Processing instrument for cellular therapy manufacturing

Abstract

The present invention is directed to a system for automated / semi -automated cell processing functions associated with one or more bioreactors during cell culture procedures, such as cell seeding, cell washing and cell harvesting.

Description

[0001] PROCESSING INSTRUMENT FOR CELLULAR THERAPY MANUFACTURING

[0002] Cross-Reference to Related Applications

[0003] This application claims priority to, and the benefit of, U.S. Provisional Application No. 63 / 599,068, filed November 15, 2023, the content of which is incorporated by reference herein in its entirety.

[0004] Technical Field

[0005] The present invention generally relates to systems and devices for cell culturing, and, more particularly, to a system including a mechanical assembly for supporting one or more bioreactors and assisting in providing automated or semi-automated cell processing functions within the one or more bioreactors during cell culture procedures.

[0006] Background

[0007] Many areas of clinical research require the isolation, preparation, and expansion of cells. For example, cell and gene therapies developed using specially engineered cells, genes, and tissues can be used for personalized and precise therapies in preventing, diagnosing, treating and / or potentially curing diseases at their source. Autologous and allogeneic cell and gene therapy workflows typically involve isolating cells from an individual, purifying and / or engineering those isolated cells, expanding and / or concentrating the isolated cells, and infusing such cells into the patient.

[0008] To ensure efficacy of the final product, it is imperative to have robust, standardized, and efficient processing in cell and gene therapy manufacturing workflows to provide consistent production of the cell therapy product. Cell viability and purity of the cell population are critical issues in the manufacturing of cell therapy products. Reduced cell viability and recovery may result from prolonged time in transit, tissue digestion, cell manipulation, operator variability, gene transduction, cryopreservation, and cell thawing. The minimal processing capacity and percentage of viable cells recovered are particularly important factors for autologous and allogeneic products because poor cell recovery may lead to a failed manufacturing run. Thus, a challenge of future success in commercializing cell therapy products is establishing a scalable manufacturing technology that can reliably reproduce the yield and quality of cell-derived products generated from small-scale R&.D methods.

[0009] Tissue cells are known to be sensitive to mechanical stresses imposed on them by agitation in bioreactors. For example, common challenges in pluripotent stem cells (PSC) aggregate formation include the influence of variables such as cell proliferation rate, cell-cell adhesion strength, cell packing density, agitation rate, and the hydrodynamic environment inside a bioreactor. Further, workflow operations typically require prolonged handling and processing times with many operators, which can negatively impact cell quality, process reliability, and cost-effectiveness. Any negative impact on cell quality (e.g., shear stress) can be magnified downstream when cells are subjected to prolonged processing. Thus, there is a need for optimizing these variables at each step in manufacturing workflows to ensure consistent production of the cell therapy product.

[0010] Summary

[0011] The present invention includes a system for providing automated / semi-automated cell processing functions associated with one or more bioreactors during cell culture procedures. More specifically, the system of the present invention includes a mechanical assembly configured to support one or more bioreactors at a time and subsequently move the one or more bioreactors in a controlled and precise manner so as to achieve a desired cell processing function, including, but not limited to, cell seeding, cell washing, and cell harvesting.

[0012] The assembly is configured to automate and replicate manual movements used during cell culture process. In particular, the assembly incorporates mechanisms for agitating the bioreactors in a specific oscillatory motion, such that the bioreactors may be gently or vigorously agitated depending on the cell culture processing needs. The assembly also incorporates rotating and tilting mechanisms to orient the bioreactors in any position required for cell processing. For example, such mechanisms allow for tilting functions so as to tilt the bioreactors to various positions to allow for filling and draining the bioreactor(s).

[0013] The assembly is configured to operate in an automated or semi-automated fashion, thereby allowing for certain cell processing functions to occur, including, but not limited to, standardized filling, cell seeding, medium exchange, mixing, agitation, concentrating, washing, and harvesting functions of cell processing. The automated / semi-automated nature of the cell processing functions provided by the system of the invention removes user-variability and increases scalability. Furthermore, the assembly is able to handle a single cell bioreactor at a time or can handle multiple cell bioreactors in a completely closed-system, thereby enabling a user to perform cell processing functions outside of the biosafety cabinet while maintaining sterility. Additionally, the ability to process multiple bioreactors at once further reduces the overall technician time required for cell processing.

[0014] As an example, the harvest function of the system allows for removal of adherent cells via mechanical agitation which is particularly useful for strongly adherent cells where use of enzymatic reagents is not permissible. The system may be used for any type of adherent cell culture and is protocol agnostic because all steps may be customized to meet specific user / cell culture protocol requirements.

[0015] Experiments comparing manual cell processing versus using the system\ of the invention show comparability in yield, cell viability, phenotype, and functionality. Output cells are of equal or better quality (as measured by viability, yield, surface marker phenotype and functionality) compared to equivalent manual process. Thus, by mechanizing bioreactor manipulation, the system of the invention provides for optimizing and standardizing variables directly influencing cell viability and the quality of the cell therapy product. Accordingly, the system of the invention provides for an improved cell therapy product.

[0016] In one aspect, the invention provides an assembly for supporting one or more bioreactors and providing automated or semi -automated cell processing functions associated with the one or more bioreactors. The assembly includes a base and supporting frame, and a moveable support structure having a first portion coupled to the supporting frame and a second portion coupled to the base. Further, the assembly includes a platform rotatably coupled to the support structure and configured to removably receive and retain one or more bioreactors thereto, and a controller configured to control movement of the platform to thereby control movement of one or more bioreactors supported by the platform based, at least in part, on one or more predefined cell processing protocols.

[0017] In some embodiments, the platform is rotatably coupled to the support structure by way of a pair of shafts. Each shaft is positioned on a respective opposing end of the platform and coupled to one of a first end and a second end of the support structure via a swivel joint, in some embodiments. Thus, the pair of shafts cooperatively define an axis of rotation of the platform. Further, in some embodiments, the first end of the support structure comprises a drive motor operably coupled to the controller. In some embodiments, the drive motor is directly coupled to a first one of the pair of shafts of the platform via a flange such that actuation of the drive motor, via input from the controller, causes rotation of the platform about the axis of rotation of the platform.

[0018] In particular embodiments, one end of the support structure is rotatably coupled to the supporting frame via a hinged connection defining an axis of rotation of the support structure. Further, in some embodiments, an underside of the support structure is coupled to a linear actuator via a pair of shafts. The linear actuator may be operably coupled to the controller such that actuation of the linear actuator, via input from the controller, causes the pair of shafts to impart force upon the underside of the support structure and causes rotation of the support structure about the axis of rotation of the support structure. This causes the platform to move between a horizontal orientation and a tilted orientation.

[0019] Further, in some embodiments, the linear actuator comprises a thread rod coupled to a stepper motor and a carriage positioned on the thread rod, the carriage coupled to the pair of shafts via a pair of shaft supports connected to swivel joints positioned at a first end of the shafts and a rotary shaft inserted horizontally through the swivel joints and the shaft supports.

[0020] In some embodiments, the underside of the support structure is coupled to the pair of shafts via a pair of shaft supports connected to swivel joints positioned at a second end of the shafts and a rotary shaft inserted horizontally through the swivel joints and the shaft supports.

[0021] When the carriage is in a home position on the thread rod, the support structure is maintained in a substantially horizontal position thereby maintaining the platform, and any bioreactors contained therein, in a corresponding horizontal position, in some embodiments.

[0022] In some embodiments, the controller is configured to align the platform in a horizontal first position and rotate the platform around the axis of rotation of the platform in a range of from 0 degrees to 360 degrees as compared to the horizontal first position. In particular embodiments, the rotation around the axis of rotation of the platform comprises oscillatory movement of the platform. In some embodiments, actuation of the stepper motor, via input from the controller, moves the carriage from the home position along the thread rod to define a tilt axis of rotation of the platform such that the platform is operable to move between a position of axial rotation around a horizontal axis of rotation of the platform to a position of axial rotation around the tilt axis of rotation of the platform. Further, the position of axial rotation around the tilt axis of rotation of the platform provides for positioning the one or more bioreactors for filling and / or draining, in some embodiments. For example, in some embodiments, the controller is operable to cause the platform to rotate around the horizontal axis of rotation and the tilt access of rotation at one or more of varying speed, varying degree of rotation, varying oscillatory motion, and varying tilt angle to provide for agitating, via a shaking and / or an oscillating motion, the contents of the one or more bioreactors.

[0023] In some embodiments, the controller interfaces with a control system architecture comprising at least one of a computer, a processor, a network, and / or a graphical user interface (GUI) for defining one or more cell processing protocols. Further, the assembly is configured to perform one or more automated or semi-automated cell processing functions in a closed and aseptic environment, in some embodiments. For example, the cell processing functions comprises at least one of cell seeding, cell washing, and cell harvesting, in some embodiments. Further, in some embodiments, the assembly is configured for removal of adherent cells via mechanical agitation with or without the use of enzymatic reagents.

[0024] In particular embodiments, the platform comprises a frame configured to hold a plurality of bioreactors in a stacked position such that an inlet port and / or an outlet port associated with each bioreactor is accessible. Further, in some embodiments, the assembly comprises one or more reservoirs operably connected to one or more pinch valves and one or more pumps configured for providing fluid flow into and out of the one or more reservoirs and one or more bioreactors retained on the platform. For example, the assembly is configured as a closed system for aseptic transfer of fluids to and from the one or more bioreactors, in some embodiments.

[0025] In some embodiments, the supporting frame comprises a plurality of supports configured for receiving and retaining one or more of tubing, a fluid reservoir, a fluid collection container, a bioprocessing bag, a pump, and a motor. Brief Description of the Drawings

[0026] FIG. 1 A is perspective view illustrating one embodiment of a system of the invention, illustrating an overall assembly in which a support structure defines a tilt axis of rotation of the platform, with the platform rotated around the tilt axis of rotation.

[0027] FIG. IB is a transparent perspective view illustrating one embodiment of a system of the invention in which the support structure and platform are each in a home position.

[0028] FIG. 1C is a perspective view illustrating one embodiment of a system of the invention.

[0029] FIG. ID is a magnified view of the fluidic connections of the embodiment of FIG. 1C, via tubing connected with one or more bioreactors retained upon the platform and one or more bags or reservoirs of the system.

[0030] FIG. 2A is a perspective view illustrating one embodiment of a platform of an assembly of the invention in which four bioreactors are received and retained upon the platform.

[0031] FIG. 2B is a perspective side view illustrating one embodiment of a platform of an assembly of the invention showing the pair of shafts defining an axis of rotation of the platform.

[0032] FIG. 2C illustrates one embodiment of a platform of an assembly of the invention showing three bioreactors each retained upon the platform via an adjustable strap.

[0033] FIG. 3 is a shaded perspective side view illustrating one embodiment of a system for automated cell processing functions of the invention.

[0034] FIG. 4 is a shaded perspective magnified view illustrating the support structure with the platform attached to the support structure on opposing ends of the support structure by way of a pair of shafts positioned on opposing ends of the platform, according to one embodiment of the invention.

[0035] FIG. 5 is a solid line perspective side view illustrating one embodiment of an assembly of the invention.

[0036] FIG. 6 illustrates a magnified perspective view of a first portion and a second portion of the support structure according to some embodiments of the invention.

[0037] FIG. 7 illustrates a perspective view of the support structure with the pair of shafts attached to the underside of the support structure according to the embodiment shown in FIG. 5.

[0038] FIG. 8 is a perspective view illustrating the carriage in a home position according to some embodiments of the invention. FIG. 9 is a shaded perspective view illustrating one embodiment of a system of the invention in which the support structure is in a home position.

[0039] FIG. 10 is a shaded perspective view illustrating the embodiment of the system of the invention shown in FIG. 9 in which the platform is rotated around the axis of rotation of the platform on a tilt axis of rotation defined by movement the axis of rotation of the support structure.

[0040] FIG. 11 illustrates a side perspective view of an assembly according to one embodiment of the invention, in which the support structure is actuated to rotate around an axis of rotation of the support structure to thereby define a tilt axis of rotation of the platform.

[0041] FIG. 12 illustrates an end perspective view of the assembly according to FIG. 11.

[0042] FIG. 13 illustrates a top perspective view of the assembly according to FIG. 11.

[0043] FIG. 14 illustrates a second end and side perspective view of the assembly according to FIG. 11.

[0044] FIG. 15 illustrates a bottom perspective view of the assembly according to FIG. 11.

[0045] FIG. 16 illustrates another embodiment of the assembly for supporting one or more bioreactors and providing automated or semi-automated cell processing functions associated with the one or more bioreactors.

[0046] FIG. 17 illustrates a tilt axis of rotation of the platform as defined by movement of the carriage along the thread rod according to the embodiment of the invention shown in FIG. 16.

[0047] FIG. 18 illustrates the platform at a tilt and at a position of axial rotation around the tilt axis such that the bioreactor is positioned for draining according to the embodiment of the invention shown in FIG. 16.

[0048] FIGs. 19A-19C illustrate Assessment of DC numbers and viability on sDP by trypan blue at day 7.

[0049] FIGs. 20A-20C illustrate Assessment of DC numbers and viability on sDP by trypan blue at day 8.

[0050] FIG. 21 illustrates an identity panel comparison of the study.

[0051] FIG. 22 illustrates an purity characterization panel of the study.

[0052] FIG. 23 illustrates assessment of sDP purity and identity, specifically the sDP Identity on Day 8. FIGs. 24A-24C illustrate sDP impurity characterization on Day 8.

[0053] FIG. 25A illustrates potency assessment.

[0054] FIG. 25B illustrates results for positive and negative controls.

[0055] FIG. 25C illustrates graphs of Assessment of sDP potency on Day 8.

[0056] FIG. 26 is a table of data which compares various characteristics of harvested cells on days 7 and 8 using the systems / devices and methods of the invention versus manual methods for four separate runs.

[0057] FIG. 27 is a table of data for run CS 26 with results measured on day 7 and 8 which compares various characteristics of harvested cells using one embodiment of systems of the invention for harvest versus manual methods for harvest.

[0058] FIG. 28 is a table of data for run CS 27 with results measured on day 7 and 8 which compares various characteristics of harvested cells using one embodiment of the invention for harvest versus manual methods for harvest.

[0059] FIG. 29 illustrates DP identity for all events, cells, live, and singlets at day 7 and 8 for manual harvest.

[0060] FIG. 30 illustrates DP identity for all events, cells, live, and singlets at day 7 and 8 for manual harvest.

[0061] FIG. 31 is a table of data for run CS 28 with results measured on day 7 and 8 which compares various characteristics of harvested cells using one embodiment of the invention for oscillation harvest versus manual methods for harvest.

[0062] FIGs. 32A and 32B are tables including data of runs CS19-CS-25 measured on Days 7 and 8 which compares various characteristics of harvested cells using one embodiment of the invention for oscillation harvest versus manual methods for harvest.

[0063] FIGs. 33 and 34 illustrate a functional allogenic assay of run no. CS 23 of D8 DC + CS 22 T cells + CD3.

[0064] Detailed Description

[0065] The present invention is directed to a system for providing cell processing functions during cell culture in a closed, aseptic manner. In particular, the present invention includes a system for providing automated / semi-automated cell processing functions associated with one or more bioreactors during cell culture procedures. The system includes a mechanical assembly configured to support one or more bioreactors at a time and subsequently move the one or more bioreactors in a controlled and precise manner so as to achieve a desired cell processing function, including, but not limited to, cell seeding, cell washing, and cell harvesting.

[0066] The assembly is configured to automate and replicate manual movements used during cell culture. In particular, the assembly incorporates mechanisms for agitating the bioreactors in a specific oscillatory motion, such that the bioreactors may be gently or vigorously agitated depending on the cell culture processing needs. The assembly also incorporates rotating and tilting mechanisms to orient the bioreactors in any position required for cell processing. For example, such mechanisms allow for tilting functions so as to tilt the bioreactors to various positions to allow for filling and draining the bioreactor(s).

[0067] The assembly is configured to operate in an automated or semi-automated fashion, thereby allowing for certain cell processing functions to occur, including, but not limited to, standardized filling, cell seeding, medium exchange, mixing, agitation, concentrating, washing, and harvesting functions of cell processing. The automated / semi-automated nature of the cell processing functions provided by the system of the invention removes user-variability and increases scalability. Furthermore, the assembly is able to handle a single cell bioreactor at a time or can handle multiple cell bioreactors, thereby enabling a user to perform cell processing functions outside of the biosafety cabinet while maintaining sterility. The automated / semi- automated system of the invention reduce the overall technician time required for cell processing due to the ability to process multiple bioreactors simultaneously or in concert with one another. The automation also removes user-variability and increases scalability. Output cells generated using the system of the invention are of equal or better quality as measured by viability, yield, surface marker phenotype, and functionality.

[0068] Overview

[0069] The invention provides a system for automating / semi-automating and standardizing cell processing functions during cell culture. The invention addresses the limitations of conventional platforms for generating a large number of cells, such as labor and materials costs, and the prolonged handling and processing times, with many operators, which can negatively impact cell quality, process reliability, and cost-effectiveness.

[0070] The system addresses the fact that various types of therapeutic cells may be sensitive to a bioreactor mixing characteristics in different ways. In particular, the hydrodynamic conditions or mixing environment within a bioreactor may significantly impact the biological performance of cells and the resulting efficiency of cell culture processes such as expansion and differentiation. For example, anchorage-dependent cells grown on microcarriers or as aggregates are more sensitive to hydrodynamic shear stress than those grown as single cells in suspension bioreactors. The system addresses these challenges and may be used for any cell culture or protocol. Thus, the system may be used for any cell processing protocol and thus are protocol agnostic.

[0071] In particular, certain embodiments the system may be used for cell processing protocols related to dendritic cell generation. The system may be used for other types of adherent and / or non-adherent cell culture processing protocols. The system may be used for cell processing protocols used to culture cell types that are cultured as aggregates, such as pluripotent stem cells (PSCs), neural stem cells, and / or mammary epithelial stem cells. The system may also be used for cell processing protocols related to the culture cells typically grown as single cells or loose clumps, such as hematopoietic stem / progenitor cells and / or immune cells like T cells and natural killer (NK) cells.

[0072] The system provides for automating / semi-automating and optimizing cell processing function such as expansion and differentiation, gas and medium exchanges, and cell harvesting for large-scale use. Thus, the system provides for a robust and scalable manufacturing process for the consistent production of allogeneic and autologous cell therapy products for patients, and reduce the percentage of cells that may be lost during various downstream processes such as harvest, wash, concentration, formulation, and fill / finish steps.

[0073] System for Automated Cell Processing Functions

[0074] The invention provides a system for automated or semi-automated cell processing functions associated with the one or more bioreactors.

[0075] FIG. 1A is perspective view illustrating one embodiment of a system 100 of the invention. As is described in more detail herein, FIG. 1A shows the support structure as defining a tilt axis of rotation of the platform, and the platform is rotated around the tilt axis of rotation.

[0076] FIG. IB is a transparent perspective outline view illustrating of the embodiment illustrated in FIG. 1A of a system 100 of the invention in which the support structure and platform are each in a home position, as is described in more detail herein.

[0077] In one aspect, the invention provides an assembly 100 for supporting one or more bioreactors 103 and providing automated or semi-automated cell processing functions associated with the one or more bioreactors. The assembly includes a base 105 and supporting frame 107, and a moveable support structure 109 having a first portion 109a coupled to the supporting frame and a second portion 109b coupled to the base 105. Thus, the support structure may have a rigid frame such that the assembly 100 includes a platform 111 rotatably coupled to the support structure 109.

[0078] The platform may be configured to removably receive and retain one or more bioreactors 103 thereto. The assembly, also interchangeably referred to as a system or device within the disclosure, includes a controller configured to control movement of the platform 111 to thereby control movement of one or more bioreactors 103 supported by the platform 111 base, at least in part, on one or more predefined cell processing protocols.

[0079] The platform 111 may be configured to retain one or more bioreactors of any size or shape. The platform may be configured to receive and retain one or more bioreactors 103 that are the same size and / or shape. The platform may be configured to receive and retain one or more bioreactors 103 that are different sizes and / or shapes. For example, the one or more bioreactors may be any bioreactor sized or shaped for a particular cell processing function, for example, Corning® CellSTACK®, Nunc™ Cell Factory™, or T-flasks. The one or more bioreactors 103 retained on the platform 111 may be detached and reattached to the assembly to allow for various functions or purposes such as incubation of the bioreactor in an incubator, or refrigeration of the bioreactor in a refrigerator. In some embodiments, the assembly may be sized and configured to fit within an incubator.

[0080] In some embodiments, the platform comprises a rigid frame configured to hold a plurality of bioreactors in a stacked position such that an inlet port and / or an outlet port associated with each bioreactors may be accessible. FIG. 1C is a perspective view illustrating one embodiment of a system 200 of the invention, in which three bioreactors 103 are retained upon the platform. The retaining mechanism may be, for example, one or more adjustable straps, clips, latches, rings, screws, slits, or slots designed to receive and retain one or more bioreactors of varying sizes and shapes.

[0081] The one or more bioreactors 103 may be connected via tubing, for example, weldable tubing, to one or more bags 113 to create a closed system for aseptic transfer of cells and fluids to and from the bioreactor, for example, from a bag 113 into the bioreactor and from the bioreactor into a bag 113. In some embodiments, valves, for example one or more pinch valves, may be used for fluid management. Fluid management may be achieved through any combination of pumps and valves as is known to persons skilled in the art. For example, fluid flow through the tubing may be controlled via one or more mechanical valves, such as a pinch valves 140 or other electronically controlled valve, as illustrated in FIGs. 1C and ID.

[0082] FIG. ID is a magnified view of the fluidic connections of the embodiment of FIG. 1C. Fluidic connections may be achieved via tubing connections, such as sterile tube welding, with one or more bioreactors retained upon the platform and one or more bags or reservoirs of the system.

[0083] The assembly may be used with, for example, any number of bioreactors. In non-limiting examples, the platform 111 may be configured to receive one, two, three, four, five, greater than five or greater than 10 bioreactors. The assembly 100 may be configured to scale up to receive and retain and any number of platforms 111 and / or bioreactors 103 to meet cell processing needs. As illustrated in FIGs. 1 A and IB, three bioreactors 103 may be received and retained on the platform 111.

[0084] FIG. 2A is a perspective view illustrating one embodiment of a platform 211 of an assembly of the invention in which four bioreactors 203 are received and retained upon the platform 211. The platform may comprise a rigid frame to hold one or more bioreactors, and a locking or latching mechanism to secure the bioreactor in place upon the platform. The platform may be configured such that one or more bioreactors retained upon the platform are securely held with substantially no movement on the platform. In this way, movement of the platform controls the movement of the bioreactors retained there upon. Referring to FIG. IB, the platform 111, in some embodiments of the assembly, may be rotatably coupled to the support structure 109 by way of a pair of shafts 115a, 115b, wherein each shaft 115a, 115b may be positioned on a respective opposing end of the platform 111 and coupled to one of a first end and a second end of the support structure 109 via a swivel joint, wherein the pair of shafts 115a, 115b cooperatively define an axis of rotation of the platform.

[0085] FIG. 2B is a perspective side view illustrating one embodiment of a platform 211 of an assembly of the invention showing the pair of shafts 215a, 215b defining an axis of rotation X of the platform 211. In non-limiting examples, and as disclosed in more detail herein, the platform 211 may rotate around the axis of rotation X from a horizontal starting point, referred to in some embodiments as a home position. The platform may rotate around the axis of rotation of the platform from any of 0 degrees to 360 degrees in full or partial rotation. The platform may rotate in a same direction. The platform may rotate in a counter direction. The platform may change rotation directions, and may rotate in an oscillatory motion. As disclosed in more detail herein, the support structure may be operable to rotate about an axis of rotation as well, such that the support structure defines a tilt access of rotation of the platform. Thus, the platform may rotate from 0 to 360 degrees around an axis of rotation that is horizontal, and / or from 0 to 360 degrees about a tilt axis of rotation defined by the axis of rotation of the support structure. The platform may rotate around the axis of rotation at one or more of varying speed, varying degree of rotation, varying oscillatory motion, and varying tilt angle to provide for agitating, shaking, and / or mixing the contents of the one or more bioreactors. The platform may rotate on the axis of rotation via a shaking and / or an oscillating motion. Accordingly, the movement of the platform may be any of rotating, shaking, agitating, mixing, and / or oscillating according to the needs of a defined cell processing protocol and / or culturing protocol. In non-limiting examples, the oscillating movement may be a gentle or vigorous back and forth movement of the one or more bioreactors on a horizontal axis of rotation and / or a tilt axis of rotation.

[0086] FIG. 2C illustrates one embodiment of a platform of an assembly of the invention showing three bioreactors each retained upon the platform via an adjustable strap.

[0087] FIG. 3 is a shaded perspective side view illustrating one embodiment of a system for automated / semi-automated cell processing functions according to the invention. As noted above, the system may include an assembly 300 for supporting one or more bioreactors 303 and providing automated or semi-automated cell processing functions associated with the one or more bioreactors 303. The assembly 300, may include a base 305 and supporting frame 307. The assembly may include a moveable support structure 309 having a first portion coupled to the supporting frame and a second portion coupled to the base. The assembly may include a platform 311 rotatably coupled to the support structure 309 and configured to removably receive and retain one or more bioreactors 303 thereto. Thus, the moveable support structure may have a rigid frame and may be operable to move the platform from a horizontal position to a tilted position above or below a horizontal position. The system may include a controller configured to control movement of the platform 311 to thereby control movement of one or more bioreactors 303 supported by the platform 311 based, at least in part, on one or more predefined cell processing protocols.

[0088] As disclosed herein, the platform 311 may be rotatably coupled to the support structure 309 by way of a pair of shafts 315a, 315b, wherein each shaft 315a, 315b may be positioned on a respective opposing end of the platform 311 and coupled to one of a first end and a second end of the support structure 309 via a swivel joint, such that the pair of shafts 315a, 315b cooperatively define an axis of rotation of the platform 311. Thus, the pair of shafts, in cooperation with the swivel j oints, allow the platform to rotate about a horizontal axis of rotation or a tilt axis of rotation. The rotatable coupling between the platform and the support structure may be any connection as is known to persons skilled in the art that provides for rotation of the platform around the axis of rotation. For example, the rotatable coupling may be a direct connection with a servo motor via a swivel joint, via u-joint and swivel joint, or other type of rotatable joint.

[0089] FIG. 4 is a shaded perspective magnified view illustrating the support structure 309 with the platform 311 attached to the support structure 309 on opposing ends of the support structure 309 by way of a pair of shafts 315a, 315b positioned on opposing ends of the platform 311, according to some embodiments of the invention. A first end of the support structure 309a may include a drive motor 317 operably coupled to the controller. In some embodiments, the drive motor 317 may be directly coupled to a first one of the pair of shafts, for example 315a, of the platform 311 via a flange such that actuation of the drive motor 317, via input from the controller, causes rotation of the platform 311 about the axis of rotation of the platform 311. Thus, the platform may be directly connected to the motor, for example a servo motor via a flange.

[0090] The motor 317 may include a motor controller and / or drive. In non-limiting examples, the motor may include an AC motor controller and drive, i.e. an electronic device that modifies the input power to the motor by adjusting the frequency of the power to the motor for the purpose of regulating the output speed and torque. The motor may include a DC motor controller and drive, i.e. an electrical device that modifies the input power by adjusting the constant or alternating current source to a pulsed, direct current output of varying pulse duration or frequency. The motor may include a servo motor controller and drive, i.e. an electronic device that modifies the input power by adjusting the constant or alternating current source to a pulsed, current output of varying pulse duration or frequency. The motor may include a stepper motor controller and drive, i.e. an electronic device that modifies the input power by adjusting the constant or alternating current source to a pulsed, or "stepped," current output. In some embodiments, the motor may be a servo motor, for example, an AC servo motor, or the motor may be a DC brushless motor. In some embodiments the motor may be an AC or DC motor.

[0091] FIG. 5 is a solid line perspective side view illustrating one embodiment of an assembly 500 of the invention. The system incorporates a mechanism to tilt to various positions the one or more bioreactors retained upon the platform. As illustrated, the support structure 509 may be rotated about an axis of rotation of the support structure 509 to cause the platform 511 to move from a horizontal orientation to a tilted orientation. The tilted orientation of the platform may be above or below a horizontal orientation. Further, as described in more detail herein, the platform may be rotated around the platform axis of rotation such that the bioreactors 503 retained therein are positioned for filling or draining functions.

[0092] In some embodiments, the mechanism to tilt the support structure includes a hinged coupling on one end of the support structure, and a means for raising and lowering the other end of the support structure. As illustrated in FIG. 5 the support structure 509 may be coupled to the supporting frame 507 and to the base 505 according to one embodiment of the invention which includes a pair of shafts, a linear actuator, and a carriage positioned on a thread rod.

[0093] For example, in some embodiments, one end of the support structure 509a may be rotatably coupled to the supporting frame 507 via a hinged connection thus defining an axis of rotation of the support structure. Accordingly, tilting of the support structure 509 from a horizontal home position to a tilted position may be possible.

[0094] FIG. 6 illustrates a magnified perspective view of a first portion 509a and a second portion 509b of the support structure 509 according to some embodiments of the invention. In some embodiments, a first portion 509a or one end of the support structure 509 may be rotatably coupled to the supporting frame 507 via one or more hinges 519. The hinge may be any type of hinge, as is known to persons skilled in the art, that provides a rotatable couple between one end of the support structure 509 and the supporting frame 507.

[0095] The rotatable couple between the support structure 509 and the supporting frame 507 defines an axis of rotation of the support structure 509. The axis of rotation provided by the coupling between the support structure and the supporting frame allows the support structure 509 to rotate around this axis such that rotation of the support structure 509 defines a tilt axis of rotation of the platform 511. In particular, the first portion 509a of the support structure 509 may remain fixed while the opposing end of the support structure raises and / or lowers to define a tilt axis of rotation of the platform.

[0096] Accordingly, referring back to FIG. 5, the platform 511 may be operable to rotate around an axis of rotation provided by the pair of shafts coupled to the support structure, and around a horizontal or tilt axis of rotation provided by the support structure axis of rotation. In some embodiments, the home position of the support structure may be a substantially horizontal position relative to the x-axis and may be defined as 0 degrees such that the support structure may be operable to rotate from about 90 degrees relative to the home position. In some embodiments, the support structure may be operable to rotate greater than 90 degrees relative to the home position. When the support structure is in a home position, the platform may also be defined as being in a home position. The platform home position may include the bioreactors positioned in a horizontal position with inlets / outlets positioned right-side up. Additionally and / or alternatively, the home position of the bioreactors may be defined as a position for filling and / or draining.

[0097] Further, in some embodiments, an underside of the support structure 509 may be coupled to a linear actuator via a pair of shafts 521a, 521b. The linear actuation may be operably coupled to the controller such that actuation of the linear actuator, via input from the controller, causes the support structure 509 to rotate around the axis of rotation such that one end of the support structure raises or lowers to define a tilt axis of rotation of the platform 511.

[0098] For example, the pair of shafts 521a, 521b may be attached to the underside of the support structure 509 via one or more shaft supports 527 and one or more swivel joints 523, for example, positioned at the end of the shafts 521a, 521b. A rotary shaft 525 may be inserted horizontally through the swivel joints 523 and the shaft supports 527.

[0099] FIG. 7 illustrates a perspective view of the support structure 509 with the pair of shafts 521a, 521b attached to the underside of the support structure 509 according to the embodiment shown in FIG. 5. The linear actuator may comprise a thread rod coupled to a stepper motor and a carriage 529 positioned on the thread rod. The opposing ends of the shafts 521a, 521b may be coupled to the carriage 529 in the same way the other ends of the shafts 521a, 521b are coupled to the underside of the support structure 509. That is, in some embodiments, the carriage 529 may be coupled to the pair of shafts 521a, 521b via a pair of shaft supports connected to swivel joints positioned at the end of the shafts, and a rotary shaft inserted horizontally through the swivel joints and the shaft supports.

[0100] Accordingly, in some embodiments, actuation of the linear actuator, via input from the controller, causes the carriage 529 to travel along the thread rod which causes the pair of shafts to impart force upon the underside of the support structure. This causes rotation of the support structure 509 about the axis of rotation of the support structure to thereby cause the platform 511 to move between a horizontal orientation and a tilted orientation. Likewise, reduction of the force by the linear actuator, by movement of the carriage along the thread rod, causes rotation of the support structure 509 such that the support structure returns to a substantially horizontal position and / or rotates to a position below horizontal.

[0101] In some embodiments, the pair of shafts may be hydraulic cylinders producing linear actuation via hydraulic pressure to impart a force on the underside of the support structure. In some embodiments the underside of the support structure may be coupled to a single shaft. In some embodiments, the underside of the support structure may be coupled to a plurality of shafts. As described in more detail herein, in some embodiments, the underside of the support structure may be not coupled to a shaft but may be actuated to define a tilt axis of rotation by a carriage coupled to a linear support structure as part of a supporting frame. FIG. 8 is a perspective view illustrating the carriage 529 in a home position according to some embodiments of the invention. When the carriage 529 is in a home position on the thread rod, the support structure 509 may be maintained in a substantially horizontal position. This position also maintains the platform 511, and any bioreactors contained therein, in a corresponding horizontal position. The platform may rotate about this horizontal axis of rotation when the platform and support structure are in a home position, as defined according to the needs of a cell processing protocol.

[0102] The home position of the carriage may be calibrated to a position at any point on the thread rod via the controller such that the support structure may be operable to move from a horizontal position to a tilt above or below the horizontal position. Accordingly, in some embodiments, the controller may be configured to align the platform 511 in a horizontal first position and rotate the platform around the axis of rotation of the platform in a range of from 0 degrees to 360 degrees as compared to the horizontal first position. The rotation around the axis of rotation of the platform 511 may be complete rotation, partial rotation, rotation in a clockwise or counterclockwise direction, and / or oscillatory movement. In some embodiments, the rotation around the axis of rotation of the platform 511 comprises oscillatory movement of the platform. Further, the controller may be further configured to align the platform in a tilt axis of rotation such that the platform rotates around the axis of rotation of the platform in a range of from 0 degrees to 360 degrees. The rotation around the axis of rotation of the platform 511 may be complete rotation, partial rotation, rotation in a clockwise or counterclockwise direction, and / or oscillatory movement. In some embodiments, the rotation around the axis of rotation of the platform 511 comprises oscillatory movement of the platform.

[0103] As described herein, in some embodiments, actuation of the stepper motor, via input from the controller, moves the carriage 529 from the home position along the thread rod to define a tilt axis of rotation of the platform 511 such that the platform may be operable to move between a position of axial rotation around a horizontal axis of rotation of the platform to a position of axial rotation around the tilt axis of rotation of the platform. For example, in some embodiments, the controller may be operable to cause the platform to rotate around the horizontal axis of rotation and the tilt access of rotation at one or more of varying speed, varying degree of rotation, varying oscillatory motion, and varying tilt angle to provide for agitating, via a shaking and / or an oscillating motion, the contents of the one or more bioreactors.

[0104] The movement of the support structure around an axis of rotation, the movement of the platform around an axis of rotation of the platform and the tilt axis of rotation defined by the support structure require several moving parts to operate with precise timing, direction, and speed. To coordinate the movement of the entire assembly according to cell processing function needs, the system may be operably associated with a computer processor that controls the movement of the support structure and the platform according to a set of defined cell processing protocols.

[0105] For example, in some embodiments, the controller interfaces with a control system architecture comprising at least one of a computer, a processor, a network, and / or a graphical user interface (GUI) for defining one or more cell processing protocols. The cell processing protocol may define the movement of the platform to achieve, in non-limiting examples, one or more of rotating, shaking, agitating, mixing, and / or oscillating according to the needs of a defined cell processing and / or culturing protocol.

[0106] A user interface for controlling one or more user inputs and displaying one or more outputs from the system, allowing a user to interact with and control the operation of the system, may be physically connected to the system or may be located remotely. The user interface may be a handheld device, e.g., a smart tablet, a smart phone, or a specialty device produced for the system. User interaction can be implemented on a computer having an I / O device, e.g., a CRT, LCD, LED, or projection device for displaying information to the user and an input or output device such as a keyboard and a pointing device, (e.g., a mouse or a trackball), by which the user can provide input to the computer. Other kinds of devices can be used to provide for interaction with a user as well. For example, feedback provided to the user may be any form of sensory feedback (e.g., visual feedback, auditory feedback, or tactile feedback), and input from the user can be received in any form, including acoustic, speech, or tactile input.

[0107] In some embodiments, the system provides for monitoring and controlling various aspects or parameters of the actuation of the support structure and rotation of the platform, such as, in non-limiting examples, speed, motion, rotation direction and extent. Monitoring and controlling various parameters of the actuation of the support structure and rotation of the platform can be performed using any type of computing device, such as a computer or programmable logic controller (PLC), that includes a processor, e g., a central processing unit, or any combination of computing devices where each device performs at least part of the process or method. The controller may include a system employing software, hardware, firmware, hardwiring, or combinations of any of these.

[0108] For example, the system may include a personal and / or portable computing device, such as a smartphone, tablet, laptop computer, or the like. In some embodiments, the computing system may be configured to communicate with a user operator via an associated smartphone or tablet. The computing system may be configured to communicate with and exchange data over a network.

[0109] The network may represent, for example, a private or non-private local area network (LAN), personal area network (PAN), storage area network (SAN), backbone network, global area network (GAN), wide area network (WAN), or collection of any such computer networks such as an intranet, extranet or the Internet (i.e., a global system of interconnected network upon which various applications or service run including, for example, the World Wide Web). In alternative embodiments, the communication path between the user interface and the system of the invention may be, in whole or in part, a wired connection.

[0110] The network may be any network that carries data. Non-limiting examples of suitable networks that may be used as network include Wi-Fi wireless data communication technology, the internet, private networks, virtual private networks (VPN), public switch telephone networks (PSTN), integrated services digital networks (ISDN), digital subscriber link networks (DSL), various second generation (2G), third generation (3G), fourth generation (4G), fifth generation (5G), and future generations of cellular-based data communication technologies, Bluetooth radio, Near Field Communication (NFC), the most recently published versions of IEEE 802.11 transmission protocol standards, other networks capable of carrying data, and combinations thereof.

[0111] In some embodiments, the network may be chosen from the internet, at least one wireless network, at least one cellular telephone network, and combinations thereof. As such, the network may include any number of additional devices, such as additional computers, routers, and switches, to facilitate communications. In some embodiments, the network may be or include a single network, and in other embodiments the network may be or include a collection of networks.

[0112] As disclosed herein, the assemblies of the invention may be used for supporting one or more bioreactors and providing automated or semi-automated cell processing functions associated with the one or more bioreactors. In some embodiments, the assembly may be configured to perform one or more automated or semi-automated cell processing functions in a closed and aseptic environment.

[0113] For example, because the assemblies of the invention incorporate a motor to gently and / or vigorously shake the bioreactor(s), and to properly orient the bioreactor(s), the system of the invention may be used for a variety of cell processing workflow steps. In some embodiments, the position of axial rotation around the tilt axis of rotation of the platform provides for positioning the one or more bioreactors for filling and / or draining. Thus, the invention provides for mechanized bioreactor manipulation to standardize cell processing function. Cell processing functions may include, in non-limiting examples, cell seeding, medium exchange, cell culturing, cell concentration, cell mixing, cell agitation, cell washing, and cell harvesting. In some embodiments, the cell processing functions comprises at least one of cell seeding, cell washing, and cell harvesting. The assemblies of the invention are protocol agnostic and may be used for any type of cell processing functions to standardize procedures and eliminate human bias and error. Because the system of the invention incorporates a mechanism to tilt to various positions the one or more bioreactors retained upon the platform, the invention may be used, in non-limiting examples, for filling and draining the bioreactor(s).

[0114] As discussed in more detail herein, the system of the invention may be programmed to run a specific cell processing protocol. The cell processing or culturing protocol may define the movement of the platform, for example a degree and / or time of, for example, rotating, shaking, agitating, mixing, and / or oscillating according to the needs of the defined cell processing and / or culturing protocol. The cell culturing protocol may include defined periods of platform movement and defined periods of platform rest. The cell culturing protocol may define a plurality of parameters associated with the cell processing function. In non-limiting examples, the parameters may be an axis of rotation of the platform, a tilt axis of the platform, a filling position of the platform, a draining position of the platform, as well as a mixing, agitating, and / or oscillating movement time, pattern, and / or intensity, depending on the cell processing function to be achieved.

[0115] The system of the invention incorporates the various axes of rotation in conjunction with the motor, to gently and / or vigorously shake the bioreactor(s) at various positions, and to properly orient the bioreactor(s) according to cell processing function needs. This allows for mechanized bioreactor manipulation to standardize cell processing functions such as cell seeding, washing, and harvesting functions. Experiments comparing manual cell processing vs the automatic / semi-automatic cell therapy oscillators of the invention show comparability in yield, cell viability, phenotype, and functionality.

[0116] The system of the invention may be used to automate / semi-automate and standardize cell seeding. In one example, the platform may be rotated to a position such that one or more bioreactors retained therein may be seeded with human peripheral blood mononuclear cells (PBMCs), and / or lysate-pulsed dendritic cells. Seeding may occur simultaneously with a plurality of bioreactors retained upon the platform, or may occur consecutively. The system of the invention may be configured to automate cell processing functions for a single bioreactor, and / or a plurality of bioreactors, individually or as a batch.

[0117] The system of the invention may be used to automate / semi-automate and standardize medium exchange. For example, medium exchange may involve removing spent medium and adding fresh medium, and multiple exchanges may be necessary to replenish nutrients, supply specific growth factors and cytokines, and eliminate metabolic wastes and other unwanted media components based on cell culture process requirements. The system provides various ways for performing medium exchange individually or simultaneously within the bioreactors retained upon the platform, for example, by pausing agitation to allow all the cells or aggregates to settle by gravity to the bottom of the bioreactor, rotating the platform to a filling position suitable for medium exchange, and / or for removing a supernatant of spent medium, and rotating the platform to a position for adding fresh medium. The system may achieve rapid and complete medium exchange for multiple bioreactors simultaneously by rapidly removing spent medium in a standardized manner.

[0118] The system of the invention may be used to automate / semi-automate and standardize harvesting cells. Separating cell aggregates and culture medium may be achieved by rotating the platform to a position for harvesting. For example, the platform, through rotation around the tilt axis of rotation and / or a horizontal axis of rotation, may be positioned at various heights and angles to remove medium from one or more specific areas of the bioreactor. In a non-limiting example, the system of the invention provides for harvest functions that allow for removal of adherent cells via mechanical agitation. This is particularly useful for strongly adherent cells where use of enzymatic reagents may be not permissible. Thus, in some embodiments, the devices / system of the invention are configured for removal of adherent cells via mechanical agitation with or without the use of enzymatic reagents.

[0119] The system of the invention may be used to automate / semi -automate and standardize one or more washing steps, for example, during the cell harvest process.

[0120] FIG. 9 is a shaded perspective view illustrating one embodiment of the system 900 of the invention in which the support structure is in a home position. The platform is also in a home position in which the platform, and thus the bioreactor(s) retained therein, are substantially horizontal and upright or right-side-up.

[0121] FIG. 10 is a shaded perspective view illustrating the embodiment of the system 900 of the invention in which the platform is rotated around the axis of rotation of the platform on a tilt axis of rotation defined by movement the axis of rotation of the support structure. In this position, the platform may be rotated around the axis of rotation of the platform such that the bioreactor(s) retained therein are rotated approximately 180 degrees from the horizontal positions for, for example, draining the bioreactor(s).

[0122] As illustrated in the embodiment of a system 900 of the invention shown in FIG. 9 and FIG. 10, the supporting frame may be configured to hold one or more reservoirs or bags 913 that may be connected (not shown) to the bioreactors retained within the platform. Thus, as disclosed herein, in some embodiments, the platform comprises a frame configured to hold a plurality of bioreactors in a stacked position such that an inlet port and / or an outlet port associated with each bioreactor may be accessible. To accommodate various cell processing functions, in some embodiments, the assembly further comprises one or more reservoirs operably connected to one or more pinch valves and one or more pumps configured for providing fluid flow into and out of the one or more reservoirs and the one or more bioreactors retained on the platform. Further, the supporting frame may be configured to support a plurality of elements associated with cell processing functions, for example tubing, fluid reservoir(s), fluid collection contained s), bioprocessing bag(s), pump(s), and / or motor(s). In some embodiments, the supporting frame comprises a plurality of supports configured for receiving and retaining one or more of tubing, one or more fluid reservoirs, one or more fluid collection containers, one or more bioprocessing bags, one or more pumps, and one or more motors. In some embodiments, the assembly may be configured as a closed system for aseptic transfer of fluids to and from the one or more bioreactors.

[0123] The system of the invention may be Grade C (ISO 7) cleanroom compatible. The assembly may be manufactured from any material suitable to meet cell processing function needs and to provide the support and stability for gentle or vigorous rotation, including oscillatory motion, of the platform with the bioreactors retained therein. For example, as is known to persons skilled in the art, the assemblies of the invention may be constructed of one or more of aluminum, an aluminum alloy, stainless steel or other alloy, plastic (such as PPE, high impact polystyrene) acrylic, and the like.

[0124] FIG. 11 illustrates a side perspective view of an assembly 1100 according to one embodiment of the invention, in which the support structure may be actuated to rotate around an axis of rotation of the support structure to thereby define a tilt axis of rotation of the platform. The platform may be further rotated around the axis of rotation of the platform to position the bioreactors retained therein for draining.

[0125] FIG. 12 illustrates an end perspective view of one end of the assembly 1100 according to FIG. 11.

[0126] FIG. 13 illustrates a top perspective view of the assembly 1100 according to FIG. 11. FIG. 14 illustrates a second end and side perspective view of the assembly 1100 according to FIG. 11.

[0127] FIG. 15 illustrates a bottom perspective view of the assembly 1100 according to FIG. 11. FIG. 16 illustrates another embodiment of the assembly 1600 for supporting one or more bioreactors and providing automated or semi-automated cell processing functions associated with the one or more bioreactors. The assembly may comprise a base 1605, and a pair of vertical support structures 1631a, 163 lb extending from the base. The assembly may include a platform 1611 movably coupled to the pair of vertical support structures 1631a, 163 lb and configured to removably receive and retain one or more bioreactors 1603 thereto. The assembly may include a controller configured to control movement of the platform 1611 to thereby control movement of one or more bioreactors 1603 supported by the platform 1611 based, at least in part, on one or more predefined cell processing protocols.

[0128] In some embodiments, the platform 1611 may be movably coupled to the pair of vertical support structures 1631a, 1631b by way of a pair of shafts 1615 positioned at respective opposing ends of the platform 1611 and coupled to a respective one of the pair of vertical support structures 1631a, 1631b, wherein the pair of shafts cooperatively define an axis of rotation of the platform.

[0129] In some embodiments, a first one of the pair of vertical support structures 1631a comprises a drive motor 1617 operably coupled to the controller, and a second one of the pair of vertical support structures 1631b comprises a linear actuator operably coupled to the controller. In some embodiments, the drive motor 1617 may be coupled to a first one of the pair of shafts 1615 of the platform 1611 via a u-joint such that actuation of the drive motor 1617, via input from the controller, causes rotation the platform 1617 about the axis of rotation of the platform 1617.

[0130] Further, in some embodiments, the linear actuator comprises a thread rod 1625 coupled to a stepper motor and a carriage 1629 positioned on the thread rod 1625, the carriage 1629 comprising a linear bearing with a carriage shaft therethrough, wherein the carriage shaft may be coupled to the second one of the shafts 1615 of the platform 1611 via a u-joint.

[0131] Further, in some embodiments, a horizontal alignment of the carriage 1629 on the thread rod 1625 with the drive motor 1617 defines a horizontal axis of rotation of the platform. Thus, as illustrated in FIG. 16, the controller may be configured to align the platform 1611 in a horizontal first position and rotate the platform 1611 around the horizontal axis of rotation in a range of from 0 degrees to 360 degrees as compared to the horizontal first position in some embodiments. For example, as illustrated in FIG. 16, the platform 1611 may be rotated 90 degrees from the horizontal first position. In some embodiments, the rotation around the horizontal axis of rotation comprises oscillatory movement of the platform.

[0132] FIG. 17 illustrates a tilt axis of rotation of the platform 1611 as defined by movement of the carriage 1629 along the thread rod 1625 according to one embodiment of the invention. In some embodiments, actuation of the stepper motor, via input from the controller, moves the carriage 1629 in a vertical direction to define a tilt axis of rotation of the platform 1611 such that the platform may be operable to move between a position of axial rotation around a horizontal axis of rotation to a position of axial rotation around the tilt axis of rotation.

[0133] FIG. 18 illustrates the platform 1611 at a tilt and at a position of axial rotation around the tilt axis such that the bioreactor(s) are positioned for draining according to one embodiment of the invention. In some embodiments, the position of axial rotation around the tilt axis of rotation provides for positioning the one or more bioreactors for filling and / or draining. As disclosed herein, the controller may be operable to cause the platform to rotate around the horizontal axis of rotation and the tilt access of rotation at one or more of varying speed, varying degree of rotation, varying oscillatory motion, and varying tilt angle to provide for agitating, via a shaking and / or an oscillating motion, the contents of the one or more bioreactors. The assembly may further comprise a constant tension spring 1633to provide vertical stabilization during rotation of the platform. A first end of the constant tension spring may be coupled to the carriage shaft and a second end of the vertical stabilizer may be coupled to the base. The constant tension spring may be an elastomeric material configured to stabilize the linear bearing and dampen vibration of the carriage shaft.

[0134] As disclosed herein, the controller may interface with a control system architecture comprising at least one of a computer, a network, and a graphical user interface (GUI) for defining one or more cell processing protocols. The assembly may be configured to perform one or more automated or semi-automated cell processing functions in a closed and aseptic environment. For example, as disclosed herein, the cell processing functions comprises at least one of cell seeding, cell washing, and cell harvesting. For example, in some embodiments, the assembly may be configured for removal of adherent cells via mechanical agitation without the use of enzymatic reagents.

[0135] As disclosed herein, the platform may comprise a stepped frame configured to hold a plurality of bioreactors in a stacked position offset from each other such that an inlet port and / or an outlet port associated with each bioreactor may be accessible. In some embodiments, the assembly further comprises one or more reservoirs operably connected to one or more pinch valves and one or more pumps configured for providing fluid flow into and out of the one or more reservoirs and one or more bioreactors retained on the platform. The assembly may be configured as a closed system for aseptic transfer of fluids to and from the one or more bioreactors.

[0136] Example: Comparability Study: NW-22 Automated Device (FACTOR) vs Manual ADCV Process

[0137] Data was collected comparing using a system of the invention as disclosed herein, referred to in the study as “FACTOR” to a manual drug process. The study focused on comparing the quality of sDP generated on day 8 of the ADCV process following BMR-4V6 using the manual process with data using processing methods of the invention using FACTOR. Two sDP processed with manual and FACTOR methods were compared in this study. The following parameters were evaluated:

[0138] • Dendritic cell (DC) numbers and viability on day 7 and 8 (manual counting using trypan blue following standard procedures)

[0139] • Identity, purity and impurity characterization using standard procedures

[0140] • Potency using standard procedures

[0141] DC Numbers and Viability on Day 7

[0142] FIGs. 19A-19C illustrate Assessment of DC numbers and viability on sDP by trypan blue at day 7. FIG. 19A illustrates assessment of DC numbers, while FIG. 19B illustrates viability measured in %. FIG. 19C illustrates the data for batch 30 and 31 with a comparison between the manual method and the automated / semi-automated processing method using FACTOR. Batch 30 comprised 125xl06DCs seeded for Day 8 cultures. Batch 31 comprised 58.54xl06DCs (dendritic cells) seeded for Day 8 cultures. There was a trend to reduced DC numbers after harvesting when the process is performed manually. This effect was more pronounced in batch 31.

[0143] DC Numbers and Viability on Day 8

[0144] FIGs. 20A-20C illustrate Assessment of DC numbers and viability on sDP by trypan blue at day 8. FIG. 20A illustrates assessment of DC numbers, while FIG. 20B illustrates viability measured in %. FIG. 20C illustrates the data for batch 30 and 31 with a comparison between the manual method and the automated processing method using FACTOR. Similar DC numbers and viability in both manual and FACTOR-processed sub-batch samples.

[0145] Identity and Impurity Characterization FIG. 21 illustrates the identity panel comparison from the study.

[0146] FIG. 22 illustrates the purity characterization panel from the study.

[0147] FIG. 23 illustrates assessment of sDP purity and identity, specifically the sDP Identity on Day 8. The identity characterization exhibited the following results:

[0148] ■ Comparable %HLA-DR+CD86+ cells observed between the manually processed subbatches and those processed using FACTOR.

[0149] ■ All samples passed the acceptance criteria >70%.

[0150] ■ Comparable %CD141+CD86+ cells observed between the manually processed subbatches and those processed using FACTOR

[0151] ■ FIGs. 24A-24C illustrate sDP impurity characterization on Day 8. FIG. 24A illustrates assessment of sDP impurities. FIG. 24B is a graph of characterization of total impurities.

[0152] FIG. 24C is a table of raw data for characterization of impurities. The data indicate a similar impurity profile of sDP processed using manual and FACTOR.

[0153] Potency assessment

[0154] FIG. 25A illustrates the potency assessment. Potency testing was carried out in an allogeneic system per standard protocols.

[0155] FIG. 25B illustrates results for positive and negative controls. Results for positive and negative controls, background control (MLR), and gating controls passed. Potency is measured in percent and defined as % T cell proliferation - % T cell proliferation in the background control. SI is defined as SI= % T cell proliferation / % T cell proliferation in the background control

[0156] FIG. 25C illustrates graphs of Assessment of sDP potency on Day 8. Specifically, the potency assay were performed for the four samples. Positive, negative and background controls passed the assay acceptance criteria. Both methods passed release criterion of SI >2. There was some variability observed in batch 30 while batch 31 was more consistent. The sample size was small.

[0157] Results:

[0158] • Trend towards better DC recovery on day 7 was observed for FACTOR processed samples.

[0159] • Similar sDP numbers and viability on day 8. • Similar sDP identity, purity and impurity profiles on day 8.

[0160] • Potency in terms of SI passed release criterion > 2 in all batches with some variability observed in batch 30.

[0161] Example: Comparability Study: Oscillation Harvest using FACTOR vs Manual Harvest

[0162] FIG. 26 is a table of data which compares various characteristics of harvested cells on days 7 and 8 using one embodiment of devices and methods of the invention versus manual methods for four separate runs.

[0163] FIG. 27 is a table of data for run CS 26 with results measured on day 7 and 8 which compares various characteristics as noted above for using one embodiment of the FACTOR system for harvest versus manual methods for harvest. Specifically, FIG. 27 also includes comparison graphs of % T cell proliferation, % potency, and stimulation index for the run.

[0164] FIG. 28 is a table of data for run CS 27 with results measured on day 7 and 8 which compares various characteristics as noted above for using one embodiment of the FACTOR system for harvest versus manual methods for harvest. Specifically, FIG. 28 also includes comparison graphs of % T cell proliferation, % potency, and stimulation index for the run.

[0165] FIG. 29 illustrates DP identity for all events, cells, live, and singlets at day 7 and 8 for manual harvest.

[0166] FIG. 30 illustrates DP identity for all events, cells, live, and singlets at day 7 and 8 for manual harvest.

[0167] FIG. 31 is a table of data for run CS 28 with results measured on day 7 and 8 which compares various characteristics as noted above for using one embodiment of the FACTOR system for oscillation harvest versus manual methods for harvest. Specifically, FIG. 31 also includes comparison graphs of % T cell proliferation, % potency, and stimulation index for the run.

[0168] FIGs. 32A and 32B are tables including data of runs CS19-CS-25 measured on Days 7 and 8 which compares various characteristics noted above for using one embodiment of the FACTOR system for oscillation harvest versus manual methods for harvest.

[0169] FIG. 33 illustrates a functional allogenic assay of run no. CS 23 of D8 DC + CS 22 T cells + CD3. FIG. 34 illustrates graphs of percent T cell proliferation, potency, and stimulation index from the functional allogenic assay of run no. CS 23. For these calculations, Potency = %Proliferating T cells (COSTIM assay) - % Proliferating T cells (MLR assay). Stimulation Index (SI) = “ / ((Proliferating T cells (COSTIM assay ) / %Proliferating T cells (MLR)

[0170] Incorporation by Reference

[0171] References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made throughout this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes.

[0172] Equivalents

[0173] Various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including references to the scientific and patent literature cited herein. The subject matter herein contains important information, exemplification and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.

Claims

Claims:

1. An assembly for supporting one or more bioreactors and providing automated or semiautomated cell processing functions associated with the one or more bioreactors, the assembly comprising: a base and supporting frame; a moveable support structure having a first portion coupled to the supporting frame and a second portion coupled to the base; a platform rotatably coupled to the support structure and configured to removably receive and retain one or more bioreactors thereto; and a controller configured to control movement of the platform to thereby control movement of one or more bioreactors supported by the platform based, at least in part, on one or more predefined cell processing protocols.

2. The assembly of claim 1, wherein the platform is rotatably coupled to the support structure by way of a pair of shafts, wherein each shaft is positioned on a respective opposing end of the platform and coupled to one of a first end and a second end of the support structure via a swivel joint, wherein the pair of shafts cooperatively define an axis of rotation of the platform.

3. The assembly of claim 2, wherein the first end of the support structure comprises a drive motor operably coupled to the controller.

4. The assembly of claim 3, wherein the drive motor is directly coupled to a first one of the pair of shafts of the platform via a flange such that actuation of the drive motor, via input from the controller, causes rotation of the platform about the axis of rotation of the platform.

5. The assembly of claim 2, wherein one end of the support structure is rotatably coupled to the supporting frame via a hinged connection defining an axis of rotation of the support structure.

6. The assembly of claim 5, wherein an underside of the support structure is coupled to a linear actuator via a pair shafts, the linear actuator being operably coupled to the controller such that actuation of the linear actuator, via input from the controller, causes the pair of shafts to impart force upon the underside of the support structure and causes rotation of the support structure about the axis of rotation of the support structure to thereby cause the platform to move between a horizontal orientation and a tilted orientation.

7. The assembly of claim 6, wherein the linear actuator comprises a thread rod coupled to a stepper motor and a carriage positioned on the thread rod, the carriage coupled to the pair of shafts via a pair of shaft supports connected to swivel joints positioned at a first end of the shafts and a rotary shaft inserted horizontally through the swivel joints and the shaft supports.

8. The assembly of claim 6, wherein the underside of the support structure is coupled to the pair of shafts via a pair of shaft supports connected to swivel joints positioned at a second end of the shafts and a rotary shaft inserted horizontally through the swivel joints and the shaft supports.

9. The assembly of claim 7, wherein when the carriage is in a home position on the thread rod, the support structure is maintained in a substantially horizontal position thereby maintaining the platform, and any bioreactors contained therein, in a corresponding horizontal position.

10. The assembly of claim 7, wherein the controller is configured to align the platform in a horizontal first position and rotate the platform around the axis of rotation of the platform in a range of from 0 degrees to 360 degrees as compared to the horizontal first position.

11. The assembly of claim 10, wherein the rotation around the axis of rotation of the platform comprises oscillatory movement of the platform.

12. The assembly of claim 9, wherein actuation of the stepper motor, via input from the controller, moves the carriage from the home position along the thread rod to define a tilt axis of rotation of the platform such that the platform is operable to move between a position of axialrotation around a horizontal axis of rotation of the platform to a position of axial rotation around the tilt axis of rotation of the platform.

13. The assembly of claim 12, wherein the position of axial rotation around the tilt axis of rotation of the platform provides for positioning the one or more bioreactors for filling and / or draining.

14. The assembly of claim 13, wherein the controller is operable to cause the platform to rotate around the horizontal axis of rotation and the tilt access of rotation at one or more of varying speed, varying degree of rotation, varying oscillatory motion, and varying tilt angle to provide for agitating, via a shaking and / or an oscillating motion, the contents of the one or more bioreactors.

15. The assembly of claim 1, wherein the controller interfaces with a control system architecture comprising at least one of a computer, a processor, a network, and / or a graphical user interface (GUI) for defining one or more cell processing protocols.

16. The assembly of claim 15, wherein the assembly is configured to perform one or more automated or semi-automated cell processing functions in a closed and aseptic environment.

17. The assembly of claim 16, wherein the cell processing functions comprises at least one of cell seeding, cell washing, and cell harvesting.

18. The assembly of claim 17, wherein the assembly is configured for removal of adherent cells via mechanical agitation with or without the use of enzymatic reagents.

19. The assembly of claim 1, wherein the platform comprises a frame configured to hold a plurality of bioreactors in a stacked position such that an inlet port and / or an outlet port associated with each bioreactor is accessible.

20. The assembly of claim 19, further comprising one or more reservoirs operably connected to one or more pinch valves and one or more pumps configured for providing fluid flow into and out of the one or more reservoirs and one or more bioreactors retained on the platform.

21. The assembly of claim 20, wherein the assembly is configured as a closed system for aseptic transfer of fluids to and from the one or more bioreactors.

22. The assembly of claim 1, wherein the supporting frame comprises a plurality of supports configured for receiving and retaining one or more of tubing, a fluid reservoir, a fluid collection container, a bioprocessing bag, a pump, and a motor.