Processing instrument for cell therapy manufacturing

Abstract

The present invention relates 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] Cross-references to related applications

[0002] This application claims priority and benefit to U.S. Provisional Application No. 63 / 599,068, filed November 15, 2023, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention generally relates to systems and apparatus for cell culture, and more particularly to a system comprising mechanical components for supporting one or more bioreactors and assisting in providing automated or semi-automated cell processing functions within one or more bioreactors during a cell culture procedure. Background Technology

[0004] 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 treatments to prevent, diagnose, treat, and / or potentially cure diseases at their source. The workflow for autologous and allogeneic cell and gene therapies typically involves isolating cells from an individual, purifying and / or engineering those isolated cells, expanding and / or concentrating the isolated cells, and infusing such cells into a patient.

[0005] To ensure the efficacy of the final product, robust, standardized, and efficient processes must be implemented in the cell and gene therapy manufacturing workflow to provide consistent production of cell therapy products. Cell viability and purity are critical issues in the manufacturing of cell therapy products. Decreased cell viability and recovery rates can be caused by excessive transport time, tissue digestion, cell manipulation, operator variations, gene transduction, cryopreservation, and cell thawing. Minimum throughput and the percentage of viable cells recovered are particularly important factors for autologous and allogeneic products, as poor cell recovery rates can lead to manufacturing run failures. Therefore, the challenge to the future commercial success of cell therapy products lies in establishing scalable manufacturing technologies that can reliably reproduce the yield and quality of cell-derived products generated by small-scale R&D methods.

[0006] Tissue cells are known to be sensitive to mechanical stresses exerted on them due to agitation within bioreactors. For example, common challenges in pluripotent stem cell (PSC) aggregate formation include the influence of variables such as cell proliferation rate, intercellular adhesion strength, cell packing density, agitation rate, and the hydrodynamic environment within the bioreactor. Furthermore, workflow operations typically require numerous operators for extended periods of handling and processing, which can negatively impact cell quality, process reliability, and cost-effectiveness. Any negative impacts on cell quality (e.g., shear stress) when cells are subjected to prolonged handling can be amplified downstream. Therefore, it is necessary to optimize these variables at each step of the manufacturing workflow to ensure consistent production of cell therapy products. Summary of the Invention

[0007] This invention includes a system for providing automated / semi-automated cell processing functions associated with one or more bioreactors during a cell culture procedure. More specifically, the system of the invention includes mechanical components configured to support one or more bioreactors at a time and subsequently move the one or more bioreactors in a controlled and precise manner to achieve desired cell processing functions, including but not limited to cell seeding, cell washing, and cell harvesting.

[0008] This component is configured to automate and reproduce manual movements used during cell culture processes. Specifically, it incorporates mechanisms for agitating the bioreactor with specific oscillating motions, allowing for gentle or vigorous agitation depending on the cell culture processing requirements. The component also incorporates rotation and tilting mechanisms to orient the bioreactor at any position required for cell processing. For example, such mechanisms allow for tilting to allow the bioreactor to be tilted to various positions to allow for filling and draining.

[0009] This component is configured to operate in an automated or semi-automated manner, thereby allowing certain cell processing functions to occur, including but not limited to standardized filling, cell seeding, culture medium exchange, mixing, agitation, concentration, washing, and harvesting functions. The automated / semi-automated nature of the cell processing functions provided by the system of this invention eliminates user variability and improves scalability. Furthermore, the component is capable of handling a single cell bioreactor at a time, or multiple cell bioreactors in a completely closed system, thereby enabling users to perform cell processing functions outside a biosafety cabinet while maintaining sterility. Additionally, the ability to handle multiple bioreactors simultaneously further reduces the overall time required for technicians to perform cell processing.

[0010] As an example, the system's harvesting function allows for the removal of adherent cells via mechanical agitation, which is particularly useful for strongly adherent cells where the use of enzymatic reagents is not permitted. The system can be used for any type of adherent cell culture and is protocol-independent, as all steps can be customized to meet specific user / cell culture protocol requirements.

[0011] Experiments comparing manual cell processing with cell processing using the system of the present invention demonstrate comparability in terms of yield, cell viability, phenotype, and functionality. The output cells exhibit equal or better quality (as measured by viability, yield, surface marker phenotype, and functionality) compared to equivalent manual processes. Therefore, by mechanizing the operation of the bioreactor, the system of the present invention provides optimization and standardization of variables directly affecting cell viability and the quality of cell therapy products. Accordingly, the system of the present invention provides an improved cell therapy product.

[0012] In one aspect, the invention provides a component for supporting one or more bioreactors and providing automated or semi-automated cell processing functionality associated with the one or more bioreactors. The component includes a base and a support frame, and a movable support structure having a first portion coupled to the support frame and a second portion coupled to the base. Further, the component includes: a platform rotatably coupled to the support structure and configured to removably receive and hold the one or more bioreactors to the platform; and a controller configured to control movement of the platform, at least in part, based on one or more predetermined cell processing protocols, thereby controlling movement of the one or more bioreactors supported by the platform.

[0013] In some embodiments, the platform is rotatably coupled to a support structure via a pair of shafts. In some embodiments, each shaft is positioned at a corresponding opposite end of the platform and coupled to one of a first end and a second end of the support structure via a rotary joint. Thus, the pair of shafts cooperatively define the axis of rotation of the platform. Further, in some embodiments, the first end of the support structure includes a drive motor operatively coupled to a controller. In some embodiments, the drive motor is directly coupled to the first shaft 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.

[0014] In a particular embodiment, one end of the support structure is rotatably connected to the support frame via a hinged connector defining the axis of rotation of the support structure. Further, in some embodiments, the lower side of the support structure is connected to a linear actuator via a pair of shafts. The linear actuator can be operatively coupled to a controller such that actuation of the linear actuator via input from the controller causes a force to be applied to the pair of shafts on the lower side of the support structure and causes rotation of the support structure about its axis of rotation. This allows the platform to move between a horizontal orientation and a tilt orientation.

[0015] Furthermore, in some embodiments, the linear actuator includes a threaded rod coupled to a stepper motor and a carriage positioned on the threaded rod, the carriage being coupled to the pair of shafts via: a pair of shaft supports connected to a rotary joint positioned at a first end of the shaft; and a rotating shaft horizontally inserted through the rotary joint and the shaft supports.

[0016] In some embodiments, the lower side of the support structure is connected to the pair of shafts via: a pair of shaft supports connected to a rotary joint positioned at a second end of the shafts; and a rotating shaft inserted horizontally through the rotary joint and the shaft supports.

[0017] In some embodiments, when the carriage is in its original position on the threaded rod, the support structure is maintained in a substantially horizontal position, thereby maintaining the platform and any bioreactors housed therein in a corresponding horizontal position.

[0018] In some embodiments, the controller is configured to align the platform in a first horizontal position and rotate the platform about its axis of rotation within a range of 0 to 360 degrees relative to the first horizontal position. In a particular embodiment, rotation about the platform's axis of rotation includes oscillating movement of the platform.

[0019] In some embodiments, actuation of the stepper motor via input from the controller moves the carriage along a threaded rod from its original position to define a tilted axis of rotation of the platform, making the platform operable between an axial rotational position about the platform's horizontal axis of rotation and an axial rotational position about the platform's tilted axis of rotation. Further, in some embodiments, the axial rotational position about the platform's tilted axis of rotation provides positioning one or more bioreactors for filling and / or discharging. For example, in some embodiments, the controller is operable to rotate the platform about the horizontal axis of rotation and the tilted rotational path at one or more of a varying speed, a varying degree of rotation, a varying oscillating motion, and a varying tilt angle to provide agitation of the contents of one or more bioreactors via rocking and / or oscillating motion.

[0020] In some embodiments, the controller interacts with a control system architecture to define one or more cell processing protocols, the control system architecture including at least one of a computer, a processor, a network, and / or a graphical user interface (GUI). Further, in some embodiments, the component is configured to perform one or more automated or semi-automated cell processing functions in a closed and sterile environment. For example, in some embodiments, the cell processing functions include at least one of cell seeding, cell washing, and cell harvesting. Further, in some embodiments, the component is configured to remove adherent cells via mechanical agitation, with or without the use of enzymatic reagents.

[0021] In a particular embodiment, the platform includes a frame configured to hold multiple bioreactors in a stacked position, allowing access to an inlet port and / or outlet port associated with each bioreactor. Further, in some embodiments, the component further includes one or more reservoirs operatively connected to one or more pinch valves and one or more pumps configured to provide fluid inflow and outflow to the one or more reservoirs and the one or more bioreactors held on the platform. For example, in some embodiments, the component is configured as a closed system for the aseptic transfer of fluid into and out of the one or more bioreactors.

[0022] In some embodiments, the support frame includes a plurality of supports configured to receive and hold one or more of pipe fittings, fluid reservoirs, fluid collection containers, bioprocessing bags, pumps, and motors. Attached Figure Description

[0023] Figure 1A This is a perspective view illustrating an embodiment of the system of the present invention, showing the overall components, wherein a support structure defines a tilting rotation axis of the platform, and the platform rotates about the tilting rotation axis.

[0024] Figure 1B This is a transparent perspective view showing an embodiment of the system of the present invention, in which the support structure and the platform are each in their original positions.

[0025] Figure 1C This is a perspective view illustrating one embodiment of the system of the present invention.

[0026] Figure 1D yes Figure 1C An enlarged view of an embodiment of a fluid connection implemented via a pipe fitting connected to one or more bags or reservoirs of one or more bioreactors and systems held on a platform.

[0027] Figure 2AThis is a perspective view illustrating one embodiment of a platform containing components of the present invention, wherein four bioreactors are received and held on the platform.

[0028] Figure 2B This is a perspective side view of an embodiment of a platform showing the components of the present invention, illustrating a pair of axes defining the axis of rotation of the platform.

[0029] Figure 2C An embodiment of the platform of the components of the present invention is shown, illustrating three bioreactors, each held on the platform by adjustable straps.

[0030] Figure 3 This is a shaded perspective side view illustrating one embodiment of the system for automating cell processing functions according to the present invention.

[0031] Figure 4 This is a shaded, enlarged perspective view of a support structure according to an embodiment of the present invention, wherein the platform is attached to the support structure at opposite ends by a pair of shafts positioned at opposite ends of the platform.

[0032] Figure 5 This is a solid-line perspective side view illustrating an embodiment of the components of the present invention.

[0033] Figure 6 Enlarged perspective views of the first and second portions of a support structure according to some embodiments of the present invention are shown.

[0034] Figure 7 Showing according to Figure 5 The illustrated embodiment shows a perspective view of the support structure, in which a pair of shafts are attached to the lower side of the support structure.

[0035] Figure 8 This is a perspective view showing a carriage in its original position according to some embodiments of the present invention.

[0036] Figure 9 This is a shaded perspective view illustrating one embodiment of the system of the present invention, with the support structure in its original position.

[0037] Figure 10 It is a demonstration Figure 9 The shaded perspective view of an embodiment of the system of the present invention shows a platform rotating about the platform's rotation axis on an inclined rotation axis defined by the movement of the rotation axis of the support structure.

[0038] Figure 11 A side perspective view of a component according to an embodiment of the present invention is shown, wherein a support structure is actuated to rotate about a rotation axis of the support structure, thereby defining a tilt rotation axis of the platform.

[0039] Figure 12 Showing according to Figure 11 The end-view stereoscopic view of the component.

[0040] Figure 13 Showing according to Figure 11 Top-down 3D view of the components.

[0041] Figure 14 Showing according to Figure 11 The second-end side perspective of the component.

[0042] Figure 15 Showing according to Figure 11 A bottom-view 3D diagram of the components.

[0043] Figure 16 Another embodiment of a component is shown for supporting one or more bioreactors and providing automated or semi-automated cell processing functions associated with the one or more bioreactors.

[0044] Figure 17 Showing according to Figure 16 The tilting rotation axis of the platform in the embodiment of the invention shown is defined by the movement of the carriage along the threaded rod.

[0045] Figure 18 Showing according to Figure 16 The platform shown in this embodiment of the invention is tilted and axially rotated about the tilt axis, such that the bioreactor is positioned for discharge.

[0046] Figures 19A to 19C The study demonstrates the assessment of the number and activity of DCs in sDP using trypan blue on day 7.

[0047] Figures 20A to 20C The study demonstrates the assessment of the number and activity of DCs in sDP using trypan blue on day 8.

[0048] Figure 21 This study presents a comparison of the group of identifying markers.

[0049] Figure 22 The set of purity characterization markers used in this study is presented.

[0050] Figure 23 An assessment of sDP purity and identification (particularly sDP identification on day 8) is presented.

[0051] Figures 24A to 24C The sDP impurity characterization on day 8 is shown.

[0052] Figure 25A The valence assessment was presented.

[0053] Figure 25B The results of the positive and negative controls are shown.

[0054] Figure 25C A chart showing the sDP valence assessment on day 8 is presented.

[0055] Figure 26 It is a data table that compares various characteristics of cells harvested on days 7 and 8 using the system / apparatus and method of the present invention versus using manual methods for four independent runs.

[0056] Figure 27 This is a data sheet for running CS 26, which has results measured on days 7 and 8, comparing various characteristics of cells harvested using an embodiment of the system of the present invention relative to those harvested using manual methods.

[0057] Figure 28 This is a data sheet for running CS 27, which has results measured on days 7 and 8, comparing various characteristics of cells harvested using an embodiment of the invention versus harvested using manual methods.

[0058] Figure 29 It demonstrates the DP identification of all events, cells, live cells, and single cells for manual harvesting on days 7 and 8.

[0059] Figure 30 It demonstrates the DP identification of all events, cells, live cells, and single cells for manual harvesting on days 7 and 8.

[0060] Figure 31 This is a data sheet for running CS 28, which has results measured on days 7 and 8, comparing various characteristics of cells harvested using an embodiment of the invention via oscillation harvesting versus harvesting using manual methods.

[0061] Figure 32A and Figure 32B It is a table that includes data measured on days 7 and 8 of running CS19 to CS-25, which compares various characteristics of cells harvested using an embodiment of the invention via oscillation harvesting versus harvesting using manual methods.

[0062] Figure 33 and Figure 34 A functional allogeneic assay of D8 DC + CS 22 T cells + CD3 running CS 23 was demonstrated. Detailed Implementation

[0063] This invention relates to a system for providing cell processing functions in a closed, aseptic manner during cell culture. Specifically, the invention includes a system for providing automated / semi-automated cell processing functions associated with one or more bioreactors during a cell culture procedure. The system includes mechanical components configured to simultaneously support one or more bioreactors and subsequently move the one or more bioreactors in a controlled and precise manner to achieve desired cell processing functions, including but not limited to cell seeding, cell washing, and cell harvesting.

[0064] This component is configured to automate and reproduce manual movements used during cell culture. Specifically, it incorporates mechanisms for agitating the bioreactor with specific oscillating motions, allowing for gentle or vigorous agitation depending on the cell culture processing requirements. The component also incorporates rotation and tilting mechanisms to orient the bioreactor at any position required for cell processing. For example, such mechanisms allow for tilting to allow the bioreactor to be tilted to various positions to allow for filling and draining.

[0065] This component is configured to operate in an automated or semi-automated manner, thereby allowing certain cell processing functions to occur, including but not limited to standardized filling, cell seeding, culture medium exchange, mixing, agitation, concentration, washing, and harvesting functions. The automated / semi-automated nature of the cell processing functions provided by the system of the present invention eliminates user variability and improves scalability. Furthermore, the component is capable of handling a single cell bioreactor at a time, or multiple cell bioreactors, thereby allowing users to perform cell processing functions outside a biosafety cabinet while maintaining sterility. The automated / semi-automated system of the present invention reduces the overall time required for technicians to perform cell processing due to its ability to process multiple bioreactors simultaneously or in synergy with each other. Automation also eliminates user variability and improves scalability. Output cells generated using the system of the present invention have equivalent or better quality, as measured by viability, yield, surface marker phenotype, and functionality.

[0066] Overview

[0067] This invention provides a system for automating / semi-automating and standardizing cell handling functions during cell culture. The invention addresses the limitations of conventional platforms for generating large numbers of cells (such as labor and material costs) and the lengthy handling and processing times resulting from the need for numerous operators, which can negatively impact cell quality, process reliability, and cost-effectiveness.

[0068] This system addresses the fact that various types of therapeutic cells can be sensitive to the mixing characteristics of a bioreactor in different ways. In particular, the hydrodynamic conditions or mixing environment within a bioreactor can significantly affect the biological properties of cells and the efficiency of resulting cell culture processes, such as expansion and differentiation. For example, adherent-dependent cells grown on microcarriers or as aggregates are more sensitive to hydrodynamic shear stress compared to cells grown as single cells in a suspension bioreactor. This system solves these challenges and can be used with any cell culture or protocol. Therefore, this system can be used with any cell treatment protocol and is thus protocol-independent.

[0069] Specifically, certain embodiments of this system can be used in cell processing protocols related to dendritic cell generation. This system can be used in other types of adherent and / or non-adherent cell culture processing protocols. This system can be used in cell processing protocols for culturing cell types cultured as aggregates, such as pluripotent stem cells (PSCs), neural stem cells, and / or mammary epithelial stem cells. This system can also be used in cell processing protocols associated with cultured cells that typically grow as single cells or loose clumps, such as hematopoietic stem cells / progenitor cells and / or immune cells, such as T cells and natural killer (NK) cells.

[0070] This system enables the automation / semi-automation and optimization of cell processing functions, such as expansion and differentiation, gas and culture medium exchange, and cell harvesting, for large-scale applications. Therefore, the system provides a robust and scalable manufacturing process for the consistent production of allogeneic and autologous cell therapy products for patients, and reduces the percentage of cells that may be lost during various downstream processes, such as harvesting, washing, concentration, formulation, and filling / finishing steps.

[0071] Systems for automating cell processing functions

[0072] The present invention provides a system for automated or semi-automated cell processing functions associated with one or more bioreactors.

[0073] Figure 1A This is a perspective view illustrating one embodiment of the system 100 of the present invention. As described in more detail herein, Figure 1A A support structure is shown that defines the tilting rotation axis of the platform, and the platform rotates about the tilting rotation axis.

[0074] Figure 1B This demonstrates the system 100 of the present invention. Figure 1A The illustrated embodiment is a transparent three-dimensional outline view, in which the support structure and platform are each in their original positions, as described in more detail herein.

[0075] In one aspect, the present invention provides a component 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 component includes a base 105 and a support frame 107, and a movable support structure 109 having a first portion 109a coupled to the support frame and a second portion 109b coupled to the base 105. Thus, the support structure can have a rigid frame such that the component 100 includes a platform 111 rotatably coupled to the support structure 109.

[0076] The platform can be configured to removably receive one or more bioreactors 103 and hold the one or more bioreactors to the platform. Components (which may also be interchangeably referred to as systems or devices within this disclosure) include a controller configured to control the movement of the platform 111 based at least in part on one or more predetermined cell treatment protocols, thereby controlling the movement of the one or more bioreactors 103 supported by the platform 111.

[0077] Platform 111 can be configured to hold one or more bioreactors of any size or shape. Platform 103 can be configured to receive and hold one or more bioreactors 103 of the same size and / or shape. Platform 103 can be configured to receive and hold one or more bioreactors 103 of different sizes and / or shapes. For example, the one or more bioreactors can be any bioreactor designed for a specific cell processing function, such as Corning® CellSTACK®, Nunc™ Cell Factory™, or T-flasks. One or more bioreactors 103 held on platform 111 can be detached and reattached to the assembly to allow for various functions or purposes, such as culturing the bioreactors in an incubator or refrigerating the bioreactors in a refrigerator. In some embodiments, the assembly can be designed to be assembled within an incubator.

[0078] In some embodiments, the platform includes a rigid frame configured to hold multiple bioreactors in a stacked position, allowing access to the inlet and / or outlet ports associated with each bioreactor.

[0079] Figure 1C This is a perspective view illustrating one embodiment of the system 200 of the present invention, wherein three bioreactors 103 are held on a platform. The holding mechanism may be, for example, one or more adjustable straps, clamps, buckles, rings, screws, slits, or slots, which are designed to receive and hold one or more bioreactors of different sizes and shapes.

[0080] One or more bioreactors 103 may be connected to one or more bags 113 via fittings (e.g., weldable fittings) to form a closed system for the aseptic transfer of cells and fluids between the bioreactor and the bags (e.g., from bag 113 to the bioreactor and from the bioreactor to bag 113). In some embodiments, valves (e.g., one or more pinch valves) may be used for fluid management. Fluid management can be achieved by any combination of pumps and valves, as known to those skilled in the art. For example, fluid flow through the fittings may be controlled via one or more mechanical valves (such as pinch valve 140 or other electronically controlled valves, such as...) Figure 1C and Figure 1D What is shown.

[0081] Figure 1D yes Figure 1C An enlarged view of the fluid connection of an embodiment. The fluid connection can be implemented via a pipe fitting (such as sterile tubing welding) to connect one or more bioreactors and one or more bags or reservoirs of a system held on a platform.

[0082] The component can be used with, for example, any number of bioreactors. In a non-limiting example, platform 111 can be configured to accommodate one, two, three, four, five, more than five, or more than ten bioreactors. Component 100 can be configured to expand to accommodate and hold any number of platforms 111 and / or bioreactors 103 to meet cell processing needs. Figure 1A and Figure 1B As shown, the three bioreactors 103 can be accommodated and held on platform 111.

[0083] Figure 2A This is a perspective view illustrating one embodiment of a platform 211 of the components of the present invention, wherein four bioreactors 203 are received and held on the platform 211. The platform may include a rigid frame for securing one or more bioreactors, and a locking or latching mechanism for securing the bioreactors in place on the platform. The platform may be configured such that one or more bioreactors held on the platform are securely held without substantial movement on the platform. In this way, movement of the platform controls movement of the bioreactors held thereon.

[0084] refer to Figure 1B In some embodiments of the component, the platform 111 may be rotatably connected to the support structure 109 via a pair of shafts 115a, 115b, wherein each shaft 115a, 115b may be positioned on a corresponding opposite end of the platform 111 and connected to one of a first end and a second end of the support structure 109 via a rotary joint, wherein the pair of shafts 115a, 115b cooperate to define the axis of rotation of the platform.

[0085] Figure 2B This is a perspective side view illustrating one embodiment of a platform 211 of the components of the present invention, showing a pair of axes 215a, 215b defining a rotation axis X of the platform 211. In a non-limiting example, and as disclosed in more detail herein, the platform 211 can rotate about the rotation axis X from a horizontal starting point (referred to in some embodiments as the original position). The platform can rotate completely or partially about the platform's rotation axis by any angle from 0 degrees to 360 degrees. The platform can rotate in the same direction. The platform can rotate in opposite directions. The platform can change its direction of rotation and can rotate in an oscillating motion. As disclosed in more detail herein, the support structure can also be operable to rotate about the rotation axis, such that the support structure defines a tilting rotation path for the platform. Thus, the platform can rotate 0 degrees to 360 degrees about a horizontal rotation axis, and / or about a tilting rotation axis defined by the rotation axis of the support structure. The platform can rotate about a rotation axis at one or more of the following: varying speed, varying degree of rotation, varying oscillating motion, and varying tilt angle, to provide agitation, shaking, and / or mixing of the contents of one or more bioreactors. The platform can rotate on the rotation axis via shaking and / or oscillating motion. Accordingly, depending on the requirements of the defined cell treatment and / or culture protocols, the movement of the platform can be any of rotation, shaking, agitation, mixing, and / or oscillation. In a non-limiting example, oscillating movement can be a gentle or vigorous reciprocating movement of one or more bioreactors on a horizontal rotation axis and / or a tilted rotation axis.

[0086] Figure 2C An embodiment of the platform of the components of the present invention is shown, illustrating three bioreactors, each held on the platform by adjustable straps.

[0087] Figure 3This is a shaded perspective side view illustrating one embodiment of a system for automated / semi-automated cell processing according to the present invention. As described above, the system may include a component 300 for supporting one or more bioreactors 303 and providing automated or semi-automated cell processing functionality associated with the one or more bioreactors 303. Component 300 may include a base 305 and a support frame 307. The component may include a movable support structure 309 having a first portion coupled to the support frame and a second portion coupled to the base. The component may include a platform 311 rotatably coupled to the support structure 309 and configured to removably receive and hold one or more bioreactors 303 to the platform. Thus, the movable support structure may have a rigid frame and may be operable to move the platform from a horizontal position to an inclined position above or below the horizontal position. The system may include a controller configured to control the movement of the platform 311, at least in part, based on one or more predetermined cell processing schemes, thereby controlling the movement of the one or more bioreactors 303 supported by the platform 311.

[0088] As disclosed herein, platform 311 can be rotatably connected to support structure 309 via a pair of shafts 315a, 315b, wherein each shaft 315a, 315b can be positioned on a corresponding opposite end of platform 311 and connected to one of a first end and a second end of support structure 309 via a rotary joint, such that the pair of shafts 315a, 315b cooperatively define the axis of rotation of platform 311. Therefore, the pair of shafts and the rotary joint cooperatively allow the platform to rotate about a horizontal or tilted axis of rotation. The rotatable connection between the platform and the support structure can be any connection known to those skilled in the art that enables the platform to rotate about an axis of rotation. For example, the rotatable connection can be a direct connection to a servo motor via a rotary joint, a connection via a universal joint and a rotary joint, or a connection via other types of rotatable joints.

[0089] Figure 4 This is a shaded, enlarged perspective view of a support structure 309 according to some embodiments of the invention, wherein a platform 311 is attached to the support structure 309 at opposite ends by a pair of shafts 315a, 315b positioned at opposite ends of the platform 311. A first end of the support structure 309a may include a drive motor 317 operatively coupled to a controller. In some embodiments, the drive motor 317 may be directly coupled via a flange to a first shaft (e.g., 315a) of the pair of shafts on the platform 311, such that actuation of the drive motor 317 via an input from the controller causes rotation of the platform 311 about its axis of rotation. Therefore, the platform may be directly connected to a motor (e.g., a servo motor) via a flange.

[0090] Motor 317 may include a motor controller and / or a driver. In a non-limiting example, the motor may include an AC motor controller and driver, i.e., electronic equipment that modifies the input power of the motor by adjusting the frequency of the motor's power to regulate output speed and torque. The motor may include a DC motor controller and driver, i.e., electrical equipment that modifies the input power by adjusting a constant current source or AC current source to a pulsed DC output with varying pulse duration or frequency. The motor may include a servo motor controller and driver, i.e., electronic equipment that modifies the input power by adjusting a constant current source or AC current source to a pulsed current output with varying pulse duration or frequency. The motor may include a stepper motor controller and driver, i.e., electronic equipment that modifies the input power by adjusting a constant current source or AC current source to a pulsed current output or a "stepping" current output. In some embodiments, the motor may be a servo motor (e.g., 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] Figure 5 This is a solid-line perspective side view illustrating one embodiment of component 500 of the present invention. The system incorporates mechanisms for tilting one or more bioreactors held on a platform to various positions. As shown, support structure 509 can be rotated about an axis of rotation of support structure 509 to cause platform 511 to move from a horizontal orientation to a tilted orientation. The tilted orientation of the platform can be above or below the horizontal orientation. Further, as described in more detail herein, the platform can be rotated about an axis of rotation of the platform such that the bioreactor 503 held therein is positioned for filling or discharging functions.

[0092] In some embodiments, the mechanism for tilting the support structure includes a hinged connector located at one end of the support structure and means for raising and lowering the other end of the support structure. For example... Figure 5 As shown, according to one embodiment of the invention, the support structure 509 can be coupled to the support frame 507 and the base 505. This embodiment includes a pair of shafts, a linear actuator, and a carriage positioned on a threaded rod.

[0093] For example, in some embodiments, one end of the support structure 509a may be rotatably connected to the support frame 507 via a hinged connector, thereby defining the axis of rotation of the support structure. Accordingly, it may be possible for the support structure 509 to tilt from its original horizontal position to an inclined position.

[0094] Figure 6Enlarged perspective views of a first portion 509a and a second portion 509b of a support structure 509 according to some embodiments of the present invention are shown. In some embodiments, the first portion 509a or an end of the support structure 509 may be rotatably connected to a support frame 507 via one or more hinges 519. The hinges may be any type of hinge as known to those skilled in the art, providing a rotatable connection between an end of the support structure 509 and the support frame 507.

[0095] The rotatable connection between the support structure 509 and the support frame 507 defines the axis of rotation of the support structure 509. The axis of rotation provided by the connection between the support structure and the support frame allows the support structure 509 to rotate about this axis, such that the rotation of the support structure 509 defines the tilt axis of rotation of the platform 511. Specifically, the first portion 509a of the support structure 509 can remain fixed, while the opposite ends of the support structure are raised and / or lowered to define the tilt axis of rotation of the platform.

[0096] Accordingly, return to reference Figure 5 Platform 511 is operable to rotate about an axis of rotation provided by the pair of axes coupled to the support structure and about a horizontal or inclined axis of rotation provided by the axis of rotation of the support structure. In some embodiments, the original position of the support structure may be a substantially horizontal position relative to the x-axis and may be defined at 0 degrees, such that the support structure is operable to rotate about 90 degrees relative to the original position. In some embodiments, the support structure may be operable to rotate more than 90 degrees relative to the original position. When the support structure is in the original position, the platform may also be defined as being in the original position. The original position of the platform may include the bioreactor positioned in a horizontal position with the inlet / outlet facing upwards. Additionally and / or alternatively, the original position of the bioreactor may be defined as a position for filling and / or discharging.

[0097] Furthermore, in some embodiments, the lower side of the support structure 509 may be coupled to a linear actuator via a pair of shafts 521a, 521b. The linear actuator may be operatively coupled to a controller such that actuation of the linear actuator via input from the controller causes the support structure 509 to rotate about a rotation axis, raising or lowering one end of the support structure to define the tilt rotation axis of the platform 511.

[0098] For example, the pair of shafts 521a, 521b can be attached to the underside of the support structure 509 via one or more shaft supports 527 and one or more rotary joints 523, for example, located at the ends of the shafts 521a, 521b. The rotating shaft 525 can be inserted horizontally through the rotary joints 523 and the shaft supports 527.

[0099] Figure 7 Showing according to Figure 5 The illustrated embodiment shows a perspective view of the support structure 509, wherein the pair of shafts 521a, 521b are attached to the underside of the support structure 509. The linear actuator may include a threaded rod coupled to a stepper motor and a carriage 529 positioned on the threaded rod. The opposite ends of the shafts 521a, 521b may be coupled to the carriage 529 in the same manner as 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 rotary joints positioned at the ends of these shafts; and a rotating shaft horizontally inserted through the rotary joints and 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 threaded rod, which applies a force to the underside of the support structure. This causes the support structure 509 to rotate about its axis of rotation, thereby moving the platform 511 between a horizontal and tilted orientation. Similarly, reducing the force by the linear actuator via the carriage's movement along the threaded rod causes rotation of the support structure 509, returning it to a substantially horizontal position and / or rotating it to a position below horizontal.

[0101] In some embodiments, the pair of shafts may be hydraulic cylinders that generate linear actuation via hydraulic pressure to apply force to 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 multiple shafts. As described in more detail herein, in some embodiments, the underside of the support structure may not be coupled to a shaft, but may be actuated by means of a carriage coupled to a linear support structure that is part of a support frame to define a tilting axis of rotation.

[0102] Figure 8 This is a perspective view showing the carriage 529 in its original position according to some embodiments of the invention. When the carriage 529 is in its original position on the threaded rod, the support structure 509 can be maintained in a substantially horizontal position. This position also maintains the platform 511 and any bioreactor housed therein in a corresponding horizontal position. When the platform and support structure are in their original positions, the platform can rotate about this horizontal axis of rotation as defined according to the requirements of the cell treatment protocol.

[0103] The initial position of the carriage can be calibrated via a controller to any point on the threaded rod, making the support structure operable to move from a horizontal position to an inclined position above or below the horizontal position. Accordingly, in some embodiments, the controller can be configured to align the platform 511 in a first horizontal position and rotate the platform about its axis of rotation within a range of 0 to 360 degrees compared to the first horizontal position. Rotation about the axis of rotation of the platform 511 can be a full rotation, a partial rotation, a clockwise or counterclockwise rotation, and / or an oscillating movement. In some embodiments, rotation about the axis of rotation of the platform 511 includes an oscillating movement of the platform. Further, the controller can be further configured to align the platform on an inclined axis of rotation, such that the platform rotates about its axis of rotation within a range of 0 to 360 degrees. Rotation about the axis of rotation of the platform 511 can be a full rotation, a partial rotation, a clockwise or counterclockwise rotation, and / or an oscillating movement. In some embodiments, rotation about the axis of rotation of the platform 511 includes an oscillating movement of the platform.

[0104] As described herein, in some embodiments, actuation of the stepper motor via input from a controller moves the carriage 529 along a threaded rod from its original position to define the tilting axis of rotation of the platform 511, making the platform operable to move between an axial rotational position about the platform's horizontal axis of rotation and an axial rotational position about the platform's tilting axis of rotation. For example, in some embodiments, the controller may be operable to rotate the platform about the horizontal axis of rotation and the tilting rotation path at one or more of a varying speed, a varying degree of rotation, a varying oscillating motion, and a varying tilt angle to provide agitation of the contents of one or more bioreactors via rocking and / or oscillating motion.

[0105] The movement of the support structure around its axis of rotation, the platform around its axis of rotation, and the tilted 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 the cell processing functional requirements, the system can be operatively associated with a computer processor that controls the movement of the support structure and platform according to a defined set of cell processing protocols.

[0106] For example, in some embodiments, the controller interacts with a control system architecture to define one or more cell processing protocols, the control system architecture including at least one of a computer, a processor, a network, and / or a graphical user interface (GUI). In a non-limiting example, the cell processing protocol may define the movement of the platform to achieve one or more of rotation, shaking, agitation, mixing, and / or oscillation as required by the defined cell processing and / or culture protocol.

[0107] A user interface used to control one or more user inputs and display one or more outputs from the system (thus allowing the user to interact with the system and control its operation) can be physically connected to the system or can be located remotely. The user interface can be a handheld device, such as a smart tablet, smartphone, or a dedicated device manufactured for the system. User interaction can be implemented on a computer with I / O devices, such as CRT, LCD, LED, or projection devices for displaying information to the user, and input or output devices such as keyboards and pointing devices (e.g., mice or trackballs) through which the user can provide input to the computer. Other types of devices can also be used to provide interaction with the user. For example, feedback provided to the user can be any form of sensory feedback (e.g., visual, auditory, or tactile feedback), and input from the user can be received in any form (including acoustic, speech, or tactile input).

[0108] In some embodiments, the system provides monitoring and control of various aspects or parameters (such as speed, motion, direction of rotation, and degree of rotation, in non-limiting examples) of the actuation of the support structure and the rotation of the platform. The monitoring and control of the various parameters of the actuation of the support structure and the rotation of the platform can be performed using any type of computing device including a processor (e.g., a central processing unit, such as a computer or programmable logic controller (PLC)) or any combination of computing devices (where each device performs at least a portion of a process or method). The controller may include a system employing software, hardware, firmware, hardwiring, or any combination thereof.

[0109] For example, the system may include personal and / or portable computing devices, such as smartphones, tablets, laptops, etc. 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 and exchange data over a network.

[0110] The network can refer to, for example, a private or non-private local area network (LAN), a personal area network (PAN), a storage area network (SAN), a backbone network, a global area network (GAN), a wide area network (WAN), or any collection of such computer networks, such as an intranet, an extranet, or the Internet (i.e., a globally interconnected network system on which various applications or services run, including, for example, the World Wide Web). In alternative embodiments, the communication path between the user interface and the system of the present invention can be wholly or partially wired.

[0111] A network can be any network that carries data. Non-limiting examples of suitable networks that can be used as networks include Wi-Fi wireless data communication technologies, the Internet, private networks, virtual private networks (VPNs), the Public Switched Telephone Network (PSTN), Integrated Services Digital Network (ISDN), Digital Subscriber Link Network (DSL), various second-generation (2G), third-generation (3G), fourth-generation (4G), fifth-generation (5G), and future-generation cellular-based data communication technologies, Bluetooth radio, Near Field Communication (NFC), the latest release of the IEEE 802.11 transport protocol standard, other networks capable of carrying data, and combinations thereof.

[0112] In some embodiments, the network may be selected from the Internet, at least one wireless network, at least one cellular telephone network, and combinations thereof. Therefore, the network may include any number of additional devices (such as additional computers, routers, and switches) to facilitate communication. 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.

[0113] As disclosed herein, the components of the present invention can be used to support one or more bioreactors and provide automated or semi-automated cell processing functions associated with one or more bioreactors. In some embodiments, the components can be configured to perform one or more automated or semi-automated cell processing functions in a closed and sterile environment.

[0114] For example, because the components of the present invention incorporate motors to gently and / or vigorously agitate and properly orient the bioreactor, the system of the present invention can be used for various cell processing workflow steps. In some embodiments, axial rotational positions about a tilted axis of rotation of the platform provide for positioning one or more bioreactors for filling and / or discharging. Thus, the present invention provides mechanized bioreactor manipulation to standardize cell processing functions. In a non-limiting example, cell processing functions may include cell inoculation, culture medium exchange, cell culture, cell concentration, cell mixing, cell agitation, cell washing, and cell harvesting. In some embodiments, cell processing functions include at least one of cell inoculation, cell washing, and cell harvesting. The components of the present invention are scheme-independent and can be used for any type of cell processing function to standardize procedures and eliminate human bias and error. Because the system of the present invention incorporates mechanisms for tilting one or more bioreactors held on the platform to various positions, in a non-limiting example, the present invention can be used for filling and discharging bioreactors.

[0115] As discussed in more detail herein, the system of the present invention can be programmed to perform a specific cell treatment protocol. The cell treatment or culture protocol can, for example, define the movement of the platform, such as the degree and / or duration of rotation, shaking, agitation, mixing, and / or oscillation, according to the requirements of the defined cell treatment and / or culture protocol. The cell culture protocol may include defined periods of platform movement and defined periods of platform stillness. The cell culture protocol may define multiple parameters associated with the cell treatment function. In a non-limiting example, depending on the cell treatment function to be achieved, the parameters may be the platform's axis of rotation, the platform's tilt axis, the platform's filling position, the platform's discharging position, and the mixing, agitation, and / or oscillation movement time, pattern, and / or intensity.

[0116] The system of this invention incorporates various rotating axes coupled with motors to gently and / or vigorously agitate the bioreactor in various positions and to correctly orient the bioreactor according to the needs of cell processing functions. This allows for mechanized bioreactor manipulation to standardize cell processing functions such as cell inoculation, washing, and harvesting. Experiments comparing manual cell processing with the automated / semi-automated cell therapy shaker of this invention demonstrate comparability in terms of yield, cell viability, phenotype, and functionality.

[0117] The system of the present invention can be used to automate / semi-automatize and standardize cell seeding. In one example, the platform can be rotated to a position such that one or more bioreactors held therein can be seeded with human peripheral blood mononuclear cells (PBMCs) and / or dendritic cells pulsed with lysates. Seeding can occur simultaneously with multiple bioreactors held on the platform, or it can occur sequentially. The system of the present invention can be configured to automate cell processing functions individually or as a batch for single and / or multiple bioreactors.

[0118] The system of this invention can be used to automate / semi-automatize and standardize culture medium exchange. For example, culture medium exchange may involve removing used culture medium and adding fresh medium, and may require multiple exchanges to replenish nutrients, supply specific growth factors and cytokines, and remove metabolic waste and other unwanted culture medium components based on cell culture process requirements. The system provides various methods for performing culture medium exchange individually or simultaneously within a bioreactor held on a platform, for example, by: pausing agitation to allow all cells or aggregates to settle to the bottom of the bioreactor under gravity; rotating the platform to a filling position suitable for culture medium exchange and / or removal of the supernatant from used culture medium; and rotating the platform to a position for adding fresh culture medium. This system can simultaneously achieve rapid and complete culture medium exchange in multiple bioreactors by rapidly removing used culture medium in a standardized manner.

[0119] The system of the present invention can be used to automate / semi-automatize and standardize cell harvesting. Separation of cell aggregates and culture medium can be achieved by rotating the platform to a position for harvesting. For example, the platform can be positioned at various heights and angles by rotation about an inclined axis of rotation and / or a horizontal axis of rotation to remove culture medium from one or more specific areas of a bioreactor. In a non-limiting example, the system of the present invention provides a harvesting function that allows for the removal of adherent cells via mechanical agitation. This is particularly useful for strongly adherent cells where the use of enzymatic reagents may not be permitted. Therefore, in some embodiments, the device / system of the present invention is configured to remove adherent cells via mechanical agitation with or without the use of enzymatic reagents.

[0120] The system of the present invention can be used, for example, to automate / semi-automate and standardize one or more washing steps during the cell harvesting process.

[0121] Figure 9 This is a shaded perspective view illustrating one embodiment of the system 900 of the present invention, with the support structure in its original position. The platform is also in its original position, in which the platform and thus the bioreactor held therein are substantially horizontal and facing upwards or frontally.

[0122] Figure 10 This is a shaded perspective view illustrating an embodiment of the system 900 of the present invention, wherein the platform rotates about the platform's rotation axis on an inclined rotation axis defined by the movement of the rotation axis of the supporting structure. In this position, the platform can rotate about the platform's rotation axis, such that the bioreactor held therein rotates approximately 180 degrees from a horizontal position to, for example, discharge the bioreactor.

[0123] like Figure 9 and Figure 10As illustrated in the embodiment of the system 900 of the present invention, the support frame can be configured to hold one or more reservoirs or bags 913 that can be connected (not shown) to a bioreactor held within the platform. Thus, as disclosed herein, in some embodiments, the platform includes a frame configured to hold multiple bioreactors in a stacked position, allowing access to an inlet port and / or outlet port associated with each bioreactor. To accommodate various cell treatment functions, in some embodiments, the components further include one or more reservoirs operatively connected to one or more pinch valves and one or more pumps configured to provide fluid inflow and outflow to the one or more reservoirs and the one or more bioreactors held on the platform. Further, the support frame can be configured to support multiple elements associated with the cell treatment function, such as fittings, fluid reservoirs, fluid collection containers, biotreatment bags, pumps, and / or motors. In some embodiments, the support frame includes multiple supports configured to receive and hold one or more of the fittings, one or more fluid reservoirs, one or more fluid collection containers, one or more biotreatment bags, one or more pumps, and one or more motors. In some embodiments, the component may be configured as a closed system for the aseptic transfer of fluids entering or exiting one or more bioreactors.

[0124] The system of this invention is compatible with Class C (ISO 7) cleanrooms. Components can be made of any material suitable for meeting the functional requirements of cell processing and providing support and stability for the platform along with the bioreactor held therein during gentle or vigorous rotation (including oscillating motion). For example, as known to those skilled in the art, the components of this invention can be constructed from one or more of aluminum, aluminum alloys, stainless steel or other alloys, plastics (such as PPE, high-impact polystyrene), acrylic resins, etc.

[0125] Figure 11 A side perspective view of a component 1100 according to an embodiment of the invention is shown, wherein a support structure can be actuated to rotate about a rotation axis of the support structure, thereby defining a tilting rotation axis of the platform. The platform can be further rotated about a rotation axis of the platform to position the bioreactor held therein for discharge.

[0126] Figure 12 Showing according to Figure 11 End view of one end of component 1100.

[0127] Figure 13 Showing according to Figure 11 Top-view perspective of component 1100.

[0128] Figure 14 Showing according to Figure 11 The second end side perspective of component 1100.

[0129] Figure 15 Showing according to Figure 11 A bottom-view stereoscopic view of component 1100.

[0130] Figure 16 Another embodiment of a component 1600 for supporting one or more bioreactors and providing automated or semi-automated cell treatment functions associated with the one or more bioreactors is shown. The component may include a base 1605 and a pair of vertical support structures 1631a, 1631b extending from the base. The component may include a platform 1611 movably coupled to the pair of vertical support structures 1631a, 1631b and configured to removably receive and hold one or more bioreactors 1603 to the platform. The component may include a controller configured to control the movement of the platform 1611, at least in part, based on one or more predetermined cell treatment protocols, thereby controlling the movement of the one or more bioreactors 1603 supported by the platform 1611.

[0131] In some embodiments, platform 1611 may be movably connected to a pair of vertical support structures 1631a, 1631b via a pair of shafts 1615, the pair of shafts being positioned at respective opposite ends of platform 1611 and connected to the respective vertical support structures in the pair of vertical support structures 1631a, 1631b, wherein the pair of shafts cooperatively define the rotation axis of the platform.

[0132] In some embodiments, the first vertical support structure 1631a of the pair of vertical support structures includes a drive motor 1617 operatively coupled to a controller, and the second vertical support structure 1631b of the pair of vertical support structures includes a linear actuator operatively coupled to a controller. In some embodiments, the drive motor 1617 may be coupled via a universal joint to a first shaft of the pair of shafts 1615 of the platform 1611, such that actuation of the drive motor 1617 via input from the controller causes the platform 1617 to rotate about its axis of rotation.

[0133] Furthermore, in some embodiments, the linear actuator includes a threaded rod 1625 coupled to a stepper motor and a carriage 1629 positioned on the threaded rod 1625. The carriage 1629 includes a linear bearing having a carriage shaft passing through it, wherein the carriage shaft can be coupled via a universal joint to a second shaft in a shaft 1615 of the platform 1611.

[0134] Furthermore, in some embodiments, the horizontal alignment of the carriage 1629 on the threaded rod 1625 with the drive motor 1617 defines the horizontal axis of rotation of the platform. Therefore, as... Figure 16 As shown, in some embodiments, the controller can be configured to align the platform 1611 in a first horizontal position and rotate the platform 1611 about a horizontal axis of rotation within a range of 0 to 360 degrees relative to the first horizontal position. For example, as Figure 16 As shown, platform 1611 can rotate 90 degrees from a first horizontal position. In some embodiments, rotation about a horizontal axis of rotation includes oscillating movement of the platform.

[0135] Figure 17 The tilting rotation axis of a platform 1611 according to an embodiment of the invention is shown, as defined by the movement of a carriage 1629 along a threaded rod 1625. In some embodiments, actuation of a stepper motor via input from a controller moves the carriage 1629 vertically to define the tilting rotation axis of the platform 1611, such that the platform is operable to move between axial rotational positions about a horizontal rotational axis and axial rotational positions about a tilting rotational axis.

[0136] Figure 18 A platform 1611 according to an embodiment of the invention is shown, the platform being tilted and in an axially rotated position about a tilted axis, such that a bioreactor is positioned for discharge. In some embodiments, the axially rotated position about the tilted axis of rotation provides positioning of one or more bioreactors for filling and / or discharge. As disclosed herein, a controller may be operable to rotate the platform about a horizontal axis of rotation and a tilted rotation path at one or more of varying speeds, varying degrees of rotation, varying oscillating motions, and varying tilt angles to provide agitation of the contents of one or more bioreactors via rocking and / or oscillating motions. The assembly may further include a constant tension spring 1633 to provide vertical stability during platform rotation. A first end of the constant tension spring may be coupled to a carriage shaft, and a second end of the vertical stabilizer may be coupled to a base. The constant tension spring may be an elastomeric material configured to stabilize a linear bearing and dampen vibrations of the carriage shaft.

[0137] As disclosed herein, the controller can interact with a control system architecture to define one or more cell processing protocols, the control system architecture including at least one of a computer, a network, and a graphical user interface (GUI). Components can be configured to perform one or more automated or semi-automated cell processing functions in a closed and sterile environment. For example, as disclosed herein, cell processing functions include at least one of cell seeding, cell washing, and cell harvesting. For example, in some embodiments, components can be configured to remove adherent cells via mechanical agitation without the use of enzymatic reagents.

[0138] As disclosed herein, the platform may include a stepped frame configured to hold multiple bioreactors in offset stacked positions, allowing access to inlet and / or outlet ports associated with each bioreactor. In some embodiments, the component further includes one or more reservoirs operatively connected to one or more pinch valves and one or more pumps configured to provide fluid inflow and outflow to the one or more reservoirs and the one or more bioreactors held on the platform. The component may be configured as a closed system for the aseptic transfer of fluids into and out of the one or more bioreactors.

[0139] Example: Comparability Study: NW-22 Automated Equipment (FACTOR) vs. Manual ADCV Process

[0140] Data were collected and compared with the system of the present invention (referred to as "FACTOR" in this study) as disclosed herein, to a manual pharmaceutical process. This study focuses on comparing the quality of sDPs generated on day 8 of an ADCV process (following BMR-4V6) using a manual process with data obtained using the treatment method of the present invention (using FACTOR). This study compares the two types of sDPs treated using the manual method and the FACTOR method. The following parameters were evaluated:

[0141] • Dendritic cell (DC) count and viability on days 7 and 8 (manual counting using trypan blue, following standard procedure)

[0142] • Identification, purity, and impurity characterization performed using standard procedures.

[0143] • Valence using standard procedures

[0144] DC quantity and activity on day 7

[0145] Figures 19A to 19C The study demonstrates the assessment of the number and activity of DCs in sDP using trypan blue on day 7. Figure 19A An assessment of the number of DCs was presented, while Figure 19B The activity was shown as a percentage. Figure 19C Data for batches 30 and 31 are presented, comparing the manual method with an automated / semi-automated processing method using FACTOR. Batch 30 contains 125 × 10⁸ cells inoculated for day 8 culture. 6 One DC. Batch 31 contains 58.54 × 10⁸ cells inoculated for culture on day 8. 6 The number of dendritic cells (DCs) decreased after harvest when the process was performed manually. This effect was more pronounced in batch 31.

[0146] DC quantity and activity on day 8

[0147] Figures 20A to 20C The study demonstrates the assessment of the number and activity of DCs in sDP using trypan blue on day 8. Figure 20A An assessment of the number of DCs was presented, while Figure 20B The activity was shown as a percentage. Figure 20C Data for batches 30 and 31 are presented, comparing the manual method with the automated processing method using FACTOR. DC counts and viability were similar in the sub-batch samples processed manually and by FACTOR.

[0148] Identification and impurity characterization

[0149] Figure 21 This study presents a comparison of the group of identifying markers.

[0150] Figure 22 The set of purity characterization markers used in this study is presented.

[0151] Figure 23 An assessment of sDP purity and identification (particularly sDP identification on day 8) is presented. Identification characterization yields the following results:

[0152] ■ Comparable %HLA-DR+CD86+ cells were observed between manually treated sub-batches and sub-batches treated with FACTOR.

[0153] ■ All samples passed the acceptance criteria of >70%.

[0154] ■ Comparable percentages of CD141+CD86+ cells were observed between manually processed sub-batches and sub-batches treated with FACTOR.

[0155] ■ Figures 24A to 24C The sDP impurity characterization on day 8 is shown. Figure 24A An evaluation of sDP impurities is presented. Figure 24B This is a chart characterizing the total impurities. Figure 24CThis is the raw data table used for impurity characterization. This data indicates that sDP processed manually and with FACTOR have similar impurity spectra.

[0156] Valence assessment

[0157] Figure 25A The valence assessment was demonstrated. Valence testing was performed in a heterologous system according to a standard protocol.

[0158] Figure 25B Results for positive and negative controls are presented. Results for positive and negative controls, background control (MLR), and gated control were all passed. Titer was measured as a percentage and defined as T cell proliferation % minus T cell proliferation % in the background control. SI was defined as SI = T cell proliferation % / T cell proliferation % in the background control.

[0159] Figure 25C A graph showing the sDP potency assessment on day 8 is presented. Specifically, potency was determined for four samples. The positive control, negative control, and background control passed the assay acceptance criteria. Both methods met the release criteria of SI > 2. Some variability was observed in batch 30, while batch 31 was more consistent. Sample sizes were small.

[0160] result:

[0161] • For samples treated with FACTOR, a better trend in DC recovery was observed on day 7.

[0162] • The number and activity of sDPs were similar on day 8.

[0163] • The sDP identification, purity, and impurity profiles were similar on day 8.

[0164] • All valences expressed in SI in all batches met the release criteria (>2), but some variability was observed in batch 30.

[0165] Example: Comparability study: Using FACTOR's oscillating harvest to compare with manual harvesting

[0166] Figure 26 It is a data table that compares various characteristics of cells harvested on days 7 and 8 using an embodiment of the apparatus and method of the present invention for four independent runs, relative to using a manual method.

[0167] Figure 27 This is a comparison of the various characteristics described above, using a harvesting method based on an embodiment of the FACTOR system versus harvesting using manual methods, with data from CS 26, measured on days 7 and 8. Specifically, Figure 27It also includes comparative charts of T cell proliferation, titer, and stimulation index for this run.

[0168] Figure 28 This is a data table for running CS 27, with results measured on days 7 and 8, comparing the aforementioned characteristics of harvesting using an embodiment of the FACTOR system versus harvesting using manual methods. Specifically, Figure 28 It also includes comparative charts of T cell proliferation, titer, and stimulation index for this run.

[0169] Figure 29 It demonstrates the DP identification of all events, cells, live cells, and single cells for manual harvesting on days 7 and 8.

[0170] Figure 30 It demonstrates the DP identification of all events, cells, live cells, and single cells for manual harvesting on days 7 and 8.

[0171] Figure 31 This is a data sheet for running CS 28, with results measured on days 7 and 8, comparing the aforementioned characteristics of oscillatory harvesting using an embodiment of the FACTOR system versus harvesting using manual methods. Specifically, Figure 31 It also includes comparative charts of T cell proliferation, titer, and stimulation index for this run.

[0172] Figure 32A and Figure 32B It is a table that includes data measured on days 7 and 8 of running CS19 to CS-25, comparing the aforementioned characteristics of oscillatory harvesting using an embodiment of the FACTOR system versus harvesting using manual methods.

[0173] Figure 33 A functional allogeneic assay of D8 DC + CS 22 T cells + CD3 running CS 23 was demonstrated.

[0174] Figure 34 The graphs show the percentage of T cell proliferation, titer, and stimulation index in the functional allogeneic assays running CS 23. For these calculations, titer = Proliferating T cells % (COSTIM assay) - Proliferating T cells % (MLR assay). Stimulation index (SI) = Proliferating T cells % (COSTIM assay) / Proliferating T cells % (MLR)

[0175] Incorporation

[0176] Throughout this disclosure, other sources, such as patents, patent applications, patent publications, journals, books, papers, and web content, have been referenced and cited. All such sources are hereby incorporated in their entirety for all purposes.

[0177] equivalent

[0178] In addition to those shown and described herein, various modifications to the invention and many other embodiments thereof will become apparent to those skilled in the art from the entire contents of this document (including references to scientific and patent literature cited herein). The subject matter of this document contains important information, examples, and guidance that can be adapted for practice in the various embodiments of the invention and their equivalents.

Claims

1. A component for supporting one or more bioreactors and providing automated or semi-automated cell processing functions associated with said one or more bioreactors, said component comprising: Base and support frame; A movable support structure having a first portion connected to the support frame and a second portion connected to the base; A platform, rotatably coupled to the support structure and configured to removably receive one or more bioreactors and hold the one or more bioreactors to the platform; as well as A controller configured to control the movement of the platform, at least in part, based on one or more predetermined cell treatment protocols, thereby controlling the movement of one or more bioreactors supported by the platform.

2. The component as claimed in claim 1, wherein, The platform is rotatably connected to the support structure via a pair of shafts, wherein each shaft is positioned at a corresponding opposite end of the platform and connected to one of a first end and a second end of the support structure via a rotary joint, such that the pair of shafts cooperate to define the rotation axis of the platform.

3. The component as claimed in claim 2, wherein, The first end of the support structure includes a drive motor operatively connected to the controller.

4. The component as claimed in claim 3, wherein, The drive motor is directly connected via a flange to the first shaft of a pair of shafts of the platform, such that actuation of the drive motor via input from the controller causes the platform to rotate about the platform's axis of rotation.

5. The component as claimed in claim 2, wherein, One end of the support structure is rotatably connected to the support frame via a hinged connector that defines the axis of rotation of the support structure.

6. The component of claim 5, wherein, The lower side of the support structure is connected to a linear actuator via a pair of shafts. The linear actuator is operatively connected to the controller such that actuation of the linear actuator via input from the controller causes the pair of shafts to apply a force to the lower side of the support structure and cause the support structure to rotate about its axis of rotation, thereby moving the platform between a horizontal orientation and an inclined orientation.

7. The component of claim 6, wherein, The linear actuator includes a threaded rod coupled to a stepper motor and a carriage positioned on the threaded rod, the carriage being coupled to the pair of shafts via: a pair of shaft supports connected to a rotary joint positioned at a first end of the shafts; and a rotating shaft inserted horizontally through the rotary joint and the shaft supports.

8. The component of claim 6, wherein, The lower side of the support structure is connected to the pair of shafts via: a pair of shaft supports connected to a rotary joint positioned at a second end of the shafts; and a rotating shaft inserted horizontally through the rotary joint and the shaft supports.

9. The component of claim 7, wherein, When the carriage is in its original position on the threaded rod, the support structure remains in a substantially horizontal position, thereby maintaining the platform and any bioreactors housed on the platform in a corresponding horizontal position.

10. The component of claim 7, wherein, The controller is configured to align the platform in a first horizontal position and rotate the platform about its axis of rotation within a range of 0 to 360 degrees relative to the first horizontal position.

11. The component of claim 10, wherein, The rotation about the axis of rotation of the platform includes the oscillating movement of the platform.

12. The component of claim 9, wherein, The stepper motor, actuated via input from the controller, moves the carriage along the threaded rod from its original position to define the tilt axis of rotation of the platform, enabling the platform to operate between an axial rotational position about the platform's horizontal axis of rotation and an axial rotational position about the platform's tilt axis of rotation.

13. The component of claim 12, wherein, The axial rotational position about the inclined axis of rotation of the platform provides a positioning for filling and / or discharging of the one or more bioreactors.

14. The component of claim 13, wherein, The controller is operable to rotate the platform about the horizontal rotation axis and the tilted rotation path at one or more of the following: varying speed, varying degree of rotation, varying oscillating motion, and varying tilt angle, to provide agitation of the contents of the one or more bioreactors via rocking and / or oscillating motion.

15. The component of claim 1, wherein, The controller interacts with a control system architecture to define one or more cell processing schemes, the control system architecture including at least one of a computer, a processor, a network, and / or a graphical user interface (GUI).

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

17. The component of claim 16, wherein, The cell processing function includes at least one of cell seeding, cell washing, and cell harvesting.

18. The component of claim 17, wherein, The component is configured to remove adherent cells via mechanical agitation, with or without the use of enzyme reagents.

19. The component of claim 1, wherein, The platform includes a frame configured to hold multiple bioreactors in a stacked position, allowing access to the inlet and / or outlet ports associated with each bioreactor.

20. The component of claim 19, further comprising one or more reservoirs operably connected to one or more pinch valves and one or more pumps configured to provide fluids flowing into and out of the one or more reservoirs and one or more bioreactors held on the platform.

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

22. The component of claim 1, wherein, The support frame includes multiple supports configured to receive and hold one or more of the following: tubing, fluid reservoir, fluid collection container, bioprocessing bag, pump, and motor.

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