Patent Publication Number: US-2023149922-A1

Title: Fluid connector

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims the benefit of U.S. Provisional Application No. 62/987,745, filed Mar. 10, 2020, U.S. Provisional Application No. 63/093,038, filed Oct. 16, 2020, the content of each of which is hereby incorporated by reference in its entirety. 
    
    
     TECHNICAL FIELD 
     Devices, systems, and methods herein relate to manufacturing cell products for biomedical applications using automated systems. 
     BACKGROUND 
     Cellular therapies based on hematopoietic stem cells (HSCs), chimeric antigen receptor (CAR) T cells, NK cells, tumor infiltrating lymphocytes (TILs), T-cell receptors (TCRs), regulatory T cells (T regs), gamma delta (γδ) T cells, and others rely on manufacturing of cell products. Manufacturing of such cell products typically involves multiple cell processing steps. Conventional solutions for manufacture of cell products rely on cumbersome manual operations performed in expensive biosafety cabinets and/or clean rooms. Skilled laboratory technicians, adequate sterile enclosures such as cleanroom facilities, and associated protocols and procedures for regulated (GMP) manufacturing are expensive. Many current manufacturing processes employ numerous manual reagent preparation and instrument manipulation steps during a manufacturing protocol, and the processes may require several days or even weeks. Even platforms described as automated cell processing in a closed system generally rely on pre-configured instrumentation and tubing sets that limit operational flexibility and do not reliably prevent process failure due to accidental operator/human error. 
     Most efforts to automate cell product manufacturing have been directed to automating individual processing steps of a cell therapy manufacturing workflow. Even systems that automate several steps lack end-to-end process flexibility, process robustness, and process scalability. These and other limitations of the previous attempts at automation of cell processing are addressed in various embodiments disclosed here. 
     SUMMARY 
     The present disclosure relates generally to methods and systems for processing cell products. By processing a cell product in a cartridge moved between instruments, some variations may achieve one or more advantages over prior cell manufacturing systems, including, for example, improved sterility, automation, lower cost of goods, lower labor costs, higher repeatability, higher reliability, lower risk of operator error, lower risk of contamination, higher process flexibility, higher capacity, higher instrument throughput, higher degree of process scalability, and shorter process duration. Variations of the disclosure may comprise a sterile enclosure, thereby reducing the costs of providing a clean room environment, and/or utilize a workcell having a smaller footprint than current manufacturing facilities. Furthermore, variations of the methods disclosed herein may, in some cases, be performed more quickly and with less risk of cell product loss. 
     In some variations, the disclosure provides a system for cell processing, comprising a plurality of instruments each independently configured to perform one or more cell processing operation upon a cartridge, and a robot capable of moving the cartridge between each of the plurality of instruments. 
     In some variations, the system may be enclosed in a workcell. In some variations, the workcell may be automated. In some variations, the plurality of instruments may be configured to interface with the cartridge to perform cell processing operations upon the cartridge. In some variations, the system may comprise a processor. The processor may be configured to control the robot and the plurality of instruments. 
     In some variations, the system may be configured to receive two or more cartridges. In some variations, the system may comprise the cartridge. In some variations, the cartridge may comprise a plurality of modules. In some variations, the cartridge may comprise a bioreactor module. In some variations, the cartridge may comprise a cell selection module. In some variations, the cell selection module may comprise a magnetic-activated cell selection module. In some variations, the cartridge may comprise a sorting module. In some variations, the sorting module may comprise a fluorescence activated cell sorting (FACS) module. In some variations, the cartridge may comprise an electroporation module. In some variations, the cartridge may comprise a counterflow centrifugal elutriation (CCE) module. 
     In some variations, the cartridge may comprise one or more sterile liquid transfer ports. In some variations, the cartridge may comprise a liquid transfer bus fluidically coupled to each module. In some variations, the cartridge may comprise a pump fluidically coupled to the liquid transfer bus. 
     In some variations, the system may comprise a pump actuator configured to interface with the pump. In some variations, the system may comprise a bioreactor instrument. In some variations, the bioreactor instrument may comprise multiple slots for cartridges. In some variations, the system may comprise a cell selection instrument. In some variations, the cell selection instrument may comprise a magnetic-activated cell selection instrument. 
     In some variations, the system may comprise a sorting instrument. In some variations, the sorting instrument may comprise a fluorescence activated cell sorting (FACS) instrument. In some variations, the system may comprise an electroporation instrument. In some variations, the system may comprise a counterflow centrifugal elutriation (CCE) instrument. In some variations, the system may comprise a reagent vault. 
     In some variations, the cartridge may comprise a bioreactor module and a selection module. In some variations, the cartridge may comprise a bioreactor module and a CCE module. In some variations, the cartridge may comprise a bioreactor module, selection module, and a CCE module. In some variations, the cartridge may comprise a bioreactor module, selection module, and an electroporation module. In some variations, the cartridge may comprise a bioreactor module, selection module, a CCE module, and an electroporation module. In some variations, the cartridge may comprise a second bioreactor module having an internal volume two or more, five or more, or ten or more times larger than the internal volume of the first bioreactor. 
     In some variations, the system may comprise an enclosure. In some variations, the enclosure may comprise an ISO7 cleanroom. In some variations, the enclosure may comprise an ISO6 cleanroom. In some variations, the enclosure may comprise an ISO5 cleanroom. In some variations, the enclosure may comprise a feedthrough. In some variations, the system may perform automated manufacturing of cell products. 
     In some variations, the disclosure provides a cartridge for cell processing, comprising a liquid transfer bus and a plurality of modules, each module fluidically coupled to the liquid transfer bus. 
     In some variations, the cartridge may comprise one or more sterile liquid transfer ports. In some variations, the cartridge may comprise a bioreactor module. In some variations, the cartridge may comprise a cell selection module. In some variations, the cell selection module may comprise a magnetic-activated cell selection module. In some variations, the cartridge may comprise a sorting module. In some variations, the sorting module may comprise a fluorescence activated cell sorting (FACS) module. In some variations, the cartridge may comprise an electroporation module. In some variations, the cartridge may comprise a counterflow centrifugal elutriation (CCE) module. 
     In some variations, the cartridge may comprise a mechanoporation module. In some variations, the cartridge may comprise a second bioreactor module having an internal volume two or more, five or more, or ten or more times larger than the internal volume of the first bioreactor. In some variations, the cartridge may comprise a bioreactor module, selection module, and a CCE module. In some variations, the cartridge may comprise a bioreactor module, selection module, and an electroporation module. In some variations, the cartridge may comprise a bioreactor module, selection module, a CCE module, and an electroporation module. 
     In some variations, the disclosure provides a method for processing cells, comprising moving a cartridge containing a cell product between a plurality of instruments inside an enclosed and automated workcell. The instruments may interface with the cartridge to perform cell processing steps on the cell product. 
     In some variations, cell processing steps may be performed on the cell product. In some variations, for each cell product, all cell processing steps in the method are performed in a single cartridge. 
     In some variations, the cell product may be split into a plurality of cell product portions. In some variations, the cell processing steps may be performed on the plurality of cell product portions in parallel. In some variations, at least two cell product portions of the plurality of cell product portions may be combined. 
     In some variations, the workcell may comprise a robot configured to move cartridges. In some variations, the workcell may comprise a processor. The processor may be configured to control the robot and the plurality of instruments. In some variations, the workcell may be configured to receive two or more cartridges. 
     In some variations, the cartridge may comprise a plurality of modules. In some variations, the cartridge may comprise a bioreactor module. In some variations, the cartridge may comprise a cell selection module. In some variations, the cell selection module may comprise a magnetic-activated cell selection module. 
     In some variations, the cartridge may comprise a sorting module. In some variations, the sorting module may comprise a fluorescence activated cell sorting (FACS) module. In some variations, the cartridge may comprise an electroporation module. In some variations, the cartridge may comprise a counterflow centrifugal elutriation (CCE) module. In some variations, the cartridge may comprise one or more sterile liquid transfer ports. In some variations, the cartridge may comprise a liquid transfer bus fluidically coupled to each module. In some variations, the cartridge may comprise a pump fluidically coupled to the liquid transfer bus. 
     In some variations, the workcell may comprise a pump actuator configured to interface with the pump. In some variations, the workcell may comprise a bioreactor instrument. In some variations, the bioreactor instrument may comprise multiple slots for cartridges. In some variations, the method may comprise performing the cell processing steps on two or more cartridges in parallel. 
     In some variations, the workcell may comprise a cell selection instrument. In some variations, the cell selection instrument may comprise a magnetic-activated cell selection instrument. 
     In some variations, the workcell may comprise a sorting instrument. In some variations, the sorting instrument may comprise a fluorescence activated cell sorting (FACS) instrument. In some variations, the workcell may comprise an electroporation instrument. In some variations, the workcell may comprise a counterflow centrifugal elutriation (CCE) instrument. In some variations, the workcell may comprise a reagent vault. 
     In some variations, the cartridge may comprise a bioreactor module and a selection module. In some variations, the cartridge may comprise a bioreactor module and a CCE module. In some variations, the cartridge may comprise a bioreactor module, selection module, and a CCE module. In some variations, the cartridge may comprise a bioreactor module, selection module, and an electroporation module. In some variations, the cartridge may comprise a bioreactor module, selection module, a CCE module, and an electroporation module. 
     In some variations, the workcell may comprise an enclosure. In some variations, the enclosure may comprise an ISO7 cleanroom. In some variations, the enclosure may comprise an ISO6 cleanroom. In some variations, the enclosure may comprise an ISO5 cleanroom. In some variations, the enclosure may comprise a feedthrough. 
     In some variations, the method may perform automated manufacturing of a cell product. In some variations, the cell product may comprise a chimeric antigen receptor (CAR) T cell product. In some variations, the cell product may comprise a natural killer (NK) cell product. In some variations, the cell product may comprise a hematopoietic stem cell (HSC) cell product. In some variations, the cell product may comprise a tumor infiltrating lymphocyte (TIL) cell product. In some variations, the cell product may comprise a regulatory T (Treg) cell product. 
     In some variations, the disclosure provides a method for processing a solution containing a cell product, performed in an automated system, the method comprising one or more cell processing steps, performed serially in any order, selected from: an enrichment step, a concentration step, a buffer exchange step, a formulation step, a washing step, a selection step, a resting step, an expansion step, a tissue-digestion step, an activation step, a transduction step, a transfection step, and a harvesting step. 
     In some variations, an enrichment step may comprise enriching a selected population of cells in the solution by conveying the solution to a CCE module of the cartridge via a liquid transfer bus, operating the robot to move the cartridge to a CCE instrument so that the CCE module interfaces with the CCE instrument, and operating the CCE instrument to cause the CCE module to enrich the selected population of cells. 
     In some variations, a washing step may comprise washing a selected population of cells in the solution by conveying the solution to the CCE module of the cartridge via the liquid transfer bus, operating the robot to move the cartridge to the CCE instrument so that the CCE module interfaces with the CCE instrument, and operating the CCE instrument to cause the CCE module to remove media from the solution, introduce media into the solution, and/or replace media in the solution. 
     In some variations, a selection step may comprise selecting a selected population of cells in the solution by conveying the solution to a selection module of the cartridge via the liquid transfer bus, operating the robot to move the cartridge to a selection instrument so that the selection module interfaces with the selection instrument, and operating the selection instrument to cause the selection module to select the selected population of cells. 
     In some variations, a sorting step may comprise sorting a population of cells in the solution by conveying the solution to a sorting module of the cartridge via the liquid transfer bus, operating the robot to move the cartridge to a sorting instrument so that the sorting module interfaces with the sorting instrument, and operating the sorting instrument to cause the sorting module to sort the population of cells. 
     In some variations, a resting step may comprise conveying the solution to a bioreactor module of the cartridge via the liquid transfer bus, operating the robot to move the cartridge to the bioreactor instrument so that the bioreactor module interfaces with the bioreactor instrument, and operating the bioreactor instrument to cause the bioreactor module to maintain the cells. 
     In some variations, an expansion step may comprise expanding the cells in the solution by conveying the solution to the bioreactor module of the cartridge via the liquid transfer bus, operating the robot to move the cartridge to the bioreactor instrument so that the bioreactor module interfaces with the bioreactor instrument, and operating the bioreactor instrument to cause the bioreactor module to allow the cells to expand by cellular replication. 
     In some variations, a tissue-digestion step may comprise conveying an enzyme reagent via the liquid transfer bus to a module containing a solution containing a tissue such that the enzyme reagent causes digestion of the tissue to release a select cell population into the solution. 
     In some variations, an activating step may comprise activating a selected population of cells in the solution by conveying an activating reagent via the liquid transfer bus to a module containing the solution containing the cell product. 
     In some variations, an electroporation step may comprise conveying the solution to an electroporation module of the cartridge via the liquid transfer bus, operating the robot to move the cartridge to an electroporation instrument so that the electroporation module interfaces with the electroporation instrument, and operating the electroporation instrument to cause the electroporation module to electroporate the selected population of cells in the presence of the vector. 
     In some variations, a transduction step may comprise conveying an effective amount of a vector via the liquid transfer bus to a module containing the solution containing the cell product, thereby transducing a selected population of cells in the solution. In some variations, a fill/finishing step may comprise conveying a formulation solution via the liquid transfer bus to a module containing the cell product to generate a finished cell product and conveying the finished cell product to one or more product collection bags. 
     In some variations, the method may comprise sterilizing, either manually or automatically, the cartridge in a feedthrough port. In some variations, the method may comprise introducing, either manually or automatically, one or more of a fluid and the cell product into the cartridge via a sterile liquid transfer port. In some variations, the method may comprise a harvesting step comprising removing, either manually or automatically, the cell product from the cartridge. In some variations, the cell product may comprise an immune cell. In some variations, in order, the enrichment step, the selection step, the activation step, the transduction step, the expansion step, and the harvesting step. 
     In some variations, the immune cell may comprise a genetically engineered chimeric antigen receptor T cell. In some variations, the immune cell may comprise a genetically engineered T cell receptor (TCR) cell. In some variations, the immune cell may comprise is a natural-killer (NK) cell. In some variations, the cell product may comprise a hematopoietic stem cell (HSC). In some variations, the method may comprise, in order, the enrichment step, the selection step, the resting step, the transduction step, and the harvesting step. In some variations, the cell product may comprise a tumor infiltrating lymphocyte (TIL). In some variations, the method may comprise, in order, the tissue-digestion step, the washing step, the activation step, the expansion step, and the harvesting step. 
     Also described here is a counterflow centrifugal elutriation (CCE) module, comprising a conical element having an internal surface and an external surface fixedly attached to a distal end of a linear member having an internal surface and an external surface, the proximal end of the linear member rotationally attached to a fulcrum to permit extension, retraction, and rotation of the linear member. 
     Also described here is a workcell comprising an enclosure, a plurality of instruments each independently configured to perform one or more cell processing operation upon a cartridge, and a robot capable of moving the cartridge between each of the plurality of instruments. 
     In some variations, the enclosure may comprise an air filtration inlet configured to maintain ISO 7 or better air quality within an interior zone of the workcell. In some variations, the workcell may be automated. In some variations, the instruments may interface with the cartridge to perform cell processing operations upon the cartridge. In some variations, the workcell may comprise a processor. The processor may be configured to control the robot and the plurality of instruments. 
     In some variations, the workcell may be configured to receive two or more cartridges. In some variations, the workcell may comprise the cartridge. In some variations, the cartridge may comprise a plurality of modules. In some variations, the cartridge may comprise a bioreactor module. In some variations, the cartridge may comprise a cell selection module. In some variations, the cell selection module may comprise a magnetic-activated cell selection module. In some variations, the cartridge may comprise a sorting module. 
     In some variations, the sorting module may comprise a fluorescence activated cell sorting (FACS) module. In some variations, the cartridge may comprise an electroporation module. In some variations, the cartridge may comprise a counterflow centrifugal elutriation (CCE) module. In some variations, the cartridge may comprise one or more sterile liquid transfer ports. In some variations, the cartridge may comprise a liquid transfer bus fluidically coupled to each module. In some variations, the cartridge may comprise a pump fluidically coupled to the liquid transfer bus. 
     In some variations, the workcell may comprise a pump actuator configured to interface with the pump. In some variations, the workcell may comprise a bioreactor instrument. In some variations, the bioreactor instrument may comprise multiple slots for cartridges. In some variations, the workcell may comprise a cell selection instrument. In some variations, the cell selection instrument may comprise a magnetic-activated cell selection instrument. In some variations, the workcell may comprise a sorting instrument. In some variations, the sorting instrument may comprise a fluorescence activated cell sorting (FACS) instrument. In some variations, the workcell may comprise an electroporation instrument. 
     In some variations, the workcell may comprise a counterflow centrifugal elutriation (CCE) instrument. In some variations, the workcell may comprise a reagent vault. In some variations, the cartridge may comprise a bioreactor module and a selection module. In some variations, the cartridge may comprise a bioreactor module and a CCE module. In some variations, the cartridge may comprise a bioreactor module, selection module, and a CCE module. In some variations, the cartridge may comprise a bioreactor module, selection module, and an electroporation module. In some variations, the cartridge may comprise a bioreactor module, selection module, a CCE module, and an electroporation module. In some variations, the cartridge may comprise a second bioreactor module having an internal volume two or more, five or more, or ten or more times larger than the internal volume of the first bioreactor. In some variations, the enclosure may comprise a feedthrough. In some variations, the workcell may perform automated manufacturing of cell products. In some variations, the system may comprise a plurality of bioreactor instruments. Each bioreactor instrument may be configured to receive a single cartridge. 
     Also described here is a rotor comprising a first side comprising a first fluid conduit, a second side comprising a second fluid conduit, the second side opposite the first side, and a cone coupled between the first fluid conduit and the second fluid conduit. 
     In some variations, the cone may comprise a bicone. In some variations, the bicone may comprise a first cone including a first base and a second cone including a second base. The first base may face the second base. In some variations, the rotor may comprise a magnetic portion. In some variations, the rotor may define a rotation axis. In some variations, at least a portion of the first fluid conduit and at least a portion of the second fluid conduit may extend parallel to the rotation axis. In some variations, at least a portion of the first fluid conduit and at least a portion of the second fluid conduit may be co-axial. 
     In some variations, the cone may comprise a volume of between about 10 ml and about 40 ml. In some variations, the cone may comprise a cone angle of between about 30 degrees and about 60 degrees. In some variations, at least a portion of the rotor may be optically transparent. In some variations, the rotor may comprise an asymmetric shape. In some variations, a first portion may comprise the cone and a second portion comprising a paddle shape. 
     In some variations, a cartridge for cell processing may comprise a liquid transfer bus and a plurality of modules. Each module may be fluidically linked to the liquid transfer bus. The cartridge may comprise a counterflow centrifugal elutriation (CCE) module comprising the rotors described herein. 
     Also described here is a rotor comprising a first fluid conduit, a first fluid conduit, a first cone coupled to the first fluid conduit. The first cone may comprise a first volume. A second fluid conduit may be coupled to the first cone. A second cone may be coupled to the second conduit. The second cone may comprise a second volume larger than the first volume. A third fluid conduit may be coupled to the second cone. 
     In some variations, the first cone may comprise a first bicone and the second cone may comprise a second bicone. In some variations, the first bicone may comprise a third cone including a first base and a fourth cone including a second base. The first base may face the second base. The second bicone may comprise a fifth cone including a third base and a sixth cone including a fourth base. The third base may face the fourth base. 
     In some variations, the rotor may comprise a magnetic portion. In some variations, at least a portion of the rotor may be optically transparent. In some variations, the first fluid conduit may comprise an inlet and the third fluid conduit comprises an outlet. 
     Also described here is a system for cell processing comprising a cartridge comprising a housing comprising a rotor configured to separate cells from a fluid, and an instrument comprising a magnet configured to interface with the cartridge to magnetically rotate the rotor. 
     In some variations, the cartridge may be configured to move between a plurality of instruments. In some variations, an air gap may be between the housing and the magnet. In some variations, the housing may enclose the rotor. In some variations, the housing may comprise a consumable component and the magnet comprises a durable component. 
     In some variations, the magnet may be releasably coupled to the housing. In some variations, the magnet may be configured to be moved relative to the housing. In some variations, the separated cells may comprise a first size and a first density and non-separated cells of the fluid comprise a second size and a second density different from the first size and the first density. Also described here is a cartridge for cell processing, comprising a liquid transfer bus and a plurality of modules. Each module may be fluidically linked to the liquid transfer bus. The cartridge may comprise a counterflow centrifugal elutriation (CCE) module comprising the rotor described here. 
     Also described here is a method of counterflow centrifugal elutriation (CCE) comprising moving a rotor towards a magnet, the rotor defining a rotational axis, flowing the fluid through the rotor, magnetically rotating the rotor about the rotational axis using the magnet while flowing the fluid through the rotor. 
     In some variations, image data of one or more of the fluid and particles in the rotor may be generated using an optical sensor. One or more of a rotation rate of the rotor and a flow rate of the fluid may be selected based at least in part on the image data. 
     In some variations, one or more of the fluid and the cells may be illuminated using an illumination source. In some variations, the method may comprise moving the rotor away from the magnet. In some variations, the method may comprising moving the rotor towards an illumination source and an optical sensor, and moving the rotor away from the illumination source and the optical sensor. 
     In some variations, moving the rotor comprises advancing and withdrawing the magnet relative to the rotor using a robot. In some variations, rotating the rotor comprises a rotation rate of up to 6,000 RPM. In some variations, flowing the fluid comprises a flow rate of up to about 150 ml/min while rotating the rotor. 
     Also described here is a method of magnetic-activated cell selection comprising flowing the fluid comprising input cells into a flow cell. A set of the cells may be labeled with magnetic-activated cell selection (MACS) reagent. The set of cells may be magnetically attracted towards a magnet array for a dwell time. The set of cells may flow out of the flow cell after the dwell time. 
     In some variations, the method may comprise incubating the MACS reagent with the input cells to label the set of cells with the MACS reagent. In some variations, the method may comprise incubating the MACS reagent may comprise a temperature between about 1° C. and about 10° C. In some variations, the method may comprise flowing the set of cells out of the flow cell may comprise flowing a gas through the flow cell. In some variations, the method may comprise flowing the fluid without the set of cells out of the flow cell after the dwell time. In some variations, the dwell time may be at least about one minute. In some variations, the magnet array may be disposed external to the flow cell. In some variations, the method may comprise moving the magnet array relative to the flow cell. In some variations, moving the magnet array may comprise moving the magnet array away from the flow cell to facilitate flowing the set of cells out of the flow cell. In some variations, a longitudinal axis of the flow cell may be perpendicular to ground. In some variations, the flow cell may be absent beads. 
     Also described here is a magnetic-activated cell selection (MACS) module comprising a flow cell comprising an elongate cavity having a cavity height, a magnet array may comprise a plurality of magnets. Each of the magnets may be spaced apart by a spacing distance. A ratio of the cavity height to the spacing distance may be between about 20:1 and about 1:20. 
     In some variations, the flow cell may comprise a set of linear channels comprising a first channel parallel to a second channel, and a third channel in fluid communication with each of the first channel and the second channel. In some variations, the first channel may comprise a first cavity height and the second channel may comprise a second cavity height. A ratio of the first cavity height to a second cavity height may be between about 1:1 to about 3:7. In some variations, the third channel may comprise a ratio of a length of the third channel to a diameter of the third channel of between about 2:1 to about 6:1. 
     In some variations, a first fluid conduit may be coupled to an inlet of the flow cell and an outlet of the flow cell. The first fluid conduit may be configured to receive the set of cells from the flow cell. A second fluid conduit may be coupled to the inlet of the flow cell and the outlet of the flow cell. The second fluid conduit may be configured to receive a fluid without the set of cells from the flow cell. 
     In some variations, a cartridge for cell processing may comprise a liquid transfer bus and a plurality of modules. Each module may be fluidically linked to the liquid transfer bus. The cartridge may comprise a magnetic-activated cell selection (MACS) module as described herein. 
     Also described here is a system for cell processing comprising a cartridge comprising a rotor configured for counterflow centrifugal elutriation of cells in a fluid. A first magnet may be configured to magnetically rotate the rotor and separate the cells from the fluid in the rotor. The cartridge may further comprise a flow cell in fluid communication with the rotor and configured to receive the cells from the rotor. A second magnet may be configured to magnetically separate the cells in the flow cell. 
     In some variations, an illumination source may be configured to illuminate the cells. An optical sensor may be configured to generate image data corresponding to the cells. In some variations, the system may comprise one or more of an oxygen depletion sensor, leak sensor, inertial sensor, pressure sensor, and bubble sensor. In some variations, the system may comprise one or more valves and pumps. 
     In some variations, the separated cells may comprise a first size and a first density and non-separated cells of the fluid comprise a second size and a second density different from the first size and the first density. 
     Also described here is an electroporation module comprising a fluid conduit configured to receive a first fluid comprising cells and a second fluid, a set of electrodes coupled to the fluid conduit, a pump coupled to the fluid conduit, andd a controller comprising a processor and memory. The controller may be configured to generate a first signal to introduce the first fluid into the fluid conduit using the pump, generate a second signal to introduce the second fluid into the fluid conduit such that the second fluid separates the first fluid from a third fluid, and generate an electroporation signal to electroporate the cells in the fluid conduit using the set of electrodes. 
     In some variations, the second fluid may comprise a gas or oil. In some variations, the controller may be configured to generate a third signal to introduce the third fluid into the fluid conduit, the third fluid separated from the first fluid by the second fluid. 
     In some variations, a cartridge for cell processing may comprise a liquid transfer bus and a plurality of modules. Each module may be fluidically linked to the liquid transfer bus. The cartridge may comprise an electroporation module as described here. 
     Also described here is a method of electroporating cells comprising receiving a first fluid comprising cells in a fluid conduit, receiving a second fluid in the fluid conduit to separate the first fluid from a third fluid, and applying an electroporation signal to the first fluid to electroporate the cells. 
     In some variations, the method may comprise receiving the third fluid in the fluid conduit separated from the first fluid by the second fluid. In some variations, the first fluid substantially static when applying the electroporation signal. 
     Also described here is a method of electroporating cells comprising receiving a first fluid comprising cells in a fluid conduit, applying a resistance measurement signal to the first fluid using a set of electrodes, measuring a resistance between the first fluid and the set of electrodes, and applying an electroporation signal to the first fluid based on the measured resistance. 
     In some variations, the method may comprise receiving a second fluid comprising a gas in the fluid conduit before applying the electroporation signal to the fluid, the first fluid separated from a third fluid by the second fluid. 
     Also described here is a bioreactor comprising an enclosure comprising a base, a top, and at least one sidewall. A gas-permeable membrane may be coupled to one or more of the base and the sidewall of the enclosure. 
     In some variations, the enclosure may comprise one or more nested surfaces curved around a longitudinal axis of the enclosure. In some variations, the one more nested surfaces may comprise a set of concentric toroids. In some variations, the enclosure may comprise a toroid shape. In some variations, the enclosure may comprise a first chamber having a first volume and a second chamber having a second volume, the first chamber separated from the second chamber, and the first volume smaller than the second volume. In some variations, the enclosure may comprise a column extending along a longitudinal axis of the enclosure. In some variations, a cavity may be between the enclosure and the gas-permeable membrane. In some variations, the gas-permeable membrane may extend along the base and the sidewall of the enclosure. In some variations, an outer surface of the gas-permeable membrane may comprise one or more projections. 
     In some variations, a base of the gas-permeable membrane may comprise an angle between about 3 degrees and about 10 degrees relative to the base of the enclosure. In some variations, the gas-permeable membrane may comprise a curved surface. In some variations, the gas-permeable membrane may comprise a set of patterned curved surfaces. In some variations, the set of patterned curved surfaces may comprise a radius of curvature of between about 50 mm and about 500 mm. 
     In some variations, a cartridge for cell processing may comprise a liquid transfer bus and a plurality of modules. Each module may be fluidically linked to the liquid transfer bus. The cartridge may comprise a bioreactor module as described here. In some variations, a system for cell processing may comprising the cartridge described here and may further comprise a bioreactor instrument configured to interface with the cartridge. The bioreactor instrument may comprise an agitator configured to couple to the bioreactor. The agitator may be configured to agitate cell culture media comprising cells. In some variations, a fluid connector may be configured to couple the bioreactor to a liquid transfer bus. The fluid connector may comprise foldable sidewalls. In some variations, the system may comprise a temperature regulator coupled to the bioreactor. In some variations, the system may comprise a gas regulator coupled to the bioreactor. 
     Also described here is a fluid connector comprising a first connector comprising a first proximal end configured to couple to a first fluid device, and a first distal end comprising a first port. A second connector may comprise a second proximal end configured to couple to a second fluid device, and a second distal end comprising a second port configured to couple to the first port. The first distal end may comprise a first lumen and the second distal end may comprise a second lumen. One of the first valve and the second valve may be configured to translate within the first lumen and the second lumen. 
     In some variations, the first valve and the second valve may be configured to transition from a closed configuration to an open configuration only when the first valve couples to the second valve. In some variations, the first port and the second port may be configured to transition between an open configuration and a closed configuration. In some variations, the first connector may comprise a first port actuator and/or the second connector comprises a second port actuator. In some variations, the second port may be coupled to the first port defines a chamber. 
     In some variations, one or more of the first connector and the second connector may comprise a sterilant port configured to couple to a sterilant source. The sterilant port may be configured to be in fluid communication with the first distal end and the second distal end when the second port is coupled to the first port. 
     In some variations, the chamber may be configured to receive one or more of a fluid and a sterilant from the sterilant port. In some variations, the sterilant port may be configured to receive a sterilant such that the sterilant sterilizes the first connector and the second connector. 
     In some variations, the first connector may comprise a first valve, and the second connector may comprise a second valve configured to couple to the first valve. In some variations, a first seal may comprise the first port coupled to the second port, and a second seal may comprise the first valve coupled to the second valve. In some variations, the sterilant may comprise one or more of vaporized hydrogen peroxide and ethylene oxide. 
     In some variations, the fluid connector may comprise one or more robot engagement features. In some variations, the first connector may comprise a first alignment feature and the second connector may comprise a second alignment feature configured to couple to the first alignment feature in a predetermined axial and rotational configuration. In some variations, one or more of the first fluid device and the second fluid device may comprise an instrument. 
     In some variations, a system may further comprise a robot configured to operate the fluid connector, and a controller comprising a memory and processor. The controller may be coupled to the robot. The controller may be configured to generate a first port signal to couple the first port to the second port using the robotic arm. In some variations, the controller may be configured to generate a first valve signal to translate the first valve relative to the second valve using the robotic arm, and generate a second valve signal to transition the first valve and the second valve to the open configuration. In some variations, the controller may be configured to generate a second port signal to decouple the first port from the second port. A sterility of the fluid connector may be maintained before coupling the first port to the second port and after decoupling the first port from the second port. 
     In some variations, a fluid pump may be coupled to the sterilant source. The controller may be configured to generate a first fluid pump signal to circulate a fluid into the chamber through the sterilant port. In some variations, the controller may be configured to generate a second fluid pump signal to circulate the sterilant into the chamber through the sterilant port to sterilize at least the chamber. 
     In some variations, the controller may be configured to generate a third fluid pump signal to remove the sterilant from the chamber. In some variations, the controller may be configured to generate a thermal sterilization signal to thermally sterilize the fluid connector. In some variations, the controller may be configured to generate a radiation sterilization signal to sterilize the fluid connector using radiation. In some variations, the robot may be configured to couple a fluid connector between at least two of the plurality of instruments and the cartridge. 
     In some variations, the fluid connector may further comprise a controller comprising a memory and processor, the controller coupled to the robot. The controller may be configured to generate a port signal to couple the first port to the second port using the robotic arm, generate a first valve signal to translate the first valve relative to the second valve using the robotic arm, and generate a second valve signal to transition the first valve and the second valve to the open configuration. 
     Also described here is a non-transitory computer-readable medium for transforming user-defined cell processing operations into cell processing steps to be executed by an automated cell processing system. The non-transitory computer-readable medium may comprise instructions stored thereon that when executed on a processor perform the steps of receiving an ordered input list of cell processing operations, and executing a transformation model on the ordered input list to create an ordered output list of cell processing steps capable of being performed by the system. 
     In some variations, the ordered output list may be capable of being performed by the system to control a robot to move one or more cartridges each containing a cell product between the instruments, and control the instruments to perform cell processing steps on each cell product. 
     In some variations, the method may comprise receiving one or more sets of cell processing parameters, each set associated with one of the cell processing operations, and each set of cell processing parameters specifying characteristics of the cell processing step to be performed by the instrument at that cell processing step. In some variations, the transformation model may comprise constraints on the ordered output list determined by configuration of the automated cell processing system. In some variations, the constraints may comprise information on the configuration of the automated cell processing system. In some variations, the constraints may comprise one or more of a type and/or a number of instruments, a type and/or a number of modules on the cartridge, a type and a number of reservoirs on the cartridge, a type and/or a number of sterile liquid transfer ports on the cartridge, and a number and a position of fluid paths between the modules, reservoirs, and sterile liquid transfer ports on the cartridge. 
     In some variations, the steps may further comprise receiving a set of more than one ordered input lists of cell processing operations to be performed on more than one cartridge on the automated cell processing system, and executing the transformation model on the sets of ordered input lists to create the ordered output list of cell processing steps. The ordered output list may be capable of being executed by the system to control the robot to move the more than one cartridges, each comprising its cell product, between the instruments, and control the instruments to perform cell processing steps on each cell product of each cartridge. 
     In some variations, an automated cell processing system may comprise the non-transitory computer-readable medium of any preceding claim. 
     In some variations, a computer-implemented method for transforming user-defined cell processing operations into cell processing steps to be executed by a processor of an automated cell processing system may comprise receiving an ordered input list of cell processing operations, and executing a transformation model on the ordered input list to create an ordered output list of cell processing steps capable of being performed by the system. 
     In some variations, the method may include controlling a robot to move one or more cartridges each containing a cell product between the instruments, and controlling the instruments to perform cell processing steps on each cell product. 
     In some variations, the method may comprise receiving one or more sets of cell processing parameters, each set associated with one of the cell processing operations, and each set of cell processing parameters specifying characteristics of the cell processing step to be performed by the instrument at that cell processing step. In some variations, the transformation model may comprise constraints on the ordered output list determined by configuration of the automated cell processing system. In some variations, the constraints may comprise information on the configuration of the automated cell processing system. 
     In some variations, the constraints may comprise one or more of a type and/or number of instruments, a type and/or number of modules on the cartridge, a type and number of reservoirs on the cartridge, a type and/or number of sterile liquid transfer ports on the cartridge, and a number and position of fluid paths between the modules, reservoirs, and sterile liquid transfer ports on the cartridge. 
     In some variations, the method may comprise receiving a set of more than one ordered input lists of cell processing operations to be performed on more than one cartridge on the automated cell processing system, executing the transformation model on the sets of ordered input lists to create the ordered output list of cell processing steps, controlling the robot to move the more than one cartridges, each comprising its cell product, between the instruments, and controlling the instruments to perform cell processing steps on each cell product of each cartridge. 
     Additional variations, features, and advantages of the invention will be apparent from the following detailed description and through practice of the invention. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG.  1 A  is a block diagram of an illustrative variation of a cell processing system. 
         FIG.  1 B  is a block diagram of an illustrative variation of a cartridge. 
         FIG.  2 A  is a block diagram of an illustrative variation of a cell processing system.  FIG.  2 B  is a perspective view of an illustrative variation of a workcell of a cell processing system.  FIG.  2 C  is a perspective view of an illustrative variation of a workcell and cartridge of a cell processing system.  FIG.  2 D  is a block diagram of an illustrative variation of a cell processing system.  FIG.  2 E  is a block diagram of another illustrative variation of a cell processing system. 
         FIG.  3    is a block diagram of another illustrative variation of a cell processing system. 
         FIG.  4 A  is a perspective view of another illustrative variation of a cell processing system. 
         FIG.  4 B  is another perspective view of another illustrative variation of a cell processing system. 
         FIG.  5    is a perspective view of another illustrative variation of a cell processing system. 
         FIG.  6    is a schematic diagram of an illustrative variation of a cartridge. 
         FIG.  7    is a schematic diagram of another illustrative variation of a cartridge. 
         FIG.  8 A  is a side view of an illustrative variation of a cartridge.  FIG.  8 B  is a top view of an illustrative variation of a cartridge.  FIG.  8 C  is a side view of an illustrative variation of a cartridge.  FIG.  8 D  is a perspective view of an illustrative variation of a cartridge. 
         FIG.  9    shows a cross-sectional side view of an illustrative variation of a cartridge. 
         FIG.  10 A  shows an illustrative variation of a rotary valve and an actuator.  FIG.  10 B  shows an illustrative variation of a rotary valve docked with an actuator. 
         FIG.  11 A  is a perspective view of an illustrative variation of a cartridge comprising a CCE module in an extended configuration.  FIG.  11 B  is a cross-sectional side view of illustrative variation of a CCE module in a retracted configuration.  FIG.  11 C  is a cross-sectional side view of an illustrative variation of a CCE module in an extended configuration. 
         FIG.  12 A  is a perspective view of an illustrative variation of a magnetic-activated cell sorting (MACS) instrument comprising a magnet in an ON configuration.  FIG.  12 B  is a perspective view of an illustrative variation of a MACS instrument comprising a magnet in an OFF configuration. 
         FIG.  13 A  is a perspective view of an illustrative variation of a cartridge and a bioreactor instrument.  FIG.  13 B  is a perspective view of an illustrative variation of a cartridge coupled to a bioreactor instrument. 
         FIG.  14    is a perspective view of an illustrative variation of a bioreactor instrument comprising a set of cartridges and cavities configured to receive cartridges. 
         FIG.  15    is a block diagram of an illustrative variation of a fluid connector system. 
         FIG.  16 A  is a schematic diagram of an illustrative variation of a fluid connector.  FIG.  16 B  is a detailed schematic diagram of the fluid connector depicted in  FIG.  16 A .  FIG.  16 C  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  in a coupled configuration.  FIG.  16 D  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  in an open port configuration.  FIG.  16 E  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  receiving a gas.  FIG.  16 F  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  receiving a sterilant.  FIG.  16 G  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  in an open valve configuration.  FIG.  16 H  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  transferring fluid between fluid devices coupled to the fluid connector.  FIG.  16 I  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  in a closed valve configuration.  FIG.  16 J  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  in a closed port configuration.  FIG.  16 K  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  in an uncoupled configuration.  FIG.  16 L  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  uncoupled from a sterilant source. 
         FIG.  17 A  is a front perspective view of a fluid connector in a closed port configuration.  FIG.  17 B  is a rear perspective view of the fluid connector depicted in  FIG.  17 A  in the closed port configuration.  FIG.  17 C  is a rear view of the fluid connector depicted in  FIG.  17 B  in the closed port configuration.  FIG.  17 D  is a front perspective view of a fluid connector in an open port configuration.  FIG.  17 E  is a rear perspective view of the fluid connector depicted in  FIG.  17 D  in the open port configuration.  FIG.  17 F  is a rear view of the fluid connector depicted in  FIG.  17 E  in the open port configuration. 
         FIG.  18 A  is a side view of a fluid connector in an uncoupled configuration.  FIG.  18 B  is a cross-sectional side view of a fluid connector in an uncoupled configuration.  FIG.  18 C  is a side view of a fluid connector in a coupled configuration.  FIG.  18 D  is a cross-sectional side view of a fluid connector in a coupled configuration.  FIG.  18 E  is a side view of a fluid connector in an open port configuration.  FIG.  18 F  is a cross-sectional side view of a fluid connector in an open port configuration.  FIG.  18 G  is a side view of a fluid connector in an open valve configuration.  FIG.  18 H  is a cross-sectional side view of a fluid connector in an open valve configuration. 
         FIG.  19    is a schematic diagram of an illustrative variation of a fluid connector system. 
         FIG.  20 A  is a schematic diagram of an illustrative variation of a fluid connector system. 
         FIGS.  20 B and  20 C  are schematic diagrams of an illustrative variation of a fluid connector connection process. 
         FIG.  21    is a block diagram of an illustrative variation of a fluid connector system. 
         FIG.  22    is a block diagram of an illustrative variation of a fluid connector system. 
         FIG.  23    is a block diagram of an illustrative variation of a fluid connector system. 
         FIG.  24 A  is a block diagram of an illustrative variation of a fluid connector system.  FIG.  24 B  is a schematic diagram of an illustrative variation of a fluid connector connection process.  FIG.  24 C  is a schematic diagram of an illustrative variation of a valve. 
         FIG.  25 A  is a block diagram of an illustrative variation of a fluid connector system.  FIG.  25 B  is a schematic diagram of an illustrative variation of a fluid connector connection process.  FIG.  25 C  is a schematic diagram of an illustrative variation of a valve. 
         FIG.  26 A  is a side view of an illustrative variation of a pump actuator and pump.  FIG.  26 B  is a side view of an illustrative variation of a pump actuator coupled to a pump. 
         FIG.  27    is a flowchart of an illustrative variation of a method of transferring fluid using a fluid connector. 
         FIG.  28    is a flowchart of an illustrative variation of a method of cell processing. 
         FIG.  29    is a flowchart of an illustrative variation of a method of cell processing. 
         FIG.  30 A  is a flowchart of an illustrative variation of a method of cell processing for autologous CAR T cells or engineered TCR cells.  FIG.  30 B  is a flowchart of an illustrative variation of a method of cell processing for allogeneic CAR T cells or engineered TCR cells. 
         FIG.  31    is a flowchart of an illustrative variation of a method of cell processing for HSC cells. 
         FIG.  32    is a flowchart of an illustrative variation of a method of cell processing for TIL cells. 
         FIG.  33    is a flowchart of an illustrative variation of a method of cell processing for NK-CAR cells. 
         FIGS.  34 A- 34 C  are flowcharts of illustrative variations of methods of cell processing for T reg  cells. 
         FIG.  35    is a flowchart of an illustrative variation of a method of cell processing. 
         FIG.  36    is a flowchart of an illustrative variation of a method of executing a transformation model. 
         FIG.  37    is an illustrative variation of a graphical user interface relating to an initial process design interface. 
         FIG.  38    is an illustrative variation of a graphical user interface relating to creating a process. 
         FIG.  39    is an illustrative variation of a graphical user interface relating to an empty process. 
         FIG.  40    is an illustrative variation of a graphical user interface relating to adding a reagent and a consumable container. 
         FIG.  41    is an illustrative variation of a graphical user interface relating to a process parameter. 
         FIG.  42    is an illustrative variation of a graphical user interface relating to a patient weight process parameter. 
         FIG.  43    is an illustrative variation of a graphical user interface relating to a preprocess analytic. 
         FIG.  44    is an illustrative variation of a graphical user interface relating to a white blood cell count preprocess analytic. 
         FIG.  45    is an illustrative variation of a graphical user interface relating to process parameter calculation. 
         FIG.  46    is an illustrative variation of a graphical user interface relating to a completed process setup. 
         FIG.  47    is an illustrative variation of a graphical user interface relating to process operations activation settings. 
         FIG.  48    is an illustrative variation of a graphical user interface relating to a filled process operations activation settings. 
         FIG.  49    is an illustrative variation of a graphical user interface relating to an initial process operations. 
         FIG.  50    is an illustrative variation of a graphical user interface relating to dragging in process operations. 
         FIG.  51    is another illustrative variation of a graphical user interface relating to dragging in process operations. 
         FIG.  52    is an illustrative variation of a graphical user interface relating to a filled process operations. 
         FIG.  53    is an illustrative variation of a graphical user interface relating to product monitoring. 
         FIG.  54    is another illustrative variation of a graphical user interface relating to product monitoring. 
         FIG.  55    is a block diagram of an illustrative variation of a manufacturing workflow. 
         FIG.  56    is a block diagram of an illustrative variation of a cell separation system. 
         FIG.  57    is a cross-sectional side view of an illustrative variation of a counterflow centrifugal elutriation (CCE) module. 
         FIG.  58    is a cross-sectional side view of an illustrative variation of a magnetic-activated cell selection (MACS) module. 
         FIGS.  59 A- 59 C  are perspective views of an illustrative variation of a CCE system.  FIG.  59 D  is a side cross-sectional view of an illustrative variation of a CCE system.  FIGS.  59 E- 59 G  are side cross-sectional views of an illustrative variation of a rotor of a CCE module. 
         FIG.  60 A  is a plan view of an illustrative variation of a rotor of a CCE module.  FIGS.  60 B and  60 C  are perspective views of an illustrative variation of a rotor of a CCE module.  FIG.  60 D  is a side view of an illustrative variation of a rotor of a CCE module.  FIG.  60 E  is a perspective view of an illustrative variation of a rotor in a housing.  FIGS.  60 F and  60 G  are plan schematic views of illustrative variations of a rotor of a CCE module.  FIG.  60 H  is a side view of an illustrative variation of a rotor of a CCE module.  FIG.  60 I  is a perspective view of another illustrative variation of a rotor of a CCE module.  FIG.  60 J  is a perspective view of yet another illustrative variation of a rotor of a CCE module.  FIG.  60 K  is a schematic plan view of another illustrative variation of rotor dimensions of a CCE module.  FIG.  60 L  is an image of a set of illustrative variations of rotors of a CCE module. 
         FIGS.  61 A- 61 C  are schematic views of an illustrative variation of a cell separation process. 
         FIG.  62 A  is a perspective view of an illustrative variation of a MACS system in a first configuration.  FIG.  62 B  is a perspective view of an illustrative variation of a MACS system in a second configuration.  FIG.  62 C  is a cross-sectional side view of an illustrative variation of a MACS system.  FIG.  62 D  is a perspective view of an illustrative variation of a MACS system in the second configuration.  FIG.  62 E  is a plan view of an illustrative variation of a flow cell and magnet array of a MACS system.  FIG.  62 F  is a plan view of an illustrative variation of a flow cell of a MACS system.  FIG.  62 G  is a schematic diagram of an illustrative variation of a flow cell and magnet array. 
         FIGS.  63 A- 63 E  are perspective views of illustrative variations of a magnet array. 
         FIG.  64 A  is a perspective view of an illustrative variation of a flow cell.  FIG.  64 B  is a cross-sectional side view of an illustrative variation of a flow cell.  FIG.  64 C  is a schematic diagram of an illustrative variation of a MACS system. 
         FIGS.  65 A- 65 C  are schematic diagrams of an illustrative variation of a flow cell. 
         FIGS.  66 A- 66 C  are schematic diagrams of an illustrative variation of a cell separation process. 
         FIGS.  67 A- 67 D  are schematic diagrams of an illustrative variation of a cell processing system. 
         FIG.  68 A  is a cross-sectional perspective view of an illustrative variation of a bioreactor. 
         FIG.  68 B  is a cross-sectional side view of an illustrative variation of a bioreactor.  FIG.  68 C  is a perspective view of an illustrative variation of an enclosure of a bioreactor.  FIG.  68 D  is a plan view of an illustrative variation of an enclosure of a bioreactor. 
         FIG.  68 E  is a perspective view of an illustrative variation of a membrane of a bioreactor. 
         FIG.  68 F  is a side view of an illustrative variation of a membrane of a bioreactor.  FIG.  68 G  is a perspective view of an illustrative variation of a membrane of a bioreactor.  FIG.  68 H  is a bottom view of an illustrative variation of a membrane of a bioreactor. 
         FIG.  69 A  is a cross-sectional side view of an illustrative variation of an enclosure of a bioreactor.  FIG.  69 B  is a cross-sectional perspective view of an illustrative variation of an enclosure of a bioreactor. 
         FIG.  70    is an exploded perspective view of an illustrative variation of a bioreactor. 
         FIG.  71 A  is a plan view of an illustrative variation of a bioreactor.  FIG.  71 B  is a cross-sectional side view of an illustrative variation of a bioreactor. 
         FIG.  72    is a schematic diagram of an illustrative variation of an electroporation system. 
         FIG.  73    is an exploded perspective view of an illustrative variation of an electroporation module. 
         FIGS.  74 A- 74 B  are schematic diagrams of illustrative variation of an electroporation process. 
         FIG.  75    is a circuit diagram of an illustrative variation of an electroporation process. 
         FIGS.  76 A- 76 D  are plots of illustrative variations of an electroporation process. 
         FIG.  77 A  is a flowchart of an illustrative variation of a method of separating cells.  FIG.  77 B  is a flowchart of an illustrative variation of a method of concentrating cells.  FIG.  77 C  is a flowchart of an illustrative variation of a method of buffer exchange. 
         FIG.  78    is a flowchart of another illustrative variation of a method of separating cells. 
         FIG.  79 A  is a flowchart of an illustrative variation of a closed-loop method of separating cells  7900 .  FIG.  79 B  is a flowchart of an illustrative variation of a closed-loop method of elutriating cells  7910 .  FIG.  79 C  is a flowchart of an illustrative variation of a closed-loop method of harvesting cells  7920 . 
         FIG.  80 A  is a flowchart of an illustrative variation of a method of separating cells.  FIG.  80 B  is a flowchart of an illustrative variation of a method of selecting cells. 
         FIG.  81    is a flowchart of another illustrative variation of a method of separating cells. 
         FIG.  82 A  is a flowchart of an illustrative variation of a method of preparing a bioreactor. 
         FIG.  82 B  is a flowchart of an illustrative variation of a method of loading a bioreactor.  FIG.  82 C  is a flowchart of an illustrative variation of a method of preparing a bioreactor.  FIG.  82 D  is a flowchart of an illustrative variation of a method of calibration for a bioreactor.  FIG.  82 E  is a flowchart of an illustrative variation of a method of mixing reagents.  FIG.  82 F  is a flowchart of an illustrative variation of a method of mixing reagents.  FIG.  82 G  is a flowchart of an illustrative variation of a method of culturing cells.  FIG.  82 H  is a flowchart of an illustrative variation of a method of refrigerating cells.  FIG.  82 I  is a flowchart of an illustrative variation of a method of taking a sample.  FIG.  82 J  is a flowchart of an illustrative variation of a method of culturing cells. 
         FIG.  82 K  is a flowchart of an illustrative variation of a method of media exchange.  FIG.  82 L  is a flowchart of an illustrative variation of a method of controlling gas.  FIG.  82 M  is a flowchart of an illustrative variation of a method of controlling pH. 
         FIG.  83    is a flowchart of an illustrative variation of a method of electroporating cells. 
         FIG.  84    is a flowchart of another illustrative variation of a method of electroporating cells. 
         FIG.  85    are schematic diagrams of an illustrative variation of a fluid connector. 
         FIG.  86    are schematic diagrams of an illustrative variation of a fluid connector port. 
         FIG.  87    is a schematic diagram of an illustrative variation of a fluid connector connection process. 
         FIG.  88    is a schematic diagram of an illustrative variation of a fluid connector connection process. 
         FIG.  89    is a schematic diagram of an illustrative variation of a fluid connector. 
         FIG.  90 A  is a side view of an illustrative variation of a fluid connector.  FIG.  90 B  is a perspective view of the fluid connector depicted in  FIG.  90 A .  FIG.  90 C  is a cross-sectional side view of the fluid connector depicted in  FIG.  90 A . 
         FIG.  91 A  is a side view of an illustrative variation of a fluid connector.  FIG.  91 B  is a perspective view of the fluid connector depicted in  FIG.  91 A .  FIG.  91 C  is a cross-sectional side view of the fluid connector depicted in  FIG.  91 A . 
         FIG.  91 D  is a side view of an illustrative variation of a fluid connector.  FIG.  91 E  is a perspective view of the fluid connector depicted in  FIG.  91 D .  FIG.  91 F  is a cross-sectional side view of the fluid connector depicted in  FIG.  91 D . 
         FIG.  92 A  is a side view of an illustrative variation of a fluid connector.  FIG.  92 B  is a transparent side view of the fluid connector depicted in  FIG.  92 A .  FIG.  92 C  is a perspective view of the fluid connector depicted in  FIG.  92 A .  FIG.  92 D  is a cross-sectional side view of the fluid connector depicted in  FIG.  92 A . 
         FIG.  93 A  is a perspective view of an illustrative variation of a fluid connector.  FIG.  93 B  is a transparent perspective view of the fluid connector depicted in  FIG.  93 A . 
         FIG.  94 A  is a perspective view of an illustrative variation of a fluid connector.  FIG.  94 B  is a transparent perspective view of the fluid connector depicted in  FIG.  94 A . 
         FIG.  95 A  is a perspective view of an illustrative variation of a fluid connector.  FIG.  95 B  is a transparent perspective view of the fluid connector depicted in  FIG.  95 A .  FIG.  95 C  is a detailed side view of a port in an open port configuration.  FIG.  95 D  is a detailed side view of a port in a closed port configuration. 
         FIG.  96 A  is a plan view of an illustrative variation of a fluid device.  FIG.  96 B  is a side view of an illustrative variation of a fluid device coupled to a robot.  FIG.  96 C  is a perspective view of an illustrative variation of a fluid device held by a robot. 
         FIG.  97 A  is a perspective view of an illustrative variation of a MACS module.  FIG.  97 B  is a cross-sectional perspective view of an illustrative variation of a MACS module.  FIG.  97 C  is a cross-sectional side view of an illustrative variation of a MACS module. 
         FIG.  98    is a flowchart of an illustrative variation of a method of cell processing. 
         FIG.  99    is a flowchart of an illustrative variation of a method of cell processing. 
         FIG.  100    is a flowchart of an illustrative variation of a method of cell processing. 
         FIG.  101    is a flowchart of an illustrative variation of a method of cell processing. 
         FIG.  102    is a schematic diagram of an illustrative variation of a cell processing system. 
         FIGS.  103 A and  103 B  are perspective views of an illustrative variation of a sterile liquid transfer device. 
     
    
    
     DETAILED DESCRIPTION 
     Systems and methods for processing and manufacturing cell products for biomedical applications are described herein. Cell processing methods and systems may comprise moving a cartridge containing a cell product between a plurality of instruments inside a workcell. One or more instruments may be configured to interface with the cartridge to perform cell processing steps on the cell product, such that the system (e.g., workcell) performs cell processing steps on the cell product. In some variations, a plurality of cell processing steps may be performed within a single cartridge. For example, a robotic arm may be configured to move a cartridge between instruments for different cell processing steps. The cartridge may comprise a plurality of cell processing devices (e.g., modules) such as a bioreactor, a counterflow centrifugal elutriation (CCE) module, a magnetic cell sorter (e.g., magnetic-activated cell selection module), an electroporation device (e.g., electroporation module), a sorting module (e.g. fluorescence activated cell sorting (FACS) module), an acoustic flowcell module, a centrifugation module, a microfluidic enrichment module, combinations thereof, and the like. In some variations, the system may process two or more cartridges in parallel. For example, the bioreactor may comprise a plurality of slots configured to interface with a plurality of cartridges concurrently, as one process step (e.g., cell culturing in a bioreactor) may typically be the rate limiting step for the operation of the cell processing system. The cell processing systems described herein may reduce operator intervention and increase throughput by automating cartridge (and cell product) movement between instruments using a robot. However, in some variations, the cartridge may be moved between instruments manually. Furthermore, throughput of the system may be increased by using a plurality of bioreactors, thereby allowing the system to simultaneously process a plurality of cartridges for a plurality of patients. Moreover, the automated cell processing system may facilitate sterile liquid transfers between the cartridge and instruments or other components of the system such as a fluid connector (e.g., sterile liquid transfer port), reagent vault, a second cartridge, a sampling vessel (e.g., sterile liquid transfer device, combinations thereof, and the like. 
     Workcell 
     In some variations, a system for cell processing (e.g., workcell) may comprise a plurality of instruments each independently configured to perform one or more cell processing operations upon a cartridge. A robot may be configured to move the cartridge between each of the plurality of instruments. The instruments may comprise one or more of a bioreactor instrument, a cell selection instrument (e.g., a magnetic-activated cell selection instrument), a sorting instrument (e.g., a fluorescence activated cell sorting (FACS) instrument), an electroporation instrument, a counterflow centrifugal elutriation (CCE) instrument, a reagent vault, and the like. The system may perform automated manufacturing of cell products. 
     A cartridge may be configured to be portable and facilitate automated and sterile cell processing using a workcell and robot. For example, the cartridge may be configured to move relative to one or more instruments of the workcell to perform different cell processing steps. In some variations, an instrument may be configured to move relative to a cartridge. In some variations, the cartridge may comprise a plurality of modules including one or more of a bioreactor module, a cell selection module (e.g., magnetic-activated cell selection module), a sorting module (e.g., fluorescence activated cell sorting (FACS) module), an electroporation module, and a counterflow centrifugal elutriation (CCE) module. The cartridge may further comprise one or more of a sterile liquid transfer port, a liquid transfer bus fluidically coupled to each module, and a pump fluidically coupled to the liquid transfer bus. 
     In some variations, a method of processing a solution containing a cell product may include the cell processing steps of digesting tissue using an enzyme reagent to release a select cell population into solution, enriching cells using a CCE instrument, washing cells using the CCE instrument, selecting cells in the solution using a selection instrument, sorting cells in the solution using a sorting instrument, differentiating or expanding the cells in a bioreactor, activating cells using an activating reagent, electroporating cells, transducing cells using a vector, and finishing a cell product. 
     Cell Selection System 
     The cell processing systems described herein may comprise a cell selection system configured to separate cells based on predetermined criteria. For example, cells may be separated based on physical characteristics such as size and/or density using, for example, a counterflow centrifugation elutriation instrument. Cells may also be separated based on the presence of predetermined antigens of a cell using, for example, a magnetic-activated cell selection instrument. In some variations, a cell selection system comprising modules for these separation methods may facilitate one or more cell processing steps including, but not limited to, cell concentration, cell dilution, cell washing, buffer replacement, and magnetic separation. The cell selection systems described herein may increase throughput and cell yields output, in a compact and portable structure. For example, prior to magnetically separating cells, a suspension of cells may be mixed with magnetic reagents in excess or at a predetermined concentration (e.g., cells/ml). Likewise, after magnetically separating cells, the cells may be washed in a solution (e.g., suitable buffered solution). 
     In some variations, a cell separation system may comprise a rotor configured for counterflow centrifugation elutriation of cells in a fluid, a first magnet configured to magnetically rotate the rotor and separate the cells from the fluid in the rotor, a flow cell in fluid communication with the rotor and configured to receive the cells from the rotor, and a second magnet configured to magnetically separate the cells in the flow cell. 
     In some variations, a CCE module may be integrated into a cartridge to enable a cell processing system to separate cells based on cell size and/or density. In some variations, a cell separation system may comprise a housing comprising a rotor configured to separate cells from a fluid (e.g., separate cells of different size and/or density from cells that remain in the fluid), and a magnet configured to magnetically rotate the rotor. The housing may be configured to move relative to the magnet or vice versa (e.g., move the magnet relative to the housing). The CCE modules described herein may provide cell separation within a compact and portable housing where the magnet may be disposed external to the housing (e.g., magnet disposed within a CCE instrument). 
     In some variations, a compact rotor that may aid cartridge integration may comprise input and output fluid conduits extending from the rotor towards opposing sides of a rotor housing. For example, a rotor may comprise a first side comprising a first fluid conduit and a second side comprising a second fluid conduit where the second side is opposite the first side. An elutriation chamber (e.g., cone) may be coupled between the first fluid conduit and the second fluid conduit. 
     In some variations, a method of separating cells from a fluid may comprise moving a rotor towards a magnet, the rotor defining a rotational axis, flowing the fluid through the rotor, rotating the rotor (e.g., magnetically) about the rotational axis using the magnet while flowing the fluid through the rotor, and moving the rotor away from the magnet. 
     In some variations, a method of separating cells from a fluid may comprise flowing the fluid comprising the cells into a flow cell. A set of the cells may be labeled with magnetic particles. The set of cells may be magnetically attracted towards a magnet array for a dwell time, and the set of cells may flow out of the flow cell after the dwell time. 
     In some variations, a flow cell may comprise an elongate cavity having a cavity height and a magnet array comprising a plurality of magnets, each of the magnets spaced apart by a spacing distance. A predetermined ratio between the cavity height to the spacing distance may optimize magnetic separation of the cells in the flow cell. 
     Electroporation 
     In some variations, an electroporation module as described herein may be configured to facilitate one or more of transduction and transfection of cells. As described in more detail herein, a volume of fluid (e.g., first batch) comprising cells may be physically separated from a subsequent volume of fluid (e.g., second batch, third batch) comprising cells by a gas (e.g., air gap). Applying an electroporation signal (e.g., voltage pulse, waveform) separately to each discrete batch of fluid may improve electroporation efficiency and thus increase throughput. In some variations, active electric field compensation may similarly improve electroporation efficiency and throughput. 
     In some variations, a cell processor may comprise a fluid conduit configured to receive a first fluid comprising cells and a second fluid (e.g., gas, oil), a set of electrodes coupled to the fluid conduit, a pump coupled to the fluid conduit, and a controller comprising a processor and memory. The controller may be configured to generate a first signal to introduce the first fluid into the fluid conduit using the pump, generate a second signal to introduce the second fluid into the fluid conduit such that the second fluid separates the first fluid from a third fluid, and generate an electroporation signal to electroporate the cells in the fluid conduit using the set of electrodes. 
     In some variations, a method of electroporating cells may comprise receiving a first fluid comprising cells in a fluid conduit, receiving a second fluid comprising a gas in the fluid conduit to separate the first fluid from a third fluid, and applying an electroporation signal to the first fluid to electroporate the cells. 
     In some variations, a method of electroporating cells may comprise receiving a first fluid comprising cells in a fluid conduit, applying a resistance measurement signal to the first fluid using a set of electrodes, measuring a resistance between the first fluid and the set of electrodes, and applying an electroporation signal to the first fluid based on the measured resistance. 
     Bioreactor 
     In some variations, a bioreactor may comprise an enclosure comprising a base and a sidewall, and a gas-permeable membrane coupled to one or more of the base and the sidewall of the enclosure. The gas-permeable membrane may aid cell culture. In some variations, a cell processing system may comprise the bioreactor and an agitator coupled to the bioreactor. The agitator may be configured to agitate the bioreactor based on orbital motion. 
     Fluid Connector 
     Currently, there is no automated, multi-use sterile fluid connector solution for cell therapy production where a set of sterile fluid connectors are capable of multiple connection and disconnection cycles with a system. For example, conventional sterile fluid connectors are typically single-use devices and are thus expensive and labor intensive. Generally, the fluid connectors described herein include a plurality of sealed enclosures between a sterile portion (e.g., fluid connector lumen or cavity) and an external (e.g., non-sterile) ambient environment, thereby facilitating aseptic control of a fluid connector and devices coupled thereto. The fluid connectors described herein may be a durable component that may be reused for multiple cycles while maintaining sterility and/or bioburden control. For example, the fluid connector may be sterilized using a sterilant without harming the cell product or other biological material. 
     In some variations, a sterile manufacturing system as described herein may utilize one or more sterile fluid connectors and have a configuration suitable to be manipulated by a robot such as a robotic arm. The sterile fluid connectors described herein enable the transfer of fluids in an automated, sterile, and metered manner for automating cell therapy manufacturing. Automating cell therapy manufacturing may in turn provide lower per patient manufacturing costs, a lower risk of process failure, and the ability to meet commercial scale patient demand for cell therapies. In some variations, sterile fluid connectors may increase one or more of sterility, efficiency, and speed by removing a human operator from the manufacturing process. An automated and integrated sterilization process as described herein may be applied to the fluid connector to maintain sterility of the system. For example, the fluid connector may maintain sterility through multiple connection/disconnection cycles between separate sterile closed volume fluid devices (e.g., enclosure, container, vessel, cartridge, instrument, bioreactor, enclosed vessel, sealed chamber). Accordingly, the systems, devices, and methods described herein may reduce the complexity of a sterilization process, reduce energy usage, and increase sterilization efficiency. 
     In some variations, a fluid connector may comprise a first connector configured to mate with a second connector (e.g., male connector and female connector). Respective proximal ends of the connectors may be configured to connect (e.g., be in fluid communication, form a fluid pathway) with respective fluid devices in order to transfer one or more of fluid (e.g., liquid and/or gas) and biological material (e.g., cell product) between the fluid devices. The distal ends of the connectors may comprise ports configured to mate with each other. The fluid connector may also comprise a sterilant port configured to facilitate sterilization of a chamber within the distal ends of the first and second connectors. The fluid connector may be sterilized before or after connection as desired to ensure sterility. In this manner, the fluid connector may be reused for multiple connection and disconnection cycles. 
     In some variations, a system (e.g., workcell) utilizing the fluid connectors described herein may comprise a robot configured to operate the fluid connector and a controller configured to control the robot to manipulate (e.g., move, connect, open, close, disconnect) the first and second connectors together (without human interaction) while maintaining sterility of the fluid connector and a plurality of fluid devices, thereby further reducing the risk of contamination. The fluid devices may be one or more of an instrument, cartridge, and the like. 
     Cell Processing Control 
     Systems and methods for manufacturing cell products for biomedical applications using automated systems are described herein. Conventional semi-automated solutions for cell processing do not allow users to define biological processes. Instead, users select from a limited set of predefined machine processes and process-control parameters. Currently, there is no scalable manufacturing solution for cell therapy production. For example, cell therapy manufacturing is conventionally executed batchwise (i.e. one product will be manufactured in a single room/suite, with required processing tools located inside). This can either be guided by a technician following a standard operating procedure (SOP), or in some cases, processing tools (e.g., Miltenyi Prodigy, Lonza Cocoon) can carry out a series of processing steps for a single patient product on a single multi-functional processing tool. However, existing solutions (e.g., Miltenyi Prodigy) do not allow users to define biological processes. Furthermore, the manual labor required of conventional solutions increases the risk of product contamination and human error. 
     In some variations, a set of cell therapy biological manufacturing processes may be transformed into a set of machine instructions suitable for automated execution using the systems described herein. For example, a method of transforming user-defined cell processing operations into cell processing steps to be executed by a processor of an automated cell processing system may comprise receiving an ordered input list of cell processing operations, and executing a transformation model on the ordered input list to create an ordered output list of cell processing steps capable of being performed by the system. As used herein, a transform model may refer to an algorithm, process, or transformation configured to translate a set of cell processing steps into a set of machine or hardware instructions for the system. In some variations, a robot may be controlled to move one or more cartridges each containing a cell product between the instruments, and the instruments may be controlled to perform cell processing steps on each cell product. In this manner, the systems and methods enable biologists to define manufacturing processes in biological terms and have the system transform this biological model (e.g., process definition) into a set of machine-executed instructions. 
     The end-to-end closed system automation described herein may reduce process failure rates and cost. For example, end-to-end automation may reduce manufacturing time (e.g., dwell times) and increase throughput as compared to conventional manual methods. For example, a plurality of processes (e.g., 10 or more) may be executed simultaneously. The methods described herein may further reduce opportunities for contamination and user error. Thus, the systems, apparatuses, and methods described herein may increase one or more of cell processing automation, repeatability, reliability, process flexibility, instrument throughput, process scalability, and reduce one or more of labor costs, and process duration. 
     I. System 
     Described here are systems and apparatuses configured to perform cell processing steps to manufacture a cell product (e.g., cell therapy product). In some variations, a cell processing system may comprise a plurality of instruments each independently configured to perform one or more cell processing operations upon a cartridge (e.g., fluid device), and a robot capable of moving the cartridge between each of the plurality of instruments. The use of a robot and controller may facilitate one or more of automation, efficiency, and sterility of a cell processing system. 
     In some variations, a system for cell processing may comprise a plurality of instruments each independently configured to perform one or more cell processing operation upon a cartridge. A robot may be capable of moving the cartridge between each of the plurality of instruments. In some variations, the system may be a workcell comprising an enclosure. 
       FIG.  1 A  is a block diagram of a cell processing system  100  comprising a workcell  110  and controller  120 . In some variations, the workcell  110  may comprise one or more of an instrument  112 , a cartridge  114  (e.g., consumable, fluid device), a robot  116  (e.g., robotic arm), a reagent vault  118 , a fluid connector  132 , a sterilant source  134 , a fluid source  136 , a pump  138 , a sensor  140 , and a sterile liquid transfer device  142 . In some variations, the controller  120  may comprise one or more of a processor  122 , a memory  124 , a communication device  126 , an input device  128 , and a display  130 . 
     In some variations, a workcell may comprise a fully, or at least partially, enclosed housing inside which one or more cell processing steps are performed in a fully, or at least partially, automated process. In some variations, the workcell may be an open system lacking an enclosure, which may be configured for use in clean room, biosafety cabinet, or other sterile location. In some variations, the cartridge  114  may be moved using the robot  116  to reduce manual labor in the cell processing steps. In some variations, the workcell may be configured to perform sterile liquid transfers into and out of the cartridge in a fully or partially automated process. For example, one or more fluids may be stored in a sterile liquid transfer device  142 . In some variations, the sterile liquid transfer device may be a portable consumable that may be moved within the system  100 . The sterile liquid transfer devices and fluid connectors described herein enable the transfer of fluids in an automated, sterile, and metered manner for automating cell therapy manufacturing. In some variations, the enclosure of the workcell may be configured to meet International Organization for Standardization (ISO) standard ISO7 or better (e.g., ISO6 or ISO5). An advantage of meeting ISO7 or better standards is that the system may be used in a facility that does not meet ISO7 standards (i.e. that lack a clean room or other sufficiently filtered air space). Optionally, the facility may be an ISO8 or ISO9 facility. In some variations, a workcell may comprise a volume of less than about 800 m 3 , less than about 700 m 3 , less than about 600 m 3 , less than about 500 m 3 , less than about 300 m 3 , less than about 250 m 3 , less than about 200 m 3 , less than about 150 m 3 , less than about 100 m 3 , less than about 50 m 3 , less than about 25 m 3 , less than about 10 m 3 , and less than about 5 m 3 , including all ranges and sub-values in-between. 
     In some variations, a robot  116  may be configured to manipulate consumable cartridges  114  and fluid connectors  132  between different instruments to perform a predetermined sequence of cell processing steps. In some variations, the same consumable cartridge  114  may be received by different instruments  112  and/or multiple cartridges  114  may be processed in parallel. 
     In some variations, a cartridge  114  may contain cell product from different donors or contain cell product intended for different recipients. The cell product from a single donor may be split between multiple cartridges  114  if necessary to generate enough product for therapeutic use, or when a donor is providing product for several recipients (e.g., for allogeneic transplant). The cell product for a single recipient may be split between multiple cartridges  114  if necessary to generate enough product for therapeutic use in that recipient. The cell product for a single recipient may be split between multiple cartridges  114  if necessary to generate several cell products with unique genetic modifications, and then optionally recombined in certain ratios for therapeutic use in that recipient. For example, a fluid connector  132  may be coupled between two or more cartridges  114  to transfer a cell product and/or fluid between the cartridges  114 . Furthermore, a fluid connector  132  may be coupled between any set of fluid-carrying components of the system  100  (e.g., cartridge  114 , reagent vault  118 , fluid source  136 , sterile liquid transfer device  142 , fluid conduit, container, vessel, etc.). For example, a first fluid connector may be coupled between a first cartridge and a sterile liquid transfer device, and a second fluid connector may be coupled between the sterile liquid transfer device and a second cartridge. 
     As illustrated in  FIG.  1 B , a cartridge  114  may comprise one or more of a bioreactor  150 , cell separation system  152 , electroporation module  160 , liquid transfer bus  162 , sensor  164 , and fluid connector  166 , as described in more detail herein. A cell separation system  152  may comprise one or more of a rotor  154 , flow cell  156 , and magnet  158 . In some variations, the magnet  158  may comprise one or more magnets and/or magnet arrays. For example, the cell separation system  152  may comprise a first magnet configured to magnetically rotate a rotor  154  and a second magnet (e.g., magnet array) configured to magnetically separate cells in flow cell  156 . 
     Workcell 
     In some variations, a workcell  110  may comprise at least a partially enclosed enclosure (e.g., housing) in which one or more automated cell processing steps are performed. For example, the workcell  110  may be configured to transfer sterile liquid into and out of a cartridge  114  in a fully or partially automated process. In some variations, a workcell  110  may not have an enclosure and be configured for use in a clean room, a biosafety cabinet, or other suitably clean or sterile location. In some variations, the workcell  100  may comprise a feedthrough access biosafety cabinet, quality control instrumentation, pump, consumable (e.g., fluid device), fluid connector, consumable feedthrough, and sterilization system (e.g., sterilant source and/or generator, fluid source, heater/dessicator, aerator). 
       FIG.  2 A  is a block diagram of a cell processing system including a workcell  203 . Workcell  203  may comprise an enclosure  202  having four walls, a base, and a roof. The workcell may be divided into an interior zone  204  with a feedthrough  206  access, and quality control (QC) instrumentation  212 . An air filtration inlet (not shown) may provide high-efficiency particulate air (HEPA) filtration to provide ISO7 or better air quality in the interior zone  204 . This air filtration may maintain sterile cell processing in an ISO8 or ISO9 manufacturing environment. The workcell  203  may also have an air filter on the air outlet to preserve the ISO rating of the room. In some variations, the workcell  203  may further comprise, inside the interior zone  104 , a bioreactor instrument  214 , a cell selection instrument  216  (e.g., MACS), an electroporation instrument (EP)  220 , a counterflow centrifugation elutriation (CCE) instrument  222 , a sterile liquid transfer instrument  224  (e.g., fluid connector), a reagent vault  226 , and a sterilization system  260 . The reagent vault  226  may be accessible by a user through a sample pickup port  228 . A robot  230  (e.g., support arm, robotic arm) may be configured to move one or more cartridges  250  (e.g., consumables) from any instrument to any other instrument and/or move one or more cartridges  250  to and from a reagent vault. In some variations, the workcell  203  may comprise one or more moveable barrier  213  (e.g., access, door) configured to facilitate access to one or more of the instruments in the workcell  203 . 
     In some variations of methods according to the disclosure, a human operator may load one or more empty cartridges  250  into the feedthrough  206  via cartridge port  207 . The cartridges  250  may be pre-sterilized, or the feedthrough  206  may sterilize the cartridge  250  using ultraviolet radiation (UV), or chemical sterilizing agents provided as a vapor, spray, or wash. The feedthrough  206  chamber may optionally be configured to automatically spray, wash, irradiate, or otherwise treat cartridges (e.g. with ethanol and/or isopropyl alcohol solutions, vaporized hydrogen peroxide (VHP)) to maintain sterility of the interior zone  204  (e.g., ISO 7 or better). The cartridge  250  may be passed to the biosafety cabinet  206 , where input cell product is provided and loaded to the cartridge through a sterile liquid transfer port into the cartridge  250 . The user (via robot  230 ) may then move the cartridge  250  back to the feedthrough  206  and initiate automated processing using a computer processor in the computer server rack (e.g., controller  120 ). The robot  230  may be configured to move the cartridge  250  in a predefined sequence to a plurality of instruments and stations, with the components of the workcell  200 . At the end of cell processing, the cartridge  250 , now containing the processed cell product, may be returned to the feedthrough  206  for retrieval by the user. In some variations, an outer surface of the enclosure  202  may comprise an input/output device  208  (e.g., display, touchscreen). 
       FIG.  2 B  is a perspective view of a workcell  205  of a cell processing system.  FIG.  2 C  is a perspective view of a cell processing system depicting a cartridge  250  (e.g., any of the cartridges described herein) introduced into a workcell  205  (e.g., any of the workcells described herein). A plurality of cartridges may be inserted into the workcell  205  simultaneously and undergo one or more cell processing operations in parallel. 
     In some variations, the workcell  205  may comprise a height of more than about a meter, between about 1 m and about 3 m, between about 1 m and about 5 m, between about 3, and about 10 m, between about 5 m and about 20 m, between about 10 m and about 30 m, between about 20 m and 100 m, and more than about 100 m, including all values and ranges in-between. In some variations, the workcell  205  may comprise one or more of a length and width of more than about 1 meter, between about 1 m and about 5 m, between about 3, and about 10 m, between about 5 m and about 20 m, between about 10 m and about 30 m, between about 20 m and 100 m, and more than about 100 m, including all values and ranges in-between. 
       FIG.  2 D  is a schematic illustration of a variation of a workcell  200 . Workcell  200  may comprise an enclosure  202  having four walls, a base, and a roof. The workcell may be divided into an interior zone  204  with a feedthrough  206  access, a biosafety cabinet (BSC)  208 , compute server rack  210  (e.g., controller  120 ), and quality control (QC) instrumentation  212 . An air filtration inlet (not shown) may provide high-efficiency particulate air (HEPA) filtration to provide ISO7 or better air quality in the interior zone  204 . This air filtration may maintain sterile cell processing in an ISO8 or ISO9 manufacturing environment. The workcell may also have an air filter on the air outlet to preserve the ISO rating of the room. In some variations, the workcell  200  may further comprise, inside the interior zone  204 , an instrument  211  (e.g., disposed in a universal instrument bay), a bioreactor instrument  214 , a cell selection instrument  216  (e.g., MACS, cell selection system), a cell sorting instrument  218  (e.g., FACS), an electroporation instrument (EP)  220 , and a counterflow centrifugation elutriation (CCE) instrument  222 , a sterile liquid transfer instrument  224  (e.g., fluid connector), a reagent vault  226 , and a sterilization system  260  comprising one or more of a sterilant source, fluid source, and a pump. The reagent vault  226  may be accessible by a user through a sample pickup port  228 . A robot  230  (e.g., support arm, robotic arm) may be configured to move one or more cartridges  250  (e.g., consumables) from any instrument to any other instrument or reagent vault. 
     In some variations, a human operator may load one or more cartridges  250  into the feedthrough  206 . The cartridges  250  may be pre-sterilized, or the feedthrough  206  may sterilize the cartridge  250  using ultraviolet radiation (UV), or chemical sterilizing agents provided as a spray or wash. The feedthrough  206  chamber may optionally be configured to automatically spray, wash, irradiate, or otherwise treat cartridges (e.g. with ethanol and/or isopropyl alcohol solutions) to maintain sterility of the interior zone  204  (e.g., ISO 7 or better) or the biosafety cabinet  208  (e.g., ISO 5 or better). The cartridge  250  may be passed to the biosafety cabinet  206 , where input cell product is provided and loaded to the cartridge using a sterile liquid transfer instrument  224  (e.g., fluid connector) into the cartridge  250 . The user may then move the cartridge  250  back to the feedthrough  206  and initiate automated processing using a computer processor in the computer server rack  210  (e.g., controller  120 ). The robot  230  may be configured to move the cartridge  250  in a predefined sequence to a plurality of instruments and stations, with the components of the workcell  200  being controlled by the computer processor of the computer server rack  210 . Additionally or alternatively, the sequence that the cartridge  250  moves within the workcell  200  may not be predefined. For example, cartridge  250  movement may not be dependent on one or more of the result of a previous step, sensor value, predetermined threshold (e.g., based on a quality control system), and the like. At the end of cell processing, the cartridge  250 , now containing the processed cell product, may be returned to the feedthrough  206  for retrieval by the user. Additionally or alternatively, the cell product  250  containing the processed cell product may be transferred (via a fluid connector) to a second cartridge (e.g., single-use cartridge) and stored in the reagent vault  226  for retrieval by the user. 
     In some variations, cells from a patient and starting reagents may be loaded into a cartridge (e.g., single-use cartridge) by a human operator in a biosafety cabinet located separate from the workcell or integrated into the workcell. In some variations, the cartridges described herein comprising a cell product and reagent may move through a non-sterile field without contamination since the cartridge is closed. The cartridge may further undergo an automated decontamination routine. For example, the cartridge may be placed within a feedthrough capable of facilitating decontamination of the cartridge before entering the ISO 7 environment in the workcell. 
       FIG.  2 E  is a plan schematic illustration of another variation of a workcell  201 . Workcells  200 ,  201 , and  203  may comprise an enclosure  202  having four walls, a base, and a roof. The workcell may be divided into an interior zone  204  with a feedthrough  206  access, a biosafety cabinet (BSC)  208 , compute server rack  210  (e.g., controller  120 ), and quality control (QC) instrumentation  212 . An air filtration inlet (not shown) may provide high-efficiency particulate air (HEPA) filtration to provide ISO7 or better air quality in the interior zone  204 . This air filtration may maintain sterile cell processing in an ISO8 or ISO9 manufacturing environment. The workcell may also have an air filter on the air outlet to preserve the ISO rating of the room. In some variations, the workcell  200  may further comprise, inside the interior zone  104 , an instrument  211  (e.g., disposed in a universal instrument bay), a bioreactor instrument  214 , a cell selection instrument  216  (e.g., MACS), a cell sorting instrument  218  (e.g., FACS), an electroporation instrument (EP)  220 , and a counterflow centrifugation elutriation (CCE) instrument  222 , a sterile liquid transfer instrument  224 , and a reagent vault  226 . The reagent vault  226  may be accessible by a user through a sample pickup port  228  (e.g., a door which may facilitate bulk loading of sterile liquid transfer instruments  224 ). A robot  230  (e.g., support arm, robotic arm) may be configured to move one or more cartridges  250  (e.g., consumables) from any instrument to any other instrument or reagent vault. 
     In some variations of methods according to the disclosure, a human operator may load one or more empty cartridges  250  into the feedthrough  206 . Additionally or alternatively, pre-filled cartridges may be loaded into the feedthrough  206 . The cartridges  250  may be pre-sterilized, or the feedthrough  206  may sterilize the cartridge  250  using ultraviolet radiation (UV), or chemical sterilizing agents provided as a spray or wash. The feedthrough  206  chamber may optionally be configured to automatically spray, wash, irradiate, or otherwise treat cartridges (e.g. with ethanol and/or isopropyl alcohol solutions) to maintain sterility of the interior zone  204  (e.g., ISO 7 or better) or the biosafety cabinet  208  (e.g., ISO 5 or better). The cartridge  250  may be passed to the biosafety cabinet  106 , where input cell product is provided and loaded to the cartridge through a sterile liquid transfer port into the cartridge  250 . The user may then move the cartridge  250  back to the feedthrough  206  and initiate automated processing using a computer processor in the computer server rack  210  (e.g., controller  120 ). The robot  230  may be configured to move the cartridge  250  in a predefined sequence to a plurality of instruments and stations, with the components of the workcell  200  being controlled by the computer processor of the computer server rack  210 . At the end of cell processing, the cartridge  250 , now containing the processed cell product, may be returned to the feedthrough  206  for retrieval by the user. 
     In some variations, one or more components of a sterilization system (e.g., sterilant source, pump) may be coupled to a workcell. For example,  FIG.  3    is a block diagram of a cell processing system  300  comprising a workcell  310 , sterilization system  320 , fluid connector  330  and fluid devices  340 . In some variations, the fluid devices  340  may comprise a main (e.g., consumable) feedthrough and a fluid device (e.g., reagent) feedthrough. The sterilization system  320  may comprise a sterilant source  322 , pump  324 , and heater (e.g., desiccant/dryer)  326 . For example, the heater  326  may be configured to aerate at a predetermined set of conditions. The sterilization system  320  may be coupled and in fluid communication with one or more of the workcell  310 , fluid connector  330 , and fluid device  340 . In some variations, a robot (not shown) may be configured to manipulate and operate the cell processing system  300 . For example, the fluid connector  330  may be coupled to one or more of the fluid devices  340  and instruments (not shown). One or more of the workcell  310 , fluid connector  330 , and fluid devices  340  may be sterilized and/or aerated by circulating one or more of a sterilant and fluid (e.g., heated air, vaporized hydrogen peroxide (VHP)) using the sterilization system  320 . In some variations, the sterilization system  320  may comprise one or more of vaporized hydrogen peroxide (VHP), electron-beam (e-beam) sterilization, dry thermal decontamination, and steam-in-place. In some variations, the sterilization system  320  may provide a sterility assurance level (SAL) of at least 10-3 SAL. 
       FIGS.  4 A and  4 B  illustrate perspective views of a cell processing system  400  comprising a cartridge  400 ,  402 , feedthrough  410 ,  412 , and fluid connector  420 ,  422  (e.g., sterile liquid transfer instrument). For example, cartridge  400  is shown in the feedthrough  410  in  FIG.  16 A  while a robot (not shown) has moved cartridge  400  to fluid connector  420 . 
     Robot 
     Generally, a robot may comprise any mechanical device capable of moving a cartridge from one location to another location. For example, the robot may comprise a mechanical manipulator (e.g., an arm) in a fixed location, or attached to a linear rail, or a 2- or 3-dimensional rail system. In a variation, the robot comprises a robotic shuffle system. In a further variation, the robot comprises a wheeled device. In some variations, the system comprises two or more robots of the same or different type (e.g., two robotic arms each independently configured for moving cartridges between instruments). The robot may also comprise an end effector for precise handling of different cartridges or barcode scanning or radio-frequency identification tag (RFID) reading. 
       FIG.  5    is a perspective view of a cell processing system  500  in which a robot arm moves consumable cartridges between slots in various instruments each configured to perform a different cell processing step. In some variations, the same consumable cartridge can be received by different instruments. The system  500  may comprise a modular design to accommodate different instrument configurations. In some variations, a plurality of cartridges may be processed in parallel. Each cartridge may contain a cell product from different donors or contain a cell product intended for different recipients. For example, a cell product from a single donor may be split between a plurality of cartridges to generate a predetermined quantity of cell product for therapeutic use such as when a donor is providing product for several recipients (e.g., for allogeneic transplant). In some variations, the cell product for a single recipient may be split between a plurality of cartridges to generate a predetermined quantity of product for therapeutic use in that recipient. In some variations, the cell product for a single recipient may be split between a plurality of cartridges to generate a predetermined quantity of several cell products with unique genetic modifications, which may be recombined in certain ratios for therapeutic use in that recipient. 
     Cartridge 
     Generally, the cell processing systems described herein may comprise one or more cartridges including one or more modules configured to interface with an instrument or instruments. A robot (e.g., robotic arm) may be configured to move a cartridge and/or instrument to perform one or more cell processing steps. For example, a cartridge may comprise a bioreactor module and/or fluid connector (e.g., sterile liquid transfer port) coupled by the robot to a bioreactor instrument of a workcell. Once a predetermined processing step has been completed, the cartridge may be moved by the robot to another instrument of the workcell, and another cartridge may be coupled to the bioreactor instrument. Thus, a portable cartridge and shareable instruments may increase the efficiency, throughput, and flexibility of a cell manufacturing process. 
     In some variations, the cartridge may optionally provide a self-contained device capable of performing one or more cell processing steps. The modules may be integrated into a fixed configuration within the cartridge. Additionally or alternatively, the modules may be configurable or moveable within the cartridge, permitting various cartridges to be assembled from shared modules. Similarly stated, the cartridge can be a single, closed unit with fixed components for each module; or the cartridge may contain configurable modules coupled by configurable fluidic, mechanical, optical, and electrical connections. In some variations, one or more sub-cartridges, each containing a set of modules, may be configured to be assembled to perform various cell processing workflows. The modules may each be provided in a distinct housing or may be integrated into a cartridge or sub-cartridge with other modules. The disclosure generally shows modules as distinct groups of components for the sake of simplicity, but may be arranged in any suitable configuration. For example, the components for different modules may be interspersed with each other such that each module is defined by the 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 variations, multiple cartridges may be used to process a single cell product through transfer of the cell product from one cartridge to another cartridge of the same or different type and/or by splitting cell product into more cartridges and/or pooling multiple cell products into fewer cartridges. 
     Generally, each of the instruments of the system interfaces with its respective module or modules on the cartridge e.g., an electroporation module on the cartridge (if present) is moved by the system to an electroporation instrument and interfaces with the electroporation instrument to perform an electroporation step on the cell product—and may also interface with common components, such as components of a fluidic bus line (e.g., pumps, valves, sensors, etc.). An advantage of such split module/instrument designs is that expensive components (e.g., motors, sensors, heaters, lasers, etc.) may be retained in the instruments of the system while multiple cartridges are processed. The use of disposable cartridges may eliminate the need, in such variations, to sterilize cartridges between use. Furthermore, the utilization of shared instruments (e.g. electroporation instrument, CCE instrument, MACS instrument, sterile liquid transfer instrument, FACS instrument, and the like) may be increased since a plurality of the instruments may be utilized simultaneously in parallel by a plurality of cell manufacturing processes. In contrast, conventional semi-automated instruments (e.g., Miltenyi Prodigy) have instrument components that sit idle and are incapable of simultaneous parallel use. 
       FIG.  6    is a schematic illustration of a cartridge  600  that may be a consumable produced from materials at a cost that make recycling or limited use practical. The cartridge  600  may comprise a liquid transfer bus  624  fluidically coupled to a small bioreactor module  614   a , a large bioreactor module  614   b , a cell selection module  616 , a cell sorting module  618 , an electroporation module  620 , and a counterflow centrifugation elutriation (CCE) module  622 . In some variations, the cell selection module  616  may be a magnetic-activated cell selection (MACS) module. The cell sorting module  618  may comprise a fluorescence activated cell sorting (FACS) module. The cartridge  600  may comprise a housing  602  that renders the cartridge self-contained, and optionally protects the contents from contamination. Sterile liquid transfer ports (SLTPs)  606   a - 606   k  may be fluidically coupled to reservoirs  607   a - 607   k , and each independently be a flexible bag or a rigid container. In some variations, flexible bags may be configured to hold large volumes and to permit transfer of fluid without replacing transferred fluid with liquid or gas to maintain the pressure in the reservoir, as the bag may collapse when fluid is transferred out and expand when fluid is transferred in. 
     In some variations, the liquid transfer bus  624  may comprise valves V 1  to V 28  and corresponding tubing that fluidically links the valves to one another and to each of the modules. Valves shown coupled to four fluidic lines are 4/2 (4 port 2 position) valves and valves shown coupled to three fluidic lines are 3/2 (3 port 2 position) valves. Internal flow paths of the valves are indicated in the legend. The cartridge may further comprise a first pump  632   a  and a second pump  632   b , each of which expose tubing on the exterior of the housing  602  to permit each pump to interface with pump actuators (e.g., rotors) in some instruments in the system (e.g., workcell). The liquid transfer bus  624  may be fluidically coupled to reservoir  607   d  and a product bag which is fluidically coupled to STLP  606   d  and to product input tubing lines  627   a - 627   b . An operator may input a cell product into reservoir  607   d  by connecting product input tubing line  627   a  or  627   b  to an external source of cells (e.g., a bag of cells collected from a donor). SLTP  606   d  may be configured to permit a system according to the disclosure (e.g., workcell  110 ) to add fluid to the reservoir  607   d  in an automated fashion. For example, one or more fluid-carrying containers such as reservoirs  607   a - 607   k , bags, etc. may receive fluid using an SLTP. Additionally or alternatively, the SLTP may be configured to periodically sample one or more of the fluid-carrying containers. The cartridge may further comprise collection bags  626   a - 626   c , fluidically coupled to the liquid transfer bus  624  via valves V 17 -V 19 . The cartridge  600  may be configured to permit an operator to remove the collection bags  626   a - 626   c  after completion of cell processing by the system. 
       FIG.  7    is a schematic diagram of another variation of a cartridge  700 . For example, cartridge  700  may comprise a reduced feature set compared to cartridge  600 . The cartridge  700  may comprise a liquid transfer bus  724  fluidically coupled to a bioreactor module  714 , a counterflow centrifugation elutriation (CCE) module  722 , and a module  716  selected from cell selection module, a cell sorting module, an electroporation module, or any other cell processing module. The cartridge  700  may comprise a housing  702  and sterile liquid transfer ports (SLTPs)  706   a - 706   f  (e.g., fluid connector) fluidically coupled to reservoirs  707   a - 707   f , which may be each independently be a flexible bag or a rigid container. SLTP  706   g  is fluidically coupled to the bioreactor module  714  to permit direct access by a system or an operator to the bioreactor. Reservoir  707   c  may be fluidically coupled to SLTP  707   c  and product input tubing line  727 . In some variations, the liquid transfer bus  724  may comprise 14 valves V 1 -V 3 , V 9 , V 11 -V 12 , V 17 -V 23  and V 28  and tubing that fluidically couples the values to one another and/or each of the modules. The cartridge may further comprise collection bags  726   a - 726   c  fluidically coupled to the liquid transfer bus  724  via valves V 17 -V 19 . The cartridge may further comprise a pump  732  which exposes the tubing on the exterior of the housing  702  to permit each pump to interface with a pump actuator in the system (e.g., workcell). 
     A side and top view of another variation of a cartridge is shown in respective  FIGS.  8 A and  8 B . In some variations, a cartridge  800  may comprise a bioreactor  814 , a pump  816 , and a counterflow centrifugation elutriation (CCE) module  822 . The cartridge  800  may comprise blanks  818 ,  819 , and  820  configured to house additional module(s) such as a cell selection module, cell sorting module, an electroporation module, a small bioreactor module, and the like. In some variations, a blank may define an empty volume of the cartridge reserved to house a module at another time. In some variations, the cartridge  800  may comprise two or more additional bioreactors and/or reservoirs in blanks  818 ,  819 ,  820 . Along the near surface of the cartridge  800  may be fluid connectors  806   a - 806   j  (e.g., SLTP) fluidically connected to reservoirs  807   a - 807   f . Reservoirs  807   b  and  807   e  may comprise fluid (e.g., buffer or media). Along the top surface are product input tubing lines  827   a - 827   d , which may be fluidically connected to reservoirs  807   a ,  807   b ,  807   e , and  807   f , respectively. A liquid transfer bus  824  may fluidically connect the STLPs, reservoirs, and product input tubing lines to the modules via tubing. 
     In some variations, the housing  802  may have external dimensions of about 225 mm× about 280 mm×385 mm, about 225 mm× about 295 mm×385 mm, and about 450 mm× about 300 mm× about 250 mm, including all values and sub-ranges in-between. In some variations, the cartridge  800  may be about 10%, about 20%, about 30% or more smaller in volume, including all ranges and sub-values in-between. In some variations, the cartridge  800  may be about 10%, about 20%, about 30%, about 50%, about 100%, about 200%, or more in volume, including all ranges and sub-values in-between. 
     In some variations, a cartridge  800  as shown in the side view of  FIG.  8 C  and perspective view of  FIG.  8 D  may comprise a MACS module  818 . For example, the bioreactor module  814  may comprise ports  815   a - 815   f  including a pH and dissolved oxygen (DO) sensors (ports  815   a  and  815   b ), a gas input line  815   c , an output line  815   d  each having a sterile filter behind the connector, and a coolant input line  815   e  and output line  815   f  from the bioreactor instrument interface when it interfaces with bioreactor module  814  (for heat exchange). For example, the gas input line  815   c  may be configured for gas transfer into a fluid (e.g., through headspace gas control or a gas-permeable membrane). 
       FIG.  9    shows a cross-sectional side view of a cartridge  900 . In some variations, a cartridge  900  may comprise an enclosure (e.g., housing), a bioreactor  914 , one or more pumps  916 , valve  930 , cell selection module  917 , and a counterflow centrifugation elutriation (CCE) module  922 . In some variations, the cell selection module  616  may be a magnetic-activated cell selection (MACS) module  917 . The cartridge may further comprise collection bags  926 . The cartridge  900  may optionally comprise blanks configured to house additional module(s) such as a cell selection module, a cell sorting module, an electroporation module  918 , and the like. In some variations, the cartridge  900  may comprise one or more bioreactors and/or reservoirs in the blanks. 
     In some variations, a cartridge may comprise one or more valves. In some variations, the valve  1000  on the cartridge may be configured to receive an actuator  1010  provided by an instrument (as shown in  FIG.  10 A ). As the cartridge is inserted into the instrument, the valve  1000  may be configured to dock with the actuator  1010  (as shown in  FIG.  10 B ), such that rotation of the actuator  1010  may cause switching of the valve  1000  from one position to another position. In some variations, the valves may be constructed to pinch a section of soft tubing. The pinch valves may comprise a closed configuration, and an external actuator may be configured to interface with the pinch valve (e.g., utilizing a solenoid with linear motion) to open or close the valve. The valves themselves may be configured to be disposable whereas the actuators may be integrated into an instrument configured to process cartridges repeatedly. 
     Reagent Vault 
     In some variations, the system comprises a reagent vault (or reagent vaults) where reagents are stored including but not limited to cell culture media, buffer, cytokines, proteins, enzymes, polynucleotides, transfection reagents, non-viral vectors, viral vectors, antibiotics, nutrients, cryoprotectants, solvents, cellular materials, and pharmaceutically acceptable excipients. Additionally or alternatively, waste may be stored in the reagent vault. In some variations, in-process samples extracted from one or more cartridges may be stored in the reagent vault. The reagent vault may comprise one or more controlled temperature compartments (e.g., freezers, coolers, water baths, warming chambers, or others, at e.g. about −80° C., about −20° C., about 4° C., about 25° C., about 30° C., about 37° C., and about 42° C.). Temperatures in these compartments may be varied during the cell manufacturing process to heat or cool reagents. In variations of the methods of the disclosure, a cartridge may be moved by the robot (or manually by an operator) to the reagent vault. The reagent vault interfaces with one or more sterile liquid transfer ports on the cartridge, and the reagent or material is dispensed into the cartridge. Optionally, fluid is added or removed from the cartridge before, during, or after reagent addition or removal. In some variations, the system comprises a sterile liquid transfer instrument, similarly configured to transfer fluid into or out of the cartridge in an automated, manual, or semi-automated fashion. An operator may stock the sterile liquid transfer station with reagents manually, or they may be supplied by a robot (e.g. from a feedthrough or other location). In some cases, a robot moves a reagent or reagents from the reagent vault to the sterile liquid transfer station. The reagent vault may have automated doors to permit access by the robot for sterile liquid transfer devices and/or other reagent vessels, optionally each under independent closed loop temperature control. The devices and vessels may be configured for pick-and-place movement by the robot. In some variations, the reagent vault may comprise one or more sample pickup areas. For example, a robot may be configured to move one or more reagents to and from one or more of the sample pickup areas. 
     Various materials can be used to construct the cartridge and the cartridge housing, including metal, plastic, rubber, and/or glass, or combinations thereof. The cartridge, its components, and its housing may be molded, machined, extruded, 3D printed, or any combination thereof. The cartridge may contain components that are commercially available (e.g., tubing, valves, fittings); these components may be attached or integrated with custom components or devices. The housing of the cartridge may constitute an additional layer of enclosure that further protects the sterility of the cell product. The operator may perform loading or unloading of the cartridge in an ISO 5 or better environment, utilizing aseptic technique to ensure that sterility of the contents of the cartridge is maintained when the cartridge is opened. In some variations, the operator may perform loading or unloading of the cartridge using manual aseptic connections (e.g., sterile tube welding). The robotic system may also perform sterile loading or unloading of liquids into and out of the cartridge through the use of the sterile liquid transfer instrument and sterile liquid transfer ports on the cartridge. 
     Counterflow Centrifugal Elutriation 
     Counterflow centrifugal elutriation (CCE) is a technique used to separate cells based on characteristics such as size and/or density. Counterflow centrifugal elutriation combines centrifugation with counterflow elutriation where centrifugation corresponds to the process of sedimentation under the influence of a centrifugal force field and counterflow elutriation corresponds to the process of separation by washing. Separation takes place in a cone (e.g., bicone, funnel) shaped elutriation chamber. Particles (e.g., cells) conveyed in a fluid into the elutriation chamber are acted upon by two opposing forces: centrifugal force driving the fluid away from an axis of rotation; and fluid velocity driving the fluid towards the axis of rotation (e.g., counterflow). By varying the flow rate and the centrifugal force, the separation of particles (e.g., cells) may be achieved. For example, as described in more detail herein, particles may be separated based on properties such as size and density. 
     Counterflow centrifugal elutriation may perform multiple operations useful for cell therapy manufacturing workflows including, but not limited to, cell washing, cell concentration, media/buffer replacement, transduction, and separation of white blood cells from other blood components (e.g., platelets, and red blood cells). In some variations, a fluid source (e.g., apheresis bag) for a cell separation process may comprise a suspension of white blood cells, red blood cells, platelets, and plasma. In order to separate immune cells of interest, white blood cells may be isolated and subsequently magnetically tagged for magnetic separation. A white blood cell separation step may be performed in a CCE module to separate cells based on size and density, while magnetic separation may be performed in a MACS module. In some variations, a CCE module may be integrated into a cartridge to enable a cell processing system to separate cells based on one or more of a progression through a cell cycle (e.g., G 1 /M phase cells being larger than G 0 , S, or G2 phase cells) and cell type (e.g., white blood cells from red blood cells and/or platelets). 
     Generally, a rotor configured to spin may comprise an elutriation chamber (e.g., cone, bicone). A fluid comprising a suspension of cells may be pumped under continuous flow into the rotor. As cells are introduced into the cone (e.g., bicone), the cells migrate according to their sedimentation rates to positions in the gradient where the effects of the two forces upon them are balanced. Smaller cells having low sedimentation rates (e.g., platelets) may be quickly washed toward the axis of rotation with increased flow velocity. Such smaller cells may be output (e.g., washed out) of the cone. Relatively larger (or denser) cells (e.g., red blood cells) flow through the cone relatively more slowly and reach equilibrium at an elutriation boundary where the centrifugal force and the drag force are in balance, and the fluid velocity is relatively low because the cone has widened. The largest or densest cells (e.g., white blood cells) remain near the inlet to the chamber where centrifugal force and fluid velocity are high. By increasing the flow rate in gradual steps, successive fractions of increasingly large or dense cells (e.g., platelets→red blood cells→white blood cells) may be output from the rotor. Continued incremental increases in fluid flow rate will eventually elutriate all cells from the cone. 
       FIG.  56    is a block diagram of a cell separation system  5600  comprising a workcell  5610  and at least one cartridge  5620 . In some variations, the workcell  5610  may comprise one or more of a counterflow centrifugal elutriation (CCE) instrument  5632  (e.g., first magnet), a magnetic-activated cell selection (MACS) instrument  5642  (e.g., magnet array, second magnet), a fluid connector  5652 , a pump  5654 , an imaging system comprising an optical sensor  5660  and an illumination source  5662 , a sensor  5664 , and a processor  5670 . In some variations, the cartridge  5620  may comprise one or more of a CCE module  5630  (e.g., rotor), a MACS module  5640  (e.g., flow cell), and a fluid connector  5650  (e.g., sterile liquid transfer port, liquid transfer bus). For example, a cartridge for cell processing may comprise a liquid transfer bus and a plurality of modules, each module fluidically linked to the liquid transfer bus. The modules may include any of the CCE modules or MACS modules described herein. In some variations, a robot (not shown) may be configured to move the cartridge  5620  between different locations within the workcell  5610  to perform different cell processing steps. 
     In some variations, the imaging system (e.g., optical sensor  5660 , illumination source  5622 ) may be configured to generate image data corresponding to one or more of the CCE module  5630  and MACS module  5640 . For example, image data of fluid flow through a rotor of a CCE module  5630  may be analyzed and used to control a flow rate of fluid and/or rotation rate of the rotor, as described in more detail herein. In some variations, the optical sensor  5660  may be a CMOS/CCD sensor having, for example a resolution of about 100 μm, a working distance of between about 40 mm and about 100 mm, and a focal length of less than about 8 mm. The optical sensor  5660  may be configured to operate synchronously with the illumination source  5662 . In some variations, the optical sensor  5660  may comprise one or more of a colorimeter, turbidity sensor, and optical density sensor. In some variations, the illumination source  5662  may operate as a strobe light configured to output light pulses synchronized to a rotation rate of a rotor of the CCE module  5630 . 
     In some variations, the sensor  5664  may comprise one or more of an optical density sensor configured to measure an intensity of fluid, a leak detector configured to detect moisture and/or leaks, an inertial sensor configured to measure vibration, a pressure sensor configured to measure pressure in a fluidic line (e.g., photoelectric sensor), a bubble sensor configured to detect the presence of a bubble in a fluid conduit, colorimetric sensor, vibration sensor, and the like. 
     In some variations, the fluid connector  5652  may comprise one or more valves, configured to control fluid flow between the workcell and the cartridge  5620 . The processor  5670  may correspond to the controller (e.g., processor and memory) described in more detail herein. The processor  5670  may be configured to control one or more of the CCE instrument  5632 , the MACS instrument  5642 , the pump  5654 , fluid connector  5652  (e.g., valves), the optical sensor  5660 , the illumination source  5662 , and the sensors  5664 . 
     In some variations, a system  5600  for cell processing may comprise a cartridge  5600  comprising a rotor of a CCE module  5630  configured for counterflow centrifugation elutriation of cells in a fluid. A first magnet of a CCE instrument  5632  may be configured to magnetically rotate the rotor and separate the cells from the fluid in the rotor. The cartridge may further comprise a flow cell of a MACS module  5640  coupled to the rotor and configured to receive the cells from the rotor. A second magnet of a MACS instrument  5642  may be configured to magnetically separate the cells in the flow cell. 
     In some variations, an illumination source  5662  may be configured to illuminate the cells. An optical sensor  5660  may be configured to generate image data corresponding to the cells. In some variations, the system  5600  may comprise one or more of an oxygen depletion sensor, leak sensor, inertial sensor, pressure sensor, and bubble sensor. In some variations, the system  5600  may comprise one or more valves and pumps. 
       FIG.  57    is a cross-sectional side view of a counterflow centrifugal elutriation (CCE) module  5700  comprising a housing  5710  (e.g., enclosure), a rotor  5720  configured to rotate within and relative to the housing  5710 , and one or more fluid ports  5730  (e.g., fluid inlet, fluid outlet). In some variations, the CCE module  5700  may be portable and configured to move within a workcell  5610  and cartridge  5620 . For example, a robot may move the CCE module  5700  between different instruments of a workcell  5610 . 
       FIG.  58    is a cross-sectional side view of a magnetic-activated cell selection (MACS) module comprising a housing  5810  (e.g., enclosure), a first fluid port  5820  (e.g., fluid inlet), a second fluid port  5830  (e.g., fluid outlet), and a flow cell  5810  coupled in between the first fluid port  5820  and the second fluid port  5830 . As described in more detail herein, the flow cell  5810  may comprise a cavity (e.g., chamber) comprising one or more channels (e.g., linear channels, laminar fluid flow channel). In some variations, the cavity of the flow cell  5810  may be substantially empty. For example, the flow cell  5810  may be absent a mesh, beads, tortuous channels, and the like. In some variations, the flow cell  5810  may have a longitudinal axis aligned perpendicular to ground. That is, the flow cell  5810  may be oriented vertically where the first fluid port  5820  is disposed at a higher elevation than the second fluid port  5830  such that gravity may aid fluid flow through the flow cell  5810 . In some variations, the MACS module  5800  may be portable and configured to move within a workcell  5610  and cartridge  5620 . For example, a robot may move the MACS module  5630  between different instruments of a workcell  5610 . 
       FIGS.  59 A and  59 B  are perspective views of a system  5900  for cell processing (e.g., CCE system) comprising a CCE module  5930  (e.g., cartridge) including a housing  5931  and a rotor  5910 , a CCE instrument  5932 , an optical sensor  5960 , and an illumination source  5962 . In some variations, the CCE instrument  5932  may comprise a magnet configured to magnetically rotate the rotor  5910  within the CCE module  5930 . One or more portions of the housing  5931  and rotor  5910  may be optically transparent to facilitate illumination by the illumination source  5962  and image data generation by the optical sensor  5960 . 
     In some variations, the system  5900  for cell processing may comprise a cartridge  5930  comprising a housing  5931  comprising a rotor  5910  configured to separate cells from a fluid. An instrument  5932  comprising a magnet may be configured to interface with the cartridge  5930  to magnetically rotate the rotor  5910 . The cartridge  5930  may be configured to move a cell product between a plurality of instruments. In some variations, the housing  5931  may enclose the rotor  5910 . In some variations, the housing  5931  may comprise one or more apertures  5937  configured to facilitate visualization (e.g., imaging) of the rotor  5910 .  FIGS.  59 A and  59 B  depict a magnet  5932  in proximity, but not attached, to housing  5931 .  FIG.  59 C  is a perspective view of the rotor  5910  and housing  5931  without the magnet  5932 , optical sensor  5960 , and illumination source  5962 . 
     In some variations, the cartridge  5930  (e.g., housing  5931 ,  5910 ) may comprise a consumable component such as a disposable component, limited use component, single use component, and the like. In some variations, the magnet  5932  may comprise a durable component that may be re-used a plurality of times. In some variations, the magnet  5932  may be releasably coupled to the housing  5931 . For example, the housing  5931  may be moved relative to the magnet  5932  to facilitate magnetic coupling between the magnet  5932  and a plurality of cartridges  5930 . Additionally or alternatively, the magnet  5932  may be configured to be moved relative to the housing  5931 . 
       FIG.  59 D  is a side cross-sectional view of a CCE module  5930 . In some variations, the housing  5931  of the rotor  5910  may comprise a first side  5933  comprising the first fluid port  5912  (e.g., first fluid conduit) and a second side  5935  comprising the second fluid port  5914  where the second side  5935  is opposite the first side  5933 . The rotor  5910  (including a cone or bicone as described in more detail herein) may be coupled between the first fluid port  5912  and the second fluid port  5914 . In some variations, the CCE module  5930  may comprise an air gap  5902  between the housing  5931  and a magnet  5932 . That is, the cartridge  5930  and magnet  5932  may couple in a non-contact manner. Consequently, the cartridge need not mechanically couple to the magnet  5932  to perform counterflow centrifugal elutriation. Therefore, the rotor  5910  may have a low alignment sensitivity with the magnet  5932 , as well as low vibration between the rotor  5910  and the magnet  5932 . Furthermore, the space between the rotor  5910  and magnet  5932  enables the second fluid port  5914  to extend toward the second side  5935  of the housing  5931 , thus allowing for fluid to flow on each side of the rotor  5910 . 
     In some variations, counterflow centrifugal elutriation may be performed by the system  5900  by moving a magnet  5932  towards a rotor  5910  (or vice versa). The rotor may define a rotational axis (e.g., coaxial with the first fluid port  5912  and the second fluid port  5914 ). Fluid may flow through the rotor via the first fluid port  5912  and the second fluid port  5914 . The magnet  5932  may magnetically rotate the rotor about the rotational axis while flowing the fluid through the rotor  5910 . The rotor may move away from the magnet. For example, moving the rotor  5910  may include advancing and withdrawing the rotor  5910  relative to the magnet  5932  using a robot (not shown). 
     In some variations, fluid may flow through first fluid port  5912  along the first side  5933  of the rotor  5910  and into the rotor  5910 . After counterflow centrifugal elutriation through the rotor  5910 , the fluid may flow out of the rotor  5910  through second fluid port  5914  along the second side  5935  of the rotor  5910 . 
     In some variations, counterflow centrifugal elutriation may be visualized by optical sensor  5960  and illumination source  5962  in order to monitor and modify cell separation in real-time based on predetermined criteria in a closed loop manner in order to maximize elutriation efficiency. In some variations, an optical sensor  5960  may be configured to image any portion of the rotor through which fluid flows (e.g., first fluid conduit, second fluid conduit, third fluid conduit, first bicone, second bicone). For example, image data of one or more of the fluid and the cells in the rotor  5910  may be generated using the optical sensor  5960 . In some variations, one or more of the fluid and the cells may be illuminated using the illumination source  5962 . For example, an output of a cone may be imaged by an optical sensor to identify non-target cells being elutriated. 
     In some variations, one or more of a rotation rate of the rotor and a flow rate of the fluid may be selected based at least in part on the image data. For example, the rotor may comprise a rotation rate of up to 6,000 RPM. For example, the fluid may comprise a flow rate of up to about 150 ml/min while rotating the rotor. In some variations, the rotor may be moved towards the illumination source  5962  and the optical sensor  5960 . Additionally or alternatively, the rotor  5910  may be moved away from the illumination source  5962  and the optical sensor  5960 . 
       FIG.  59 E  is a side cross-sectional view of a rotor  5910  including a first fluid port  5912  (e.g., fluid conduit, inlet) and a second fluid port  5914  (e.g., fluid conduit, outlet). In some variations, the first fluid port  5912  and the second fluid port  5914  may extend in parallel with each other and/or a rotational axis of the rotor  5910 . In some variations, the first fluid port  5912  and the second fluid port  5914  may be disposed on opposite sides of the rotor  5910 , which may simplify fluid routing, cartridge design, and also reduce manufacturing costs. For example, the fluidic seals may be simplified since they contain only a single lumen each. Conventionally, a complicated fluid flow path (including inlet and outlet) is formed on a first side of a rotor due to a fixed mechanical coupling of a drive motor to a second side of the rotor.  FIGS.  59 F and  59 G  are cross-sectional side views of a rotor  5910  disposed within housing  5931 . 
       FIG.  60 A  is a plan view of a rotor  6000  that may be used with any of the CCE systems, CCE modules, cartridges, housings, combinations thereof, and the like described herein. The rotor  6000  may comprise a first fluid conduit  6010 , a cone  6020  (e.g., bicone), a second fluid conduit  6030 , a magnetic portion  6040  (e.g., magnet), and housing  6050 . Fluid may flow sequentially through the first fluid conduit  6010 , the cone  6020 , and the second fluid conduit  6030 . In some variations, the magnetic portion  6040  may comprise one or more magnets. In some variations, the rotor  6000  may define a rotation axis  6060 . In some variations, at least a portion of the first fluid conduit  6010  and at least a portion of the second fluid conduit  6030  may extend parallel to the rotation axis (e.g., into and out of the page with respect to  FIG.  60 A ). In some variations, at least a portion of the first fluid conduit  6010  and at least a portion of the second fluid conduit  6030  may be co-axial. 
     In some variations, the cone  6020  may comprise a bicone having a first cone including a first base and a second cone including a second base such that the first base faces the second base. In some variations, a bicone may comprise a cylinder (or some other shape) between and/or in fluid communication with the first cone and the second cone. For example, one or more cones of a rotor may comprise a generally stepped shape. For example, one or more cones may comprise stacked circular steps. In some variations, a cone of a rotor may comprise a single cone. 
     In some variations, at least a portion of the rotor may be optically transparent to facilitate visualization and/or imaging of the rotor  6000  and/or fluid (e.g., cells) in the rotor  6000 . For example, the cone  6020  may be transparent, as well as portions of the first fluid conduit  6010  and the second fluid conduit  6030 . 
     In some variations, the cone may comprise a volume of between about 10 ml and about 40 ml. In some variations, the cone may comprise a cone angle of between about 40 degrees and about 60 degrees. 
     In some variations, a cone may comprise a first cone (e.g., distal cone) and a second cone (e.g., proximal cone) where the first cone is larger than the second cone. In some variations, a first cone length may be between about 60 mm and about 90 mm. In some variations, a proximal cone length may be between about 15 mm and about 40 mm. In some variations, a cone diameter (e.g., maximum diameter of the cone) may be between about 15 mm and about 40 mm. 
     In some variations, the rotor  6000  may comprise an asymmetric shape. In some variations, a first portion (e.g., first end) of the rotor  6000  may comprise the cone  6020  and a second portion (e.g., second end) may comprise a paddle shape. 
     In some variations, the cone may comprise a length of at least about 4 cm (e.g., between about 9 cm and about 12 cm), a cone diameter of about 5 cm or less (e.g., between about 3 cm and about 5 cm), a fluid flow rate of up to about 100 ml/min (e.g., between about 60 ml/min and about 100 ml/min), and a rotation rate of less than about 3000 RPM. The shape of the first cone and the second cone may be generally linear (as opposed to convex or concave). 
       FIGS.  60 B and  60 C  are perspective views, and  FIG.  60 D  is a side view of a rotor  6002  comprising a first fluid conduit  6012 , a cone  6022 , a second fluid conduit  6032 , and a housing  6052 .  FIG.  60 E  is a perspective view of the rotor  6002  disposed in a housing  6090 . 
       FIG.  60 F  is a plan view of a rotor  6004  having two cones (e.g., two bicones) configured to elutriate cells (e.g., red blood cells, leukapheresis product) in a second cone in order to recirculate a buffer for reuse. The rotor  6004  may comprise a housing  6052 , a first fluid conduit  6012 , a first cone  6022  coupled to the first fluid conduit  6012 , a second fluid conduit  6023  coupled to the first cone  6022 , and a second cone  6024  coupled to the second conduit  6023 , and a third fluid conduit  6032  coupled to the second cone  6024 . The first cone  6022  may comprise a first volume, and the second cone  6024  may comprise a second volume larger than the first volume. In some variations, a ratio of a second volume to a first volume may be between about 2:1 to about 5:1. Fluid may flow sequentially through the first fluid conduit  6012 , the first cone  6022 , the second fluid conduit  6023 , the second cone  6024 , and the third fluid conduit  6032 . In some variations, the rotor  6004  may comprise a magnetic portion  6042 . 
     In some variations, the first cone  6022  may comprise a first bicone and the second cone  6024  may comprise a second bicone. In some variations, the first bicone may comprise a third cone including a first base and a fourth cone including a second base such that the first base faces the second base. In some variations, the second bicone may comprise a fifth cone including a third base and a sixth cone including a fourth base such that the third base faces the fourth base. 
     In some variations, a portion of the rotor  6004  may be optically transparent, such as first cone  6022 , second cone  6024 , and at least a portion of first fluid conduit  6012 , second fluid conduit  6023 , and third fluid conduit  6032 . In some variations, the first fluid conduit  6012  may comprise an inlet and the third fluid conduit  6032  may comprise an outlet. 
     In some variations, cells may enter the first cone  6022  and red blood cells (RBCs)  6030  may be elutriated into the second cone  6024 . Since the second cone  6024  is further out from an axis of rotation (center of housing  6052 ), the RBCs  6030  may be concentrated at an inlet  6025  of the second cone  6024  due to centrifugation. The larger volume of the second cone  6024  may further reduce the velocity of fluid (e.g., buffer), thereby reducing the force on RBCs  6030  within the second cone  6024 . By recirculating the fluid (e.g., buffer), a higher concentration of RBCs may be elutriated with less fluid (e.g., buffer). In some variations, white blood cells  6040  may be harvested from the first cone  6022 . An optical sensor may be configured to image the first cone  6022  to generate imaging data used to identify a boundary between the WBCs  6040  and RBCs  6030 . In some variations, the recirculating fluid may be passed through a filter to remove small particles (e.g., platelets) with less fluid (e.g., buffer). 
       FIG.  60 G  is a plan view and  FIG.  60 H  is a side view of a rotor  6005  having two cones (e.g., two bicones) configured to elutriate cells (e.g., red blood cells) in a second cone. A rotor having two cones may facilitate recirculation of buffer for reuse. The rotor  6006  may comprise a housing  6052 , a first fluid conduit  6012 , a first cone  6022  coupled to the first fluid conduit  6012 , a second cone  6024  coupled to the first cone  6022 , and a fluid conduit  6032  (e.g., outlet) coupled to the second cone  6024 . 
       FIG.  60 I  is a perspective view of a rotor  6006  comprising a cone  6024  and housing  6054 .  FIG.  60 J  is a perspective view of a rotor  6007  comprising a cone  6026  and housing  6056 .  FIG.  60 K  is a schematic plan view of rotor  6008  and corresponding dimensions.  FIG.  60 L  is an image of a set of rotors having varying dimensions. 
       FIGS.  11 A- 11 C  depict another variation of the counterflow centrifugal elutriation (CCE) module  1100 .  FIG.  11 A  is a perspective view of a cartridge  1110  comprising a CCE module  1100  in an extended configuration configured to receive a CCE instrument.  FIGS.  11 B and  11 C  are cross-sectional side views of a CCE module  1100  in respective retracted and extended configurations. In some variations, a CCE module may comprise a conical element having an internal surface and an external surface fixedly attached to a distal end of a linear member having an internal surface and an external surface. The proximal end of the linear member may be rotationally attached to a fulcrum in order to enable extension, retraction, and/or rotation of the linear member. For example,  FIG.  11 C  depicts a linear member extended outside the housing of the cartridge and then rotated to generate a centrifugal force. A cell product may be conveyed between the internal surface and external surface of the linear member (optionally in tubing) to the conical element and fed into an opening at the distal end of the internal surface of the conical element, such that the flow of the cell product may run counter to the centrifugal force generated by rotation of the linear member. Cells in the cell product may be separated based on the ratio of their hydrodynamic cross section to their mass, due to the counterflow of the solution and sedimentation of cells subject to centrifugal force. The flow rate may then be increased and/or the rotation of the linear member may be decreased to permit cells to selectively return through the void in the interior surface of the linear member to the proximal end of the linear member. The selected cells may be directed into a tube that returns the selected cells to the cartridge. After an enrichment/washing step is performed, the linear member may be retracted into the housing to the retracted configuration as shown in  FIG.  11 B . 
     Magnetic Cell Selection 
     Generally, the systems and methods described herein may select cells on the basis of magnetically labeled cells corresponding to cells having a predetermined antigen. For example, a cell suspension of interest may be immunologically labeled with magnetic particles (e.g., magnetic beads) configured to selectively bind to the surface of the cells of interest. The labeled cells may generate a large magnetic moment when the cell suspension is flowed through a flow cell. The flow cell may be disposed in proximity to a magnet array (e.g., permanent magnets, electromagnet) generating a magnetic field having a gradient across the flow cell to attract the labeled cells for separation, capture, recovery, and/or purification. The magnet array may be configured to generate non-uniform magnetic fields at the edges and the interfaces of the individual magnets so as to cover the full volume of the flow cell such that a magnetophoretic force equals a drag force exerted by the fluid flowing through the flow cell. 
       FIG.  61 A- 61 C  are schematic views of a magnetic cell separation (e.g., magnetic-activated cell selection) system and process. A magnetic cell separation system may comprise a flow cell  6110  comprising an inlet  6130  and an outlet  6132 , a magnet array  6120 , a first fluid source  6140  (e.g., input sample source), a second fluid source  6142  (e.g., buffer source), a third fluid source  6150  (e.g., target cell reservoir), a fourth fluid source  6152  (e.g., waste reservoir), and a set of valves  6134 . As shown in step  6100 , a set of cells  6160 ,  6170  may comprise labeled cells  6160  (e.g., magnetically labeled cells) and non-labeled cells  6170  may flow into the flow cell  6110 . For example, a set of the cells  6160  may be labeled with a magnetic-activated cell selection (MACS) reagent. A MACS reagent may be incubated with the set of cells to label (e.g., attach, couple) the cells to the MACS reagent. As described in more detail herein, the magnet array  6120  may be disposed external to the flow cell  6110  such that the magnet array  6120  may be moveable relative to the flow cell  6110 . For example, the magnet array  6120  may move away from the flow cell  6110  to facilitate flowing the set of cells  6160  out of the flow cell  6110 . Conventional flow cells comprise tortuous paths including meshes and/or beads to capture cells. However, recovery of labeled cells from conventional flow cell configurations is difficult, By contrast, the flow cells  6110  described herein may lack tortuous paths such as beads, meshes, and the like, and therefore enable serial separations to be performed efficiently using either positive selection or negative selection. In some variations, the flow cells may comprise generally laminar channels as described in more detail herein. 
     At step  6102 , the magnet array  6120  may magnetically attract the set of cells  6160  towards the magnet array  6120  for a predetermined dwell time and/or based on a measured quantity of magnetically separated cells. In some variations, the dwell time may be at least one minute (e.g., at least two minutes, at least three minutes, at least five minutes). The non-labeled cells  6170  are not magnetically attracted to the magnet array  6120  and may flow out of the outlet  6132  of the flow cell  6110  and into the fourth fluid source  6152 . In some variations, the fluid (e.g., cells  6160 ,  6170 ) within the flow cell may be held statically within the flow cell  6110  for a dwell time before the fluid (e.g., cells  6170 ) flow from outlet  6132 . In some variations, a longitudinal axis of the flow cell  6110  may be oriented substantially perpendicular to ground in order for fluid flow through the flow cell  6110  to be aided by gravity. At step  6104 , the magnetic coupling between the magnet array  6120  and the cells  6160  may be released after the dwell time, and the cells  6160  may flow into the third reservoir  6150 . 
     In some variations, stiction may cause cells to remain attached to a surface of a flow cell even after removal of a magnet array  6120 . Therefore, a gas may be flowed through the flow cell  6110  to aid cell collection into the third reservoir  6150 . Gas flow through the flow cell may provide improved cell recovery over liquid flushing through the flow cell. An interface generated by a gas (e.g., bubble, air gap) may be maintained by gravity, thereby enabling implementation of a relatively wide flowcell that further improves cell recovery relative to a horizontally oriented flow cell. The MACS modules described herein may be configured for positive selection and/or negative selection by modifying the sequence of steps. 
     Additionally or alternatively, an optical sensor may be configured to image a flow cell to generate imaging data used to identify a quantity of cells magnetically attracted to the magnet array. Fluid containing labeled cells may be flowed out of the flow cell when a predetermined quantity of cells have been measured by the optical sensor. 
       FIG.  62 A  is a perspective view of a MACS module  6200  in a first configuration. The MACS module  6200  (as well as any of the MACS modules described herein) may be a component of any of the cartridges described herein. For example, a cartridge for cell processing may comprise a liquid transfer bus and a plurality of modules with each module fluidically linked to the liquid transfer bus. The MACS module  6200  may comprise a flow cell  6210  comprising an elongate cavity having a cavity height, an inlet  6230 , and an outlet  6232 . The MACS module  6200  may further comprise a magnet array  6220  comprising a plurality of magnets. Each of the magnets may be spaced apart by a spacing distance, such as illustrated in  FIGS.  62 G,  63 D, and  63 E , although  FIGS.  62 A- 62 E  illustrate a magnet array  6220  with magnets in contact with adjacent magnets. 
       FIG.  62 G  is a schematic diagram of the flow cell  6210  and magnet array  6220 . In some variations, the flow cell  6210  may comprise a cavity height  6202  and a cavity width  6204 . Fluid may be configured to flow through the flow cell  6210  in a first direction  6206 . The magnet array  6220  may comprise a plurality of magnets with each magnet comprising a respective width  6222 . In some variations, adjacent magnets may be separated by a predetermined spacing distance  6224 . Each magnet pair may have the same or different spacing distance  6224 . As shown in  FIG.  62 G , an orientation (e.g., poles) of the magnets in the magnet array  6220  may comprise a predetermined pattern. 
     In some variations, a ratio of the cavity height  6202  to the spacing distance  6224  is between about 20:1 and about 1:20, between about 10:1 and about 1:10, between about 5:1 and about 1:5, and between about 3:1 and about 1:3, including all values and sub-ranges in-between. In some variations, an actuator  6240  (e.g., linear, rotary) may be configured to move the magnet array  6220  relative to the flow cell  6210 . In some variations, an orientation (e.g., poles) of the magnets in the magnet array  6220  may comprise a predetermined pattern (e.g., Halbach array). 
     In some variations, the magnet array  6220  may move relative to the flow cell  6210  or vice versa.  FIG.  62 A  illustrates the MACS module  6200  in an open configuration and  FIG.  62 B  illustrates the MACS module  6200  in a closed configuration.  FIG.  62 B  is a perspective view of the MACS system  6200  in a second configuration where labeled cells may be magnetically attracted towards the magnet array  6220 . In the second configuration, the magnetic field lines generated by the magnet array traverse the flow channel exerting a magnetophoretic force on magnetically tagged cells that are injected into the channel.  FIG.  62 C  is a cross-sectional side view of the MACS system  6200  including the magnet array  6220 .  FIG.  62 D  is a perspective view of a MACS system  6200  in the second configuration.  FIG.  62 E  is a plan view of a flow cell  6210  and magnet array  6220  of a MACS system.  FIG.  62 F  is a plan view of a flow cell  6210  of a MACS system. 
       FIG.  63 A- 63 E  are perspective views of a set of magnet arrays  6300 ,  6310 ,  6320 ,  6330 ,  6340 . One or more of the size, strength, shape, spacing, and orientation of the magnets in a magnet array may be set to generate a magnetic field to attract magnetically-labeled cells. Additionally or alternatively, a magnet array may comprise a high-magnetic permeability material configured to enhance or reduce the field strength and field gradients within the flow cell. The material may be disposed between a magnet and flowcell. Additionally or alternatively, the material may be disposed within the flowcell and/or on one or more sides of the flowcell. 
       FIGS.  64 A and  64 B  are respective perspective and cross-sectional side views of a MACS module  6400  comprising a flow cell  6410  and a magnet array  6420 . The flow cell  6410  may comprise a set of linear channels  6412 ,  6414 ,  6416  comprising a first channel  6412  parallel to a second channel  6414 , and a third channel  6416  in fluid communication with each of the first channel  6412  and the second channel  6416 . As shown in  FIG.  64 B , the third channel  6416  may be disposed between the first channel  6412  and the second channel  6416  and define a volume where fluid from the first channel  6412  and the second channel  6416  interact (e.g., mix). In some variations, the flow cell  6410  may comprise a first inlet  6430  coupled to the first channel  6412  and configured to receive a first fluid  6460  (e.g., cells). A second inlet  6431  may be coupled to the second channel  6414  and configured to receive a second fluid  6470  (e.g., buffer). The flow cell  6410  may comprise a first outlet  6432  coupled to the first channel  6412  and a second outlet  6433  coupled to the second channel  6414 . 
     The magnet array  6420  may be disposed external to the flow cell  6400  and may be moved relative to the flow cell  6400  as described herein. In some variations, a longitudinal axis of the flow cell  6410  may be perpendicular to ground such that fluid flows in a generally vertical direction. 
     In some variations, the first channel  6412  may have different dimensions form the second channel  6414 . For example, a first cavity height of the first channel  6412  may be larger than a second cavity height of the second channel  6414 . For example, a ratio of the first cavity height to a second cavity height may be between about 1:1 to about 3:7, between about 1:1 to about 2:3, and between about 2:3 to about 3:7, including all values and sub-ranges in-between. Fluid flowing through the first channel  6412  may have a slower flow rate relative to the second channel  6414  due to the larger cavity height of the first channel  6412  relative to the second channel  6414 . In some variations, the third channel  6416  may comprise a ratio of a length of the third channel  6416  to a diameter of the third channel  6416  of between about 2:1 to about 6:1, between about 2:1 to about 3:1, between about 3:1 to about 4:1, between about 4:1 to about 5:1, between about 5:1 to about 6:1, and between about 3:1 to about 5:1, including all values and sub-ranges in-between. 
     As shown in  FIG.  64 B , a first fluid  6462  may flow through the flow cell  6410  generally following a first direction. The magnetically-labeled cells  6416  within the first fluid  6462  may separate from the rest of the first fluid  6462  within the third channel  6416  as the magnetic attractive forces generated by magnet array  6420  pulls the cells  6416  away from the first channel  6412  and towards the second channel  6414  (e.g., towards the magnet array  6420 ). Similarly, a second fluid  6470  (e.g., buffer) may flow through the second channel  6414 . As the cells  6416  flow towards the magnet array  6420 , they displace the second fluid  6470  flowing through the third channel  6416  such that a portion of the second fluid  6470  may flow into the first channel  6412 . In this manner, magnetically-labeled cells  6416  may be magnetically separated from a first fluid  6462  and the second fluid  6470  may aid removal of the first fluid  642  not including the cells  6416 . 
     In some variations, a set of fluidic loops may be coupled to the flow cell to enable a plurality of cell separation cycles.  FIG.  64 C  is a schematic diagram of a MACS module comprising a flow cell  6410 , a first fluid conduit  6480  coupled to an inlet  6430  of the flow cell  6410  and an outlet  6432  of the flow cell  6410 . The first fluid conduit may  6480  may be configured to receive the set of cells from an outlet  6432  of the flow cell  6410  for recovery and/or recirculation through the inlet  6430  of the flow cell  6410 . A second fluid conduit  6490  may be coupled to the inlet  6431  of the flow cell  6410  and the outlet  6433  of the flow cell  6410  to recirculate fluid such as buffer and unrecovered magnetically-labeled cells. The second fluid conduit  6490  may be configured to receive a fluid without the set of cells from the flow cell  6410 . Higher purities of labeled cells may be recovered based on a number of cycles performed. For example, a single cell separation cycle may yield about 80% cell purity, a second cell separation cycle may yield about 96% cell purity, a third cell separation cycle may yield about 99.2% cell purity, and a fourth cell separation cycle may yield about 99.84% cell purity. 
     In some variations, applying a centrifugal force to a magnetic cell separation process may further attract labeled cells toward a magnetic array independently of fluid flow rate so as to maintain throughput.  FIGS.  65 A- 65 C  are schematic diagrams of a MACS module  6500  utilizing centrifugal force to aid a cell separation process.  FIG.  65 A  depicts a flattened flow cell  6510  configured to be wrapped to form a generally cylindrical shape  6512 . The flow cell  6510  may comprise a curved flow path  6520 . 
       FIG.  65 B  illustrates a cylindrical flow cell  6510  concentrically surrounded by (e.g., nested within) a cylindrical magnet array  6530 . In  FIG.  65 B , only a cross-section of the magnet array  6530  is shown for the sake of clarity. The flow cell  6510  may be spaced apart from the magnet array  6530  by a predetermined spacing distance. Accordingly, the flow cell  6510  may be configured to rotate  6550  about a longitudinal axis to generate a centrifugal force on the fluid  6540  within the flow path  6520  in an outward direction towards the magnet array  6530 . During a cell separation process, the fluid may be subject to set of forces depicted in  FIG.  65 C  including a bulk fluidic force  6560  in an axial (e.g., bulk flow) direction, a centrifugal force  6570  in a radially outward direction from a center of rotation (e.g., proportional to a net particle system buoyancy), and a magnetic force  6580  extending radially outward from a center of rotation (e.g., proportional to net particle system magnetic attractiveness). In some variations, labeled cells may comprise a higher density than non-labeled cells. Therefore, centrifugal force may preferentially push the labeled cells towards the magnet  6530 , further increasing the specificity and efficiency of cell separation. 
       FIGS.  66 A- 66 C  are schematic diagrams of a cell separation system and process. A magnetic cell separation system may comprise a flow cell  6610  comprising a flow path  6620  (shown schematically flattened for sake of clarity), and a magnet array  6630 . As shown in step  6600 , a set of cells  6640 ,  6642  may comprise labeled cells  6640  (e.g., magnetically labeled cells) and non-labeled cells  6642  may flow into the flow path  6620  of flow cell  6610 . For example, a set of the cells  6640  may be labeled with a magnetic-activated cell selection (MACS) reagent. The magnet array  6630  may be disposed external to the flow cell  6610  such that the magnet array  6630  may be moveable relative to the flow cell  6610 . For example, the magnet array  6630  may move away from the flow cell  6610  to facilitate flowing the set of cells  6640  out of the flow cell  6610 . 
     At step  6602 , the flow cell  6650  may be rotated to generate centrifugal force to push the cells  6640 ,  6642  toward the magnet array  6630 . In some variations, a longitudinal axis of the flow cell  6610  may be oriented substantially perpendicular to ground in order for fluid flow through the flow cell  6610  to be aided by gravity. At step  6604 , the magnet array  6630  may magnetically attract the set of cells  6640  towards the magnet array  6630  for a predetermined dwell time as described herein. The non-labeled cells  6642  are not magnetically attracted to the magnet array  6630  and may flow out of the flow cell  6610  into, for example, a waste vessel. In some variations, the fluid (e.g., cells  6160 ,  6170 ) within the flow cell may be held statically within the flow cell  6110  for a dwell time before the fluid (e.g., cells  6170 ) flow from outlet  6132 . In some variations, the magnetic coupling between the magnet array  6630  and the cells  6640  may be released after the dwell time, and the cells  6640  may be recovered. 
       FIGS.  12 A and  12 B  illustrate the magnet of the MACS instrument  1200  comprising a magnet and a MACS module  1210 . The magnet is shown in  FIG.  12 A  in an ON configuration and shown in  FIG.  12 B  in an OFF configuration. 
     Bioreactor 
     The bioreactors described herein may comprise a vessel configured to culture mammalian cells. Generally, cell and gene therapy products may be grown in a bioreactor to produce a clinical dose which may subsequently be administered to a patient. A number of biological and environmental factors may be controlled to optimize the proliferation speed and success of cell growth. The bioreactor modules described herein enable one or more of monitoring, adjusting, and/or controlling of cell growth (e.g., to facilitate consistent and efficient cellular proliferation). 
       FIG.  67 A  is a schematic diagram of a cell processing system  6700  (e.g., bioreactor module) comprising one or more of a bioreactor  6710 , one or more sensors  6720 , an agitator  6730 , a temperature regulator  6740 , and a gas regulator  6750 . In some variations, the sensor  6720  may be configured to monitor (e.g., measure, sense, determine) one or more characteristics of the bioreactor module  6700  and cells in the bioreactor  6710 . For example, the sensor  6720  may comprise one or more of a pH sensor, a dissolved oxygen (DO) sensor, a temperature sensor, a glucose sensor, a lactose sensor, a cell density sensor, a humidity sensor, combinations thereof, and the like. One or more of the sensors may be a non-invasive optical sensor. 
       FIGS.  67 B- 67 D  are schematic diagrams of a cell processing system comprising a workcell  6760 , a bioreactor system  6700  (e.g., bioreactor instrument), a cartridge  6770 , an agitator  6730 , and a fluid connector  6780 . In some variations, a cartridge  6770  for cell processing may comprise a liquid transfer bus and a plurality of modules (e.g., bioreactor module, CCE module, MACS module, EP module). Each module may be fluidically linked to the liquid transfer bus. The bioreactor module may comprise at least one bioreactor. 
     The bioreactor instrument  6700  may be configured to interface with the cartridge  6770 . In some variations, the bioreactor instrument  6700  may comprise the agitator  6730  configured to couple to the bioreactor. The agitator may be configured to agitate cell culture media comprising cells. In some variations, the fluid connector  6780  may be configured to couple the bioreactor system  6700  and workcell  6760 . 
       FIG.  67 B  depicts a cartridge  6770  comprising a bioreactor disposed within a workcell  6760 . The bioreactor  6700  may be uncoupled from the workcell  6760 . Once the fluid connector  6780  couples (e.g., to create a sterile flow path) the workcell  6760  to the bioreactor  6700 , the cartridge  6770  may be moved into the bioreactor  6700 , as shown in  FIG.  67 C . For example, the cartridge  6770  may be coupled to (e.g., disposed on) an agitator  6730  and then agitated, as shown in  FIG.  67 D . In some variations, the fluid connector  6780  may comprise a set of foldable sidewalls (e.g., like an accordion) configured to receive and dissipate the agitation of the agitator  6730  without transmitting such motion to the workcell  6760 . That is, the fluid connector  6780  may function as a bellows to maintain the connection between the workcell  6760  and bioreactor  6700  without agitating the workcell  6760 . In some variations, the fluid connector  6780  may couple the bioreactor (e.g., of cartridge  6770 ) to a liquid transfer bus. 
     In some variations, an agitator may be configured to generate motion (e.g., orbital, rotary, linear) to the bioreactor in order to mix the culture in instances where it is required to encourage interactions with a reagent and cells. For example, orbital motion may be used to create a homogenous culture volume such that a small sample taken from the culture may be representative of the culture at large. In some variations, the agitator  6730  may comprise one or more impellers. The agitator  6730  may be configured to provide variable-intensity mixing during culture at defined periods. 
     In some variations, orbital motion may encourage increased interactions within the cell culture, such as in the toroidal bioreactors described herein that comprise a geometry that may encourage the continuous and gentle flow of fluid around the bioreactor, thereby aiding homogenous mixing with minimal shear stress transferred to the cells. 
     In some variations, the temperature regulator  6740  may be configured to control a temperature of a bioreactor and corresponding processes. The temperature regulator  6740  may be coupled to the bioreactor. For example, the temperature regulator  6740  may control a temperature of a cell culture to be between about 2° C. and about 40° C. and thereby ensure that a culture is heated to physiological conditions and cooled to slow metabolic processes (e.g., to keep cells in a dormant state) as desired. For example, the thermal regulator  6740  may comprise a circulating coolant coupled to a heat exchanger coupled to a thermal interface (e.g., heating/cooling plate). 
     In some variations, the gas regulator  6750  may be coupled to the bioreactor and configured to control a gas composition of a bioreactor and corresponding processes using one or more of Clean Dry Air (CDA), carbon dioxide, and nitrogen. The gas regulator  6750  may be coupled to the bioreactor. For example, the sensors  6720  and gas regulator  6750  may provide closed-loop gas control of the bioreactor module  6700 . In some variations, CDA may comprise oxygen such as pure oxygen. In some variations, the gas regulator may comprise a manifold coupled to one or more gas sources. The manifold may include a solenoid coupled to a valve (e.g., restrictive orifice) configured to control gas flow through the bioreactor  6710 . The solenoid may be configured to pulse to control a quantity and composition of gas received through the manifold. Additionally or alternatively, one or more of a proportional valve and Mass Flow Controller (MFC) may be configured to meter and control the flow of gas to a manifold. In some variations, the gas regulator  6750  may comprise one or more sensors to measure the gas mixture and/or flow rate. Additionally or alternatively, the sensors may be configured for closed-loop control of gas flow through the gas regulator. 
     In some variations, measured pH from a pH sensor may be used to control a pH of the bioreactor  6710  using the gas regulator  6750 . For example, in response to the measured pH, gas regulator  6750  may control a CO 2  concentration of the gas contacting the cell culture to control the free hydrogen ions and pH of the culture. In some variations, a pH of the bioreactor  6710  may be between about 5.5 and about 8.5. One or more of CO 2  composition of the gas in the bioreactor  6710 , buffer, and reagents (e.g., acid, base) may be used to regulate pH. In some variations, a dissolved oxygen concentration of the bioreactor  6710  may be between about 0% and about 21%. Nitrogen composition of the gas in the bioreactor  6710  may be used to regulate the dissolved oxygen concentration. For example, control of both the agitator in the bioreactor and the flow rate and composition of the gas contacting the cell culture may regulate the dissolved carbon dioxide concentration. 
     In some variations, measured dissolved oxygen from a dissolved oxygen sensor may be used to control an oxygen concentration (e.g., below atmospheric levels) of the bioreactor  6710  using the gas regulator  6750 . For example, gas regulator  6750  may control a nitrogen concentration of the gas contacting the cell culture to create hypoxic conditions. 
       FIGS.  68 A and  68 B  are cross-sectional perspective views of a bioreactor  6800  comprising an enclosure  6810  comprising a base  6812 , a sidewall  6814 , and a top  6816 . A gas-permeable membrane  6820  may be coupled to one or more of the base  6812  and the sidewall  6814  of the enclosure  6810 . In some variations, the enclosure  6810  may comprise a first chamber  6830  having a first volume and a second chamber  6832  having a second volume, the first chamber  6830  separated from the second chamber  6832 , and the first volume smaller than the second volume. In some variations, the first chamber  6830  may be concentrically nested within the second chamber  6832 . For example, nesting the chambers may enable larger overall working volume ranges (e.g.,  100 : 1 ). The first chamber  6830  may comprise a well shape with an angled base surface to promote fluid pooling at a center of the first chamber  6830  during aspiration. In some variations, the base  6812  may be disposed on a thermal regulator (not shown) such as a thermoelectric element. In some variations, the enclosure  6810  may be composed of a thermally conductive material such as a metal (e.g., aluminum). 
     In some variations, the bioreactor  6800  may be coupled to a gas regulator (not shown) to facilitate gas transfer through the gas-permeable membrane  6820  (e.g., into and out of the culture). The gas-permeable membrane  6820  may be configured to hold a cell culture. Gas may diffuse through the surfaces of the culture that contact the gas-permeable membrane to enable increased oxygenation of the cell culture and removal of gaseous metabolic byproducts of the cell culture, and thus increase the potential for metabolic activity. For example, the gas-permeable membrane  6820  enables dissolved oxygen to diffuse into the culture in close proximity to a cell bed where the oxygen may be consumed. In some variations, the bioreactor may be coupled to both a first gas regulator to facilitate gas transfer through the gas-permeable membrane and a second gas regulator to facilitate control of headspace gas composition. 
     In addition to gas transfer, the bioreactors described herein may be configured to efficiently control a temperature of a cell culture using a conductive thermal interface (e.g., gas-permeable membrane  6820 , enclosure  6810 ) along both a base and sidewall of the bioreactor. 
     In some variations, the first chamber  6830  may comprise a working volume of between about 10 ml and about 100 ml. In some variations, the first chamber  6830  may comprise a total volume of between about 10 ml and about 130 ml. In some variations, the second chamber  6832  may comprise a working volume of between about 100 ml and about 1000 ml. In some variations, the second chamber  6832  may comprise a total volume of between about 100 ml and about 1400 ml. In some variations, the first chamber  6830  may comprise a diameter of between about 10 mm and about 100 mm, and a height of between about 10 mm and about 100 mm. In some variations, the second chamber  6832  may comprise a diameter of between about 100 mm and about 250 mm, and a height of between about 10 mm and about 100 mm. 
     As shown in  FIG.  68 B , a base  6822  of the gas-permeable membrane  6820  may comprise an angle between about 3 degrees and about 10 degrees relative to the base  6812  of the enclosure  6810 . Similarly,  FIGS.  69 A and  69 B  depict a sloped base. For example, due to a slope of the base  6822 , the chambers  6830 ,  6832  are deeper towards a center of the bioreactor  6800 . This may encourage cell growth towards a center of the bioreactor  6800 , which may aid one or more of cell sampling, cell transfer, cell recovery, and the like. In some variations, orbital motion of the bioreactor  6800  may promote cell congregation toward a center of the bioreactor  6800 , thereby increasing interaction between the cells. 
     In some variations, the gas-permeable membrane  680  may comprise a curved surface. In some variations, the gas-permeable membrane may comprise a set of patterned curved surfaces. For example, the set of patterned curved surfaces may comprise a radius of curvature of between about 50 mm and about 500 mm. 
     In some variations, the bioreactor may be configured to facilitate monitoring (e.g., temperature, pH, dissolved oxygen) and fluid flow (e.g., gas composition, fluid transfer) between the chambers. As shown in  FIG.  68 C , the enclosure  6810  may comprise one or more nested surfaces curved around a longitudinal axis (e.g., center) of the enclosure  6810 . For example, the nested surfaces may comprise a set of concentric toroids. The enclosure  6810  may comprise a toroid shape.  FIG.  68 C  is a perspective view and  FIG.  68 D  is a bottom view of enclosure  6810  comprising a set of apertures  6818  (e.g., holes, openings, slits, slots). In some variations, the apertures  6818  may enable gas and/or heat transfer between the components and chambers of the bioreactor  6800 . Additionally or alternatively, one or more sensors may be coupled to the apertures  6818 . For example, the apertures  6818  may be coupled to a non-contact sensor (e.g., pH, DO) such as an optical sensor (not shown) configured to determine a fluorescent spot disposed on a surface of the bioreactor. In some variations, one or more of a sensor and fluid connector may be introduced through the apertures  6818 . 
     In some variations, the gas-permeable membrane extends along the base  6812  and the sidewall  6814  of the enclosure  6810 , as shown in  FIG.  68 B . In some variations, the gas-permeable membrane extends only along the base  6812  of the enclosure  6810 .  FIG.  68 E  is a perspective view and  FIG.  68 F  is a side view of the gas-permeable membrane  6820  where an outer surface of the gas-permeable membrane  6820  comprises one or more projections  6824  (e.g., projections, spacers, ribs). The projections  6824  are also depicted in the perspective view of  FIG.  68 G  and bottom view of  FIG.  68 H . The projections  6824  contact the enclosure  6810  and define a cavity between the enclosure  6810  and the gas-permeable membrane  6820 . That is, the projections  6824  may be configured to mechanically space away the enclosure  6810  from a portion of the gas-permeable membrane  6820  to facilitate thermal transfer from the enclosure  6810  to the cell culture. In some variations the gas-permeable membrane may comprise polydimethylsiloxane (PDMS) (e.g., silicone), fluorinated ethylene propylene (FEP), polyolefin (PO), polystyrene (PS), ethyl vinyl acetate (EVA) and have a thickness of between about 0.1 mm and about 0.4 mm, between about 0.2 mm and about 0.3 mm, and about 0.25 mm, including all ranges and sub-values in-between. 
       FIG.  69 A  is a cross-sectional side view of an enclosure  6910  of a bioreactor comprising a first chamber  6912 , a second chamber  6914 , and a column  6916  extending along a longitudinal axis of the enclosure  6910 .  FIG.  69 B  is a cross-sectional perspective view of the enclosure  6910  showing the nested curves of the enclosure  6910 . The column  6916  may be configured to promote cell culture in combination with agitation such as orbital motion. 
       FIG.  70    is an exploded perspective view of a bioreactor  7000  comprising an enclosure  7010 , a gas-permeable membrane  7020 , and a top  7030 . The top  7030  may be composed of a material such as polyethylene. 
       FIG.  71 A  is a plan view of a bioreactor  7100  comprising a first chamber  7110  and a second chamber  7120 .  FIG.  71 B  is a cross-sectional side view of the bioreactor  7100 . 
       FIGS.  13 A and  13 B  are perspective views of a cartridge  1300  and bioreactor instrument interface  1310 . The bioreactor instrument interface  1310  is coupled to the cartridge  1300  in  FIG.  13 B . 
       FIG.  14    is a perspective view of a bioreactor instrument  1410  comprising a set of cartridges  1400 ,  1402 ,  1404  and cavities  1420 ,  1422 ,  1424  configured to receive a respective cartridge. In some variations, each cartridge may be docked to enable simultaneous expansion, culturing, or resting steps. 
     Electroporation Module 
     In some variations, an electroporation module may be configured to facilitate intracellular delivery of macromolecules (i.e., transfection by electroporation). An electroporation module may contain a continuous flow or batch mode chamber and one or more sets of electrodes for applying direct or alternating current to the chamber. An electrical discharge from one or more capacitors, or current sources, may generate sufficient current in the chamber to promote transfer of a polynucleotide, protein, nucleoprotein complex, or other macromolecule into the cells in the cell product. As with other modules described herein, one or more components used for the process step (here, electroporation) may be provided on the cartridge or in the instrument to which the cartridge interfaces. For example, the capacitor(s) and/or batteries may be provided in the module on the cartridge or in the instrument. The electroporation module may, in some variations, be configured to apply an electric field to a cell suspension under continuous flow in a microfluidic device, e.g., as described in Garcia et al. Sci. Rep. 6:21238 (2016). 
     Additionally or alternatively, intracellular delivery of macromolecules may also be achieved by other methods, such as mechanoporation. It should be understood that throughout the disclosure variations comprising an electroporation module may instead or in addition comprise a mechanoporation module, or another module configured to perform any suitable method of delivering macromolecules into cells. Mechanoporation can be achieved by, for example, applying transient, fluidic pressure to a solution containing cells, or by applying physical pressure to the cells (e.g., by microneedles). Illustrative methods of mechanoporation by passing a cell suspension through a constriction are provided, e.g., in International Patent Publication No. WO 2017/041051 and WO 2017/123663, and are incorporated by reference herein. Mechanoporation can also be achieved by applying a vortex to a cell suspension in a microfluidic device. 
       FIG.  72    is a schematic diagram of an electroporation module  7200  (e.g., electroporation system) comprising an electroporation chamber  7210  (which may comprise a fluid conduit), a pump  7220 , an inlet  7230 , an outlet  7232 , a set of pinch valves  7234 , a first fluid source  7240  (e.g., fluid reservoir, cell reservoir), a second fluid source  7242  (e.g., vent, gas source), a set of sensors  7250  (e.g., bubble sensors), and a controller (e.g., processor and memory) configured to control the module  7200 , and a signal generator  7270  configured to deliver an electroporation signal (e.g., voltage pulse) to the electroporation chamber  7210 . 
     In some variations, the fluid conduit  7210  may be configured to receive a first fluid comprising cells and a second fluid. A set of electrodes may be coupled to the fluid conduit  7210 . A pump may be coupled to the fluid conduit  7210 . The controller  7260  may be configured to generate a first signal to introduce the first fluid into the fluid conduit  7210  using the pump  7220 , generate a second signal to introduce the second fluid into the fluid conduit  7210  such that the second fluid separates the first fluid from a third fluid, and generate an electroporation signal to electroporate the cells in the fluid conduit  7210  using the set of electrodes. 
     In some variations, the second fluid may comprise a gas or oil. In some variations, the controller may be configured to generate a third signal to introduce the third fluid into the fluid conduit  7210 . The third fluid may be separated from the first fluid by the second fluid. In some variations, a cartridge for cell processing may comprise a liquid transfer bus and a plurality of modules such as the electroporation module  7200 . Each module may be fluidically linked to the liquid transfer bus. 
     The set of sensors  7250  may be configured to measure fluid changes in a fluid conduit such as a change from a first fluid to a second fluid (e.g., liquid to air) in the fluid conduit. The module  7200  may further comprise a set of valves configured to ensure fluid does not backflow into the electroporation chamber  7210  and/or fluid source  7240 . The electroporation chamber  7210  may comprise a cavity configured to hold a fluid to be electroporated and a set of electrodes to apply an electroporation signal to the fluid. For example, the signal generator  7270  may generate a square valve pulse as described in more detail herein. 
     In some variations, the electroporation module  7200  (e.g., valves  7234 , pump  7220 , sensors  7250 , and controller  7260 ) may be configured to control fluid flow through the electroporation chamber  7210  in a discontinuous (e.g., batch process) manner. For example, a first batch of cells may undergo electroporation and be physically separated from a second batch of cells by an intermediate fluid such as air or fluid such as oil. Separating cell batches may reduce mixing of transfected and non-transfected cells, and further ensure fixed batch volume. That is, a fluid gap may form a visually verifiable boundary between cell batches to reduce diffusion and mixing between electroporated and non-electroporated cells. Separating cell batches may reduce the duration of time that cells are exposed to certain cytotoxic reagents (e.g., electroporation buffer), thereby increasing performance. 
     In some variations, a batch of cells may be electroporated when substantially static (e.g., substantially no fluid flow state). By contrast, conventional continuous flow electroporation has an upper fluid flow rate limit correlated to a transfection efficiency. In the batch processing described herein, cell batches may be transferred into and out of the electroporation chamber  7210  at a predetermined rate to increase the overall throughput of the system  7200  without a decrease in electroporation efficiency. Furthermore, the electroporation system  7200  does not utilize a precisely controlled flow rate/pulse rate such as those needed for continuous flow electroporation systems. 
       FIG.  73    is an exploded perspective view of an electroporation module  7300  may comprise ah electrode  7310 , a fluid conduit  7320  (e.g., electroporation chamber), a substrate  7330  (e.g. alloy busbar), a housing  7340 , and a fastener  7350 . In some variations, the fluid conduit  7320  may be configured to hold a volume of fluid between about 0.4 ml and about 3.5 ml. The electroporation module  7300  is a parallel-plate design. In some variations, the electrodes may comprise stainless steel and may be separated by an insulating gasket. In some variations, the electrodes may be polished and/or coated with nonreactive materials (e.g., gold, platinum) to reduce gradual buildup of biological matter (e.g., charged molecules, DNA, proteins) on the electrode surface. 
     Generally, a method of electroporating cells may comprise receiving a first fluid comprising cells in a fluid conduit, receiving a second fluid in the fluid conduit to separate the first fluid from a third fluid, applying an electroporation signal to the first fluid to electroporate the cells. In some variations, the third fluid may be received in the fluid conduit separated from the first fluid by the second fluid. In some variations, the first fluid may be substantially static when applying the electroporation signal. 
       FIGS.  74 A- 74 B  are schematic diagrams of variations of an electroporation process  7400 ,  7402 . A method  7400  may include loading cells  7410  into an electroporation chamber  7450 . For example, at step  7412 , a first fluid may be pumped into the electroporation by opening valve v 1  and the pump generating negative pressure (valves v 2  and v 3  are closed). At step  7414 , a second fluid (e.g., gas, oil) may separate the first fluid from a third fluid to create a first batch of cells to electroporate. For example, valves v 1  and v 3  may be closed with valve v 2  open and the pump generating negative pressure. In some variations, a loading volume may be between about 1 ml and about 3 ml with a pumping time of between about 8 seconds and about 15 seconds (at a rate of about 20 ml/min). At step  7420 , the cells of the first fluid may be electroporated with each of the valves closed and the pump off. At step  7430 , the cells of the first fluid may be flowed out of the electroporation chamber  7450  to output where valves v 1  and v 2  are closed, valve v 3  is open, and the pump generates positive pressure. 
       FIG.  74 B  depicts another configuration where a pump is disposed between an input and the electroporation chamber such that the pump may be configured to pump in a single direction. A method  7402  may include loading cells  7411  into an electroporation chamber  7450 . For example, at step  7416 , a first fluid may be pumped into the electroporation by opening valve v 1  and v 4 , and the pump generating positive pressure (valves v 2  and v 3  are closed). At step  7418 , a second fluid (e.g., gas, oil) may separate the first fluid from a third fluid to create a first batch of cells to electroporate. For example, valves v 1  and v 3  may be closed with valves v 2  and v 4  open, and the pump generating positive pressure. At step  7422 , the cells of the first fluid may be electroporated with each of the valves closed and the pump off. At step  7432 , the cells of the first fluid may be flowed out of the electroporation chamber  7450  to output where valves v 1  and v 4  are closed, valves v 2  and v 3  are open, and the pump generates positive pressure. 
     In some variations, an impedance/resistance across electrodes of an electroporation system may increase over time due to electrode passivation/degradation due to charged biological matter (e.g., charged molecules, DNA, proteins) attaching to the electrode surface. Active electrical field compensation may be applied to ensure a consistent electrical field strength applied to cells over multiple batches of cells. This may reduce the need for electrode surface modification to reduce passivation. 
       FIG.  75    is a circuit diagram of a resistor divider network for an electroporation process  7500 . For example, a set of cells may be introduced into an electroporation chamber  7510  to which a voltage V chip  may be applied. Fluid resistance R b  corresponds to a fluid (e.g., cell mixture) resistance. Assuming a uniform cell distribution, the fluid resistance R b  should be consistent, also assuming the same volume of each fluid batch being electroporated. R i  corresponds to a resistance between fluid and electrode, which increases over time through the electroporation process. In a conventional electroporation process, voltage V ps  is constant. However, due to the increasing R i  over time, the voltage applied to the fluid will decrease over time, leading to lower electrical field strength. 
     Due to variations in fluid resistance R b  and the low number of pulses that may be applied, interpolation to compensate for reduced electrical field strength may not accurately compensate for electrode passivation. 
     In some variations, a method of electroporating cells may comprise receiving a first fluid comprising cells in a fluid conduit, applying a resistance measurement signal to the first fluid using a set of electrodes, measuring a resistance between the first fluid and the set of electrodes, and applying an electroporation signal to the first fluid based on the measured resistance. In some variations, a second fluid comprising a gas may be received in the fluid conduit before applying the electroporation signal to the fluid. The first fluid may be separated from a third fluid by the second fluid. 
       FIGS.  76 A- 76 D  are plots  7600 ,  7602 ,  7604 ,  7606  of measurement waveforms and electroporation waveforms.  FIG.  76 A  depicts a first resistance measurement pulse  7620  with a low voltage and a wide pulse width.  FIG.  76 B  depicts a second resistance measurement pulse  7622  with a high voltage and a short pulse width.  FIG.  76 C  depicts a third resistance measurement pulse  7624  with a continuous low voltage waveform to monitor an impedance change continuously over time.  FIG.  76 D  depicts a fourth resistance measurement pulse  7626  with a low AC voltage waveform to monitor an impedance change continuously over time. Each of the resistance measurement pulses avoid inducing electroporation in the cells by reducing voltage and/or pulse width. By monitoring the voltage current of the applied resistance measurement pulse, a change in resistance may be measured and the electroporation pulse applied to a cell batch may be compensated accordingly. 
     In some variations, an electroporation signal may comprise between about 1 pulse and about 50 pulses, a voltage of between about 100 V and about 700 V, a pulse width of between about 100 μs and about 1 ms, a pulse spacing between about 5 second to about 30 seconds, a resistance pulse voltage of between about 10 V and about 40 V, and a resistance pulse width of between about 10 μs and about 50 μs. 
     For example, an eight-batch electroporation run may receive one electroporation pulse per batch. Each electroporation pulse may have an electrical field strength between about 0.5 kV/cm and about 2.0 kV/cm. The resistance measurement pulse applied before each batch may have an electrical field strength less than about 0.2 kV/cm such that electroporation is not induced by the resistance measurement pulse. 
     Sterile Liquid Transfer Device 
     Generally, the sterile liquid transfer devices described herein may be configured to store fluid for transfer to another component of a cell processing system such as a cartridge, bioreactor, and the like. In some variations, the sterile liquid transfer device may comprise a portable consumable configured to be moved using a robot. For example, a robot may be configured to move a sterile liquid transfer device from a reagent vault to an ISO 7 space to a sterile liquid transfer instrument within a cell processing system. The sterile liquid transfer device enables the transfer of fluids in an automated, sterile, and metered manner for automating cell therapy manufacturing. 
       FIGS.  103 A and  103 B  are perspective views of a sterile liquid transfer device  10300  comprising a fluid cavity  10310  (e.g., container, vessel), fluid connector  10320  (e.g., fluid connector), and pump  10330 . Fluid stored within fluid cavity  10310  may be transferred in and out of the sterile liquid transfer device  10300  through the fluid connector  10320  using the pump  10330 . In some variations, the sterile liquid transfer device  10300  may comprise an engagement feature  10340  (e.g., robot mount) to facilitate robotic arm control. 
     Fluid Connector 
     Generally, the aseptic fluid connectors described herein may form a sterile fluid pathway between at least two fluid devices to enable fluid transfer that may be one or more of sterile, fully automated, and precisely metered (e.g., precise control of a transferred fluid volume). In some variations, the robot may be configured to couple a fluid connector between at least two of the plurality of instruments and one or more cartridge. In some variations, the robot may be configured to operate the fluid controller to open and close a set of ports and valves of the fluid connector. The use of a robot and controller to operate the fluid connector may facilitate automation and sterility of a cell processing system. 
     In some variations, a system may comprise a robot configured to operate a fluid connector as described herein, and a controller comprising a memory and processor. The controller may be coupled to the robot. The controller may be configured to generate a port signal to couple the first port to the second port using the robotic arm, generate a first valve signal to translate the first valve relative to the second valve using the robotic arm, and generate a second valve signal to transition the first valve and the second valve to the open configuration. 
     In some variations, a fluid pump may be coupled to the sterilant source, and the controller may be configured to generate a first fluid signal to circulate a fluid into the chamber through the sterilant port. The controller may be configured to generate a second fluid signal to circulate the sterilant into the chamber through the sterilant port to sterilize at least the chamber. The controller may be configured to generate a third fluid signal to remove the sterilant from the chamber. 
     In some variations, the controller may be configured to generate a port signal to couple the first port to the second port using the robotic arm, generate a first valve signal to translate the first valve relative to the second valve using the robotic arm, and generate a second valve signal to transition the first valve and the second valve to the open configuration. 
     The fluid connector may further allow for a plurality of connection cycles in a sterile system and may be controlled without human intervention. For example, the fluid connector may comprise one or more of engagement features to facilitate robotic arm control and alignment features to ensure proper connection between connector components.  FIG.  15    is a block diagram of an illustrative variation of a fluid connector system  1500  comprising a fluid connector  1510 , first fluid device  1520 , second fluid device  1522 , sterilant source  1530 , fluid source  1532 , robot (e.g., robotic arm)  1540 , and controller  1550 . The fluid connector  1510  may be removably coupled (e.g., connected/disconnected, attached/detached) to each of the first fluid device  1520 , second fluid device  1522 , sterilant source  1532 , fluid source  1532 , and robot  1540 . In some variations, a fluid device may comprise one or more of a cartridge and sterile liquid transfer device. For example, a sterile liquid transfer device may be in fluid communication with a cartridge via the fluid connector. As described in more detail herein, separate portions (e.g., male connector, female connector) of the fluid connector  1510  may be removably coupled to each other. The robot  1540  may be configured to physically manipulate (e.g., removably couple) one or more of the fluid connector  1510 , first fluid device  1520 , second fluid device  1522 , sterilant source  1530 , and fluid source  1532  in a predetermined manner. For example, the robot  1540  may connect the fluid connector  1510  between the first fluid device  1520  and the second fluid device  1522 . The robot  1540  may also connect the sterilant source  1530  and/or fluid source  1532  to a sterilant port of the fluid connector  1510 . In some variations, the robot  1540  may control one or more valves and/or ports of the fluid connector  1510 , and thereby initiate a sterilization process for one or more portions of the fluid connector  1510  using, for example, sterilant from the sterilant source  1530 . The controller  1550  may be coupled to one or more of the robot  1540 , sterilant source  1530 , and fluid source  1532  to control one or more of fluid transfer and sterilization. 
       FIG.  16 A  is a schematic diagram of an illustrative variation of a fluid connector  1600 . The fluid connector  1600  may comprise a lumen extending along its length and be disposed between a first fluid device  1630  and a second fluid device  1640  to enable fluid flow through the fluid connector  1600 . In some variations, the first fluid device  1630  and second fluid device  1640  may be aseptically connected and disconnected using the fluid connector  1600 . The fluid devices  1630 ,  1640  may comprise a closed sterile device, and may be the same or different types of fluid devices. For example, the fluid devices  1630 ,  1640  may comprise one or more of a sterile liquid transfer device and consumable. In some variations, the fluid connector  1600  may comprise a first connector  1610  including a first proximal end  1612  and a first distal end  1614 . The first proximal end  1612  may be configured to couple to the first fluid device  1630 . The first distal end  1614  may include a first port  1616 , first housing  1617 , and a first valve  1618 . The first housing  1617  may be configured to receive the first port  1616  in a closed configuration as described in more detail herein. 
     The fluid connector  1600  may further comprise a second connector  1620  including a second proximal end  1622  and a second distal end  1624 . The second proximal end  1622  may be configured to couple to the second fluid device  1640 . The second distal end  1624  may include a second port  1626 , second housing  1627 , and a second valve  1628 . The second housing  1627  may be configured to receive the second port  1626  in a closed configuration. In  FIG.  16 A , the first connector  1610  comprises a sterilant port  1650  configured to couple to a sterilant source (not shown). Additionally or alternatively, the second connector  1620  may comprise the sterilant port  1650 . The sterilant port  1650  may be configured to be in fluid communication with the first distal end  1614  and the second distal end  1624  when the second port  1626  is coupled to the first port  1616  as described in more detail herein. 
     In some variations, a fluid device  1630 ,  1640  may comprise a sterilant chamber and a sterilant port configured to receive a sterilant. The sterilant chamber may enclose a fluid device connector (not shown) configured to couple to a proximal end of a first connector  1610  or second connector  1620 . The fluid device  1630 ,  1640  may receive a sterilant in a similar manner as the fluid connector  1600 . 
       FIG.  16 B  is a detailed schematic diagram of the first connector  1610  including a first port housing  1617  and a chamber  1615 . The chamber  1615  may be defined by the cavity enclosed by one or more of the distal ends  1614 ,  1624 . For example, the chamber  1615  in  FIG.  16 B  may comprise the portion of the first connector  1610  between the first valve  1618  and the first port  1616  in the closed configuration (e.g., the first distal end  1614 ). In some variations, the first chamber  1615  may comprise a volume of between about 1 cm 3  and about 5 cm 3 . When the first connector  1610  is coupled to the second connector  1620  and the ports  1616 ,  1626  are in an open configuration (as shown in  FIG.  16 D ), the chamber  1616  may comprise the portion of the fluid connector  1600  between the first valve  1618  and the second valve  1628  (e.g., the first distal end  1614  and second distal end  1624 ). The chamber  1615  may comprise an enclosed volume configured to receive a fluid such as a sterilant from the sterilant port  1650 . In some variations, the sterilant port  1650  may comprise an inlet  1652  and outlet  1654 . Methods of using a fluid connector are described in more detail with respect to  FIGS.  16 C- 16 L and  27   . 
     In some variations, the fluid connector  1600  may comprise one or more alignment features and robot engagement features configured to facilitate robotic manipulation, as described in more detail herein. In some variations, the fluid connector  1600  may be coupled to one or more sensors, pumps, and valves to facilitate fluid transfer and monitoring. 
     In some variations, the components of the fluid connector in contact with fluid may be USP Class VI compatible for cell processing and/or GMP applications. In some variations, the components of the fluid connector may be composed of a material including, but not limited to, one or more of cyclic olefin copolymer (COC), polychlorotrifluoroethylene, polyetherimide, polysulfone, polystyrene, polycarbonate, polypropylene, silicone, polyetheretherketone, polymethylmethacrylate, nylon, acrylic, polyvinylchloride, vinyl, phenolic resin, petroleum-derived polymers, glass, polyethylene, terephthalate, metal, stainless steel, titanium, aluminum, cobalt-chromium, chrome, silicates, glass, alloys, ceramics, carbohydrate polymer, mineraloid matter, and combinations or composites thereof. 
       FIGS.  17 A- 18 D  depict external and internal views of variations of a fluid connector.  FIG.  17 A  is a front perspective view of a fluid connector  1700  in a closed port configuration.  FIG.  17 B  is a rear perspective view and  FIG.  17 C  is a rear view of the fluid connector  1700 . Generally, the fluid connector may comprise a plurality of internal seals to reduce contamination and aid sterilization, as well as alignment features to aid proper registration of the fluid connector components. 
     The fluid connector  1700  may comprise a lumen extending along its length. In some variations, the fluid connector  1700  may comprise a first connector  1710  including a first proximal end  1712  and a first distal end  1714 . The first proximal end  1712  may be configured to couple to a first fluid device (not shown for the sake of clarity). The first proximal end  1712  may comprise a Luer connector or any other suitable connector. The first distal end  1714  may include a first port  1716  and first housing  1717 . The first housing  1717  is shown in  FIG.  17 A  holding the first port  1716  in a closed configuration. The first connector  1710  further comprises a sterilant port  1750 ,  1752  configured to couple to a sterilant source (not shown for the sake of clarity). In some variations, the sterilant port may comprise an inlet and outlet. In some variations, the sterilant port may optionally comprise one or more of a check valve and particle filter configured to reduce contamination into the sterilant port when not connected to a robot or actuator. The first connector  1710  may comprise a first alignment feature  1760  such as a set of protrusions on the first distal end  1714  of the first connector  1710 . The alignment features may ensure that small positioning errors due to robotic manipulation do not impact the operation of the fluid connector. 
     The fluid connector  1700  may further comprise a second connector  1720  including a second proximal end  1722  and a second distal end  1724 . The second proximal end  1722  may be configured to couple to the second fluid device (not shown for the sake of clarity). The second proximal end  1722  may comprise a Luer connector or any other suitable connector. The second distal end  1724  may include a second port  1726  and second housing  1727 . The second housing  1727  is shown in  FIG.  17 A  holding the second port  1726  in the closed configuration. The second connector  1720  may comprise a second alignment feature  1762  such as a set of holes on the second distal end  1724  of the second connector  1720 . The second alignment feature  1762  may be configured to couple to the first alignment feature  1760  in a predetermined axial and rotational configuration to aid mating of the first connector  1710  and the second connector  1720 . 
     The first port  1716  and the second port  1726  retained within respective first housing  1717  of the first distal end  1714  and second housing  1727  of the second distal end  1724  facilitates robotic control as the ports  1716 ,  1726  are not separable from the fluid connector  1700 , and therefore reduces the risk of failure of automated handling by a robot. 
     In some variations, the first connector  1710  may comprise a first robot engagement feature  1770  and the second connector  1720  may comprise a second robot engagement feature  1772 . The robot engagement features  1770 ,  1772  may be configured to be manipulated by a robot (e.g., robot  1540 ) such a robotic arm. In some variations, the robot engagement features  1770 ,  1772  may be operatively coupled to a respective first port  1716  and second port  1726  and configured to actuate the ports  1716 ,  1726  between a closed port configuration and an open port configuration, as shown in  FIGS.  17 A- 17 F . Additionally or alternatively, a user may manually actuate the robot engagement features  1770 ,  1772  to actuate respective ports  1716 ,  1726 . 
       FIG.  17 D  is a front perspective view of the fluid connector  1700  in an open port configuration.  FIG.  17 E  is a rear perspective view and  FIG.  17 F  is a rear view of the fluid connector  1700  in the open port configuration. In the open port configuration, the first valve  1718  of the first connector  1710  and the second valve  1728  of the second connector  1720  are shown in  FIG.  17 D . 
       FIG.  18 A  is a side view and  FIG.  18 B  is a cross-sectional side view of a fluid connector  1800  in an uncoupled configuration. In some variations, the fluid connector  1800  may comprise a first connector  1810  including a first housing  1817  comprising a first port  1816 , a sterilant port  1850  configured to couple to a sterilant source (not shown), a first alignment feature  1860  configured to couple to a corresponding alignment feature (not shown) of the second connector  1820 . The fluid connector  1800  may comprise a second connector  1820  including a second housing  1827  comprising a second port  1826 . The first connector  1810  and second connector  1820  may be axially aligned and alignment features may aid rotational alignment of the first connector  1810  to the second connector  1820 . The first valve  1818  may comprise a first valve stem  1819  and the second valve  1828  may comprise a second valve stem  1829 . 
       FIG.  18 C  is a side view and  FIG.  18 D  is a cross-sectional side view of the fluid connector  1800  in a coupled configuration where the first housing  1817  and the second housing  527  are brought together but where the first connector  1810  and the second connector  1820  are not in fluid communication since the first port  1816  and the second port  1826  are both in the closed configuration. The first alignment features on each connector  1810 ,  1820  may be configured to ensure axial and/or rotational alignment between the first connector  1810  and the second connector  1820 . 
       FIG.  18 E  is a side view and  FIG.  18 F  is a cross-sectional side view of the fluid connector  1800  in an open port configuration. Each of the first port  1817  and the second port  1827  are transitioned from the closed configuration to an open configuration. This creates a closed internal volume within respective distal ends of each connector  1810 ,  1820 . Each of first valve  1818  and second valve  1828  is in a closed configuration such that fluid flow is inhibited between the first connector  1810  and the second connector  1820 . still restricted on each half on account of the auto-shutoff valves in both sides. 
       FIG.  18 G  is a side view and  FIG.  18 H  is a cross-sectional side view of the fluid connector  1800  in an open valve configuration where the first valve  1818  is coupled to the second valve  1828 . For example, the second valve  1828  may be translated along a longitudinal axis of the second connector  1820  towards the first valve  1818 . As shown in  FIGS.  18 G and  18 H , the second connector  1820  may be axially compressed to translate the second valve  1828  towards the first valve  1818 . The first valve  1818  coupled to the second valve  1828  may form a radial seal, and the first valve stem  1819  and the second valve stem  1829  may be in contact to enable fluid communication between the first connector  1810  and the second connector  1820 . 
       FIGS.  19 - 26 B  are schematic diagrams of variations of fluid connector systems for coupling fluid devices. In some variations, a fluid connector may comprise a first connector configured to couple to any one of a plurality of second connectors.  FIG.  19    is a schematic diagram of an illustrative variation of a fluid connector system  1900  comprising a first connector  1910 , a plurality of second connectors  1920 ,  1921 ,  1922 , a first fluid device  1930  (e.g., sterile liquid transfer device), a second fluid device  1940  (e.g., consumable), and a robot  1960  (e.g., robotic arm, 3 DOF robot). The first connector  1910  may be coupled in fluid communication with the first fluid device  1930 , and the second connectors  1920 ,  1921 ,  1922  may be coupled in fluid communication with the second fluid device  1940 . The first connector  1910  and the second connectors  1920 ,  1921 ,  1922  may each comprise a port  1916  configured to couple to a corresponding port as described in more detail herein. The robot  1960  may comprise one or more end effectors  1962 ,  1964  configured to manipulate and/or couple to one or more of the first fluid device  1930  and first connector  1910 . For example, the first connector  1910  may comprise one or more sterilization ports  1950  configured to couple to an end effector  1962  (e.g., gripper). Similarly, the first fluid device  1930  may comprise one or more fluid ports  1952  configured to couple to an end effector  1964 . 
     In some variations, the robot  1960  may be configured to couple to one or more of a sterilant source, fluid source, and pump in order to facilitate efficient and shared fluidic connections between the fluid device, fluid connector, and a sterilization system. For example,  FIG.  96 A  is a plan view of a fluid device  9600  (e.g., sterile liquid transfer device) comprising a fluid port  9610  configured to couple to a fluid source (not shown) and a sterilization port  9620  configured to couple to a sterilant source (not shown). The  FIGS.  96 B and  96 C  are respective side and perspective views of a fluid device  9600  coupled to a robot  9650 . In some variations, the robot  9650  may comprise one or more fluid conduits  9660  configured to couple to one or more of the fluid port  9610  and sterilization port  9620  of the fluid device  9600 . 
     In some variations, a fluid connector may comprise a third connector disposed between a first connector and a second connector.  FIG.  20 A  is a schematic diagram of an illustrative variation of a fluid connector system  2000  comprising a first connector  2010 , a plurality of second connectors  2020 ,  2021 ,  2022 , a third connector  2070  (e.g., instrument, sterilization enclosure), a first fluid device  2030  (e.g., sterile liquid transfer device), a second fluid device  2040  (e.g., consumable), and a robot  2060  (e.g., robotic arm, 3 DOF robot, 1 DOF robot). The first connector  2010  may be coupled in fluid communication with the first fluid device  2030 , and the second connectors  2020 ,  2021 ,  2022  may be coupled in fluid communication with the second fluid device  2040 . The third connector  2070  may be coupled between the first connector  2010  and one of the second connectors  2020 ,  2021 ,  2022 . The third connector  2070  may comprise a lumen configured to receive and circulate a sterilant through one or more portions of the first connector  2010 , second connector  2020 ,  2021 ,  2022 , and third connector  2070 . In some variations, the sterilization port  2052  may be non-removably coupled to a sterilant source and/or fluid source, thereby simplifying one or more of the first fluid device  2030  and first connector  2010 . 
     The robot  2060  may comprise one or more end effectors  2062 ,  2064 ,  2066  configured to manipulate and/or couple to one or more of the first fluid device  2030 , first connector  2010 , and third connector  2070 . For example, the first fluid device  2030  may comprise one or more fluid ports  2050  configured to couple to an end effector  2062 . Similarly, the third connector  2070  may comprise one or more sterilization ports  2052  configured to couple to robot  2060  (e.g., end effector  2064 ). In some variations, the robot  2060  may be configured to couple to one or more of a sterilant source, fluid source, and pump in order to facilitate efficient and shared fluidic connections between the fluid device, fluid connector, and a sterilization system. 
       FIGS.  20 B and  20 C  are schematic diagrams of a fluid connector connection process. In  FIG.  20 B , a third connector  2070  may be coupled to a distal end of a first connector  2010 , at  2002 . A distal end of the second connector  2020  may be coupled to the third connector  2070 , at  2004 . The second connector  2020  may be translated through the third connector  2070  to directly couple the second connector  2020  to the first connector  2010 , at  2006 . 
     In  FIG.  20 C , a third connector  2070  may be coupled to a distal end of the first connector  2010  and a distal end of the second connector  2020 , at  2002 . Each of the first connector  2010  and the second connector  2020  may be translated toward each other through the third connector  2070 , at  2005 . The second connector  2020  may be further translated towards the first connector  2010  to directly couple the first connector  2010  to the second connector  2010 , at  2007 .  FIG.  20 C  further illustrates a first port  2090  and a second port  2092  that may transition between a closed port configuration and an open port configuration. 
     In some variations, a fluid connector may comprise a third connector disposed between a first connector and a second connector. The third connector may be coupled to a second robot different from a first robot coupled to the first connector.  FIG.  21    is a block diagram of an illustrative variation of a fluid connector system  2100  comprising a first connector  2110 , a plurality of second connectors  2120 ,  2121 ,  2122 , a third connector  2170  (e.g., instrument, sterilization enclosure), a first fluid device  2130  (e.g., sterile liquid transfer device), a second fluid device  2140  (e.g., consumable), a first robot  2160 , and a second robot  2166 . The first connector  2110  may be coupled in fluid communication with the first fluid device  2130 , and the second connectors  2120 ,  2121 ,  2122  may be coupled in fluid communication with the second fluid device  2140 . The third connector  2170  may be coupled between the first connector  2110  and one of the second connectors  2120 ,  2121 ,  2122 . The third connector  2170  may comprise a lumen configured to receive and circulate a sterilant through one or more portions of the first connector  2110 , second connector  2120 ,  2121 ,  2122 , and third connector  2170 . In some variations, the third connector  2170  may be non-removably coupled to a sterilant source and/or fluid source, thereby simplifying one or more of the first fluid device  2130  and first connector  2110 . 
     The first robot  2160  may comprise one or more end effectors  2162 ,  2164  configured to manipulate and/or couple to one or more of the first fluid device  2130  and first connector  2110 . For example, the first fluid device  2130  may comprise one or more fluid ports  2150  configured to couple to an end effector  2162 . The third connector  2170  may be coupled to a second robot  2166  (e.g., 3 DOF robot). In some variations, the robot  2160 ,  2166  may be configured to couple to one or more of a sterilant source, fluid source, and pump in order to facilitate efficient and shared fluidic connections between the fluid device, fluid connector, and a sterilization system. 
     In some variations, a fluid connector may comprise a sterilant source coupled to a plurality of second connectors.  FIG.  22    is a block diagram of an illustrative variation of a fluid connector system  2200  comprising a first connector  2210 , a plurality of second connectors  2220 ,  2221 ,  2222 , a first fluid device  2230  (e.g., sterile liquid transfer device), a second fluid device  2240  (e.g., consumable), a robot  2260 , a sterilant source  2290  comprising one or more valves, and a sterilant switch  2292 . The first connector  2210  may be coupled in fluid communication with the first fluid device  2230 , and the second connectors  2220 ,  2221 ,  2222  may be coupled in fluid communication with the second fluid device  2240 . The robot  2260  may comprise one or more end effectors  2262 ,  2264  configured to manipulate and/or couple to one or more of the first fluid device  2230  and first connector  2210 . For example, the first fluid device  2230  may comprise one or more fluid ports  2250  configured to couple to an end effector  2262 . In some variations, the sterilant source  2290  may be coupled to the switch  2292 . The switch  2292  may be coupled to each of the second connectors  2220 ,  2221 ,  2222  in order to facilitate efficient and shared fluidic connections between the fluid device, fluid connector, and sterilization system. In some variations, a sterilant conduit may be routed from the switch  2292  through the second fluid device  2240  to a respective second connector  2220 ,  2221 ,  2222 . 
     In some variations, a fluid device may comprise one or more sterilant valves coupled to a plurality of second connectors.  FIG.  23    is a block diagram of an illustrative variation of a fluid connector system.  FIG.  23    is a block diagram of an illustrative variation of a fluid connector system  2300  comprising a first connector  2310 , a plurality of second connectors  2320 ,  2321 ,  2322 , a first fluid device  2330  (e.g., sterile liquid transfer device), a second fluid device  2340  (e.g., consumable), a robot  2360 , a set of sterilant valves  2390  disposed within a housing of the second fluid device  2340 , and a sterilant switch  2392 . The first connector  2310  may be coupled in fluid communication with the first fluid device  2330 , and the second connectors  2320 ,  2321 ,  2322  may be coupled in fluid communication with the second fluid device  2340 . The robot  2360  may comprise one or more end effectors  2362 ,  2364  configured to manipulate and/or couple to one or more of the first fluid device  2330  and first connector  2310 . For example, the first fluid device  2330  may comprise one or more fluid ports  2350  configured to couple to an end effector  2362 . In some variations, the sterilant valves  2390  may be coupled to the switch  2392 . The switch  2392  may be coupled to each of the second connectors  2320 ,  2321 ,  2322  via the sterilant valves  2390  in order to facilitate efficient and shared fluidic connections between the fluid device, fluid connector, and sterilization system. In some variations, a sterilant conduit may be routed from the switch  2392  through the second fluid device  2340  to a respective second connector  2320 ,  2321 ,  2322 . 
     In some variations, a fluid connector may comprise a sterilant source coupled to a plurality of second connectors each having a sterilant port (e.g., sterilant valve) and a sterilant conduit through a fluid device.  FIG.  24 A  is a block diagram of an illustrative variation of a fluid connector system  2400  comprising a first connector  2410 , a plurality of second connectors  2420 ,  2421 ,  2422 , a first fluid device  2430  (e.g., sterile liquid transfer device), a second fluid device  2440  (e.g., consumable), a robot  2460 , and a sterilant switch  2492  coupled to a sterilant source (not shown). The first connector  2410  may be coupled in fluid communication with the first fluid device  2430 , and the second connectors  2420 ,  2421 ,  2422  may be coupled in fluid communication with the second fluid device  2440 . The robot  2460  may comprise one or more end effectors  2462 ,  2464  configured to manipulate and/or couple to one or more of the first fluid device  2430  and first connector  2410 . For example, the first fluid device  2430  may comprise one or more fluid ports  2450  configured to couple to an end effector  2462 . 
     In some variations, each of the second connectors  2420 ,  2421 ,  2422 , may comprise a respective sterilant port  2494 ,  2496 ,  2498  comprising a valve coupled to a distal end of the second connector  2420 ,  2421 ,  2422 . In some variations, a sterilant conduit may be routed from the switch  2492  through the second fluid device  2440  to a respective sterilant port  2494 ,  2496 ,  2498 . In some variations, a sterilant source (not shown) may be coupled to the switch  2492 . The switch  2492  may be coupled to each of the second connectors  2420 ,  2421 ,  2422  via the sterilant ports  2494 ,  2496 ,  2498  in order to facilitate efficient and shared fluidic connections between the fluid device, fluid connector, and sterilization system. 
       FIG.  24 B  are schematic diagrams of a fluid connector connection process  2402 ,  2404 ,  2406  where a first connector  2410  is coupled to a second connector  2420 . For example, the sterilant port  2494  is in a closed valve configuration when the first connector  2410  and the second connector  2420  are separated and uncoupled  2402 .  FIG.  24 C  is a detailed schematic diagram of the sterilant valve  2494 . In some variations, the valve  2494  may transition to an open valve configuration when the first connector  2410  is coupled to the second connector  2420 , at  2404  and  2406 . 
     In some variations, a plurality of second connectors may comprise one or more pneumatic sterilant valves and a sterilant path through a fluid device.  FIG.  25 A  is a block diagram of an illustrative variation of a fluid connector system  2500  comprising a first connector  2510 , a plurality of second connectors  2520 ,  2521 ,  2522 , a first fluid device  2530  (e.g., sterile liquid transfer device), a second fluid device  2540  (e.g., consumable), a robot  2560 , and a sterilant switch  2592  coupled to a sterilant source (not shown). The first connector  2510  may be coupled in fluid communication with the first fluid device  2530 , and the second connectors  2520 ,  2521 ,  2522  may be coupled in fluid communication with the second fluid device  2540 . 
     In some variations, each of the second connectors  2520 ,  2521 ,  2522 , may comprise a respective pneumatic sterilant port  2594 ,  2596 ,  2598  comprising a valve coupled to a distal end of the second connector  2520 ,  2521 ,  2522 . In some variations, a sterilant conduit may be routed from the switch  2592  through the second fluid device  2540  to a respective sterilant port  2594 ,  2596 ,  2598 . In some variations, a sterilant source (not shown) may be coupled to the switch  2592 . The switch  2592  may be coupled to each of the second connectors  2520 ,  2521 ,  2522  via the sterilant ports  2594 ,  2596 ,  2598  in order to facilitate efficient and shared fluidic connections between the fluid device, fluid connector, and sterilization system. 
     The robot  2560  may comprise one or more end effectors  2562 ,  2564  configured to manipulate and/or couple to one or more of the first fluid device  2530 , first connector  2510 , and sterilant ports  2594 ,  2596 ,  2598 . For example, the first fluid device  2530  may comprise one or more fluid ports  2550  configured to couple to an end effector  2562 . Similarly, sterilant ports  2594 ,  2596 ,  2598  may be configured to couple to the end effector  2562  to pneumatically actuate the sterilant ports  2594 ,  2596 ,  2598 . A pneumatically actuated sterilant port may enable the sterilant conduit to be formed with a fewer number of check valves between the sterilant ports  2594 ,  2596 ,  2598  and switch  2592 . 
       FIG.  25 B  are schematic diagrams of a fluid connector connection process  2502  and  2504 , where a first connector  2510  is coupled to a second connector  2520 . For example, the sterilant port  2594  is in a closed valve configuration when the first connector  2510  and the second connector  2520  are separated and uncoupled  2502 .  FIG.  25 C  is a detailed schematic diagram of the sterilant valve  2594 . In some variations, the valve  2594  may transition to an open valve configuration when the first connector  2510  is coupled to the second connector  2520  and the valve  2594  is pneumatically actuated, at  2504 . 
     Liquid Transfer Bus 
     Generally, to permit transfer of one or more of a cell product (that is, solution(s) containing cell product), fluids, and reagents between the modules, the modules of the cartridge may be fluidically coupled to one another either directly or via one or more liquid transfer buses. In some variations, a liquid transfer bus may comprise a portion of the cartridge configured to control the flow and distribution of the cell product between modules and reservoirs. A liquid transfer bus may comprise one or more of a fluid manifold, fluid conduit (e.g., tubing), and one or more valves (including but not limited to 2/2 valves, 3/2 valves, 3/3 valves, 4/2 valves, and rotary selector valves). 
     Transfer of the cell product, reagents, or fluids within the cartridge may be achieved by any pump or other structure that generates a pressure differential between fluid in one portion of the cartridge and fluid in another portion of the cartridge. For example, the cartridge may comprise one or more pump; the cartridge may be pre-loaded with pressurized fluid contained behind a valve; the cartridge may be connected to a fluid source or a fluid sink. The cartridge may contain one or more mechanical pumps (e.g., linear pump, peristaltic pump, gear pump, screw pump, plunger pump) or portions of a pump (i.e. the pump may interface with a pump actuator). External pressure may be applied to the cartridge, to tubing within the cartridge, or to a bag within the cartridge (that is, applying pressure either to the liquid in the bag or to headspace gas of the bag). In some variations, an arrangement of the components of the cartridge may facilitate gravity-based fluid transfer within the cartridge (e.g., gravity-fed pumping). Although one advantage of the disclosed variations may be reduced operator intervention, the systems and methods of the disclosure may use manual operation in the designed workflow or as an adjunct to automated operation in case of imperfect automated system operation. For example, a process step may include manual intervention, such as fluid input or output. An operator may intervene in an automated process to correct device operation, (e.g. manually compressing a bag to flush remaining fluid into the system). Fluid may comprise liquid and/or gas, as compressed gases supplied externally or provided in pressurized chambers may be used to generate liquid flow, e.g., transfer of solution containing a cell product from one module to another. 
     In some variations, the liquid transfer bus may be configured to deliver the cell product(s) to each of a series of modules in an order set by the design of the cartridge, or in an order determined by operation of the system by the processor or processors. Similarly stated, some variations of the cartridge may have the advantage that the order of cell processing steps as well as the process parameters for any of the cell therapy processing steps may not be set by the cartridge but rather are controlled by the controller. In some variations, the liquid transfer bus may be controlled to deliver the cell product to the modules in any of various sequences, or to bypass one or more modules (e.g., by configuring the state of the valve(s) attached to the fluidic bus). In some variations, a module may be used more than once in a method of cell processing. Optionally, the method may comprise performing one or more wash steps. For example, a counterflow centrifugal elutriation (CCE) module may be used more than once. In an illustrative method, the method comprises culturing the cell product in a first bioreactor module, transferring the cell product to the CCE module to enrich for a desired cell type, transferring the cell product to a second bioreactor module for a second culturing step, washing the CCE module using a wash solution, and transferring the cell product to the CCE module for a second enrichment step. 
     In some variations, the liquid transfer bus or the liquid transfer buses may be fluidically coupled to multiple bags or reservoirs used to provide solutions or reagents, store cell products, or to collect waste solutions or reagents. 
     In some variations, the cartridge may comprise one or more pumps, which may be fluidically coupled to the liquid transfer bus and/or one or more modules. The pump(s) may include a motor operatively coupled to control circuits and a power source (e.g. a battery or electrical connectors for an off-cartridge power source). In some variations, the pump may be divided into a pump on the cartridge and pump actuators on one or more instruments of the system. The pump may be an opening in the cartridge with tubing arranged around the circumference of the opening and configured to receive a pump actuator (e.g., a peristaltic rotor). By dividing components of the pump that contact the cell product (i.e. tubing) from components of the pump that perform operations of the cell product, (i.e. the pump actuator, e.g., peristaltic rotor), the cartridge may be compact and simplified. For example,  FIG.  26 A  and  FIG.  26 B  illustrate a pump head  2610  and a pump  2610  of a cartridge in an uncoupled configuration ( FIG.  13 A ) and a coupled configuration. 
     In some variations, one or more pumps  146  (e.g., fluid pump) may generate a predetermined fluid flow rate to circulate a sterilant and/or fluid. In some variations, a pump may comprise one or more of a positive displacement pump (e.g., peristaltic pump, diaphragm pump, syringe pump), centrifugal pump, combinations thereof, and the like. One or more fluid sources may be coupled to the pump. 
     In some variations, the pump may be configured to receive a pump signal (generated by a controller) configured to circulate a sterilant for a dwell time sufficient to sterilize at least a portion of a fluid connector. For example, the pump may be configured to circulate the sterilant for at least 10 seconds. In some variations, the pump may be configured to receive a pump signal configured to circulate a non-sterilant gas (e.g., inert gas, air) to remove the sterilant. 
     In some variations, a discontinuous flow pump (e.g., peristaltic pump) may generate pulsatile flow as, for example, a tube contracts and relaxes between rollers. In some variations, closed loop feedback from a flow sensor may be used to compensate for pulsatile flow to generate a substantially continuous flow rate. For example, a flow sensor may be coupled to a fluid conduit to measure the flow rate. A controller may receive the measured flow rate and generate a pump signal to the pump based on a proportional correction function configured to reduce the “ripples” measured by the flow sensor. Additionally or alternatively, a controller may apply periodic error correction to a pump signal to reduce periodic error that may be unique to each pump. For example, a flow sensor may measure and determine a periodic error of a pump. A pump signal comprising the periodic error correction may correspond to a waveform comprising an inverse shape of the error. The resulting pump flow may correct for fluctuations in flow rate. 
     Controller 
     In some variations, a system  100  may comprise a controller  120  (e.g., computing device) comprising one or more of a processor  122 , memory  124 , communication device,  126 , input device  128 , and display  130 . The controller  120  may be configured to control (e.g., operate) the workcell  110 . The controller  120  may comprise a plurality of devices. For example, the workcell  110  may enclose one or more components of the controller  120  (e.g., processor  122 , memory  124 , communication device  126 ) while one or more components of the controller  120  may be provided remotely to the workcell  110  (e.g., input device  128 , display  130 ). 
     Processor 
     The processor (e.g., processor  122 ) described here may process data and/or other signals to control one or more components of the system (e.g., workcell  110 , controller  120 ). The processor may be configured to receive, process, compile, compute, store, access, read, write, and/or transmit data and/or other signals. Additionally, or alternatively, the processor may be configured to control one or more components of a device and/or one or more components of controller (e.g., console, touchscreen, personal computer, laptop, tablet, server). 
     In some variations, the processor may be configured to access or receive data and/or other signals from one or more of workcell  110 , server, controller  120 , and a storage medium (e.g., memory, flash drive, memory card, database). In some variations, the processor may be any suitable processing device configured to run and/or execute a set of instructions or code and may include one or more data processors, image processors, graphics processing units (GPU), physics processing units, digital signal processors (DSP), analog signal processors, mixed-signal processors, machine learning processors, deep learning processors, finite state machines (FSM), compression processors (e.g., data compression to reduce data rate and/or memory requirements), encryption processors (e.g., for secure wireless data transfer), and/or central processing units (CPU). The processor may be, for example, a general purpose processor, Field Programmable Gate Array (FPGA), an Application Specific Integrated Circuit (ASIC), a processor board, and/or the like. The processor may be configured to run and/or execute application processes and/or other modules, processes and/or functions associated with the system. The underlying device technologies may be provided in a variety of component types (e.g., metal-oxide semiconductor field-effect transistor (MOSFET) technologies like complementary metal-oxide semiconductor (CMOS), bipolar technologies like emitter-coupled logic (ECL), polymer technologies (e.g., silicon-conjugated polymer and metal-conjugated polymer-metal structures), mixed analog and digital, and the like. 
     The systems, devices, and/or methods described herein may be performed by software (executed on hardware), hardware, or a combination thereof. Hardware modules may include, for example, a general-purpose processor (or microprocessor or microcontroller), a field programmable gate array (FPGA), and/or an application specific integrated circuit (ASIC). Software modules (executed on hardware) may be expressed in a variety of software languages (e.g., computer code), including structured text, typescript, C, C++, C#, Java®, Python, Ruby, Visual Basic®, and/or other object-oriented, procedural, or other programming language and development tools. Examples of computer code include, but are not limited to, micro-code or micro-instructions, machine instructions, such as produced by a compiler, code used to produce a web service, and files containing higher-level instructions that are executed by a computer using an interpreter. Additional examples of computer code include, but are not limited to, control signals, encrypted code, and compressed code 
     Memory 
     The cell processing systems and devices described here may include a memory (e.g., memory  124 ) configured to store data and/or information. In some variations, the memory may include one or more of a random access memory (RAM), static RAM (SRAM), dynamic RAM (DRAM), a memory buffer, an erasable programmable read-only memory (EPROM), an electrically erasable read-only memory (EEPROM), a read-only memory (ROM), flash memory, volatile memory, non-volatile memory, combinations thereof, and the like. In some variations, the memory may store instructions to cause the processor to execute modules, processes, and/or functions associated with the device, such as image processing, image display, sensor data, data and/or signal transmission, data and/or signal reception, and/or communication. Some variations described herein may relate to a computer storage product with a non-transitory computer-readable medium (also may be referred to as a non-transitory processor-readable medium) having instructions or computer code thereon for performing various computer-implemented operations. The computer-readable medium (or processor-readable medium) is non-transitory in the sense that it does not include transitory propagating signals per se (e.g., a propagating electromagnetic wave carrying information on a transmission medium such as space or a cable). The computer code (also may be referred to as code or algorithm) may be those designed and constructed for the specific purpose or purposes. In some variations, the memory may be configured to store any received data and/or data generated by the controller and/or workcell. In some variations, the memory may be configured to store data temporarily or permanently 
     Input Device 
     In some variations, the display may include and/or be operatively coupled to an input device  128  (e.g., touch screen) configured to receive input data from a user. For example, user input to an input device  128  (e.g., keyboard, buttons, touch screen) may be received and processed by a processor (e.g., processor  122 ) and memory (e.g., memory  124 ) of the system  100 . The input device may include at least one switch configured to generate a user input. For example, an input device may include a touch surface for a user to provide input (e.g., finger contact to the touch surface) corresponding to a user input. An input device including a touch surface may be configured to detect contact and movement on the touch surface using any of a plurality of touch sensitivity technologies including capacitive, resistive, infrared, optical imaging, dispersive signal, acoustic pulse recognition, and surface acoustic wave technologies. In variations of an input device including at least one switch, a switch may have, for example, at least one of a button (e.g., hard key, soft key), touch surface, keyboard, analog stick (e.g., joystick), directional pad, mouse, trackball, jog dial, step switch, rocker switch, pointer device (e.g., stylus), motion sensor, image sensor, and microphone. A motion sensor may receive user movement data from an optical sensor and classify a user gesture as a user input. A microphone may receive audio data and recognize a user voice as a user input. 
     In some variations, the cell processing system may optionally include one more output devices in addition to the display, such as, for example, an audio device and haptic device. An audio device may audibly output any system data, alarms, and/or notifications. For example, the audio device may output an audible alarm when a malfunction is detected. In some variations, an audio device may include at least one of a speaker, piezoelectric audio device, magnetostrictive speaker, and/or digital speaker. In some variations, a user may communicate with other users using the audio device and a communication channel. For example, a user may form an audio communication channel (e.g., VoIP call). 
     Additionally or alternatively, the system may include a haptic device configured to provide additional sensory output (e.g., force feedback) to the user. For example, a haptic device may generate a tactile response (e.g., vibration) to confirm user input to an input device (e.g., touch surface). As another example, haptic feedback may notify that user input is overridden by the processor. 
     Communication Device 
     In some variations, the controller may include a communication device (e.g., communication device  126 ) configured to communicate with another controller and one or more databases. The communication device may be configured to connect the controller to another system (e.g., Internet, remote server, database, workcell) by wired or wireless connection. In some variations, the system may be in communication with other devices via one or more wired and/or wireless networks. In some variations, the communication device may include a radiofrequency receiver, transmitter, and/or optical (e.g., infrared) receiver and transmitter configured to communicate with one or more devices and/or networks. The communication device may communicate by wires and/or wirelessly. 
     The communication device may include RF circuitry configured to receive and send RF signals. The RF circuitry may convert electrical signals to/from electromagnetic signals and communicate with communications networks and other communications devices via the electromagnetic signals. The RF circuitry may include well-known circuitry for performing these functions, including but not limited to an antenna system, an RF transceiver, one or more amplifiers, a tuner, one or more oscillators, a digital signal processor, a CODEC chipset, a subscriber identity module (SIM) card, memory, and so forth. 
     Wireless communication through any of the devices may use any of plurality of communication standards, protocols and technologies, including but not limited to, Global System for Mobile Communications (GSM), Enhanced Data GSM Environment (EDGE), high-speed downlink packet access (HSDPA), high-speed uplink packet access (HSUPA), Evolution, Data-Only (EV-DO), HSPA, HSPA+, Dual-Cell HSPA (DC-HSPDA), long term evolution (LTE), near field communication (NFC), wideband code division multiple access (W-CDMA), code division multiple access (CDMA), time division multiple access (TDMA), Bluetooth, Wireless Fidelity (WiFi) (e.g., IEEE 802.11a, IEEE 802.11b, IEEE 802.11g, IEEE 802.11n, and the like), voice over Internet Protocol (VoIP), Wi-MAX, a protocol for e-mail (e.g., Internet message access protocol (IMAP) and/or post office protocol (POP)), instant messaging (e.g., extensible messaging and presence protocol (XMPP), Session Initiation Protocol for Instant Messaging and Presence Leveraging Extensions (SIMPLE), Instant Messaging and Presence Service (IMPS)), and/or Short Message Service (SMS), EtherCAT, OPC Unified Architecture, or any other suitable communication protocol. In some variations, the devices herein may directly communicate with each other without transmitting data through a network (e.g., through NFC, Bluetooth, WiFi, RFID, and the like). 
     In some variations, the systems, devices, and methods described herein may be in communication with other wireless devices via, for example, one or more networks, each of which may be any type of network (e.g., wired network, wireless network). The communication may or may not be encrypted. A wireless network may refer to any type of digital network that is not connected by cables of any kind. Examples of wireless communication in a wireless network include, but are not limited to cellular, radio, satellite, and microwave communication. However, a wireless network may connect to a wired network in order to interface with the Internet, other carrier voice and data networks, business networks, and personal networks. A wired network is typically carried over copper twisted pair, coaxial cable and/or fiber optic cables. There are many different types of wired networks including wide area networks (WAN), metropolitan area networks (MAN), local area networks (LAN), Internet area networks (IAN), campus area networks (CAN), global area networks (GAN), like the Internet, and virtual private networks (VPN). Hereinafter, network refers to any combination of wireless, wired, public and private data networks that are typically interconnected through the Internet, to provide a unified networking and information access system. 
     Cellular communication may encompass technologies such as GSM, PCS, CDMA or GPRS, W-CDMA, EDGE or CDMA2000, LTE, WiMAX, and 5G networking standards. Some wireless network deployments combine networks from multiple cellular networks or use a mix of cellular, Wi-Fi, and satellite communication. 
     Display 
     Image data may be output on a display e.g., display  130 ) of a cell processing system. In some variations, a display may include at least one of a light emitting diode (LED), liquid crystal display (LCD), electroluminescent display (ELD), plasma display panel (PDP), thin film transistor (TFT), organic light emitting diodes (OLED), electronic paper/e-ink display, laser display, and/or holographic display. 
     II. Methods 
     Generally, the systems and devices described herein may perform one or more cell processing steps to manufacture a cell product.  FIG.  28    is a flowchart of a method of cell processing  2800 . The method  2800  may include enriching a selected population of cells in a solution (e.g., fluid)  2802 . For example, the solution may be conveyed to a CCE module of a cartridge via a liquid transfer bus. A robot may be operated to move the cartridge to a CCE instrument so that the CCE module interfaces with the CCE instrument. The CCE instrument may be operated to cause the CCE module to enrich the selected population of cells. Additionally or alternatively, the cell product may be introduced into and out of the cartridge via a sterile liquid transfer port (either manually or automatically) for any of the steps described herein. In some variations, the cartridge may be sterilized in a feedthrough port (either manually or automatically). 
     In some variations, a selected population of cells in the solution may be washed  2804 . For example, the solution may be conveyed to the CCE module of the cartridge via the liquid transfer bus. A robot may be operated to move the cartridge to the CCE instrument so that the CCE module interfaces with the CCE instrument. The CCE instrument may be operated to cause the CCE module to remove media from the solution, introduce media into the solution, and/or replace media in the solution. 
     In some variations, a population of cells in the solution may be selected  2806 . For example, the solution may be conveyed to a selection module of the cartridge via the liquid transfer bus. The robot may be operated to move the cartridge to a selection instrument so that the selection module interfaces with the selection instrument. The selection instrument may be operated to cause the selection module to select the selected population of cells. 
     In some variations, a population of cells in the solution may be sorted  2808 . For example, the solution may be conveyed to a sorting module of the cartridge via the liquid transfer bus. The robot may be operated to move the cartridge to a sorting instrument so that the sorting module interfaces with the sorting instrument. The sorting instrument may be operated to cause the sorting module to sort the population of cells. 
     In some variations, the solution may be conveyed to a bioreactor module of the cartridge via the liquid transfer bus to rest  2810 . For example, the robot may be operated to move the cartridge to a bioreactor instrument so that a bioreactor module interfaces with the bioreactor instrument. The bioreactor instrument may be operated to cause the bioreactor module to maintain the cells at a set of predetermined conditions. 
     In some variations, the cells may be expanded in the solution  2812 . For example, the solution may be conveyed to the bioreactor module of the cartridge via the liquid transfer bus. The robot may be operated to move the cartridge to the bioreactor instrument so that the bioreactor module interfaces with the bioreactor instrument. The bioreactor instrument may be operated to cause the bioreactor module to expand the cells by cellular replication. 
     In some variations, tissue may be digested by conveying an enzyme reagent via the liquid transfer bus to a module containing a solution containing a tissue such that the tissue releases a select cell population into the solution  2814 . 
     In some variations, a selected population of cells in the solution may be activated by conveying an activating reagent via the liquid transfer bus to a module containing the solution containing the cell product  2816 . 
     In some variations, the solution may be conveyed to an electroporation module of the cartridge via the liquid transfer bus and receive an electroporation signal to electroporate the cells in the solution  2818 . For example, the robot may be operated to move the cartridge to an electroporation instrument so that the electroporation module interfaces with the electroporation instrument. The electroporation instrument may be operated to cause the electroporation module to electroporate the selected population of cells in the presence of genetic material. 
     In some variations, an effective amount of a vector may be conveyed via the liquid transfer bus to a module containing the solution containing the cell product, thereby transducing a selected population of cells in the solution  2820 . 
     In some variations, a formulation solution may be conveyed via the liquid transfer bus to a module containing the cell product to generate a finished cell product  2822 . For example, the finished cell product may be conveyed to one or more product collection bags. In some variations, finishing a cell product may comprise one or more steps of washing cells, concentrating cells, exchanging a buffer of the cells with a formulation buffer, and dosing cells in the formulation buffer in predetermined quantities into one or more product collection bags and/or vessels. 
     In some variations, the cell product may be removed, either manually or automatically, from the cartridge to harvest the cells  2824 . 
     In some variations, the cell product may comprise one or more of an immune cell genetically engineered chimeric antigen receptor T cell, a genetically engineered T cell receptor (TCR) cell, a hematopoietic stem cell (HSC), and a tumor infiltrating lymphocyte (TIL). In some variations, the immune cell may comprise a natural-killer (NK) cell. 
     Methods of cell processing may include a subset of cell processing steps in any suitable order. For example, the method of cell processing may include, in order, the enrichment step  2802 , the selection step  2806 , the activation step  2816 , the transduction step  2820 , the expansion step  2812 , and the harvesting step  2824 . In some variations, the method of cell processing may include, in order, the enrichment step  2802 , the selection step  2806 , the resting step  2810 , the transduction step  2820 , and the harvesting step  2824 . In some variations, the method of cell processing may include, in order, the tissue-digestion step  2820 , the washing step  2804 , the activation step  2816 , the expansion step  2812 , and the harvesting step  2824 . 
     Generally, the methods described herein may offload the complex steps performed in cell processing operation to a set of instruments, thereby reducing the cost of the cartridge (which may be a consumable). In some variations, the cartridge may contain the cell product (e.g., solution containing cells) throughout a manufacturing process, with different instruments interfacing with the cartridge at appropriate times to perform one or more cell processing steps. For example, a cell processing step may comprise conveying cells and reagents to each of the modules within the cartridge. A set of instruments interfacing with a cartridge facilitates process flexibility where a workcell may be customized with a predetermined set of instruments for a predetermined cell therapy product. For example, the order of cell processing steps may be customized for each cell product as described in more detail herein with respect to  FIGS.  35 - 55   . 
     In some variations, a cell product may be retained within the cartridge throughout a manufacturing process (e.g., workflow). Additionally or alternatively, the cell product may be removed from the cartridge for one or more cell processing steps, either manually by an operator, or automatically through a fluid connector (e.g., SLTP) or other access ports on the cartridge. The cell product may then be returned to the same cartridge, transferred to another cartridge, or split among several cartridges. In some variations, one or more cell processing steps may be performed outside the cartridge. In some variations, processing within the workcell may facilitate sterile cell processing within the cartridge. 
       FIG.  29    is a flowchart of a method of cell processing and illustrates cell processing steps performed on a cartridge (e.g., consumable) within a workcell including a CCE instrument module, a sterile liquid transfer (SLT) instrument module, and a bioreactor instrument module. The consumable may be configured to interface with any of the CCE instrument module, SLT instrument module, and bioreactor instrument module to perform one or more cell processing steps. For example, a robot (or operator) may be configured to move a cartridge between any of the modules of the workcell. A pump head in an instrument may engage the consumable cartridge in order to convey fluids between the modules of the cartridge, into or out of various reservoirs in the cartridge, and/or through ports that permit reagents to be added or removed from the cartridge. 
     In some variations, the CCE instrument module may comprise a pump and centrifuge configured to interface with a cartridge (e.g., consumable). The SLT instrument module may comprise one or more fluid connectors be configured to interface with one or more of a bag and bioreactor of a cartridge. The bioreactor instrument module may comprise one or more sensors, temperature regulators, pumps, agitators, and the like, and be configured to interface with the cartridge. In some variations, the cell product may be contained within the cartridge throughout cell processing. 
     A method of cell processing depicted in  FIG.  29    may include moving a fluid (e.g., cells in solution) in a product bag to a CCE module (e.g., rotor) of a cartridge (e.g., consumable) using a pump  2910 . In some variations, the fluid may be enriched using the CCE module  2912 . For example, blood constituents may be collected in a waste bag  2913 . In some variations, the fluid may be washed using the CCE module  2914 . For example, buffer may be collected in a waste bag  2915 . In some variations, media may be exchanged using the CCE module  2916 . For example, one or more of buffer (e.g., formulation buffer) and media may be collected in a waste bag  2917 . In some variations, fluid may be moved to a bioreactor of the cartridge  2918 . 
     In some variations, a fluid connector may fill a bag with a reagent  2920 . In some variations, a reagent (e.g., bead, vector) may be added to a bioreactor of a cartridge  2922 . In some variations, a fluid connector removes waste from a bag  2924 . In some variations, a fluid connector may optionally remove a sample from a bioreactor. 
     In some variations, cells may be moved to a bioreactor  2930 . In some variations, the cells may undergo activation or genetic modification  2932 . In some variations, the cells may undergo incubation  2934 . In some variations, the cells may undergo perfusion using a pump  2936 . For example, spent media may be collected in a waste bag  2937 . In some variations, cells may undergo expansion  2938 . In some variations, cells may be harvested after media exchange  2940 . 
       FIG.  30 A  is a flowchart of a method of cell processing for autologous CAR T cells or engineered TCR cells. The method  3000  may comprise the steps of enrichment, selection, activation, genetic modification, expansion, harvest/formulation, and cryopreservation.  FIG.  30 B  is a flowchart of a method of cell processing for allogeneic CAR T cells or engineered TCR cells. The method  3010  may comprise the steps of enrichment, activation, genetic modification (e.g., transduction, transfection), alpha/beta T cell depletion, expansion, harvest/pool/formulation, and cryopreservation. 
       FIG.  31    is a flowchart of a method of cell processing for hematopoietic stem cell (HSC) cells. The method  3100  may comprise the steps of enrichment, selection, rest, genetic modification, harvest/formulation, and cryopreservation. 
       FIG.  32    is a flowchart of a method of cell processing for tumor infiltrating lymphocyte (TIL) cells. The method  3200  may comprise the steps of tissue digestion, washing, selection, activation, expansion, harvest/formulation, and cryopreservation. 
       FIG.  33    is a flowchart of a method of cell processing for natural killing (NK) CAR cells. The method  3300  may comprise the steps of enrichment, selection, activation, genetic modification, expansion, harvest/formulation, and cryopreservation. 
       FIGS.  34 A- 34 C  are flowcharts of methods of cell processing for regulatory T (T reg ) cells. The method  3400  may comprise the steps of enrichment, selection, harvest/formulation, cryopreservation. The method  3402  may comprise the steps of enrichment, selection, activation, genetic modification, expansion, selection (optionally), harvest/formulation, and cryopreservation. The method  3404  may comprise the steps of introducing feeder cell culture for enrichment, selection, activation/expansion, and harvest/irradiation. Another set of cells may undergo enrichment, selection, co-culture with the processed feeder cells, harvest, and cryopreservation. 
       FIGS.  98 - 101    are flowcharts of methods of cell processing for cell therapy workflows comprising split (e.g., parallel) processing. The method  9800  may comprise the steps of enrichment, selection, activation, genetic modification, expansion, formulation, and cryopreservation. For example, a cell processing method  9800  (e.g., workflow) may comprise splitting a cell product into two or more portions after an enrichment step. The split portions may be processed in parallel within a single cartridge. In some variations, one or more split portions may be transferred to two or more cartridges and processed in parallel. One or more cell processing parameters (e.g., timing of process steps, types of reagents added, transfection constructs, and the like) may be configured independently for each split portion of the cell product. In some variations, the split portions may be pooled after the expansion step. 
     The method  9900  may comprise the steps of enrichment, selection, activation, genetic modification, expansion, formulation, and cryopreservation. For example, a cell processing method  9900  (e.g., workflow) may comprise splitting a cell product into two or more portions after an activation step. The split portions may be processed in parallel within a single cartridge. In some variations, one or more split portions may be transferred to two or more cartridges and processed in parallel. One or more cell processing parameters (e.g., timing of process steps, types of reagents added, transfection constructs, and the like) may be configured independently for each split portion of the cell product. In some variations, the split portions may be pooled after the expansion step and/or the genetic modification step. 
     The method  10000  may comprise the steps of enrichment, selection, activation, genetic modification, expansion, formulation, and cryopreservation. For example, a cell processing method  10000  (e.g., workflow) may comprise splitting a cell product into two or more portions after a selection step. The split portions may be processed in parallel within a single cartridge. In some variations, one or more split portions may be transferred to two or more cartridges and processed in parallel. One or more cell processing parameters (e.g., timing of process steps, types of reagents added, transfection constructs, and the like) may be configured independently for each split portion of the cell product. In some variations, the split portions may not be pooled. 
     The method  10100  may comprise the steps of enrichment, selection, activation, genetic modification, expansion, formulation, and cryopreservation. For example, a cell processing method  10100  (e.g., workflow) may comprise splitting a cell product into two or more portions as starting materials. The separate products may remain segregated and processed in parallel as split portions within a single cartridge or a plurality of cartridges. One or more cell processing parameters (e.g., timing of process steps, types of reagents added, transfection constructs, and the like) may be configured independently for each split portion. In some variations, the split portions may be pooled after the expansion step. 
       FIG.  102    is a schematic diagram of a cell processing system  10200  configured for split processing within a single cartridge. For example, the methods  9800 - 10100  described with respect to  FIGS.  98 - 101    may be performed within the cartridge  10210 . In some variations, the system  10200  may comprise a sterile liquid transfer device  10220  comprising a reagent  10222 , and a cartridge  10210  comprising a plurality of bioreactor modules  10230 , a pump module  10240 , a thermal module  10245 , a pressure driven flow module  10250 , a MACS module  10255 , an electroporation module  10260 , a FACS module  10265 , a CCE module  10270 , and a blank module  10280 . The cartridge  10210  may further comprise a reagent storage  10285 , a plurality of product bags  10290 , and a liquid transfer bus  10295 . The liquid transfer bus  10295  may be configured to couple the components of the cartridge  10210  for fluid communication. 
     In some variations, loading and removing of cell product into and out of the cartridge may be performed in the system or outside the system. In some variations, the cartridge is loaded bedside to the patient or donor and then delivered to a cell processing system in or near the hospital, or shipped to a facility where the cell processing system is installed. Likewise, the cell product may be removed from the cartridge after processing either at a facility or closer to the intended recipient of the cell product (the patient). Optionally the cell product is frozen before, during, or after the methods of the disclosure-optionally after addition of one or more cryoprotectants to the cell product. In some variations, the system comprises a freezer and/or a liquid nitrogen source. In some variations, the system comprises a water bath or a warming chamber containing gas of controlled temperature to permit controlled thawing of the cell product, e.g. a water bath set to between about 20° C. and about 40° C. In some variations, the cartridge is made of materials that resistant mechanical damage when frozen. 
     Automated Cell Processing 
     Described here are methods of transforming user-defined cell processing operations into cell processing steps using the automated cell processing systems and devices described herein. In some variations, cell processing operations are received and transformed into cell processing steps to be performed by the system given a set of predetermined constraints. For example, a user may input a set of biologic process steps and corresponding biologic process parameters to be executed by a cell processing system. Optionally, process parameters may be customized for each cartridge or sets of cartridges. 
       FIG.  35    is a flowchart that generally describes a variation of a method of automated cell processing. The method  3500  may include receiving an ordered input list of cell processing operations  3502 . For example, a set of more than one ordered input list of cell processing operations may be received to be performed on more than one cartridge on an automated cell processing system. For example, as shown in the GUI  4900  of  FIG.  49    and described in more detail herein, one or more biologic process inputs (e.g., available operations) such as enrichment, MACS selection, activation, transduction, transfection, expansion, and inline analysis may be selected as an ordered input list of cell processing operations. Furthermore, GUI  5200  of  FIG.  52    illustrates a complete ordered input list of cell processing operations (e.g., set of selected operations)  5220  selected by a user. 
     In some variations, one or more sets of cell processing parameters may be received  3504 . Each set of cell processing parameters may be associated with one of the cell processing operations. Each set of cell processing parameters may specify characteristics of the cell processing step to be performed by the instrument at that cell processing step. For example, the GUI  4000  of  FIG.  40    illustrates reagent and container parameters, the GUI  4200  of  FIG.  42    illustrates an example of a process parameter, the GUI  4400  of  FIG.  44    illustrates an example of a preprocess analytic, and the GUI  4800  of  FIG.  48    illustrates an example of a set of activation settings. 
     In some variations, a transformation model may be executed on the ordered input list  3506 . In some variations, the transformation model may comprise constraints on the ordered output list determined by a predetermined configuration of the automated cell processing system. For example, the constraints may comprise information on the configuration of the automated cell processing system. 
     In some variations, the constraints may comprise one or more of a type and/or number and/or state of instruments, a type and/or number and/or state of modules on the cartridge, a type and/or number of reservoirs on the cartridge, a type and/or number of sterile liquid transfer ports on the cartridge, and number and position of fluid paths between the modules, reservoirs, and sterile liquid transfer ports on the cartridge. 
     In some variations, a set of predetermined constraints may be placed on a set of the process control parameters. For example, the volume and/or the type of reagents used may be constrained based on the size of the system and/or products manufactured. Other process parameter constraints may include, but is not limited to, one or more or temperature, volume, time, pH, cell size, cell number, cell density, cell viability, dissolved oxygen, glucose levels, volumes of onboard reagent storage and waste, combinations thereof, and the like. For example, the GUI  4000  of  FIG.  40    depicts that a reagent has a volume per unit of 30 ml and a required volume of 54 ml, and a consumable container has a volume per unit of 75 ml. The GUI  4800  of  FIG.  48    depicts that an activation concentration is 12 mg/L, an activation culture time is 1600 seconds, activation temperature is 18° C., and a gas mix includes 21% oxygen, 78.06% nitrogen, and 0.04% of carbon dioxide. These constraints may be applied by a transformation model to generate an ordered output list of cell processing steps that affect how one or more of the robot, instrument, and cartridge are operated and the cell product manufactured. 
     In some variations, the order of operations may be constrained based on hardware constraints. For example, the robot may be limited to moving one cartridge at a time. Similarly, an instrument may be constrained to operating on a predetermined number of cartridges at once. 
     In some variations, as illustrated in the GUI  4900  of  FIG.  49   , a load product operation must be the first operation performed, and may be performed once for each process. A fill and finish operation may always be the last operation performed before product completion, and may be performed once for each process. 
     In some variation, the system may prevent the user from executing a set of operations in an order that cannot be performed by the system. 
     In some variations, a notification (e.g., warning, alert) may be output if a user orders a set of operations in a “non-standard” manner. For example, a notification may be output if the same type of operation is repeated sequentially (e.g., enrichment immediately followed by enrichment). Similarly, a notification may be output if an operation (e.g., selection, activation) is used two times or more within a given process when such an operation is typically used just once in a given process. 
     In some variations, an output of the transformation model may correspond to an ordered output list of cell processing steps capable of being performed by the system  3508 . For example, the transformation model may be executed on the sets of ordered input lists to create the ordered output list of cell processing steps. The output list of cell processing steps may control a robot, cartridge, and one or more instruments. 
     In some variations, the ordered output list is performed by the system to control a robot to move one or more cartridges each containing a cell product between the instruments  3510 . For example, the MACS selection process selected by the user may correspond to the robot  230  of  FIG.  2    moving the cartridge  250  to the cell selection instrument  216  from, for example, another instrument. In some variations, the ordered output list may comprise instructions for a robot to load a cartridge (e.g., single use consumable) into the cell processing system (e.g., workcell). Furthermore, the robot may be configured to move the cartridge to a first instrument position. 
     In some variations, the ordered output list is further performed by the system to control one or more of the instruments to perform one or more cell processing steps on one or more cell products  3512  of a respective cartridge. For example, the compute server rack  210  (e.g., controller  120 ) may be configured to control an electroporation module  220  configured to apply a pulsed electric field to a cell suspension of a cartridge  250 . In some variations, the ordered output list may comprise instructions for an instrument (e.g., bioreactor) to process the product (e.g., transfer the cell product from a small bioreactor module to a large bioreactor module). Furthermore, the instrument may be further configured to operate under a set of process parameters (e.g., 9 hour duration, pH of 6.7, temperature between 37.3° C. and 37.8° C., mixing mode  3 ). As another example, the ordered output list may comprise instructions to operate a sterile liquid transfer module to perform one or more of removing waste from a cartridge, adding media to the cartridge, and adding a MACS reagent to the cartridge. 
     In some variations, one or more electronic batch records may be generated  3514  based on the process parameters and data collected from sensors during process execution. Batch records generated by the system may include process parameters, time logging, sensor measurements from the instruments, QC parameters determined by QC instrumentation, and other records. 
       FIG.  36    is a flowchart that generally describes a variation of a method of executing a transformation model  3600 . In some variations, one or more biological functions may be generated and output to a user. For example, a set of configurable biological function blocks may be displayed on a graphical user interface for user selection. The GUI may enable a user to select and order the biological function blocks and define biological control parameters. One or more control parameters of the biologic function blocks may be modified by a user if desired. In some variations, one or more biologic function templates may be generated comprising a predefined sequence of biological function blocks. One or more biological control parameters of the biologic function templates may be modified by a user if desired. 
     In some variations, a cell processing system may be configured to receive and/or store one or more biologic function (e.g., process) inputs from the user  3604 . For example, a user may select one or more predefined biological function templates. 
     In some variations, a biologic process model (e.g., process definition) may be generated based on the biologic process inputs  3606 . In some variations, a biologic process model may include one or more of enrichment, isolation, MACS selection, FACS selection, activation, genetic modification, gene transfer, transduction, transfection, expansion, formulation (e.g., harvest, pool), cryopreservation, T cell depletion, rest, tissue digestion, washing, irradiation, co-culture, combinations thereof, and the like. 
     In some variations, the biologic process model may be transformed into an instrument execution process model  3608 . For example, each biological function block in the biological process model may correspond to an ordered list of cell processing system operations with corresponding hardware control parameters. The instrument execution process model may comprise the sequence of hardware operations corresponding to the biologic process model. As described herein, the transformation model may comprise one or more constraints. 
     Optionally, in some variations, a cell processing system may be configured to receive and/or store one or more instrument execution process inputs from the user  3610 . For example, a user may modify the transformed instrument execution process model if desired. The user may select specific hardware components to perform certain steps, modify timing parameters, and the like. 
     In some variations, the instrument execution process may be executed to generate the cell product  3612 . For example, the cell processing system at run-time may process the cell product through the system as defined by the instrument execution process model. 
     In some variations, an instrument execution process may be executed  3612 . In some variations, an instrument execution process model may be transformed back into a biologic process model  3614 . This progress of the biologic process model may be output (e.g., displayed) to a user for monitoring. For example, the instrument execution process model may comprise one or more references (e.g., pointers) back to the biological process model so that run-time execution progress may be reported against the biological process model. 
     In some variations, a cell product may be monitored  3616 . For example, the GUIs  5300  and  5400  of respective  FIGS.  53  and  54    illustrate sensor data monitored by the system for a plurality of products. For example, a number of viable cells and a status of a process (e.g., as a function of percentage completion) may be graphically illustrated for a user. 
     In some variations, an electronic record may be generated based on the monitored data  3618 . For example, one or more electronic batch records may be generated in compliance with, for example, 21 CFR regulations. 
       FIG.  55    is a block diagram of an illustrative variation of a manufacturing workflow  5500  comprising a processing platform  5520  (e.g., system  100 , workcell  110 ,  200 ,  201 ) configured to generate a plurality of cell products (e.g., first product, second product, third product) in parallel. For example, a first workflow  5510  for a first product may include a plurality of biologic processes  5512  executed in a predetermined sequence using corresponding elements  5522  (e.g., hardware) of the platform  5520 . Simultaneously, a second workflow  5530  for a second product may execute a predetermined sequence of biologic processes  5530  using corresponding elements  5524  of the platform  5520 . In this manner, hardware resources of the platform  5520  may be efficiently utilized to increase throughput. In some variations, about 2, 3, 4, 5, 6, 7, 8, 9, 10, or more cell products may be manufactured simultaneously on the platform  5520 . The transformation model may include hardware constraints that eliminate scheduling conflicts to ensure that, for example, the same instrument is not used for different products at the same time. 
     Graphical User Interface 
     In some variations, a graphical user interface (GUI) may be configured for designing a process and monitoring a product.  FIG.  37    is a variation of a GUI  3700  comprising an initial process design interface. For example, GUI  3700  may be a process design home page. The GUI  3700  may indicate that no processes have been selected or loaded. A create icon  3710  (e.g., “Create a Process”) may be selectable for a user to begin a process design process. In some variations, one or more of the GUIs described herein may include a search bar. 
       FIG.  38    is a variation of a GUI  3800  relating to creating a process. GUI  3800  may be displayed following selection of the create icon  3710  in  FIG.  37   . For example, GUI  3800  may comprise a process creation window  3810  allowing a user to input and/or select one or more of a process name, process description, and template. In some variations, a user may select from a list of predetermined templates. For example, a user may create a process and save it as a template for later selection. 
       FIG.  39    is a variation of a GUI  3900  comprising relating to an empty process. GUI  3900  may be displayed following confirmation in GUI  3800  that a process is to be created. GUI  3900  may indicate the process name (e.g., Car T Therapy) and may highlight Process Setup icon  3910  and allow process specific parameters to be added such as process reagents and containers, process parameters, and preprocess analytics. GUI  3900  may further comprise an Add Process Reagents and Containers icon  3920 , Add Process Parameters icon  3930 , and Add Preprocess Analytics icon  3940 . Once process setup is completed, one or more process elements may be specified. 
     In some variations, the GUI  3900  may comprise one or more predetermined templates for a set of biological processes (e.g., CAR-T, NK cells, HSC, TIL, etc.). For example, the templates may aid process development and be validated starting points for process development. The templates may be further modified (e.g., customized) based on user requirements. 
       FIG.  40    is a variation of a GUI  4000  comprising relating to adding a reagent and a consumable container. GUI  4000  may be displayed following selection of an Add Process Reagents and Containers icon  3920  in  FIG.  39   . For example, GUI  4000  may comprise an Add Reagent and Container window  4010  enabling a user to input and/or select one or more reagents comprising a reagent kind, manufacturer, part number, volume per unit, required volume and required reagent inputs (e.g., lot number, expiration date, requires container transfer). Add Reagent and Container window  4010  may comprise one or more of an input field, selection box, drop-down selector, and the like. Furthermore, the Add Reagent and Container window  3810  may enable a user to input and/or select one or more consumable containers comprising a manufacturer, part number, volume per unit, and required container inputs (e.g., lot number, expiration date). In some variations, a user may select from a list of predetermined templates. For example, a user may create a process and save it as a template. 
       FIG.  41    is a variation of a GUI  4100  comprising relating to a process parameter. GUI  4100  may be displayed following selection of an Add Process Reagents and Containers icon  3930  in  FIG.  39   . For example, GUI  4100  may comprise an Add Process Parameter window  4110  enabling a user to input and/or select one or more parameters comprising a name, parameter identification, description, data type, units, and parameter type. Add Process Parameter window  4010  may comprise one or more of an input field, selection box, drop-down selector, and the like. In some variations, a user may select from a list of predetermined templates. For example, a user may create a parameter and save it as a template.  FIG.  42    is a variation of a GUI  4200  comprising relating to a patient weight process parameter. For example, GUI  4200  may comprise an Add Process Parameter window  4110  having filled in parameter information including patient weight, data type (e.g., integer), units (e.g., kg), and parameter type (e.g., input). 
       FIG.  43    is a variation of a GUI  4300  relating to a preprocess analytic. GUI  4300  may be displayed following selection of an Add Preprocess analytics icon  3940  in  FIG.  39   . For example, GUI  4300  may comprise an Add Preprocess Analytic window  4310  enabling a user to input and/or select one or more parameters comprising a name, identifier, description, data type, and display group. Add Preprocess Analytic window  4310  may comprise one or more of an input field, selection box, drop-down selector, and the like. In some variations, a user may select from a list of predetermined templates. For example, a user may create a parameter and save it as a template. 
       FIG.  44    is a variation of a GUI  4400  relating to a white blood cell count preprocess analytic. For example, GUI  4400  may comprise an Add Preprocess Analytic window  4410  having filled in preprocess analytic information including name (e.g., CBC White Blood Cell Count), identifier (e.g., CBC-white-blood-cell-count), description (e.g., Number of white blood cells in a sample), data type (e.g., float), and display group (e.g., WBC). 
       FIG.  45    is a variation of a GUI  4500  relating to a process parameter calculation. GUI  4500  may be displayed following selection of an Add Preprocess analytics icon  3940  in  FIG.  39    and selection of a “Calculation” parameter type. For example, GUI  4500  may comprise an Add Preprocess Analytic window  4510  enabling a user to input and/or select one or more parameters comprising a name, identifier, description, data type, display group, units, and parameter type. Furthermore, a Calculation Builder may enable a user to define a formula (e.g., algorithm, equation) to perform a predetermined calculation. For example, a Calculation Builder may comprise one or more of a set of available parameters (e.g., patient weight), constant value, equation, and operands. 
       FIG.  46    is a variation of a GUI  4600  relating to a completed process setup. For example, GUI  4600  may comprise a Process Setup window  4610  having a filled in process reagents, containers, process parameters, and preprocess analytics. Once process setup is completed, one or more process elements may be specified. 
       FIG.  47    is a variation of a GUI  4700  relating to process operations activation settings. GUI  4700  may be displayed following selection of a Process elements icon  4620  in  FIG.  46   . For example, GUI  4700  may comprise an Activation settings window  4710  allowing a user to input and/or select one or more of activation concentration (e.g., mg/L), activation culture time (e.g., seconds), activation temperature (e.g., ° C.), and gas mix mode. In some variations, a user may select from a list of predetermined templates. For example, a user may create a set of activation settings and save it as a template for later selection. 
       FIG.  48    is a variation of a GUI  4800  relating to a filled process operations activation settings. For example, GUI  4800  may comprise an Activation settings window  4810  having filled in Activation setting information. In some variations, a set of gases (e.g., O 2 , N 2 , CO 2 ) and corresponding concentrations may be specified. 
       FIG.  49    is a variation of a GUI  4900  relating to a process operations interface. The GUI  4900  may comprise an Available Operations window  4910  and a Selected Operations window  4920 . The available options for selection may include one or more biologic process inputs as described herein including, but not limited to, enrichment, MACS selection, activation, transduction, transfection, expansion, and inline analysis. One or more of the operations may be selected and dragged into the Selected Operations window  4920 . The selected operations may be reordered within the Selected Operations window  4920 . 
       FIG.  50    is a variation of a GUI  5000  relating to dragging process operations. The GUI  5000  may comprise an Available Operations window  5010 , a Selected Operations window  5020 , and a selected (e.g., dragged) operation  5030  that may be drag and dropped between the Available Operations window  5010  and the Selected Operations window  5020 . The Selected Operations window  5020  may comprise a plurality of selected operations. 
       FIG.  51    is a variation of a GUI  5100  relating to dragging process operations. The GUI  5100  may comprise an Available Operations window  5110 , a Selected Operations window  5120 , and a selected (e.g., dragged) operation  5130  that may be drag and dropped between the Available Operations window  5110  and the Selected Operations window  5120 . The Selected Operations window  5120  may comprise a plurality of selected operations. 
       FIG.  52    is a variation of a GUI  5200  relating to a filled process operations. For example, the GUI  5200  may comprise an Available Operations window  5210  and a Selected Operations window  5220  comprising a completed set of selected operations. In some variations, the settings (e.g., parameters) of each operation may be selectively modified by the user by selecting a corresponding icon (e.g., gear icon). 
       FIGS.  53  and  54    are variations of a GUI  5300  and  5400  relating to product monitoring. The GUI  5300  and  5400  may comprise respective monitoring windows  5310 ,  5410 . For example, the GUI  5310  may monitor a plurality of products  5320  and output one or more product characteristics  5330  including, but not limited to, a summary, process data, online analytics, imaging, process audit logs, process parameters, and process schedule. The monitoring window  5410  may monitor one or more product characteristics of one or more products. For example, the product characteristics may include, but is not limited to, one or more of a process name, identification, process identification, progress, estimated completion, current step, and message. 
       FIG.  77 A  is a flowchart of a method of separating cells  7700  using a CCE module.  FIG.  77 B  is a flowchart of a method of concentrating cells  7710  using a CCE module.  FIG.  77 C  is a flowchart of a method of buffer exchange  7720  using a CCE module. 
       FIG.  78    is a flowchart of a method of separating cells  7800 . A method of counterflow centrifugal elutriation (CCE)  7800  may comprise the step moving a rotor towards a magnet  7802 . The rotor may define a rotational axis. In some variations, moving the rotor comprises advancing and withdrawing the magnet relative to the rotor using a robot. The rotor may be optionally moved towards an illumination source and an optical sensor  7804 . Fluid may be flowed through the rotor  7806 . In some variations, flowing the fluid comprises a flow rate of up to about 150 ml/min while rotating the rotor. The rotor may be magnetically rotated about the rotational axis using the magnet while flowing the fluid through the rotor  7808 . In some variations, rotating the rotor comprises a rotation rate of up to 6,000 RPM. One or more of the fluid and the cells may be optionally illuminated using an illumination source  7810 . Image data of one or more of the fluid and biological material (e.g., particles, cellular material) in the rotor may optionally be generated using an optical sensor  7812 . One or more of a rotation rate of the rotor and a flow rate of the fluid may optionally be selected based at least in part on the image data  7814 . The fluid may be flowed out of the rotor  7816 . The rotor may be moved away from the magnet  7818 . The rotor may optionally be moved away from the illumination source and the optical sensor  7820 . 
       FIG.  79 A  is a flowchart of a closed-loop method of separating cells  7900 .  FIG.  79 B  is a flowchart of a closed-loop method of elutriating cells  7910 .  FIG.  79 C  is a flowchart of a closed-loop method of harvesting cells  7920 . 
       FIG.  80 A  is a flowchart of a method of separating cells  8000 .  FIG.  80 B  is a flowchart of a method of selecting cells  8010 . 
       FIG.  81    is a flowchart of a method of separating cells  8100 . A method of magnetic-activated cell selection (MACS) may comprise labeling cells with a reagent  8102 . In some variations, a magnetic-activated cell selection (MACS) reagent may be incubated with the input cells to label the set of cells with the MACS reagent. In some variations, incubating the MACS reagent comprises a temperature between about 1° C. and about 10° C. The fluid comprising input cells may be flowed into a flow cell  8104 . A set of the cells are labeled with the MACS reagent. In some variations, the magnet array may optionally be moved relative to the flow cell  8106 . In some variations, the set of cells may be magnetically attracted towards a magnet array for a dwell time  8108 . In some variations, the dwell time may be at least about one minute. In some variations, the magnet array may be disposed external to the flow cell. In some variations, a longitudinal axis of the flow cell is perpendicular to ground. In some variations, the flow cell may be absent beads. In some variations, the magnet array may optionally be moved away from the flow cell to facilitate flowing the set of cells out of the flow cell  8110 . The set of cells may be flowed out of the flow cell after the dwell time  8112 . For example, flowing the set of cells out of the flow cell may comprise flowing a gas through the flow cell. The fluid without the set of cells may optionally be flowed out of the flow cell after the dwell time  8114 . 
       FIG.  82 A  is a flowchart of a method of preparing a bioreactor  8200 .  FIG.  82 B  is a flowchart of a method of loading a bioreactor  8210 .  FIG.  82 C  is a flowchart of a method of preparing a bioreactor  8220 .  FIG.  82 D  is a flowchart of a method of calibration for a bioreactor  8230 .  FIG.  82 E  is a flowchart of a method of mixing reagents  8240 .  FIG.  82 F  is a flowchart of a method of mixing reagents  8250 .  FIG.  82 G  is a flowchart of a method of culturing cells  8260 .  FIG.  82 H  is a flowchart of a method of refrigerating cells  8270 .  FIG.  82 I  is a flowchart of a method of taking a sample  8270 .  FIG.  82 J  is a flowchart of a method of culturing cells  8280 .  FIG.  82 K  is a flowchart of a method of media exchange  8290 .  FIG.  82 L  is a flowchart of a method of controlling gas  8292 .  FIG.  82 M  is a flowchart of a method of controlling pH  8294 . 
       FIG.  83    is a flowchart of a method of electroporating cells  8300  using an electroporation module. In some variations, an electroporation module may comprise a fluid conduit configured to receive a first fluid comprising cells and a second fluid, a set of electrodes coupled to the fluid conduit, a pump coupled to the fluid conduit, and a controller comprising a processor and memory. 
     A method of electroporating cells may optionally comprise generating a first signal to introduce the first fluid into the fluid conduit using the pump  8302 . A first fluid comprising cells in a fluid conduit may be received  8304 . In some variations, a second signal may optionally be generated to introduce the second fluid into the fluid conduit such that the second fluid separates the first fluid from a third fluid  8306 . In some variations, the second fluid may comprise a gas or oil. A second fluid in the fluid conduit may be received to separate the first fluid from a third fluid  8308 . An electroporation signal may optionally be generated to electroporate the cells in the fluid conduit using the set of electrodes  8310 . An electroporation signal may be applied to the first fluid to electroporate the cells  8312 . In some variations, the first fluid may be substantially static when applying the electroporation signal. In some variations, a third signal may optionally be generated to introduce the third fluid into the fluid conduit  8314 . The third fluid may be separated from the first fluid by the second fluid. The third fluid may optionally be received in the fluid conduit separated from the first fluid by the second fluid  8316 . 
       FIG.  84    is a flowchart of a method of electroporating cells  8400 . A method of electroporating cells may comprise receiving a first fluid comprising cells in a fluid conduit  8402 . A resistance measurement signal may be applied to the first fluid using a set of electrodes  8404 . A resistance may be measured between the first fluid and the set of electrodes  8406 . An electroporation signal may be applied to the first fluid based on the measured resistance  8408 . In some variations, a second fluid comprising a gas may optionally be received in the fluid conduit before applying the electroporation signal to the fluid. The first fluid may be separated from a third fluid by the second fluid. 
     Fluid Connector 
     A method of transferring fluid using a fluid connector  2700  is described in the flowchart of  FIG.  27    and illustrated schematically in the corresponding steps depicted in  FIGS.  16 B- 16 L . The method  2700  may comprise the step of coupling a sterilant source to a fluid connector  2702 . For example, as shown in  FIG.  16 B , the inlet  1652  and outlet  1654  is coupled to a sterilant source to form a fluid pathway or connection. In some variations, a robot may be configured to couple and decouple the sterilant source to the sterilant port  1650  using a fluid conduit such as a tube. In some variations, the fluid connector  1600  may comprise a plurality of sterilant ports  1650 . As described herein, in some variations, a sterilant port may optionally comprise one or more of a check valve and a particle filter configured to reduce ingress of debris (e.g., after disconnecting the fluid connector). In some variations, the sterilant source may comprise or be coupled to a pump configured to circulate a sterilant through the sterilant port  1650 . In some variations, the sterilant port  1650  may be coupled to one or more of a sterilant source and a fluid source such as a heated air source. For example, a first sterilant port may be configured to couple to a first sterilant source, a second sterilant port may be configured to couple to a second sterilant source, and a third sterilant source may be configured to couple to an air source. 
     The separate portions of the fluid connector  1600  may be brought together and mated. The method  2700  may comprise coupling a first port of a first connector to a second port of a second connector  2704 .  FIG.  16 C  is a schematic diagram of the fluid connector  1600  where the first port  216  and second port  226  are in a coupled configuration (e.g. docked position) that forms a first seal. In some variations, the first connector  1610  and the second connector  1620  may be axially and/or rotationally aligned, and one or more of the connectors  1610 ,  1620  may be translated to couple the connectors  1610 ,  1620  together. In  FIG.  16 C , the first port  1616  and the second port  1626  are each in a closed configuration where the lumens of the respective first connector  1610  and second connector  1620  are sealed from the external environment to maintain sterility of the lumen of the fluid connector  1600 . Furthermore, the first valve  1618  and the second valve  1628  are each in a closed configuration that seals the proximal and distal ends of the connectors from each other. For example, the first valve  1618  in the closed configuration forms a seal (e.g., barrier) between the first proximal end  1612  and the first distal end  1614 . Similarly, the second valve  1628  in the closed configuration forms a seal between the second proximal end  1622  and the second distal end  1624 . In this manner, even if a portion of a connector is contaminated (e.g., first distal end  1614 ), then the other portions of the fluid connector  1600  (e.g., first proximal end  1612 , second connector  1620 ) may remain sterile by virtue of one or more of the port seals and valve seals. 
     The ports may be transitioned to an open configuration such that a distal end of the connectors may be in fluid communication. The method  2700  may comprise transitioning the ports to an open configuration  2706 .  FIG.  16 D  is a schematic diagram of the fluid connector  1600  where the first port  1616  and the second port  1626  are transitioned into an open port configuration to create a shared volume between the valves  1618 ,  1628  that is isolated from the external environment. In  FIG.  16 D , the first valve  1618  and the second valve  1628  are in the closed configuration such that the chamber  1615  defines the volume (e.g., cavity) of the fluid connector  1600  between the first valve  1618  and the second valve  1628 . That is, the first distal end  1614  is in fluid communication with the second distal end  1624 . The ports  1616 ,  1627  may be received and/or held in respective housings  1617 ,  1627  in the closed configuration. In some variations, a robot may be configured to transition the ports  1616 ,  1626  between the open configuration and the closed configuration as described in more detail herein. Additionally or alternatively, the first port  1616  and second port  1626  may automatically transition (e.g., mechanically actuate) from the closed configuration to the open configuration upon mating the first port  1616  to the second port  1626 . 
     In some variations, a fluid may be flowed into the fluid connector to aid sterilization. The method  2700  may comprise flowing fluid (e.g., liquid, gas) into the fluid connector through the sterilant port  2708 .  FIG.  16 E  is a schematic diagram of the fluid connector  1600  where the first chamber  1615  receives a fluid such as air at a predetermined temperature, pressure, and/or humidity. In some variations, one or more portions of the fluid connector  1600  may be dehumidified. For example, pressurized hot air may optionally be circulated within chamber  1615  in order to remove residual fluid, moisture, and raise a temperature of the inner surfaces of the chamber  1615 . The circulated fluid may flow through housings  1617 ,  1627  and over inner and/or outer surfaces of the ports  1616 ,  1626 . 
     Generally, sterilization of a fluid connector may comprise one or more steps of dehumidification, conditioning, decontamination, and aeration (e.g., ventilation). Dehumidification may include removing moisture from the fluid connector. Conditioning may include heating the surfaces of the fluid connector to be decontaminated in order to prevent condensation and aid sterilization. Decontamination may include circulating a sterilant through the fluid connector at a predetermined concentration, rate, and exposure time. Aeration may include removing the sterilant from the fluid connector by circulating a gas (e.g., sterile air) through the fluid connector. 
     A sterilant may be flowed into the fluid connector to sterilize one or more portions of the fluid connector. As described in more detail herein, the sterilant may be, for example, vaporized hydrogen peroxide (VHP) and/or ionized hydrogen peroxide (IHP). The method  2700  may comprise flowing a sterilant into the fluid connector through the sterilant port  2710 .  FIG.  16 F  is a schematic diagram of the fluid connector  1600  where the first chamber  1615  receives the sterilant for a predetermined amount of time (e.g., dwell time). For example, the sterilant may be circulated within the chamber  1615  to sterilize the chamber  1615  of the fluid connector  1600  and any contents disposed therein (e.g., other fluid, biological material). In some variations, the dwell time may be up to about 10 minutes, and between about 1 minute to about 10 minutes, including all ranges and sub-values in-between. In some variations, the vaporized hydrogen peroxide may comprise a concentration between about 50% and about 70%, including all ranges and sub-values in-between. Additionally or alternatively, one or more of the first valve  1618  and the second valve  1628  may be in the open configuration such that the sterilant may be circulated through other portions of the fluid connector  1600  such as first proximal end  1612  and second proximal end  1622 . 
     In some variations, the valves may be translated relative to each other. The method  2700  may comprise translating a first valve relative to a second valve  2712 .  FIG.  16 G  is a schematic diagram of the fluid connector  1600  where the first valve  1618  and second valve  1628  are coupled to each other (e.g., transfer position). The first valve  1618  coupled to the second valve  1628  forms a second seal between the first connector  1610  and the second connector  1620 . 
     The valves may be transitioned to an open configuration such that each end of the fluid connector is in fluid communication. The method  2700  may comprise transitioning the first valve and the second valve from a closed configuration to an open configuration  2714 . In some variations, the first valve and the second valve may comprise a spring-loaded shutoff configured to actuate to the open configuration, thereby allowing for fluidic communication between the sterile lumens of the first connector  1610  and the second connector  1620 . In some variations, each of the first valve  1618  of a first connector  1610  and the second valve  1628  of a second connector  1620  may comprise an engagement feature such as threading configured to facilitate coupling between the first valve  1618  and the second valve  1628 . For example, once the second valve  1628  is translated to contact the first valve  1618 , the engagement features of the valves  1618 ,  1628  may be coupled (e.g., locked) by rotating (e.g., twisting) one of the first valve  1618  and the second valve  1628  to engage their respective threads to each other. Conversely, one of the first valve  1618  and the second valve  1628  may be rotated in the opposite direction to uncouple (e.g., unlock) the first valve  1618  from the second valve  1628 . 
     In some variations, fluid may flow through the fluid connector  2716 .  FIG.  16 H  is a schematic diagram of the fluid connector depicted in  FIG.  16 A  transferring fluid between fluid devices coupled to the fluid connector. For example, the contents (e.g., fluid, biological material) of the first fluid device  1630  and the second fluid device  1640  may be transferred through the fluid connector  1600 . In some variations, one or more of a pump, gravity feed, and the like may aid transfer through the fluid connector  1600 . 
     In some variations, another fluid may be flowed into the fluid connector after fluid transfer between a first fluid device and a second fluid device has been completed. The method  2700  may comprise flowing fluid (e.g., liquid, gas, sterilant) into the fluid connector through the sterilant port  2708  to remove a fluid and/or biological material from the fluid connector  2718 . For example, flowing an inert gas into the fluid connector may reduce drops of liquid from forming when the first connector and second connector are separated. If a sterilant is flowed through the fluid connector, another fluid such as an inert gas may be flowed to aerate the fluid connector and ensure that the sterilant is removed. 
     To begin decoupling the fluid connector, the valves may be translated away from each other. The method  2700  may comprise decoupling the first connector and the second connector  2720 . In some variations, a robot may be configured to manipulate the fluid connector  1600  to transition the valves  1618 ,  1628  to a closed configuration and to translate the valves  1618 ,  1628  away from each other, which may occur simultaneously or independently. The valves  1618 ,  1628  in the closed configuration inhibit fluid flow between the first connector  1610  and the second connector  1620 .  FIG.  16 I  is a schematic diagram of the fluid connector  1600  in a closed valve configuration where the second valve  1628  is translated away from the first valve  1618 . Accordingly, the fluid connector  1600  returns to the docked position. For example, the first valve  1618  and the second valve  1628  may be configured to engage their respective spring-loaded shutoff features to form a seal and reduce drips and/or leaks. In some variations, one or more of a fluid and sterilant may optionally be configured to circulate through the chamber  1615  to remove moisture and/or sterilize the chamber  1615 . 
       FIG.  16 J  is a schematic diagram of the fluid connector  1600  where the first port  1616  and the second port  1626  are transitioned from the open port configuration to the closed port configuration. In some variations, a robot may be configured to manipulate the fluid connector  1600  to transition the ports  1616 ,  1626  to a closed position to seal a lumen of the first connector  1610  from a lumen of the second connector  1620 . In some variations, the ports  1616 ,  1626  may be configured to automatically transition to the closed port configuration when the first valve  1618  separates from the second valve  1628 . 
       FIG.  16 K  is a schematic diagram of the fluid connector  1600  where the second connector  1620  is translated away from the first connector  1610 . In some variations, a robot may be configured to manipulate the fluid connector  1600  to separate the first connector  1610  from the second connector  1620 .  FIG.  16 K  depicts the fluid connector  1600  in a disengaged configuration. 
       FIG.  16 L  is a schematic diagram of the fluid connector  1600  decoupled from the sterilant source. In some variations, a robot may be configured to manipulate the fluid connector  1600  and/or sterilant source to separate the sterilant source  1650  from the sterilant source. In some variations, the sterilant source may be decoupled from the fluid connector  1600  at any point after completing a sterilization process. 
     In some variations, the cartridge comprises one or more Sterile Liquid Transfer Ports (SLTPs) configured for use with a Sterile Liquid Transfer Device (SLTD). In some variations, the SLTP comprises one or more of a cap, a fitting, and a tube fluidically coupled to the fitting. The cap may be removable or pierceable. The fitting may be a push-to-connect fitting (PTCF) or a threaded fitting. PTCF include male-to-female, female-to-male, and androgynous fittings. Illustrative SLTPs and SLTDs suitable for use in the systems of the disclosure may include, for example, AseptiQuik® S connectors, Lynx® CDR connectors, Kleenpak™ connectors, Intact™ connectors, GE LifeScience® ReadyMate connectors. 
     When the disclosure refers to sterile liquid transfer devices, sterile liquid transfer ports, and sterile liquid transfer, the word “sterile” should be understood as a non-limiting description of some variations—an optional feature providing advantages in operation of certain systems and methods of the disclosure. Maintaining sterility is typically desirable for cell processing but may be achieved in various ways, including but not limited to providing sterile reagents, media, cells, and other solutions; sterilizing cartridge(s) and/or cartridge component(s) after loading (preserving the cell product from destruction); and/or operating the system in a sterile enclosure, environment, building, room, or the like. Such operator performed or system performed sterilization steps may make the cartridge or cartridge components sterile and/or preserve the sterility of the cartridge or cartridge components. 
     III. Examples 
       FIGS.  85 - 96 D  are diagrams of other variations of a fluid connector.  FIG.  85    depicts a fluid connector  8500  comprising a first connector  8510  including a first cap  8516  and a second connector  8520  including a second cap  8526 . Fluid connector  8500  may comprise a male connector and a female connector, each with a removable cap and internal self-shutoff valve configured to reduce leaks and drips. The first cap  8516  and the second cap  8526  may be removable from their respective connectors  8510 ,  8520 . 
     In some variations, the fluid connector may be used with a self-sterilizing cap and decap tool  8600  depicted in  FIG.  86   . The cap/decap tool  8600  may be configured to facilitate a sterile environment (e.g., ISO5) where the caps may be removed and the connectors pressed together, first sealing the connectors to each other, and then pressed further to transition the internal self-shutoff valves to an open configuration. 
     In some variations, the tool  8600  may be configured to remove and re-apply caps to the fluid connector  8500 , and to provide a sterile volume for aseptic connection and disconnection of the fluid connector  8500  pair. In some variations, a method of using the tool  8600  may comprise inserting both capped connectors in a first configuration (e.g., where the caps approach the closed shutters) such that the fluid connectors form a seal within a lumen of the decap tool  8600 . In some variations the shutters may be opened to ensure a decap mechanism is retracted. Both capped connectors may be pushed to form a second configuration. The decap mechanism may be engaged to lock into features on the caps. Both capped connectors may be retracted to the first configuration where the caps are retained in the decap mechanism. The decap mechanism may be retracted such that the caps are held within a recess in the tool  8600 . The internal volume may optionally be decontaminated with sterilant or heat. Both connectors may be advanced to connect and perform the transfer. The steps described herein may be sequentially reversed. 
       FIG.  87    depict a coupling sequence for a self-sealing fluid connector  8700  comprising a first connector  8710  and a second connector  8720 . The fluid connector  8700  may be configured to reduce leaks and drips and may facilitate smoother fluid flow path by removing spring elements from contact with fluid. 
       FIG.  88    depict a coupling sequence for a self-sealing fluid connector  8800  comprising a first connector  8810  and a second connector  8820 . 
     In some variations, a fluid connector may transfer fluids in a sterile manner using a retractable needle.  FIG.  89    depicts a fluid connector  8900  comprising a first connector  8910  and a second connector  8920 . The first connector  8910  may comprise a first cap  8916  configured to removably couple to a distal end of the first connector  8910 . The first connector  8910  may comprise a first elastomeric member  8970  (e.g., sealing septum) and a first thermal member  8972  (e.g., thermally resealable septum) disposed at a distal end of the first connector  8910 . The first connector  8910  may further comprise a needle  8990  and a spring  8992  coupled to the first elastomeric member  8970  and the needle  8990 . The second connector  8920  may comprise a second cap  8926  configured to removably couple to a distal end of the second connector  8920 . The second connector  8920  may comprise a second elastomeric member  8980  (e.g., sealing septum) and a second thermal member  8982  (e.g., thermally resealable septum) disposed at a distal end of the second connector  8920 . 
     In some variations, the needle  8990  may be advanced through each of the first elastomeric member  8970 , first thermal member  8972 , second thermal member  8982 , and second elastomeric member  8980  to form a fluid pathway between the first connector  8910  and the second connector  8920 . Fluid may flow through the first connector  8910  and into the second connector  8920  via a lumen of needle  8990 . Each of the elastomeric members  8970 ,  8980  and thermal members  8972 ,  8982  may seal once the needle  8990  is withdrawn from a distal end of the first connector  8910 . For example, the thermal member  8972 ,  8982  may be configured to thermally seal at a predetermined temperature and the elastomeric members  8970 ,  8980  may self-seal once the needle  8990  has been withdrawn. In some variations, the fluid connector  8900  may be thermally decontaminated and resealed after fluid transfer. For example, the fluid connector  8900  (e.g., thermal members  8972 ,  8982 ) may be heated using one or more of a laser, contact heating, heated air, combinations thereof, and the like. 
     In some variations, a fluid connector may comprise a port comprising an actuator configured to transition the port between a closed port configuration and an open port configuration. In some variations, the actuator may comprise a spring such as an external spring, a rotary spring, and a linear spring, as described in more detail with respect to  FIGS.  90 A- 96 D . 
       FIGS.  90 A- 90 C  depict a fluid connector having an external spring actuator.  FIG.  90 A  is a side view,  FIG.  90 B  is a perspective view, and  FIG.  90 C  is a cross-sectional side view of a fluid connector  9000  comprising a first connector  9010  and second connector  9020 . The first connector  9010  may comprise a first port  9016  comprising a first spring  9036 , and the second connector  9020  may comprise a second port  9026  comprising a second spring  9046 . The springs  9036 ,  9046  may be configured to actuate respective ports  9016 ,  9026  between a closed port configuration and an open port configuration. Although not shown in  FIG.  90 C , springs  9036 ,  9046  may be coupled in an extended configuration to the pin in the open port configuration. 
       FIGS.  91 A- 91 F  depict a fluid connector having a linear spring actuator.  FIG.  91 A  is a side view,  FIG.  91 B  is a perspective view, and  FIG.  91 C  is a cross-sectional side view of the fluid connector  9100  in an open port configuration. The fluid connector  9100  may comprise a first connector  9110  and second connector  9120 . The first connector  9110  may comprise a first port  9116  comprising a first spring  9136 , and the second connector  9120  may comprise a second port  9126  comprising a second spring  9146 . The springs  9136 ,  9146  may be configured to actuate respective ports  9116 ,  9126  between a closed port configuration and an open port configuration.  FIG.  91 D  is a side view,  FIG.  91 E  is a perspective view, and  FIG.  91 F  is a cross-sectional side view of the fluid connector  9100  in a closed configuration. 
       FIGS.  92 A- 92 D  depict a fluid connector having a rotary spring actuator.  FIG.  92 A  is a side view,  FIG.  92 B  is a transparent side view,  FIG.  92 C  is a perspective view, and  FIG.  92 D  is a cross-sectional side view of a fluid connector  9200  comprising a first connector  9210  and second connector  9220 . The first connector  9210  may comprise a first port  9216  comprising a first spring  9236 , and the second connector  9220  may comprise a second port  9226  comprising a second spring  9246 . The springs  9236 ,  9246  may be configured to actuate respective ports  9216 ,  9226  between a closed port configuration and an open port configuration.  FIG.  92 B  shows the ports  9216 ,  9226  in an open port configuration and  FIG.  92 D  shows the ports  9216 ,  9226  in a closed port configuration. 
       FIGS.  93 A- 94 B  depict fluid connectors having ports enclosed within a housing (e.g., enclosure).  FIG.  93 A  is a perspective view and  FIG.  93 B  is a transparent perspective view of a fluid connector  9300  comprising a first connector  9310  having a first housing  9338  and first actuator  9336 , and a second connector  9320  having a second housing  9348  and a second actuator  9346 .  FIG.  93 B  shows a first port  9316  enclosed within first  9338  housing. The first port  9316  is coupled to the first actuator  9336  configured to transition the first port  9316  between an open port configuration (shown in  FIG.  93 B ) and a closed port configuration. 
       FIG.  94 A  is a perspective and  FIG.  94 B  is a transparent perspective view of a fluid connector  9400  comprising a first connector  9410  having a first housing  9438  and a first actuator  9436 , and a second connector  9420  having a second housing  9448  and a second actuator  9446 .  FIG.  94 B  shows a first port  9416  enclosed within first  9438  housing. The first actuator  9436  coupled to the first port  9416  may be configured to transition the first port  9416  between an open port configuration (shown in  FIG.  94 B ) and a closed port configuration. 
       FIG.  95 A  is a perspective view and  FIG.  95 B  is a transparent perspective view of a fluid connector  9500  comprising a first connector  9510  having a first housing  9538 , first port  9516 , and a first actuator  9536 . A second connector  9520  may comprise a second housing  9548 , second port  9526 , and a second actuator  9546 .  FIG.  95 B  shows the first port  9516  and the second port  9526  each in an open port configuration. For example, the first actuator  9536  coupled to the first port  9516  may be configured to transition the first port  9516  between an open port configuration and a closed port configuration.  FIG.  95 C  is a detailed side view of the first port  9516  and first actuator  9536  in an open port configuration, and  FIG.  95 D  is a detailed side view of the first port  9516  and first actuator  9536  in a closed port configuration. 
       FIG.  97 A  is a perspective view of a MACS module.  FIG.  97 B  is a cross-sectional perspective view of a MACS module.  FIG.  97 C  is a cross-sectional side view of a MACS module. 
     As used herein, sterile should be understood as a non-limiting description of some variations, an optional feature providing advantages in operation of certain systems and methods of the disclosure. Maintaining sterility is typically desirable for cell processing but may be achieved in various ways, including but not limited to providing sterile reagents, media, cells, and other solutions; sterilizing cartridge(s) and/or cartridge component(s) after loading (preserving the cell product from destruction); and/or operating the system in a sterile enclosure, environment, building, room, or the like. Such user or system performed sterilization steps may make the cartridge or cartridge components sterile and/or preserve the sterility of the cartridge or cartridge components. 
     All references cited are herein incorporated by reference in their entirety. 
     As used herein, the singular forms “a”, “an” and “the” include plural referents unless the context clearly dictates otherwise. “And” as used herein is interchangeably used with “or” unless expressly stated otherwise. 
     All embodiments of any aspect of the disclosure can be used in combination, unless the context clearly dictates otherwise. 
     Unless the context clearly requires otherwise, throughout the description and the claims, the words ‘comprise’, ‘comprising’, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”. Words using the singular or plural number also include the plural and singular number, respectively. Additionally, the words “herein,” “above,” “below,” and words of similar import, when used in this application, shall refer to this application as a whole and not to any particular portions of the application. 
     While embodiments of the present invention have been shown and described herein, those skilled in the art will understand that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.