b may be configured for bidirectional fluid flow, thereby allowing fluid to flow from the end panel to the bridge 900, or vice versa.

[0085] The first bridge 900 can be securely connected to the central panel and / or end panels. For example, the first bridge 900 may have a plurality of openings configured to receive mechanical fasteners configured to connect the first bridge 900 to the central panel, and additionally or alternatively, a plurality of openings configured to receive mechanical fasteners configured to connect the first bridge 900 to the end panels. In some modifications, the first bridge may have a plurality of central panel fastener openings, such as a first central panel fastener opening 940a, a second central panel fastener opening 940b, a third central panel fastener opening 940c, and so on. Thus, mechanical fasteners (e.g., screws, nails, bolts) can extend from or through the central panel to the central panel fastener openings to securely connect the first bridge to the central panel. In some modifications, additional or alternative means, such as adhesive (e.g., glue), may be used to connect the first bridge to the central panel. Similarly, the first bridge may have multiple end panel fastener openings, such as a first end panel fastener opening 930a, a second end panel fastener opening 930b, and a third end panel fastener opening 930c. Mechanical fasteners (e.g., screws, nails, bolts) can be extended from or through the end panel to the end panel fastener openings to securely connect the first bridge to the end panel. In some modifications, additional or alternative means, such as adhesive (e.g., glue), may be used to connect the first bridge to the end panel.

[0086] The first bridge 900 may have one or more fluid paths. For example, Figures 9D and 9E show variations of the fluid paths of the first bridge 900. The fluid paths of the first bridge 900 may be defined by grooves, recesses, or channels. The fluid paths may have cross-sectional shapes suitable for transporting fluids, such as circular, square, triangular, trapezoidal, or combinations thereof. For example, Figure 9D shows an exemplary variation of the first fluid path 950a of the first bridge 900. The first fluid path 950a may extend from the first central transfer port 910a to the first end panel transfer port 920a. The first fluid path 950a may have one or more bends and have a circular cross-section. Thus, the central panel can be fluidly connected to the end panel via the fluid path 910a. In another example, Figure 9E shows an exemplary variation of the second fluid path 950b of the first bridge 900. A second fluid path 950b may extend from a second central transfer port 910b to a second end panel transfer port 920b. The second fluid path 950b may have one or more bends and have a circular cross-section. Thus, the central panel can be fluidically connected to the end panel via the fluid path 910b. The fluid paths 950a and 950b may be used simultaneously, or in some modifications, they may not be used simultaneously. The fluid paths 950a and 950b may be fluidically isolated from each other, so that the fluid does not mix between the fluid paths 950a and 950b.

[0087] Figures 10A to 10E show exemplary variations of the second bridge 1000. The second bridge 1000 may have multiple fluid transfer ports similar to those described with reference to Figures 9A to 9E. For example, the second bridge 1000 may have multiple central panel transfer ports, such as a first central panel transfer port 1010a and a second central panel transfer port 1010b. The descriptions of the central panel transfer ports 1010a and 1010b may correspond to the descriptions provided for the central panel transfer ports 910a and 910b with reference to Figures 9A to 9E. In another example, the second bridge 1000 may have multiple end panel transfer ports, such as a first end transfer port 1020a and a second end transfer port 1020b. The descriptions of the end panel transfer ports 1020a and 1020b may correspond to the descriptions provided for the end panel transfer ports 920a and 920b with reference to Figures 9A to 9E.

[0088] The second bridge 1000 can be securely connected to the central panel and / or end panels in a manner similar to that described for the first bridge 900. For example, the second bridge 1000 may have a plurality of openings configured to receive mechanical fasteners configured to connect the second bridge 1000 to the central panel, and additionally or alternatively, may have a plurality of openings configured to receive mechanical fasteners configured to connect the second bridge 1000 to the end panels. That is, in some modifications, the second bridge 1000 may have a plurality of central panel fastener openings, such as a first central panel fastener opening 1040a, a second central panel fastener opening 1040b, a third central panel fastener opening 1040c, and so on. The description of the central panel fastener openings 1040a to 1040c may correspond to the description provided for the central panel fastener openings 940a to 940c with reference to Figures 9A to 9E. Similarly, the second bridge 1000 may have multiple end panel fastener openings, such as a first end panel fastener opening 1030a, a second end panel fastener opening 1030b, and a third end panel fastener opening 1030c. The description of the end panel fastener openings 1030a to 1030c may correspond to the description provided for the end panel fastener openings 930a to 930c with reference to Figures 9A to 9E.

[0089] The second bridge 1000 may have one or more fluid paths. Figures 10D and 10E show variations of the fluid paths of the second bridge 1000. For example, the second bridge may have a first fluid path 1050a and a second fluid path 1050b. A description of the first and second fluid paths 1050a and 1050b may correspond to the description provided for the first and second fluid paths 950a and 950b with reference to Figures 9D and 9E.

[0090] II. Fluid Control Methods In general, the fluid control systems described herein can facilitate one or more cell processing methods in an automated cell processing work cell by controlling the flow of fluid to one or more modules of the cell processing work cell. Fluid control can be performed according to a predetermined workflow. The workflow can be pre-programmed by the user via the work cell controller. A fluid manifold can be controlled to deliver fluid to one or more modules according to any workflow. The fluid manifold can be fluidically coupled to a plurality of fluid containers used for providing solutions or reagents, storing cell products, or collecting waste liquids or reagents.

[0091] Figure 11 provides a flowchart of an exemplary method for controlling the flow of fluid in an automated working cell. As illustrated, method 1101 may include supplying fluid to a fluid manifold of cartridge 1110. For example, the fluid may include a solution (e.g., cell solution, cell suspension) having one or more of cells, culture medium, buffer, and reagent. In some modifications, the fluid may be supplied to a fluid conduit (e.g., tubing) fluidically connected to a valve on a first or second end panel. In further modifications, the fluid may be supplied to a fluid conduit fluidically connected to a fluid transfer port on a central panel. The fluid conduit may fluidly connect the fluid manifold (e.g., first end panel, second end panel, central panel, degassing module, aeration manifold) to another module of the cartridge or working cell. For example, the first end panel may be fluidically connected to a first bioreactor module of the bioreactor module, and / or the second end panel may be fluidically connected to a second bioreactor module of the bioreactor module. Fluid conduits can be configured to optimally reduce their length, number, and / or diameter, thereby minimizing entanglement of the fluid conduits.

[0092] Method 1101 may further include flowing fluid through one or more fluid paths among a first end panel, a second end panel, and a central panel 1120. The fluid path may be fluidically connected to a first bridge, which may be fluidically connected to a central panel. Thus, fluid may flow from a valve in the first end panel to the central panel. In some modifications, the fluid may flow in the opposite direction, so that the fluid may flow from the central panel through the fluid paths of the first bridge and the first end panel, and exit the fluid manifold through a valve in the first end panel. The second end panel may similarly be fluidically connected to the central section via a second bridge. In some modifications, the central panel may be in direct fluid communication with the first and / or second end panels, so that its fluid path can be fluidically connected without using the first and / or second bridge.

[0093] Method 1101 may further include performing a degassing process 1130. The degassing process may be performed by a degassing module. The degassing process may include removing a gas (e.g., air) from a liquid (e.g., a cell suspension). The gas may be dissolved in the liquid. In some modifications, the gas can be removed from the solution by a change in temperature. For example, the fluid may enter the fluid manifold at a first temperature (e.g., about 4°C) and then rise to a second temperature (e.g., about 20°C) as the fluid flows through the fluid manifold. When the gas is removed from the solution, bubbles may form in the liquid, which could improperly trigger it and / or one or more sensors that obstruct the flow of fluid through one or more fluid paths in the fluid manifold. Thus, a degassing process can remove the gas to facilitate the flow of fluid through the fluid manifold.

[0094] Method 1101 may further include performing an aeration process 1140. The aeration process may be performed by an aeration manifold. The aeration manifold may be configured to transport a gas (e.g., sterile air) from an external environment (e.g., outside the cartridge and inside the working cell) and transport this gas to one or more fluid pathways in the aeration manifold and / or central panel. The gas may be contained within fluid channels (e.g., fluid cavities) of the aeration manifold, as a result of the gas being selectively released into the fluid pathways to release any particles that may have clogged the filters of the aeration manifold and / or to remove any condensed fluid in the fluid pathways. Advantageously, the transported gas may increase the flow velocity of the fluid flowing through one or more fluid pathways.

[0095] Method 1101 may further include measuring one or more parameters through a window in a first end panel 1150. For example, the window may be configured for optical detection, thereby facilitating fluid measurement. For example, the measurement may include a bubble count value. The bubble count value may be compared to a predetermined condition, thereby determining the response of the fluid manifold. In some modifications, the response may include routing the fluid through a bubble trap in the first end panel if the comparison indicates that the bubble count value satisfies or exceeds a predetermined condition. In some modifications, a second end panel may have a similar window performing a similar function.

[0096] Method 1101 may further include measuring one or more parameters via sensors on a central panel 1160. The sensors may include pressure sensors. For example, a pressure sensor can measure the pressure value of a fluid flowing through one or more fluid paths on a central panel. The fluid may correspond to a fluid pump, and as a result, the pressure sensor can indicate the performance of the pump when transferring fluid to or from the central panel. That is, the pressure sensor can indicate the fluid velocity. For example, a pressure sensor may include a strain gauge that can bend as the fluid flows across the pressure sensor, and as a result, the bending can change the resistance value of the strain gauge. The resistance value can be correlated with the pressure value. The pressure value may be transmitted to a flex circuit, which can compile pressure values ​​from multiple pressure sensors. The flex circuit can communicate with a controller of a working cell.

[0097] Method 1101 may further include extracting fluid through a fluid extraction port 1170. A fluid extraction port, which may have a needleless injection port, may provide an extraction route that can be used to quickly remove fluid from a fluid manifold in an emergency, etc. An emergency may include a loss of power, leakage elsewhere in the fluid manifold, blockage of fluid in the fluid manifold and / or cartridge or elsewhere, or interruption of another workflow. The extraction process may be performed by opening (e.g., puncturing) the fluid extraction port, extracting the fluid, and closing the fluid extraction port. In some modifications, the fluid extraction port may be closed when removing an instrument, such as a syringe, that may have been used to initially open the fluid extraction port.

[0098] Method 1101 may further include transferring fluid to another module of the cartridge 1180. For example, a fluid manifold may be fluidically connected to one or more modules of the cartridge. The fluid connection may facilitate one or more cell processing steps that may be performed within one or more modules. Thus, the fluid manifold may control the timing, sequencing, and / or duration of any cell processing steps by controlling the fluid to or from the module in which the cell processing steps are performed. The fluid manifold may automatically control the fluid according to a predetermined workflow, which may be determined in advance by a controller of the working cell.

[0099] Throughout this application, the term “approximately” is used to indicate that a value includes inherent variations in error in the device or method employed to determine that value, or variations present between samples being measured. Unless otherwise stated or evident from the context, “approximately” means within plus or minus 10 percent of a reported number (except where such a number is greater than 100% of a possible value or less than 0%). When used in conjunction with a range or set of values, the term “approximately” applies to each of the endpoints of the range or the values ​​listed in the set of values, unless otherwise indicated. As used herein, the terms “approximately” and “about” are used interchangeably.

[0100] While embodiments of the present invention have been shown and described herein, those skilled in the art will understand that such embodiments are provided only as examples. Those skilled in the art will recall numerous variations, modifications, and substitutions without departing from the present invention. It should be understood that various substitutes for the embodiments of the present invention described herein may be used in carrying out the invention. The following claims define the scope of the present invention, and the methods and structures within these claims, as well as their equivalents, are intended to be encompassed thereby.

Claims

1. A cartridge for processing cells, A fluid manifold comprising a first end panel, a second end panel, and a central panel connecting the first and second end panels, A cartridge in which each of the first and second end panels comprises a plurality of fluid paths formed therein and a plurality of valves for controlling the flow of fluid through the plurality of fluid paths.

2. The cartridge according to claim 1, wherein the first end panel further comprises at least one window portion configured for optical detection.

3. The cartridge according to claim 2, wherein the at least one window portion includes a bubble sensing window portion.

4. The cartridge according to claim 1, wherein the first end panel further comprises a bubble trap.

5. The cartridge according to claim 1, wherein the second end panel further comprises at least one fluid extraction port.

6. The cartridge according to claim 5, wherein the at least one fluid extraction port comprises a needle-free injection port.

7. The cartridge according to claim 1, wherein at least one of the plurality of valves is a pinch valve.

8. The cartridge according to claim 1, wherein the central panel comprises a plurality of fluid paths.

9. The cartridge according to claim 8, wherein the plurality of fluid paths of the central panel are fluidly connected to the plurality of fluid paths of the first and second end panels, respectively.

10. The cartridge according to claim 8, wherein the central panel further comprises at least one pressure sensor configured to monitor the flow of fluid through the plurality of fluid paths of the central panel.

11. The cartridge according to claim 8, wherein the plurality of fluid paths of the central panel are fluidly connected to one or more pumps.

12. The cartridge according to claim 8, wherein the central panel is connected to a ventilation manifold configured to supply sterile air to the plurality of fluid paths of the central panel.

13. The cartridge according to claim 8, wherein the central panel is connected to a degassing module having an air-permeable membrane.

14. The cartridge according to claim 1, wherein the central panel connects the first end panel and the second end panel via first and second bridges.

15. The cartridge according to claim 1, wherein the fluid manifold is fluidly connected to one or more modules of the cartridge.

16. The cartridge according to claim 15, wherein one or more modules of the cartridge are selected from the group consisting of an elutriation module, an electroporation module, a spinoculation module, and a cell sorting module.

17. The cartridge according to claim 1, wherein the first end panel is fluidly connected to a first bioreactor module, and the second end panel is fluidly connected to a second bioreactor module.

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