Material transfer devices, and related systems and methods

Abstract

The present invention provides a material transfer device for use in conjunction with bioprocessing, as well as related systems and methods. [Solution] A method comprising: receiving microcarriers or cell aggregates in used culture medium from a first bioreactor within a material transfer device; holding the microcarriers or cell aggregates on a first side of the filter of the material transfer device while flowing the used culture medium through the filter in a first direction; reversing the flow through the filter by flowing the fresh culture medium through the filter in a second direction opposite to the first direction in order to resuspend the microcarriers or cell aggregates in fresh culture medium; and flowing the microcarriers or cell aggregates in the fresh culture medium from the material transfer device to a second bioreactor.

Description

Technical Field

[0001] Related Applications Section This application claims the benefit and priority of U.S. Provisional Patent Application No. 62 / 704,473 filed on May 12, 2020, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0002] The present invention relates generally to bioprocessing equipment, and in particular to material transfer devices and related systems and methods for use with bioprocessing. Background Art

[0003] Biomanufacturing processes for therapeutic cells grown in suspension culture in single-use bioreactors have been developed for a wide range of cell and gene therapy applications. Due to their types and characteristics, many of these therapeutic cells rapidly proliferate either as aggregated clumps or while adhered to the surface of microcarriers (MC). Addressing the unique process requirements of these cell aggregates or MC is essential for the scalable manufacturing of cell therapy products and gene therapy products for treating patients with indications of severe diseases, since cell aggregates and MC are larger and heavier than suspended single cells.

[0004] As part of the cell culture process, it is necessary to exchange the liquid medium in which cell aggregates or MC are suspended, that is, to remove spent medium and add fresh medium. In cell proliferation steps, this exchange can be used to replenish nutrients and eliminate metabolic waste. In multi-stage directed differentiation procedures, rapid and efficient medium exchange between each stage is important for removing previously used differentiation factors and preventing undesired heterogeneous differentiation.

[0005] There are various known techniques for performing medium changes during cell proliferation or differentiation of therapeutic cells in a bioreactor. One common method is to pause agitation and allow all cell aggregates or MCs to settle to the bottom of the bioreactor by gravity. Once a bed of settled cell aggregates or MCs has formed, the supernatant of the used medium is removed from the bed of settled cell aggregates or MCs, fresh medium is added, and agitation is resumed to resuspend the cell aggregates or MCs. This sedimentation method has two potential problems. Firstly, especially during long-term medium change processes in large bioreactors, a temporary cessation of mixing can lead to cell damage through unwanted aggregation, nutrient starvation, and deviations from key process parameters such as temperature, pH, and dissolved oxygen levels. Secondly, it is difficult to completely remove the used medium without losing cells near the bed of settled cell aggregates or MCs, and as a result, unwanted residual differentiation factors and metabolic waste may be carried over to the used medium. Furthermore, these problems worsen as the bioreactor volume increases, requiring longer periods for more cell aggregates or MCs to settle during the medium exchange process, and necessitating the removal and replenishment of larger amounts of medium.

[0006] The large-scale production of therapeutic cells grown on microclusters or as cell aggregates has not yet been demonstrated, and the need to enable production technologies is becoming more urgent as more therapeutic cell candidates approach clinical trials. A potential obstacle to large-scale cell culture processes is the current lack of reliable and robust methods for large-scale medium changes, particularly for differentiating pluripotent stem cells (PSCs) grown as suspended aggregates in bioreactors. [Overview of the project]

[0007] The disclosed embodiments relate to material transfer devices and associated systems that efficiently filter cells, cell aggregates, and / or associated microcarriers (MCs) from used culture media, while reducing stress on cells during the filtration and / or transfer process. Thus, the disclosed embodiments minimize the time therapeutic cells spend outside the ideal fluid environment of a bioreactor (consisting of nutrients, agitation, temperature, etc.), thereby addressing current obstacles to commercial-scale production. The material transfer devices and systems are intended to form sterile connections to other devices used in the cell culture process (e.g., bioreactors). This ensures that therapeutic cells remain in a completely closed system throughout the cell culture process to minimize the risk of contamination from the external environment. The material transfer devices and systems may also be automated and can provide relatively rapid cycles. Thus, the material transfer devices may have a relatively small volume compared to a bioreactor, while providing medium exchange for larger bioreactor volumes.

[0008] To this end, the material transfer device includes a housing comprising first and second housing portions separated by a screen and having corresponding first and second ports. The first pair of ports can be used to transport culture media containing cell aggregates or microcarriers into and out of the housing, the screen can be used to separate the cell aggregates or microcarriers from the culture media, and the second pair of ports can be used to transport culture media that does not contain cell aggregates or microcarriers into and out of the housing.

[0009] For example, one of the first pair of ports may be used to transport used culture medium containing cell aggregates or microcarriers into the first housing portion, and one of the second pair of ports may be used to transport used culture medium out of the housing. The other of the second pair of ports may then be used to transport fresh culture medium into the housing to rehydrate the cell aggregates and microcarriers by passing through the screen, and the other of the first pair of ports may be used to transport fresh culture medium with suspended cell aggregates or microcarriers out of the first housing portion. The screen, housing, and / or ports may be arranged in different configurations. For example, the screen and / or housing portion may be parallel to the horizontal plane and / or angled to the horizontal plane. Furthermore, the ports may be arranged in different configurations so that used culture medium can be transported downward or upward through the material transfer device.

[0010] According to the first embodiment, the material transfer device includes a housing comprising a first housing portion and a second housing portion, and a screen. The first housing portion includes a first inlet port, a first outlet port, and a first transfer opening. The second housing portion has a second inlet port, a second outlet port, and a second transfer opening. The first transfer opening is disposed adjacent to and in communication with the second transfer opening. The screen is disposed between the housing portions adjacent to the first and second transfer openings.

[0011] According to a second embodiment, a method is used using a material transfer device having a first housing portion and a second housing portion, with a screen disposed between the housing portions, each housing portion including an inlet port, an outlet port, and corresponding fluid tubes connected to the inlet port and the outlet port, the method comprising pumping spent culture medium containing cell aggregates or microcarriers through the inlet port of the first housing portion. The method comprises filtering the cell aggregates or microcarriers using a screen and passing the spent culture medium through the screen into the second housing portion. The method comprises pumping the spent culture medium out of the outlet port of the second housing portion and pumping fresh culture medium into the first housing portion or the second housing portion. The method comprises resuspending the cell aggregates or microcarriers. The method comprises pumping fresh culture medium and suspending the cell aggregates or microcarriers out of the outlet port of the first housing portion.

[0012] According to a third embodiment, a control system for controlling fluid flow through a material transfer device having a first housing portion and a second housing portion, with a screen disposed between the housing portions, wherein each housing portion includes an inlet port, an outlet port, and corresponding fluid tubes connected to the inlet port and the outlet port, and the control system includes a base, and a plurality of valves and a plurality of pumps. The base is adapted to define a receptacle and support the material transfer device, and the plurality of valves and the plurality of pumps are adapted to control the flow of culture medium through the inlet port and the outlet port, respectively, of the transfer device.

[0013] According to the fourth embodiment, the material transfer device includes a housing and a screen. The housing includes a first housing portion and a second housing portion, each housing portion including an inlet port and an outlet port. The screen is disposed between the housing portions. The inlet port of the first housing portion is configured to allow used culture medium containing cell aggregates or microcarriers to flow into the first housing portion, and the screen is adapted to prevent cell aggregates or microcarriers from passing through the screen, while allowing the used culture medium to pass through the screen and into the second housing portion and towards the outlet port of the second housing portion. The inlet port of the second housing portion is configured to allow fresh culture medium to flow into the second housing portion, and the screen is adapted to allow the fresh culture medium to flow through the screen and resuspend cell aggregates or microcarriers in the first housing portion.

[0014] According to the fifth embodiment, the material transfer device includes a mesh screen, a first housing portion, and a second housing portion. The first housing portion has a cell inlet and a cell outlet, and the second housing portion has a culture medium outlet and a culture medium inlet and is coupled to the first housing portion. The mesh screen is positioned between the first housing portion and the second housing portion and is adapted to prevent cell aggregates or microcarriers from passing through the screen and to allow used culture medium to pass through the screen and flow to the fluid outlet. The mesh screen is also adapted to allow fresh culture medium to pass through the screen and move to the cell outlet.

[0015] According to the sixth embodiment, the method includes receiving microcarriers or cell aggregates in used culture medium from a first bioreactor within a material transfer device; retaining the microcarriers or cell aggregates on the first side of the filter of the material transfer device while flowing the used culture medium through a filter in a first direction; reversing the flow through the filter by flowing fresh culture medium through a second direction opposite to the first direction, thereby resuspending the microcarriers or cell aggregates in the fresh culture medium; and flowing the microcarriers or cell aggregates in the fresh culture medium from the material transfer device to a second bioreactor.

[0016] Further according to the first, second, third, fourth, fifth, and / or sixth embodiments described above, the apparatus and / or method may similarly further include one or more of the following embodiments.

[0017] In one embodiment, a first inlet port of a first housing portion is arranged to transport used culture medium containing cell aggregates or microcarriers into the first housing portion, and a screen is adapted to prevent cell aggregates or microcarriers from passing through the screen, while allowing the used culture medium to pass through the screen and into the second housing portion and toward the second outlet port of the second housing portion. Furthermore, a second inlet port of the second housing portion is arranged to transport fresh culture medium into the second housing portion, and a screen is adapted to allow the fresh culture medium to flow through the screen and resuspend the cell aggregates or microcarriers in the first housing portion.

[0018] According to another embodiment, the first outlet port of the first housing portion is positioned to allow fresh medium containing cell aggregates or microcarriers to flow out of the first housing portion.

[0019] In another embodiment, the screen is a flexible screen that defines multiple pores that allow material to flow through it.

[0020] According to another embodiment, the material transfer device further comprises a support operably coupled to the housing and disposed between the first housing portion and the second housing portion and adjacent to the screen to provide structural support for the screen.

[0021] According to another embodiment, the support comprises at least one of a first support extending across the first transfer opening and a second support extending across the second transfer opening.

[0022] According to another embodiment, the first support is coupled to the first housing portion adjacent the first transfer opening, and the second support is coupled to the second housing portion adjacent the second transfer opening.

[0023] According to another embodiment, the support comprises a lattice structure.

[0024] According to another embodiment, the first housing portion and the second housing portion are each a rigid or semi-rigid structure.

[0025] According to another embodiment, the first housing portion and the second housing portion each comprise or are made of a flexible material.

[0026] According to another embodiment, the first housing portion is removably coupled to the second housing portion or is integrally formed with the second housing portion.

[0027] According to another embodiment, the first housing portion is an upper housing portion and the second housing portion is a lower housing portion.

[0028] According to another embodiment, the material transfer device comprises a seal at an interface between the first housing portion and the second housing portion.

[0029] According to another embodiment, the first inlet port and the first outlet port are centrally disposed within the first housing portion, and the second inlet port and the second outlet port are centrally disposed within the second housing portion.

[0030] According to another embodiment, when the material transfer device is in use, the screen is disposed horizontally relative to a horizontal plane.

[0031] According to another embodiment, the first housing portion is a lower housing portion and the second housing portion is an upper housing portion.

[0032] According to another embodiment, the first housing portion comprises a funnel shape leading to the first outlet port.

[0033] According to another embodiment, the first housing portion further comprises an internal barrier extending across the width of the first housing portion and adapted to prevent accumulated cell aggregates or microcarriers from covering the first inlet port of the first housing portion.

[0034] According to another embodiment, the second outlet port of the second housing portion is adapted to be positioned lower than the second inlet port of the second housing portion.

[0035] According to another embodiment, the first inlet port of the first housing portion is positioned relative to the screen to allow spent culture medium containing cell aggregates or microcarriers to flow tangentially along the surface of the screen.

[0036] According to another embodiment, the screen and the housing are at an angle relative to a horizontal plane when the material transfer device is in use.

[0037] According to another embodiment, the method further comprises passing fresh medium through the screen into the first housing portion.

[0038] In another embodiment, passing the used culture medium through the screen includes passing the used culture medium through a first transfer opening in a first housing portion and a second transfer opening in a second housing portion that is adjacent to and in communication with the first transfer opening.

[0039] According to another embodiment, the method includes supporting a screen using a support operably coupled to a housing and disposed between a first housing portion and a second housing portion.

[0040] In another embodiment, passing the used culture medium through a screen includes passing the used culture medium through a screen positioned horizontally with respect to a horizontal plane.

[0041] In another embodiment, passing the used culture medium through a screen includes passing the used culture medium through a screen positioned at a certain angle to a horizontal plane.

[0042] Another embodiment further includes a material transfer device disposed on or otherwise mounted on a base.

[0043] In another embodiment, the valve and pump are configured to (1) pump used culture medium containing cell aggregates or microcarriers through the inlet port of the first housing portion, allowing the cell aggregates or microcarriers to be filtered by a screen and sent through the screen into the second housing portion; (2) pump the used culture medium out of the outlet port of the second housing portion; (3) pump fresh culture medium into the second housing portion, allowing the fresh culture medium to flow through a screen into the first housing portion and resuspend the cell aggregates or microcarriers; and (4) pump the fresh culture medium with suspended cell aggregates or microcarriers out of the outlet port of the first housing portion.

[0044] In another embodiment, the base further includes a wall that is coupled to the base and carries two of the valves and two of the corresponding pumps, the base further carries two of the valves and two of the corresponding pumps.

[0045] According to another embodiment, the base includes a first side opposite to a second side, each side defining a channel through which at least one fluid tube is adapted.

[0046] According to another embodiment, the control system includes a flow meter adapted to determine the flow rate of used culture medium entering and leaving a second portion of the housing, and the control system is adapted to change the pump flow rate that pumps the used culture medium out of the outlet port of the second housing portion of the transfer device in response to the determined flow rate that satisfies a threshold flow valve.

[0047] According to another embodiment, the control system includes a user interface adapted to allow inputs to be received by the control system and to control the flow sequence through the transport device.

[0048] In another embodiment, the valve is a pinch valve.

[0049] According to another embodiment, the control system includes a heater adapted to heat the material transfer device.

[0050] According to another embodiment, the method includes discharging the used medium from the material transfer device before reversing the flow through the filter by flowing the fresh medium through the filter in a second direction.

[0051] According to another embodiment, receiving microcarriers or cell aggregates in spent culture medium from a first bioreactor into a material transfer device includes pumping the microcarriers or cell aggregates in spent culture medium into the material transfer device.

[0052] According to another embodiment, receiving microcarriers or cell aggregates in spent culture medium from a first bioreactor into a material transfer device includes, based on gravity, flowing the microcarriers or cell aggregates in spent culture medium into the material transfer device.

[0053] According to another embodiment, receiving microcarriers or cell aggregates in spent culture medium from a first bioreactor into a material transfer device includes flowing the microcarriers or cell aggregates in spent culture medium into the material transfer device based on the pressure difference between the first bioreactor and the material transfer device.

[0054] Another embodiment further includes heating the material transfer device.

[0055] Another embodiment further includes rotating the material transfer device to facilitate the movement of microcarriers or cell aggregates away from the screen.

[0056] In another embodiment, the first bioreactor, the material transfer device, and the second bioreactor are a closed system for providing a sterile environment.

[0057] In another embodiment, the first bioreactor, the material transfer device, and the second bioreactor are aseptically connected as part of a completely closed system. [Brief explanation of the drawing]

[0058] [Figure 1] A schematic diagram of an exemplary control system as taught in this disclosure is shown. [Figure 2] Figure 1 is a cross-sectional view of an embodiment of the material transfer device. [Figure 3] Figure 2 is an enlarged view of the material transfer device. [Figure 4] This is a cross-sectional view of another embodiment of the material transfer device shown in Figure 1. [Figure 5]Figure 4 is a front view of the material transfer device, showing the funnel-shaped portion of the first housing section. [Figure 6] Figure 1 is an isometric view of an embodiment of the control system. [Figure 7] Figure 6 is an isometric view of the control system, in which the material transfer device is located inside the receptacle and / or rests on the upper surface of the base. [Modes for carrying out the invention]

[0059] The following text discloses a detailed description of exemplary methods, apparatus, and / or products, but it should be understood that the legal scope of the property rights is defined by the claims language set out at the end of this patent. Therefore, the following detailed description should be interpreted as examples only, and does not describe all possible examples, as it would be impractical, if not impossible, to describe all possible examples. Many alternative examples may be implemented using either the current art or art developed after the filing date of this patent. Such alternative examples are assumed to still be within the scope of the claims.

[0060] The exemplary methodologies, apparatus, and / or products differ from seemingly similar methods and devices that utilize mesh filters to restrict MCs in a fluid. Commercial devices with porous mesh filters (such as cell strainers) are intended to capture MCs while separating cells from the MCs, and then allow the desired cells or target cell products to flow through the pores. The desired cells or products remain in the same fluid after passing through the filter, while the MCs are collected on one side of the filter and subsequently discarded. Typically, the pore size is selected so that the cells are separated from the surface of the MCs before entering the device, and so as to restrict the passage of the MCs. Alternatively, if the purpose of the cells is to produce proteins or similar cell-based products intended to pass through the mesh, the cells may remain attached to the surface of the MCs, and MCs with attached cells are still discarded together. Aggregates of therapeutic cells are typically similar in size to or even larger than the MCs. However, since the cells themselves are the desired products, the cell aggregates must be recovered during the medium exchange process and cannot be handled in the same way as the disposable MCs in the cell retention device.

[0061] In contrast, the following exemplary methodologies, apparatus, and / or products are distinguished by holding MCs or cell aggregates on one side of a mesh filter and then recovering and transferring them by inverting the flow of new medium through the mesh filter into a new bioreactor filled with fresh medium and pre-treated. This process using the exemplary device allows for a rapid and complete medium exchange. When the therapeutic cells of interest are grown on the surface of the MCs or as aggregates, they are not intended to be discarded in any way. Rather, MCs with cells or cell aggregates grown on their surface enter the exemplary device (outside the cell culture device such as a bioreactor) in the used medium and are then collected and recovered on one side of the mesh filter. Only undesirable used medium flows rapidly through the filter and is discarded as waste. At a given time, fresh medium flows from the opposite side of the filter (the same as the waste side) to remove the MCs and aggregates from the filtration pores and suspend them in the new medium. Fresh culture medium containing MCs (microclusters) with the desired cells growing on its surface, or MCs with the cells growing as aggregates, exits the exemplary device from the opposite side of the mesh (the waste side) into a new bioreactor filled with fresh medium and pre-treated.

[0062] Furthermore, to facilitate the complete recovery of MCs or cell aggregates, the device can be rotated to allow gravity to drop the MCs or aggregates from the mesh filter during the backflow of the culture medium. This can be done quickly before fresh medium is introduced into the device to ensure a complete medium exchange. Additionally, the rotatable device may incorporate a heating element to minimize any temperature differences experienced by cells during short periods when they are not suspended in heated medium.

[0063] Figure 1 illustrates a schematic diagram of an exemplary control system 100 as taught in this disclosure. The control system 100 can be used to transfer cell aggregates or microcarriers (MCs) from a first bioreactor 102 to a second bioreactor 104. The control system 100 can also be used to transfer MCs from one or more first bioreactors to one or more second bioreactors. The MCs may be suspended in a culture medium that can be replaced by the control system 100.

[0064] In the embodiments shown, the control system 100 includes a receptacle 105 for receiving or otherwise transporting the material transfer device 106, and in part includes a plurality of valves 108, 110, 112, 114, a plurality of pumps 116, 118, 120, 122, a flow meter 123, a heater 124, and a controller 125. In some implementations, the first bioreactor 102, the material transfer device 106, and the second bioreactor 104 are a closed system that provides a sterile environment. The sterile environment may be a sterile environment free from contaminants, etc. The controller 125 is electrically and / or communicatively coupled to valves 108, 110, 112, 114, pumps 116, 118, 120, 122, flow meter 123, and heater 124, and is adapted to cause the valves 108, 110, 112, 114, pumps 116, 118, 120, 122, flow meter 123, and heater 124 to perform various functions as disclosed herein. The valves 108-114 may be pinch valves, and the pumps 116-122 may be peristaltic pumps. However, other types of valves may prove suitable, other types of pumps may prove suitable, and / or one or more of the valves 108-114 and / or pumps 116-122 may be omitted as further described below.

[0065] The material transfer device 106 and valves 108-112 function as a four-way fluid control device, controlling the flow of used culture medium through the housing 126 of the material transfer device 106, the flow from the first bioreactor 102 to the waste receptacle 127, and the flow of fresh culture medium from the fresh culture medium source 128 through the housing 126 of the material transfer device 106 to the second bioreactor 104. Pumps 116, 118, 120, and 122 are used to pump used or fresh culture medium through fluid tubes 130, 131, 132, and 133 that fluid-couple the material transfer device 106, the first bioreactor 102, the waste receptacle 127, the fresh culture medium source 128, and the second bioreactor 104.

[0066] In the illustrated embodiment, the material transfer device 106 includes a screen 129 disposed between a first portion 134 and a second portion 135 of the housing 126. The housing portions 134 and / or 135 may be detachably coupled to each other or formed integrally together, and each may be a rigid structure, a semi-rigid structure, and / or made of a flexible material.

[0067] The first housing portion 134 includes a first inlet port 136, a first outlet port 137, and a first transfer opening 138. Similarly, the second housing portion 135 includes a second outlet port 140 adjacent to and communicating with the first transfer opening 138, a second inlet port 142, and a second transfer opening 143. As a result of the communication between the transfer openings 138 and 143, a fluid (e.g., culture medium) can flow between the first transfer opening 138 and the second transfer opening 143, and through the screen 129 adjacent to the first transfer opening 138 and the second transfer opening 143.

[0068] In embodiments where the first housing portion 134 and the second housing portion 135 are detachably coupled together, a seal 145 may be disposed at the interface between the first housing portion 134 and the second housing portion 135 to prevent fluid from leaking out of the housing 126. The seal 145 may also be an external seal that wraps around the housing 126 at the interface between the housing portions 134 and 135. Furthermore, in such versions, the housing portions 134 and 135 may be coupled together with one or more fasteners, clamps, magnets, etc.

[0069] During the MC transfer / culture medium exchange process, valves 108 and 110 may be open, and pump 116 may pump the used culture medium containing the MC through the first inlet port 136 and into the first housing section 134. Screen 129 allows the used culture medium to pass through screen 129 and enter the second housing section 135, while preventing the MC from passing through screen 129. The used culture medium may flow into the second housing section 138 based on gravity or based on the fill level of the used culture medium in the housing 126, for example, when the first housing section 134 is below the second housing section 135. As the used culture medium is collected in the second housing section 135, or thereafter, pump 118 may pump the used culture medium from the second housing section 135 to the waste receptacle 127.

[0070] As MC is collected on the screen 129, the flow rate of spent medium into and / or from the material transfer device 106 may decrease. To compensate for the change in flow rate through the screen 129, the controller 125 may change the pump speed of the pump 116. In this example, a flow meter 123 is used to determine the flow rate of spent medium leaving the second housing section 135. The controller 125 may change the pump flow rate of the pump 116 in response to the determined flow rate that satisfies the threshold flow valve. Thus, the control system 100 may dynamically change the pump speed based on the flow rate into, through, and / or from the material transfer device 106.

[0071] In some embodiments, after a threshold amount of MC is collected on screen 129 and / or after used medium is pumped out of the second housing section 135, the controller 125 may close valves 108, 110 and open valves 112, 114, causing the pump 120 to transport fresh medium from the fresh medium source 128 through the second inlet port 142 into the second housing section 135. Alternatively, the second inlet port 142 may be coupled to the first housing section 134, so that the fresh medium flows directly into the first housing section 134. The fresh medium entering the housing 124 may be preheated and / or a heater 124 may heat the fresh medium and / or the contents of the material transfer device 106 to reduce the temperature fluctuations to which the MC is exposed. The heater 124 may heat a section of the housing 126 or the entire housing 126.

[0072] After the second housing portion 135 is filled with fresh medium, the fresh medium passes through the screen 129 and resuspends the MC in the first housing portion 134, for example by removing the MC from the screen 129. As the first housing portion 134 is filled with fresh medium, and / or thereafter, the pump 122 can pump the fresh medium suspending the MC through the first outlet port 137 and to the second bioreactor 104. The second bioreactor 104 may be pre-treated with fresh medium so as to allow any spent medium contained in the housing 126 to be diluted in the larger total volume of the second bioreactor 104 after the pump 118 has pumped the spent medium from the material transfer device 106. Subsequently, if additional MC is transferred between the bioreactors 102, 104, the controller 125 can close valves 112, 114 and open valves 108, 110. Although the above embodiment discloses the transfer of MC from one bioreactor to another, the MC may be transferred from one or more first bioreactors to one or more other second bioreactors. For example, the MC may be transferred from two bioreactors to a single bioreactor.

[0073] The first inlet port 136 and the first outlet port 137 may be positioned toward the top of the material transfer device 106, on the side of the material transfer device 106, and / or toward the bottom of the material transfer device 106. If the first inlet port 136 and the first outlet port 137 are positioned toward the top of the material transfer device 106, MC may accumulate on the upper surface of the screen 129, and the spent medium may exit toward the bottom of the material transfer device 106. If the first inlet port 136 and the first outlet port 137 are positioned toward the bottom of the material transfer device 106, MC may accumulate on the lower surface of the screen 129 and / or otherwise within the lower portion of the housing 126, and the spent medium may exit toward the top of the material transfer device 106. Regardless of the orientation of ports 136, 137, 140, and 142, the control system 100 can allow the fluid through the material transfer device 106 to flow in any direction.

[0074] In some implementations, the control system 100 also includes a rotor 143 configured to rotate the material transfer device 106 to facilitate the MC moving away from the screen 129 and being resuspended. The rotor 143 may include a cradle defining a receptacle 105, actuators, etc. The cradle may be journaled to allow the cradle to move between different positions.

[0075] In the above embodiments, pumps 116-122 are used to pump the fluid through the material transfer device 106, but one or more pumps 116-122 may be omitted. If pumps 116-122 are omitted, the MC may be less likely to be damaged during the transfer process. In such examples, the culture medium and / or MC may be moved based on the differential pressure between the material transfer device 106 and any of the external devices 102, 104, 127, or 128. In an embodiment, the first bioreactor 102 may be pressurized to allow used culture medium containing MC to flow into the first housing portion 134 after valve 108 is opened. In another embodiment, the culture medium transfer device 106 may be pressurized to allow fresh culture medium containing MC to flow into the second bioreactor 104 after valve 114 is opened. Other fluid flow options may prove appropriate, such as using gravity.

[0076] Referring again to the material transfer device 106, the material transfer device 106 includes a support 144 for providing structural support to the screen 129, which is operably coupled to the housing 126 and is disposed between the first housing portion 134 and the second housing portion 135, and adjacent to the screen 129. In the shown embodiment, the support 144 includes a first support 146 extending across the first transfer opening 138, and a second support 148 extending across the second transfer opening 143. The support 146 and / or 148 may include a lattice structure or another structure.

[0077] Referring to controller 125, in the illustrated embodiment, controller 125 includes a user interface 150, a communication interface 152, one or more processors 154, and memory 156 for storing instructions that can be executed by one or more processors 154 to perform various functions, including those in the disclosed embodiment. The user interface 150, the communication interface 152, and the memory 156 are electrically and / or communicatively coupled to one or more processors 154.

[0078] In the embodiment, the user interface 150 is adapted to receive input from the user, such as a desired flow sequence, and to provide the user with information related to the operation of the control system 100. The received input may allow the controller 125 to customize a program to automate the control of valves 108, 110, 112, 114 and / or pumps 116, 118, 120, and / or 122, depending on the desired flow sequence and / or material transfer device 106 being used. The user interface 150 may include a touchscreen, display, keyboard, speaker, mouse, trackball, and / or voice recognition system. The touchscreen and / or display may show a graphical user interface (GUI).

[0079] In the embodiment, the communication interface 152 is adapted to enable communication between the control system 100 and a remote system (e.g., a computer) over a network. The network may include an intranet, a local area network (LAN), a wide area network (WAN), etc. Some of the communications provided to the remote system may be associated with flow sequences, transport data, filtration data, flow rate data, etc., which are generated by or otherwise acquired by the control system 100.

[0080] One or more processors 154 and / or control systems 100 may include one or more processor-based systems or microprocessor-based systems. In some embodiments, one or more processors 154 and / or control systems 100 include reduced instruction set computers (RISCs), application-specific integrated circuits (ASICs), field-programmable gate arrays (FPGAs), field-programmable logic devices (FPLDs), logic circuits, and / or other logic-based devices that perform a variety of functions, including those described herein.

[0081] Memory 156 may include one or more of the following: hard disk drives, flash memory, read-only memory (ROM), erasable programmable read-only memory (EPROM), electrically erasable programmable read-only memory (EEPROM), random access memory (RAM), non-volatile RAM (NVRAM) memory, compact discs (CDs), digital general-purpose discs (DVDs), caches, and / or any other storage devices or storage disks in which information is stored for any period of time (e.g., permanently, temporarily, for a long period, for buffering, for caching).

[0082] Figure 2 is a cross-sectional view of an embodiment of the material transfer device 106 of Figure 1. The material transfer device 106 may be a disposable device intended to maintain a closed system having bioreactors 102, 104, a waste receptacle 127, and / or a fresh culture medium source 128 to which fluid tubes 130, 131, 132, 133 are sterilely connected. In the embodiment shown, the material transfer device 106 includes a first housing portion 134 oriented as an upper housing portion 158 and a second housing portion 135 oriented as a lower housing portion 160. The screen 129 is positioned horizontally (e.g., parallel) between the housing portions 134, 135 with respect to the horizontal plane 162. The supports 146, 148 are also positioned horizontally with respect to the horizontal plane 162. Based on the orientation of the upper housing 158 and the lower housing 160, the spent culture medium may flow through the material transfer device 106 in the direction generally indicated by arrow 164, based on gravity. However, alternatively, the spent culture medium may flow through the material transfer device 106 in a direction generally opposite to the direction generally indicated by arrow 164. Furthermore, the upper and lower housings 158, 160 and / or screen 129 may have different orientations (see, for example, Figures 4 and 5).

[0083] Still referring to the embodiment in Figure 2, the first inlet port 136 and the first outlet port 137 are centrally located within the first housing portion 134, and the second outlet port 140 and the second inlet port 142 are centrally located within the second housing portion 135. However, any one of the ports 136, 137, 140, and 142 may have a different arrangement. For example, the ports 136, 140, and 142 may have axes parallel to the central plane of the material transfer device 106 (see, for example, Figures 4 and 5).

[0084] Figure 3 is an enlarged view of the material transfer device 106 of Figure 2. In the embodiment shown, the screen 129 contains a number of pores 165 that allow the flow of culture medium through it but filter out MC. The pores 165 allow MC to accumulate on the top surface 166 of the screen 129 when used culture medium flows through the screen 129 in the direction indicated by arrow 168 and / or when fresh culture medium flows through the screen 129 in the direction indicated by arrow 168 and rehydrates the MC. Each of the pores 165 may be the same size, or some or all of the pores 165 may be of different sizes. The size of the pores 165 may correspond to the MC to be filtered. In the embodiment, the pores 165 are about 70 microns (μm) to about 75 μm.

[0085] Supports 146, 148 may be formed as a grid structure 170 adapted to provide structural support on both sides (e.g., top and bottom) of the screen 129 to reduce the possibility of the screen 129 bending and / or sagging in response to the weight of the culture medium flowing through the screen 129 and / or the MC accumulating on the screen 129. Supports 146, 148 and / or associated grid structure 170 may be arranged to reduce the number of pores 165 blocked by the structures 146, 148 and / or 170. Thus, more culture medium can freely pass through the screen 129 in the direction generally indicated by arrow 168 and / or in the direction generally opposite to the direction indicated by arrow 168. In embodiments, the screen 129 may include a number of pores, taking into account a number of pores 165 that may become clogged with MC and a number of pores 165 that remain unclogging and allow liquid to flow rapidly through the pores 165.

[0086] Figure 4 is a cross-sectional view of another embodiment of the material transfer device 106 of Figure 1. The housing 126, screen 129, and supports 146, 148 of the material transfer device 106 in Figure 4 are similar to the housing 126, screen 129, and supports 146, 148 of the material transfer device 106 in Figure 2. However, in contrast to the material transfer device in Figure 2, the ports 136, 137, 140, 142 of the material transfer device 106 in Figure 4 are arranged differently, with the first housing portion 134 oriented as the lower housing portion 172 and the second housing portion 135 oriented as the upper housing portion 173. However, as stated above, any of the ports 136, 137, 140, 142 can be used to introduce used culture medium into the material transfer device 106, and any of the ports 136, 137, 140, 142 can be used to introduce fresh culture medium into the material transfer device 106.

[0087] In some embodiments, some of the axes of ports 136, 140, and 142 are positioned substantially parallel to the central plane 176 of the material transfer device 106, with the first inlet port 136 positioned higher than the first outlet port 137 and the second outlet port 140 positioned lower than the second inlet port 142. In the embodiments shown, the screen 129 and housing 126 are positioned at an angle to the horizontal plane 162 when the material transfer device 106 is in use. However, the screen 129 and housing 126 may be positioned in different orientations.

[0088] During operation, the first inlet port 136 flows the spent medium containing MC into the first housing portion 134, flowing tangentially along the surface 175 of the screen 129. The spent medium passes through the screen 129 into the second housing portion 135 and exits through the second outlet port 140. The funnel-shaped portion 174 of the first housing portion 134 connects to the first outlet port 137 and is positioned to accumulate MC within the first housing portion 134. In the shown embodiment, the first housing portion 134 includes an internal barrier 177 that extends across the width of the first housing portion 134 and is adapted to prevent MC accumulating in the funnel-shaped portion 174 from covering the first inlet port 136 of the first housing portion 134. The internal barrier 177 may alternatively extend across a portion of the width of the first housing portion 134, and / or the internal barrier 177 may be omitted.

[0089] Figure 5 is a front view of the material transfer device 106 of Figure 4, showing the funnel-shaped portion 174 of the first housing portion 134. Fluid tubes 130, 131, 132, and 133 are shown toward the bottom of the material transfer device 106, and fluid tube 132 is shown toward the top of the material transfer device 106.

[0090] Figure 6 is an isometric view of an embodiment of the control system 100 of Figure 1. In the embodiment shown, the control system 100 includes a base 178 defining a receptacle 105 and carrying two of the valves 110, 112 and two of the corresponding pumps 118, 120, and a wall 180 coupled to the base 178 and carrying two of the valves 108, 114 and two of the corresponding pumps 116, 122. The base 178 includes a receptacle 105 having a recessed region 181 that provides space for fluid tubes 131, 132 to bend from the material transfer device 106 and to be coupled to the corresponding valves 110 and / or 112, and / or pumps 118 and / or 120. A heater 124 is shown to be located within the recessed region 181. However, the heater 124 may be located in a different place and may still be positioned to maintain the MC in the material transfer device 106 at a relatively consistent temperature. The base 178 also includes a first side 182 opposite to the second side 184, and each side 182, 184 defines channels 186, 188 through which at least one of the fluid tubes 131, 132 passes.

[0091] Figure 7 is an isometric view of the control system 100, in which the material transfer device 106 is located within the receptacle 105 and / or rests on the upper surface 190 of the base 178. During operation, the control system 100 can activate one or more of the valves 108, 110, 112, 114 and / or pumps 116, 118, 120, 122 to replace used culture medium with fresh culture medium and to transfer MC between the first bioreactor 102 and the second bioreactor 104. Figure 7 shows the bioreactors 102, 104, the waste receptacle 127, and the fresh culture medium source 128 coupled to the corresponding ports 136, 137, 140, 142, although the bioreactors 102, 104 may be coupled to ports 140, 142 and the waste receptacle 127, and the fresh culture medium source 128 may be coupled to ports 136, 137. In such an arrangement, the culture medium that carries MC may enter and exit the material transfer device 106 from the bottom, and the culture medium that does not carry MC may enter and exit the material transfer device 106 from the top.

[0092] Furthermore, although several embodiments are disclosed herein, any feature from any embodiment can be combined with or replaced with other features from other embodiments. Moreover, although several embodiments are disclosed herein, modifications can be made to the disclosed embodiments without departing from the claims.

Claims

1. It is a method, Within the material transfer device, microcarriers or cell aggregates in used culture medium are received from the first bioreactor. The microcarriers or cell aggregates are held on the first side of the filter of the material transfer device, while the used culture medium is flowed through the filter in the first direction. In order to resuspend the microcarriers or cell aggregates in fresh medium, the flow through the filter is reversed by flowing the fresh medium through the filter in a second direction opposite to the first direction. A method comprising flowing the microcarriers or cell aggregates in the fresh culture medium from the material transfer device to a second bioreactor.

2. The method according to claim 1, further comprising discharging the used culture medium from the material transfer device before reversing the flow through the filter by flowing the fresh culture medium through the filter in the second direction.

3. The method according to any one of the prior claims, wherein receiving microcarriers or cell aggregates in the spent culture medium from the first bioreactor into the material transfer device includes pumping the microcarriers or cell aggregates in the spent culture medium into the material transfer device.

4. The method according to claim 1 or 2, wherein receiving microcarriers or cell aggregates in the spent culture medium from the first bioreactor into the material transfer device includes, on the basis of gravity, flowing the microcarriers or cell aggregates in the spent culture medium into the material transfer device.

5. The method according to claim 1 or 2, wherein receiving microcarriers or cell aggregates in the spent culture medium from the first bioreactor into the material transfer device includes flowing the microcarriers or cell aggregates in the spent culture medium into the material transfer device based on a pressure difference between the first bioreactor and the material transfer device.

6. The method according to any one of the prior claims, further comprising heating the material transfer device.

7. The method according to any one of the prior claims, further comprising rotating the material transfer device to facilitate the movement of the microcarriers or the cell aggregates away from the screen.

8. The method according to any one of the prior claims, wherein the first bioreactor, the material transfer device, and the second bioreactor are aseptically connected as part of a completely closed system.

9. A material transfer device, A housing including a first housing portion and a second housing portion, The first housing portion comprises a first inlet port, a first outlet port, and a first transfer opening, The housing comprises a second housing portion having a second inlet port, a second outlet port, and a second transfer opening, and the first transfer opening being adjacent to and in communication with the second transfer opening. A material transfer device comprising: a screen disposed between the housing portion adjacent to the first and second transfer openings.

10. The first inlet port of the first housing portion is arranged to transport used culture medium containing cell aggregates or microcarriers into the first housing portion, and the screen is adapted to prevent the cell aggregates or microcarriers from passing through the screen, while allowing the used culture medium to pass through the screen and into the second housing portion and toward the second outlet port of the second housing portion. The material transfer device according to claim 9, wherein the second inlet port of the second housing portion is arranged to transport fresh culture medium into the second housing portion, and the screen is adapted to allow the fresh culture medium to flow through the screen to resuspend the cell aggregates or the microcarriers in the first housing portion.

11. The material transfer device according to claim 10, wherein the first outlet port of the first housing portion is arranged to allow the fresh culture medium suspending the cell aggregate or the microcarrier to flow out of the first housing portion.

12. The material transfer device according to any one of claims 9 to 11, wherein the screen is a flexible screen that defines a plurality of pores that allow material to flow through it.

13. The material transfer device according to any one of claims 9 to 12, further comprising a support operably coupled to the housing and disposed between the first housing portion and the second housing portion and adjacent to the screen, to provide a structural support for the screen.

14. The material transfer device according to claim 13, wherein the support comprises at least one of a first support extending across the first transfer opening and a second support extending across the second transfer opening.

15. The material transfer device according to claim 14, wherein the first support is coupled to the first housing portion adjacent to the first transfer opening, and the second support is coupled to the second housing portion adjacent to the second transfer opening.

16. The material transfer device according to any one of claims 13 to 15, wherein the support has a lattice structure.

17. The material transfer device according to any one of claims 9 to 16, wherein the first housing portion and the second housing portion are each rigid or semi-rigid structures.

18. The material transfer device according to any one of claims 9 to 17, wherein the first housing portion is detachably coupled to the second housing portion or is integrally formed with the second housing portion.

19. The material transfer device according to any one of claims 9 to 18, wherein the first housing portion is the upper housing portion and the second housing portion is the lower housing portion.

20. The material transfer device according to any one of claims 9 to 19, further comprising a seal at the interface between the first housing portion and the second housing portion.

21. The material transfer device according to any one of claims 9 to 20, wherein the first inlet port and the first outlet port are centrally located within the first housing portion, and the second inlet port and the second outlet port are centrally located within the second housing portion.

22. The material transfer device according to any one of claims 9 to 21, wherein when the material transfer device is in use, the screen is arranged horizontally with respect to the horizontal plane.

23. The material transfer device according to any one of claims 9 to 18 and 20 to 22, wherein the first housing portion is the lower housing portion and the second housing portion is the upper housing portion.

24. The material transfer device according to claim 23, wherein the first housing portion includes a funnel shape connected to the first outlet port.

25. The material transfer device according to claim 23 or 24, wherein the first housing portion further comprises an internal barrier extending across the width of the first housing portion and adapted to prevent accumulated cell aggregates or microcarriers from covering the first inlet port of the first housing portion.

26. The material transfer device according to any one of claims 23 to 25, wherein the second outlet port of the second housing portion is adapted to be positioned lower than the second inlet port of the second housing portion.

27. The material transfer device according to any one of claims 23 to 26, wherein the first inlet port of the first housing portion is positioned relative to the screen such as to allow the used culture medium containing the cell aggregates or microcarriers to flow tangentially along the surface of the screen.

28. The material transfer device according to any one of claims 23 to 27, wherein the screen and the housing are at an angle with respect to the horizontal plane when the material transfer device is in use.

29. A method of using a material transfer device comprising a first housing portion and a second housing portion, wherein each housing portion comprises an inlet port, an outlet port, and a corresponding fluid pipe connected to the inlet port and the outlet port, The used culture medium containing cell aggregates or microcarriers is pumped through the inlet port of the first housing portion, Filtering the cell aggregates or microcarriers using a screen, The used culture medium is passed through the screen and placed into the second housing section. The used culture medium is pumped out from the outlet port of the second housing portion, The fresh culture medium is pumped into the first housing portion or the second housing portion, Resuspending the cell aggregates or the microcarriers, A method comprising pumping the fresh culture medium in which the cell aggregates or microcarriers are suspended from the outlet port of the first housing portion.

30. The method according to claim 29, wherein passing the used culture medium through the screen includes passing the used culture medium through a first transfer opening of the first housing portion and a second transfer opening of the second housing portion which is adjacent to the first transfer opening and is disposed in communication with the first transfer opening.

31. The method according to claim 29 or 30, further comprising supporting the screen using a support operably coupled to the housing and disposed between the first housing portion and the second housing portion.

32. The method according to any one of claims 29 to 31, wherein passing the used culture medium through the screen includes passing the used culture medium through the screen which is arranged horizontally with respect to a horizontal plane.

33. The method according to any one of claims 29 to 31, wherein passing the used culture medium through the screen includes passing the used culture medium through the screen which is arranged at a certain angle with respect to a horizontal plane.

34. The method according to any one of claims 29 to 33, further comprising passing the fresh culture medium through the screen and allowing it to flow into the first housing portion.

35. A control system for controlling fluid flow through a material transfer device having a first housing portion and a second housing portion, wherein each housing portion includes an inlet port, an outlet port, and corresponding fluid pipes connected to the inlet port and the outlet port, A base is provided to define the receptacle and to support the material transfer device, A control system comprising a plurality of valves and a plurality of pumps adapted to control the flow of culture medium through the inlet port and the outlet port, respectively, of the transfer device.

36. The control system according to claim 35, further comprising the material transfer device disposed on the base or otherwise mounted on the base.

37. The valve and the pump (1) pump used culture medium containing cell aggregates or microcarriers through the inlet port of the first housing portion, allowing the cell aggregates or microcarriers to be filtered by the screen, allowing the used culture medium to pass through the base screen and be sent into the second housing portion, (2) pump the used culture medium out of the outlet port of the second housing portion, and (3) fresh The control system according to claim 35 or 37, which is adapted to pump culture medium into the second housing portion, pass the fresh culture medium through the screen and allow it to flow into the first housing portion, allowing the cell aggregates or the microcarriers to be resuspended, and (4) pump the fresh culture medium in which the cell aggregates or the microcarriers are suspended out of the outlet port of the first housing portion.

38. The control system according to any one of claims 35 to 37, further comprising a wall coupled to the base and carrying two of the valves and two of the corresponding pumps, wherein the base further carries two of the valves and two of the corresponding pumps.

39. The control system according to any one of claims 35 to 38, wherein the base includes a first side opposite to a second side, and each side defines a channel through which at least one fluid tube is adapted.

40. The control system according to any one of claims 35 to 39, further comprising a flow meter adapted to determine the flow rate of the used culture medium entering and leaving the second portion of the housing, wherein the control system is adapted to change the pump flow rate for pumping the used culture medium out of the outlet port of the second housing portion of the transfer device in response to the determined flow rate that satisfies a threshold flow valve.

41. The control system according to any one of claims 35 to 40, further comprising a user interface adapted to allow input to be received by the control system and to control the flow sequence through the transport device.

42. The control system according to any one of claims 35 to 40, wherein the valve is a pinch valve.

43. The control system according to any one of claims 35 to 42, further comprising a heater adapted to heat the material transfer device.

Images