Automated cell culture medium filling system

Abstract

A cell culture system for culturing cells is provided. The cell culture system includes a cell culture vessel having a cell culture chamber for culturing cells, an inlet configured for fluid flow to fill the cell culture vessel, and an outlet configured for fluid exit from the cell culture vessel. The system further includes at least one fill sensor positioned to detect when fluid in the cell culture vessel reaches a fill level at a predetermined location in the cell culture vessel during filling of the cell culture vessel, the at least one fill sensor being capable of generating a detection signal when the fluid in the cell culture vessel reaches the fill level. The system also includes an actuator for changing the orientation of the cell culture vessel in response to the detection signal.

Description

Cross - reference to related applications

[0001] This application claims the benefit of priority under 35 U.S.C. § 119 to U.S. Provisional Patent Application No. 63 / 058796, filed July 30, 2020, and all disclosures of this provisional application are hereby incorporated by reference and made a part of this specification.

Technical Field

[0002] The present disclosure relates to a multi - position support for a cell culture device, and more particularly to a multi - position support having two arrangement settings: an upright arrangement setting and a tilting arrangement setting.

Background Art

[0003] Many types of cell culture articles are configured to be stacked cell culture units or cell culture units that can be stacked. For example, usually, T - flasks are made so that their upper and lower surfaces are flat surfaces, whereby T - flasks can be stacked and used in a space - saving manner. Also, in order to reduce the time and labor involved in filling and emptying, there are improved T - flasks with a plurality of culture surfaces arranged in parallel within the flask. Also, as other culture devices, there are multi - element assemblies having a plurality of culture surfaces arranged in parallel or stacked. In many such stacked culture assemblies, the culture layers are isolated from each other to reduce the hydrostatic pressure on the lower culture layer. This is because as the number of stacked layers increases, the influence of hydrostatic pressure may increase.

[0004] One example of a cell culture material is Corning's HYPERStack® system. The HYPERStack system consists of multiple modules formed from multiple individual stackette layers, which can be interconnected by flexible tubes connected to tube connectors. This interconnection of modules is intended for filling and draining the HYPERStack system. Furthermore, devices such as valves can be used to control the inflow and outflow of fluid into and out of the HYPERStack system. However, the use of these devices such as valves is cumbersome and could potentially create points where leaks may occur.

[0005] The current filling and draining process for the HYPERStack system is inconsistent. This is because the current filling and draining protocol requires tilting the HYPERStack system at various stages to achieve better results. Furthermore, the lack of accessories to support this protocol has forced users to make do with readily available items in their research facilities, such as tube clamps, tube racks, and door stoppers. Therefore, a multi-position support is needed that can be used to manipulate the cell culture apparatus during the filling and draining procedure and to reliably position the cell culture apparatus at multiple tilt angles.

[0006] Furthermore, even with improved accessories or multi-position supports, manual filling and draining still requires careful attention from the user. For example, filling the unit too quickly or filling it with too much culture medium can put excessive stress on the container or clog the exhaust filter. These problems can lead to leaks or contamination of the container during use. This issue is further amplified if the user attempts to fill or use multiple cell culture devices at once. [Overview of the project] [Problems that the invention aims to solve]

[0007] To avoid the aforementioned problems, a semi-automated or fully automated cell culture system is needed that can monitor the units while filling them with culture medium. [Means for solving the problem]

[0008] A cell culture system is provided according to several embodiments of the present disclosure. The cell culture system comprises a cell culture vessel having a cell culture chamber for culturing cells, an inlet configured for the flow of liquid to fill the cell culture vessel, and an outlet configured for the exit of the cell culture vessel. The system further comprises at least one filling sensor positioned to detect when the liquid in the cell culture vessel has reached a filling level at a predetermined position in the cell culture vessel during filling of the cell culture vessel. The at least one filling sensor can generate a detection signal when the liquid in the cell culture vessel has reached the filling level. Furthermore, the system further comprises an actuator for changing the orientation of the cell culture vessel in response to the detection signal.

[0009] In further embodiments of the multiple embodiments of the present disclosure, at least one filling sensor may include a first filling sensor. The first filling sensor is configured to detect when the liquid in the cell culture vessel reaches a first filling level and can generate a first detection signal when the liquid in the cell culture vessel reaches the first filling level. The actuator can then change the orientation of the cell culture vessel from a first orientation to a second orientation in response to the first detection signal.

[0010] At least one filling sensor may further include a second filling sensor. The second filling sensor is positioned to detect when the liquid in the cell culture vessel reaches a second filling level and can generate a second detection signal when the liquid in the cell culture vessel reaches the second filling level. The system can then stop filling the cell culture vessel in response to the second detection signal. The second filling level is different from the first filling level. In one embodiment of several designs, the cell culture vessel is a multilayer cell culture vessel.

[0011] In an additional embodiment, the system may further include a control device for receiving detection signals from at least one sensor and controlling an actuator in response to the detection signal. The system may also further include a pump fluid-connected to the inlet of a cell culture vessel. The control device can control the flow rate of the pump.

[0012] According to several aspects of several embodiments, the cell culture vessel comprises a filling side and a support-facing side. An inlet and an outlet are located on the filling side. The support-facing side is the side adjacent to the filling side, and in the first position, the support-facing side faces a substantially horizontal support member located beneath the cell culture vessel. In the first position, the support-facing side forms a first angle with respect to the support member, the outlet is at a first distance from the support member, and the inlet is closer to the support member than the outlet. In the second position, the support-facing side forms a composite angle with respect to the support member. This composite angle includes (i) a second angle between the length of the support-facing side and the support member, and (ii) a third angle between the width of the support-facing side and the support member. Also in the second position, the outlet is at a second distance from the support member, and the second distance is greater than the first distance.

[0013] According to several aspects of some embodiments, a cell culture vessel may also include a plurality of cell culture chambers, also called a stack or layer. The cell culture chambers are stacked on top of each other, and the bottom surface of each cell culture chamber includes a cell culture surface. The cell culture vessel may have a top surface, a bottom surface, and four sides extending from the top surface to the bottom surface. The first side includes part of an inlet access column (filling column) and part of an outlet access column (exhaust column). The third side is the side opposite to the first side. The second side is adjacent to the first and third sides and includes part of an inlet access column (filling column) along the end adjacent to the first side. The fourth side is adjacent to the first and third sides and opposite to the second side and includes part of an outlet access column (exhaust column) along the end adjacent to the first side. Inlet ports (filling ports) and outlet ports (exhaust ports) may be located on the top surface of the cell culture vessel along one side of the cell culture vessel. The inlet port (filling port) and outlet port (exhaust port) can be located at both corners (both ends) of the corresponding side of the cell culture vessel. The cell culture vessel may further include an inlet access column (filling column) that communicates with each cell culture chamber and the inlet port (filling port). The cell culture vessel may further include an outlet access column (exhaust column) that communicates with each cell culture chamber and the outlet port (exhaust port). The inlet access column (filling column) extends vertically from the inlet port (filling port) at the top of the cell culture vessel to the lowest chamber of the cell culture vessel and can be located at the corner of the cell culture vessel. The outlet access column (exhaust column) extends vertically from the outlet port (exhaust port) at the top of the cell culture vessel to the lowest chamber of the cell culture vessel. The inlet access column (filling column) can be placed in the corner of the cell culture vessel, and the outlet access column (exhaust column) can be placed in the opposite corner of the vessel, on the same side surface as the inlet access column (filling column). In the initial orientation, the cell culture vessel is in the incubation position where the side with the inlet port and outlet port is the uppermost surface.In the first filling position, the cell culture vessel is rotated 90 degrees from its initial position so that the outlet port is vertically above the inlet port and the top surface faces outward rather than upward. In the second filling position, the cell culture vessel is rotated 90 degrees from the first filling position so that the side opposite the inlet and outlet columns is in the same horizontal plane as the surface of the support member. The outlet and inlet ports are in the same horizontal plane spaced apart from the surface of the support member, and the top surface faces outward.

[0014] In some embodiments, in further aspects, at least one fill sensor is mounted outside the cell culture vessel to detect the fill water level through the wall of the cell culture vessel. The cell culture vessel may comprise a plurality of cell culture chambers and a manifold connecting the plurality of cell culture chambers, and at least one fill sensor may be mounted on the manifold. The cell culture vessel may comprise a plurality of cell culture chambers and an inlet access column (fill column) connecting the plurality of cell culture chambers, and at least one fill sensor may be mounted on the inlet access column. In some embodiments, at least one fill sensor is removable and reusable from the cell culture vessel. At least one fill sensor may include at least one of an optical sensor, a through-beam sensor, or a photoelectric sensor.

[0015] In a further embodiment of the multiple embodiments, the system further comprises a flow controller positioned to detect the fluid pressure in the piping of the system. The flow controller can reduce the flow rate of the system in response to the fluid pressure in the piping being above a predetermined pressure value.

[0016] In an additional embodiment, the system further comprises a valve at the outlet of the cell culture vessel. The valve can stop the filling of the cell culture vessel by closing the fluid path at or near the outlet of the cell culture vessel. The valve may be a pinch valve located outside the tube connected to the outlet. The valve may be designed to close the fluid path in response to at least one of a detection signal from one of at least one filling sensors and / or the fluid pressure in the piping.

[0017] In some further embodiments of the system, the system further comprises a multi-position support for supporting cell culture vessels. An actuator can be attached to the multi-position support to change the orientation of the multi-position support with respect to the horizontal support surface on which the multi-position support is positioned.

[0018] In some additional embodiments of the system, the actuator has an adjustable length including a first length and a second length. When the actuator is set to the first length, the cell culture vessel is in a first position, and when it is set to the second length, the cell culture vessel is in a second position. The first length can be greater than the second length. The actuator may comprise at least one of a solenoid, a piezoelectric material, a pneumatic piston, and a hydraulic piston.

[0019] In some additional embodiments of the system, the system may include a third filling sensor positioned to detect when the liquid in the cell culture vessel reaches a third filling level. The third filling sensor can generate a third detection signal when the liquid in the cell culture vessel reaches a third filling level different from the first and second filling levels. Based on the third detection signal from the third filling sensor, the control device may be configured to slow down the filling rate of the cell culture vessel.

[0020] In one embodiment of several embodiments, when filling a cell culture vessel with liquid, the liquid reaches a first filling level before a second filling level. In some embodiments, when filling a cell culture vessel with liquid, the liquid reaches a third filling level after the first filling level but before the second filling level.

[0021] In one embodiment of multiple configurations, the control device can stop filling the cell culture vessel based on a second detection signal from a second filling sensor.

[0022] In some further embodiments of the multiposition support, the multiposition support has a main base that is placed on a support member when set upright, a support surface provided at a position vertically offset from the main base when set upright, which supports the cell culture vessel when the cell culture vessel is on the support surface, and an intermediate surface extending from the main base to the support surface, which connects to the main base at a boundary portion that extends at an angle inclined with respect to the side surface of the main base. The multiposition support has a tilted configuration in which, while supporting the cell culture vessel on the support surface, the multiposition support is rotated around the boundary portion to bring the support surface closer to the support member than when set upright.

[0023] The multi-position support may further have a sub-base that rests on the support member when in an upright position. In some embodiments, the aforementioned support surface is the first support surface, and the multi-position support further has a second support surface located between the main base and the sub-base, the second support surface supporting the cell culture vessel when the cell culture vessel is on the second support surface. The first and second support surfaces may be substantially on the same plane, inclined with respect to the main base. The multi-position support may further have support flanges that engage with the filled side of the cell culture vessel to restrain the cell culture vessel on the first and second support surfaces. The multi-position support may further have other support flanges extending from the first support surface that engage with the rear side of the cell culture vessel opposite to the filled side. The second support surface may be spaced apart from the support member. In some aspects of some embodiments, the support surfaces are connected to intermediate surfaces at other boundaries that are inclined with respect to the side of the main base. The inclination angles of the two boundary sections relative to the sides of the main base can be made approximately the same.

[0024] An additional embodiment provides a method for changing the filling angle of a cell culture apparatus comprising a cell culture vessel including a plurality of cell culture modules fluidly connected to each other by a fluid manifold and an air manifold. The method includes the steps of: connecting the cell culture apparatus to a multi-position support having tilted and upright configurations; filling the cell culture apparatus with the multi-position support supporting the cell culture apparatus in either the upright or tilted configuration; detecting that the cell culture apparatus has been filled to a first filling level using a first filling sensor provided outside the cell culture module; changing the multi-position support from one of the tilted and upright configurations to the other based on the detection by the first filling sensor; detecting that the cell culture apparatus has been filled to a second filling level different from the first filling level using a second filling sensor provided outside the cell culture module; and stopping the filling based on the detection by the second filling sensor.

[0025] In some aspects of several embodiments of the present method, each of the plurality of cell culture modules includes a plurality of layers of cell culture chambers.

[0026] In a further aspect of some embodiments of the present method, the multi-position support can have a main base portion that is placed on the support member in the upright arrangement setting, a support surface provided at a position vertically offset from the main base portion in the upright arrangement setting, the support surface supporting the cell culture device in a state where the cell culture device is on the support surface, and an intermediate surface extending from the main base portion to the support surface, the intermediate surface being connected to the main base portion at a boundary portion extending at an oblique angle with respect to the side surface of the main base portion. In the tilted arrangement setting, the multi-position support is rotated about the boundary portion while supporting the cell culture device on the support surface, and the support surface is brought closer to the support member than in the upright arrangement setting. <000009-0>

[0027] In some aspects of several embodiments of the present method, the above-described filling step includes a step of filling the cell culture device in a state where the cell culture device is supported on the multi-position support in the upright arrangement setting. The method can further include a step of tilting the cell culture device using the multi-position support by rotating the multi-position support about the boundary portion. In a further aspect of the present method, the step of changing the multi-position support from one of the tilted arrangement setting and the upright arrangement setting to the other includes a step of moving an actuator attached to the multi-position support.

Advantages of the Invention

[0028] [[ID=1,7]] This specification describes a cell culture medium filling system that semi-automates or fully automates the filling and draining of a cell culture system by automatically controlling the orientation and / or flow rate of the cell culture vessel when filling or draining liquid culture medium into the cell culture vessel, and which can switch the orientation of the cell culture vessel between an upright position setting and a tilted position setting to position it at different angles relative to the horizontal. By positioning the cell culture vessel at different angles, the results of filling and draining can be improved in a way that enhances reliability, consistency, and efficiency. Furthermore, in the tilted position setting, the filling side (front) of the cell culture vessel is positioned at a high position at a composite angle that lifts the air manifold in both the vertical and front-to-back directions. [Brief explanation of the drawing]

[0029] [Figure 1] Perspective view of a cell culture apparatus equipped with a manifold according to one or more embodiments illustrated and described herein. [Figure 2] Schematic diagram of multiple stacket layers for use in the cell culture apparatus shown in Figure 1, according to one or more embodiments illustrated and described herein. [Figure 3] A side view showing the cell culture apparatus shown in Figure 1 supported by a multi-position support in an upright configuration, according to one or more embodiments illustrated and described herein. [Figure 4] A perspective view of the multiposition support shown in Figure 3, according to one or more embodiments illustrated and described herein. [Figure 5] Plan view of a multiposition support shown in Figure 4, according to one or more embodiments illustrated and described herein. [Figure 6] A side view showing the multi-position support shown in Figure 3 in a tilted configuration, according to one or more embodiments illustrated and described herein. [Figure 7] An end view showing the multi-position support shown in Figure 6 in a tilted configuration, according to one or more embodiments illustrated and described herein. [Figure 8]A perspective view showing an automated cell culture medium filling system in a first position according to one or more embodiments illustrated and described herein. [Figure 9] Figure 8 is a perspective view showing the automated cell culture medium filling system in its second orientation. [Figure 10] Perspective view of a cell culture apparatus according to one or more embodiments illustrated and described herein. [Figure 11] A side view showing the cell culture apparatus shown in Figure 10 in a first filled position, according to one or more embodiments illustrated and described herein. [Figure 12] A side view showing the cell culture apparatus shown in Figure 10 in a second filled position, according to one or more embodiments illustrated and described herein. [Figure 13] A side view showing the cell culture apparatus shown in Figure 10 in the incubation position, according to one or more embodiments illustrated and described herein. [Modes for carrying out the invention]

[0030] Please note that the drawings are not necessarily drawn to scale. Similar numbers used within the drawings indicate similar components, processes, etc. However, when a component is indicated by a certain number in one drawing, it should be understood that this is not intended to limit that component to other drawings that use the same number. Similarly, when components are indicated by different numbers, this is not intended to indicate that these components with different numbers cannot be identical or similar.

[0031] The following detailed description refers to the accompanying drawings, which constitute part of this specification and illustrate several specific embodiments of the devices, systems, and methods. However, it should be understood that other embodiments may be conceived and implemented without departing from the scope or spirit of this disclosure. Therefore, the following detailed description should not be construed as restrictive.

[0032] All scientific and technical terms used herein have their meanings as commonly used in the art, unless otherwise specified. The definitions provided herein are for the purpose of facilitating the understanding of certain terms that are used repeatedly herein and are not intended to limit the scope of this disclosure.

[0033] In this specification and the appended claims, the singular forms “a,” “an,” and “the” shall also include embodiments having the plural form unless it is clear from the context that the plural form is not included. In this specification and the appended claims, the term “or” shall generally be used to mean “and / or” unless it is clear from the context that “and / or” is not included.

[0034] In this specification, "have," "including," and "comprise" (have, having, include, including, comprise, comprising, etc.) are used in an open-ended sense, generally meaning "including, but not limited to."

[0035] This disclosure describes a cell culture system capable of automating the filling and / or draining of liquid culture medium into cell culture vessels. Embodiments of this system are intended to be comprised of various combinations of the components described herein. Such components include one or more of the following: cell culture vessels, filling sensors for detecting the water level in the cell culture vessels, actuators for changing the orientation of the cell culture vessels during filling, multi-position supports for supporting the cell culture vessels, control devices, pressure sensors, and various connections, fittings, tubes, and manifolds. The embodiments described herein monitor the water level of the liquid culture medium using the filling sensors during filling or draining of the vessels and change the orientation of the cell culture vessels or adjust the filling speed according to the water level. The systems and methods disclosed herein enable semi-automatic or fully automatic filling and / or draining of cell culture vessels. As a result, a cell culture system and method is provided that reduces the risk of leakage or contamination in the cell culture system and the risk of other stresses being applied to the cell culture system, while reducing the degree of monitoring and attention required by the user during the filling or draining procedure.

[0036] As described above, this disclosure relates to a multi-position support for a multilayer cell culture apparatus. The multi-position support can be formed by bending a plate to provide a main base that is placed on a support member when the apparatus is in an upright position. A support surface is provided at a position vertically offset from the main base when the apparatus is in an upright position, and this support surface supports the multilayer cell culture apparatus at an angle to the surface of the support member or at an angle to the horizontal. The multi-position support further has an intermediate surface that extends from the main base to the support surface. The intermediate surface is connected to the main base by a boundary that extends at an angle inclined with respect to the side surface of the main base. The multi-position support also has a tilted position setting. The tilted position setting is a position setting in which, while the multilayer cell culture apparatus is supported on the support surface, the multi-position support is rotated around a boundary on the support surface so that the support surface is closer to the support member than when the apparatus is in an upright position.

[0037] A multilayer cell culture apparatus comprises cell culture modules. Each cell culture module has multiple growth (culture) surfaces within a cell culture chamber, which are connected to each other by a manifold to form a cell culture device. Cell culture modules can also be further connected to other cell culture modules by manifolds to form stacked cell culture devices. Multiple culture surfaces can also be stacked to create a multilayer configuration. The manifold may include an integrated columnar structure formed as a monolithic component relative to the manifold. The columnar structure has an inlet port, and provides at least a portion of a fluid channel extending from the inlet port, providing fluid communication with the individual cell culture chambers within the cell culture module. The manifold and its associated columnar structure provide a closed system that, when the cell culture apparatus is in use, allows the columnar structure to be connected to a flexible tube to isolate the cell culture chamber from the environment.

[0038] In several embodiments of this disclosure, one or more sensors can be used to measure the water level in a cell culture vessel or manifold. Because there may be some variation in filling rates between cell culture devices, if a user attempts to fill multiple vessels at once, the sensors in each cell culture device can determine the appropriate time for changing the vessel's orientation or altering the fluid flow for each specific vessel.

[0039] Referring to Figure 1, the cell culture apparatus 10 comprises three cell culture modules 12, 14, and 16, each containing multiple layers of cell culture chambers 18, which are stacked in sequence to form the multilayer cell culture apparatus 10. Each cell culture module 12, 14, and 16 utilizes two manifolds 20 and 22. Liquid can enter and exit the cell culture modules 12, 14, and 16 via the first manifold 20. Therefore, the first manifold 20 can be called a fluid manifold. Air can enter and exit the cell culture modules 12, 14, and 16 via the second manifold 22. Therefore, the second manifold 22 can be called an air manifold.

[0040] As shown in Figure 2, each cell culture module 12, 14, and 16 can be equipped with multiple stacket layers 24. By stacking these stacket layers 24, multiple cell culture chambers 18 can be formed so that there are airway spaces 25 between the cell culture chambers 18. Figure 2 is a schematic diagram showing how multiple stacket layers 24 are stacked to form a stacked state of cell culture chambers 18 and cell culture surfaces 26. The cell culture surface 26 is equipped with, for example, a membrane 28 that is gas permeable and liquid impermeable. The stacket layers 24 have airway spaces 25, which allow gas to move between the cell culture chambers 18 and the outside of the cell culture apparatus 10. Referring again to Figure 1, spacers 31, 33, and 35 can separate the cell culture modules 12, 14, and 16 from each other. The spacers 31, 33, and 35 can serve as structural supports for the individual cell culture modules 12, 14, and 16. In some embodiments, the total number of cell culture chambers 18 can be increased by replacing either or both of the spacers 31, 33 with an additional stacket layer 24. Furthermore, instead of allowing air to remain in the cell culture chambers 18, a riser volume can be provided above the cell culture module 12 to capture residual air.

[0041] The cell culture modules or parts thereof described herein may be formed from any suitable material. However, materials intended to come into contact with cells or culture media are preferably compatible with cells and media. Typically, cell culture modules are formed from polymer materials. Examples of suitable polymer materials include polystyrene, polymethyl methacrylate, polyvinyl chloride, polycarbonate, polysulfone, polystyrene copolymer, fluoropolymer, polyester, polyamide, polystyrene-butadiene copolymer, fully hydrogenated styrene polymer, polycarbonate-PDMS copolymer, and polyolefins such as polyethylene, polypropylene, polymethylpentene, polypropylene copolymer, and cyclic olefin copolymer.

[0042] In some embodiments, the culture module includes a membrane 28 that is gas-permeable and liquid-impermeable, thereby allowing gas to move between the cell culture chamber 18 and ultimately outside the cell culture assembly. Such a culture module may include spacers or spacer layers positioned adjacent to the membrane outside the chamber, thereby allowing air to flow between the stacked units. An example of a commercially available cell culture apparatus including such a stacked gas-permeable culture unit is Corning's "HYPERStack" cell culture apparatus. Examples of suitable gas-permeable polymer materials useful for membrane formation include polystyrene, polyethylene, polycarbonate, polyolefin, ethylene vinyl acetate, polymethylpentene, polypropylene, polytetrafluoroethylene (PTFE), or compatible fluoropolymers, silicone rubber or silicone copolymers, poly(styrene-butadiene-styrene), or combinations thereof. However, a variety of polymer materials can be used within a range acceptable for manufacturability and compatibility with cell proliferation. The membrane thickness is preferably such that gas can efficiently move through the membrane. For example, polystyrene membranes can be used for cell proliferation at various thicknesses, but can be as thick as approximately 0.003 inches (approximately 75 micrometers). Thus, the membrane can be of any thickness, preferably approximately 25 to 250 micrometers, or approximately 25 to 125 micrometers. The membrane allows for free gas exchange between the assembly chamber and the external environment and can take on any size and shape. Preferably, the membrane is durable for the manufacture, handling, and operation of the apparatus.

[0043] As described above, the cell culture modules 12, 14, and 16 can be connected to each other by manifolds 20 and 22. Manifold 20 comprises a side wall base structure 30 and a column structure 32. The column structure 32 is formed as a monolithic component relative to the side wall base structure 30, thereby obtaining an integrated manifold 20. The column structure 32 is equipped with a receptacle structure 34, which provides at least a portion of a fluid channel extending from the receptacle structure 34 and providing fluid communication with the individual cell culture chambers 18 in the cell culture modules 12, 14, and 16. Manifold 20 can be configured to allow filling and draining of the cell culture chambers 18.

[0044] The manifold 22 also comprises a side wall base structure 30' and a column structure 32'. The column structure 32' is formed as a monolithic component relative to the side wall base structure 30', thereby obtaining an integrated manifold 22. The column structure 32' is equipped with a hanging structure 34', which provides at least a portion of the fluid flow path extending from the individual cell culture chambers 18 in the cell culture modules 12, 14, and 16 to the hanging structure 34'. The manifold 22 can be configured to allow the cell culture chambers 18 to be filled and drained by allowing air to enter and exit the cell culture apparatus 10. In some embodiments, the column structure 32' can be positioned offset from the illustrated position to control the flow of culture medium to the column structure 32'.

[0045] In a normal filling procedure, the cell culture device 10 can be placed with its left side facing the support surface or tray side, with the left side facing downwards. In this position, the front of the cell culture device 10, where the manifolds 20 and 22 are located, is tilted downwards, becoming the first filling position at the start of filling (see side view in Figure 3). Then, the inflow of liquid culture medium into the cell culture container begins. For example, the culture medium may be pumped to the lower column structure 32 via the support structure 34 using a peristaltic pump, or the container may be filled using a culture medium flow utilizing gravity. When the water level of the liquid culture medium in the cell culture device 10 rises to the first filling water level, which is a predetermined position, the position of the cell culture device 10 (and the filling tray, if one is used) is changed to the second filling position. In this second filling position, filling can be continued until the liquid culture medium reaches the final filling water level of the cell culture device 10. Once the final filling level is reached, the culture medium flow can be stopped, and the inlets and outlets from manifolds 20 and 22 can be closed or tightened to create a closed system. At this point, the cell culture apparatus 10 is ready for use in cell culture.

[0046] As described above, several embodiments of this disclosure include a type of packing tray, namely a multi-position support. Details of the multi-position support are provided below. This will allow for a better understanding of the packing and / or draining process of the cell culture apparatus 10, including what the first and second packing positions of the cell culture apparatus 10 are and how the cell culture apparatus 10 is moved to change its position.

[0047] Referring to Figure 3, by using the multi-position support 50, the cell culture apparatus 10 can be filled and drained while tilted horizontally with the side surface 40 facing downwards, as shown in Figure 3. By using the multi-position support 50, the cell culture apparatus 10 can be reliably positioned with the side surface 40 facing downwards at a predetermined inclination angle θ1 (for example, about 10 to 12 degrees) with respect to the support member 42 or the horizontal. The side surface 40 closest to the fluid manifold 20 is placed on the multi-position support 50, and the fluid manifold 20 is positioned lower than the air manifold 22. As will be described in more detail below, by tilting the multi-position support 50 and switching between an upright position (shown in Figure 3) and a tilted position, the cell culture apparatus 10 can be positioned at different angles with respect to the horizontal.

[0048] Referring to Figures 4 and 5, the multi-position support 50 is shown in a detached state. The multi-position support 50 is formed as a monolithic bent plate having a bottom 52, an upper part 54, both ends 56, 58, and both sides 60, 62. The multi-position support 50 is provided with position tabs 64, 66 on the side 62. The position tabs 64, 66 engage with the bottom edge 68 (Figure 3) of the cell culture apparatus 10 when the multi-position support 50 is in an upright position, and help to hold the cell culture apparatus 10 in a predetermined position with the multi-position support 50 facing downwards. In some embodiments, recesses 71, 73 can be provided in the bottom edge 68 of the cell culture apparatus 10, with a size and position to accommodate the position tabs 64, 66. The position tabs 64, 66 may have a bent portion 75. By gripping the bottom edge 68 using the bent portion 75, it is possible to prevent the cell culture apparatus 10 from moving laterally and coming off the multi-position support 50.

[0049] The multi-position support 50 has a main base 70 which is placed on a support member (e.g., a table or laboratory bench) when the multi-position support 50 is in an upright configuration as shown in the figure. A main support surface 72 for supporting the cell culture apparatus 10 is provided at a position vertically offset from the main base 70 in the upright configuration. The multi-position support 50 also has an intermediate surface 74 extending from the main base 70 to the main support surface 72. The intermediate surface 74 is connected to the main base 70 by a boundary portion 76 formed as a bent portion extending at an angle inclined with respect to the sides 60, 62 of the multi-position support 50. The intermediate surface 74 is also connected to the main support surface 72 by a boundary portion 77 formed as a bent portion extending at an angle inclined with respect to the sides 60, 62. In some embodiments, the inclination angles of the boundary portion 76 and the boundary portion 77 may be approximately the same angle (e.g., a difference of 5 degrees or less) with respect to the sides 60, 62, or they may be different angles.

[0050] Furthermore, the multi-position support 50 has a sub-base 79 which is placed on a support member when the multi-position support 50 is in an upright position. In addition, a sub-support surface 78 that supports the cell culture apparatus 10 is provided at a position vertically offset from the sub-base 79 in the upright position. The sub-support surface 78 and the main support surface 72 are angled with respect to the horizontal and are on the same plane, inclined with respect to the main base 70 and the sub-base 79. Furthermore, the multi-position support 50 has another intermediate surface 80 that extends from the sub-base 79 to the sub-support surface 78. The intermediate surface 80 is a boundary portion 82 formed as a bent portion that extends perpendicularly to the sides 60, 62 of the multi-position support 50 and is connected to the sub-base 79. Another intermediate surface 84 extends from the main base 70 to the sub-support surface 78. The intermediate surface 84 is formed as a bent portion 86 that extends perpendicularly to the side surfaces 60 and 62, and is connected to the main base 70. A handle portion 88 is provided at the end 56. The handle portion 88 may also be provided with a support flange 90 that is positioned vertically offset from the sub-base 79 in an upright configuration to support the cell culture apparatus 10. A support flange 94 is provided at the end 58 that extends vertically from the main support surface 72. The support flange 94 is used to hold the cell culture apparatus 10 on the main support surface 72.

[0051] Figure 3 shows the cell culture apparatus 10 supported on a multi-position support 50 in an upright configuration. In the upright configuration, the cell culture apparatus 10 is positioned at an angle of θ1 (for example, approximately 10 to 12 degrees) with respect to the horizontal, with the rear part 100 being higher than the front part 102. However, the vertical angle is parallel to the horizontal (0 degrees). In this upright configuration, the cell culture apparatus 10 can be positioned as a filling start position for initiating the filling of the cell culture apparatus 10. The filling start position is a position in which a gentle filling angle is obtained by lowering the front part 102 to the rear part 100. This filling angle reduces the generation of bubbles in the fluid and promotes the discharge of air through the air manifold and filter connected to the cell culture apparatus 10.

[0052] When filling the cell culture apparatus 10 with the multi-position support 50 in the upright position, the liquid level inside the cell culture apparatus 10 rises toward the air manifold 22 and, consequently, toward the filter connected to the air manifold. If the filter gets wet, the outflow rate of air from the cell culture apparatus 10 decreases, which can increase the internal pressure of the cell culture apparatus 10 and create an undesirable environment inside the cell culture apparatus 10. Therefore, to reduce the possibility of the fluid reaching the filter, the multi-position support 50 is provided with a tilted position setting, which is a position setting in which the multi-position support 50 is rotated together with the cell culture apparatus 10. Rotation to the tilted position setting can be done without lifting either the multi-position support 50 or the cell culture apparatus 10. By applying a force F to the rear corner 110 of the cell culture apparatus 10 and rotating the multi-position support 50 and the cell culture apparatus 10 around the boundary 76, the multi-position support 50 can be easily tilted manually together with the cell culture apparatus 10. Since the boundary portion 76 extends at an angle inclined with respect to the sides 60 and 62 of the multi-position support 50, tilting it changes both the front-to-back angle and the up-to-down angle, raising the position of the upper part of the air manifold to which the filter is connected. According to an embodiment described later, this tilting operation can also be performed by an automated cell culture system without manually applying a force F. However, the same multi-position support 50 shown and described can be used for both manual and automated tilting operations.

[0053] Referring to Figure 6, the multi-position support 50 and the cell culture device 10 are shown in a tilted configuration. In the tilted configuration, the front part 102 is higher than the rear part 100, at an angle of θ2 (for example, 11 to 13 degrees) with respect to the horizontal. As shown in the figure, in the tilted configuration, the corner 112 between the side 40 and the rear part 100 of the cell culture device 10 is placed on the support member. Referring to Figure 7, the upper part 116 is higher than the bottom part 114, at an angle of θ3 (7 to 9 degrees) with respect to the horizontal. Therefore, in the tilted configuration, the multi-position support 50 and the cell culture device 10 are positioned at a composite angle of θ2 (front to back) and θ3 (up and down), which can be called the filling completion position. Once the cell culture device 10 is filled, the side 60 of the multi-position support 50 closest to the upper part 116 of the cell culture device 10 can be rotated upward to position the cell culture device 10 upright. Therefore, all operations of the cell culture device 10 in the filling process can be performed solely using the multi-position support 50, and there is no need to lift the cell culture device 10 from the multi-position support 50 during operation. Drainage from the cell culture device 10 can be performed in the reverse order.

[0054] By using the multi-position support described above, it becomes possible to operate the cell culture apparatus without having to handle the cell culture apparatus separately from the multi-position support during filling or draining operations. Therefore, the multi-position support improves processing efficiency and saves the user time by allowing for increased filling and draining speeds and easy and quick angle changes. Furthermore, the multi-position support allows for clear and concise control protocols, reducing errors and the possibility of defective and / or damaged products. In addition, because the oscillating motion is performed using a fixed tilt angle method with the multi-position support, angle variations can be reduced. Moreover, by providing a composite tilt angle on the multi-position support, the possibility of wetting the filter attached to the air manifold is reduced. In some embodiments, the multi-position device can be made of stainless steel, which improves durability and allows for compliance with good manufacturing practice (GMP) standards. Manufacturing costs can also be reduced by forming the multi-position device with sheet material layered on a metal brake. In this case, it becomes possible to modify the device without reorganizing the equipment at great cost.

[0055] Referring to Figures 8 and 9, a cell culture system 200 is shown that enables automatic filling of the cell culture apparatus 210. Although not shown in detail in Figures 8 and 9, the structure and features of the cell culture apparatus 210 can be the same as those of the cell culture apparatus described elsewhere in this disclosure. Similarly, the multi-position support 250 shown in Figures 8 and 9 is the same as the multi-position support described elsewhere in this disclosure. Therefore, the following description focuses on additional components and features shown in Figures 8 and 9, and thus related to the automatic cell culture medium filling system 200.

[0056] The cell culture apparatus 210 comprises three cell culture modules 212, 214, and 216, each containing multiple layers of cell culture chambers (see Figure 2), which are stacked sequentially to form the multilayer cell culture apparatus 210. Each cell culture module 212, 214, and 216 is equipped with two manifolds 220 and 222. Liquid can enter and exit the cell culture modules 212, 214, and 216 via the first manifold 220. Therefore, the first manifold 220 can be called a fluid manifold. Air can enter and exit the cell culture modules 212, 214, and 216 via the second manifold 222. Therefore, the second manifold 222 can be called an air manifold.

[0057] System 200 includes a first filling sensor 302 positioned to detect the presence of liquid culture medium at a first filling level 303 within the cell culture apparatus 210. A second filling sensor 304 may also be provided to detect the presence of liquid culture medium at a second filling level 305 within the cell culture apparatus 210. Optionally, a third or further filling sensor (not shown) can be provided to obtain additional functionality. For example, a third filling sensor may be provided to detect the presence of liquid culture medium at a third filling level 307 within the cell culture apparatus 210. This third filling level 307 can be used, for example, to change (e.g., slow down) the rate at which liquid culture medium is filled into the cell culture apparatus 210. As shown in Figures 8 and 9, the positions of the first filling level 303, the second filling level 305, and the third filling level 307 are located on the second manifold 222. By arranging them on the second manifold 222 in this way, the filling water levels 303, 305, and 307 are located at the top of the cell culture apparatus 210 during filling, thus indicating filling water levels that gradually approach the final filling water level of the cell culture apparatus 210. Therefore, the positions of these filling water levels 303, 305, and 307 are useful and accessible for measuring the filling water level of the cell culture apparatus 210. Furthermore, since the liquid culture medium level can be captured with an unobstructed, clear view on the manifold 222, the liquid culture medium level can be easily detected with various types of sensors. In some embodiments, the second filling water level 305 indicates the final filling water level of the cell culture apparatus 210. The filling sensors 302 and 304 can be any type of sensor known in the art that is capable of detecting when the liquid culture medium has reached a specific position within the cell culture apparatus 210. In some embodiments, the sensors include at least one of an optical sensor, a through-beam sensor, or a photoelectric sensor.

[0058] The cell culture system 200 also further includes an actuator 300 positioned to move the cell culture apparatus 210 from a first position (e.g., Figure 8) to a second position (e.g., Figure 9) during filling. The plane 400 in Figures 8 and 9 represents the plane of a support member (e.g., a stand, a workbench, or another generally horizontal surface supporting the system 200). In some preferred embodiments, when the first filling sensor 302 detects that the liquid culture medium has reached a first filling level 303, the actuator 300 moves the cell culture apparatus 210 from the first position shown in Figure 8 (with the piston 301 of the actuator 300 extended) to the second position shown in Figure 9 (with the piston 301 of the actuator retracted). In further aspects of some embodiments, when another filling sensor detects that the liquid culture medium is at a predetermined filling level, the actuator 300 may also move the cell culture apparatus 210 to yet another position. If necessary, the actuator 300 may be equipped with a solenoid or other mechanism (e.g., a piezoelectric, pneumatic or hydraulic piston, or a motor) to extend (e.g., Figure 8) and retract (e.g., Figure 9) the actuator 300 to achieve a desired posture. The actuator 300 may be integrated with the multi-position support 250, or may be configured to be attachable to the multi-position support 250.

[0059] The filling sensors 302, 304 and actuator 300 can be connected to a control device (not shown). Specifically, the filling sensors 302, 304 can transmit a detection signal to the control device when they detect that the liquid culture medium has reached a predetermined filling level (e.g., 303, 305). The control device can then control the actuator 300 to change the orientation of the cell culture apparatus 210, or control other components of the system (such as pumps and valves) to control the flow of liquid culture medium to the cell culture apparatus 210 and control the filling speed. Furthermore, by connecting the filling sensors to a timer, the cell culture system 200 can be made to function with a delay by using a timer that is started by a signal from the filling sensors.

[0060] In some embodiments, the detection signal from the first filling sensor 302 can temporarily stop the filling process, and the filling can be resumed once the cell culture apparatus is in a second position, which is the desired position.

[0061] Furthermore, the cell culture system 200 may further include a filling port 310 (e.g., a fluid conduit such as a tube) connected to the inlet of the first manifold 220. The filling port 310 can be connected to means such as a pump for delivering fluid into the cell culture apparatus 210. Additionally, the second manifold 222 is connected to an exhaust port 312 for discharging fluid (e.g., air) from the outlet of the second manifold 222 during filling.

[0062] Furthermore, the cell culture system 200 may be equipped with various other valves and sensors to assist in monitoring and controlling the filling process. For example, one or more inlet valves 314 may be provided on or inside the filling port 310, and one or more outlet valves 316 may be provided on or inside the exhaust port 312. The inlet valves 314 can control the flow of liquid to the cell culture apparatus 210 by closing or restricting the filling port 310. The outlet valves 316 can effectively stop the filling process by closing the exhaust port 312 to prevent fluids such as air from leaking out of the cell culture apparatus. In some embodiments, the inlet valves 314 and outlet valves 316 are pinch valves for closing the inlet and outlet, respectively. For example, the inlet valves 314 and outlet valves 316 may be solenoid pinch valves. Since the pinch valves can be used on the outside of the tube, they are easily reusable. Furthermore, the pinch valve can be made small enough so as not to interfere with the disposable clamp, allowing the operator to loosen the pinch valve with only a simple pre-processing step of tightening the pipe with a disposable clamp to disconnect the connection.

[0063] Furthermore, various flow sensors and pressure sensors can be used in the cell culture system 200. As shown in Figures 8 and 9, a flow control sensor 318 can be provided in the filling port 310. In some embodiments, the flow control sensor 318 is an in-pipe pressure sensor for monitoring the pressure of the liquid culture medium during filling. The flow control sensor 318 is connected to a control device that controls the pump, and the flow rate of the liquid culture medium can be adjusted based on feedback from the flow control sensor 318. For example, if the pressure is too high, the pump speed can be automatically reduced. Such a pressure sensor may have a surface such as a stainless steel foil surface that can be cleaned after use, thereby making the pressure sensor reusable.

[0064] In some embodiments, the automated filling systems, apparatus, and methods described herein can be used with cell culture vessels having multiple layers or stacks. For example, in some embodiments, the cell culture vessel may be a CellSTACK® culture vessel available from Corning Incorporated, Corning, New York. Such a cell culture vessel 1000 is shown in Figures 10 to 13. Figure 10 is a perspective view of a cell culture apparatus (or cell culture vessel) 1000 according to one or more embodiments illustrated and described herein. Figure 11 is a side view showing the cell culture apparatus 1000 shown in Figure 10 in a first filling position 1060 according to one or more embodiments illustrated and described herein. Figure 12 is a side view showing the cell culture apparatus 1000 shown in Figure 10 in a second filling position 1070 according to one or more embodiments illustrated and described herein. Figure 13 is a side view showing the cell culture apparatus 1000 shown in Figure 10 in an incubation position 1080 according to one or more embodiments illustrated and described herein.

[0065] As shown in Figures 10 to 13, a cell culture vessel 1000, such as a "CellSTACK" culture vessel, has a flat bottom surface 1010, a flat top surface 1020 opposite to the bottom surface 1010, and four side walls 1011, 1013, 1015, and 1017 extending vertically from the bottom surface 1010 to the top surface 1020. Multiple layers (cell culture chambers or stacks) 1030 can be placed between the bottom surface 1010 and the top surface 1020. Each layer (cell culture chamber) 1030 has a culture bottom surface 1035, and one side surface 1011 of each cell culture chamber 1030 communicates with a packing column 1040 and an exhaust column 1050. The cell culture vessel can be equipped with any appropriate number of cell culture chambers 1030. While not intended to be limiting, for example, a cell culture vessel may consist of one stack (culture chamber), two stacks (culture chambers), five stacks (culture chambers), ten stacks (culture chambers), or forty stacks (culture chambers).

[0066] The packing column 1040 and packing port 1045, and the exhaust column 1050 and exhaust port 1055, allow direct access to the chamber bottom 1035, enabling highly flexible and sterile filling and draining of a completely sealed system by injection, pipetting, or tubing. The packing column 1040 and exhaust column 1050 may have access ports 1045 and 1055 on the upper surface 1020 of the cell culture vessel 1000, and the access ports may be threaded to engage with a cap or tubing. In some embodiments, the packing port 1045 and exhaust port 1055 are configured to engage with a cap by appropriate connections such as threading. In some embodiments, the cap may have a porous and / or hydrophobic membrane to allow gas exchange with minimal risk of contamination. In some embodiments, the cap may include a packing cap that is sealed integrally with the tubing to allow direct sterile transfer of culture medium and cells by pumping or gravity. The packing column 1040 and the exhaust column 1050 can be positioned at both ends of the same side surface 1011 of the cell culture vessel 1000, respectively. The packing column 1040 extends vertically from the bottom surface 1010 of the cell culture vessel to the top surface 1020 and communicates with each layer (culture chamber) 1030 within the cell culture vessel. The packing column 1040 is equipped with a packing port 1045 on the top surface 1020 of the cell culture vessel 1000, and a desired cap and / or tube can be attached to the packing port 1045. The exhaust column 1050 extends vertically from the bottom surface 1010 of the cell culture vessel to the top surface 1020 and communicates with each layer (culture chamber) 1030 within the cell culture vessel. The exhaust column 1050 is equipped with an exhaust port 1055 on the upper surface 1020 of the cell culture vessel 1000, and a desired cap and / or tube can be attached to the exhaust port 1055.

[0067] In some embodiments, the chambers 1030 of the cell culture vessel 1000 can be filled by an automated filling system as described in several embodiments disclosed herein. In other embodiments, each chamber of the cell culture vessel can be filled by any suitable means such as gravity, a peristaltic pump, or injection. In some embodiments, the cell culture vessel 1000 can be attached to an automated filling system and components disclosed herein, such as a system that may include a filling sensor, to enable automated filling of the cell culture chambers 1030 of the cell culture vessel 1000 with minimal user intervention. Aseptic connections can also be made in a laminar flow hood or cleanroom environment. When using a closed-system filling system, such as a gravity or pump-assisted filling system, the cell culture vessel can also be moved to a non-sterile environment.

[0068] When filling the cell culture vessel 1000, the cap, such as a normal exhaust cap, placed over the filling port 1045 can be replaced with a filling cap. The cell culture vessel 1000 can then be moved to the first filling position 1060. In the first filling position 1060, the cell culture vessel 1000 is placed (supported) on the work surface 1090 with its side 1011 facing downwards, and the filling port 1045 is located near the bottom of the cell culture vessel 1000. In some embodiments, the normal exhaust cap placed over the exhaust port 1055 can be replaced with an exhaust filling cap equipped with an exhaust filter to reduce back pressure. The tube of the filling cap can then be connected to the tube of a sterile dispensing container containing the cell suspension. In some embodiments, the cell suspension can be pumped from the dispensing container into the cell culture vessel to fill the cell culture chambers. The filling of the culture medium into the chambers may not be uniform at first, but afterwards, the water level of the culture medium becomes uniform in each chamber (stack) of the cell culture vessel.

[0069] Once filling is complete, the cell culture vessel can be moved to a second filling position 1070. In the first filling position 1060, the cell culture vessel is placed with its side 1011 facing downwards and the filling port 1045 near the bottom. In the second filling position 1070, the cell culture vessel is rotated 90 degrees so that the side 1011, which has the filling port 1045 and the exhaust port 1055, is at the top of the vessel. In the second filling position 1070, the side 1015 opposite to the side 1011 is placed (supported) on the surface (work surface) 1090 of the support member. The filling cap covering the filling port 1045 can then be replaced with an exhaust cap or a solid cap.

[0070] Subsequently, the cell culture vessel can be placed in incubation position 1080. In incubation position 1080, the bottom surface 1010 and top surface 1020 of the cell culture vessel 1000 and the culture bottom surface 1035 of the culture chamber 1030 are horizontal. For example, the cell culture vessel 1000 can be positioned so that its top surface 1020 and bottom surface 1010 are parallel to the work surface 1090 on which the cell culture vessel is placed, resulting in a flat, horizontal position. Therefore, by moving the cell culture vessel 1000 to incubation position 1080, gravity can cause the surface of each chamber (stack) within the cell culture vessel to be covered with cell culture medium. In some embodiments, the user or manipulator can assist in covering the surface of each cell culture chamber with medium by carefully tilting the cell culture vessel 1000 so as not to cause the medium to flow over the edges of each cell culture chamber into the access column. In some embodiments, the user manually operates (positions) the cell culture chamber to various positions such as a first filling position 1060, a second filling position 1070, and an incubation position 1080. In some embodiments, each step of positioning the cell culture vessel for filling is automated and performed by a manipulator. In several embodiments, the manipulator has a size and configuration that accommodates one or more cell culture vessels and can operate (position) the cell culture vessel for filling automatically or with minimal user manual involvement. In some embodiments, the manipulator can be a commercially available manipulator from Shikoku Kogyo Co., Ltd. (Tokushima City, Japan).

[0071] Subsequently, the cell culture vessel can be placed on a horizontal, flat surface within the incubator or greenhouse that supports the mass of the cell culture vessel. After proper incubation, the cell culture vessel may be completely drained, or it may require a change of culture medium. If the cell culture vessel is to be completely drained, the filling port cap can be replaced with a filling cap. The cell culture vessel can then be connected to a sterile recovery container, and the culture medium can be transferred from the cell culture vessel to the recovery container by gravity or pump pressure.

[0072] Embodiments of an automated cell culture medium filling system and related methods have been disclosed above. However, those skilled in the art will understand that the automated cell culture medium filling system and methods described herein can also be implemented in embodiments other than those disclosed. The disclosed embodiments are presented for illustrative purposes only and not for limitation.

[0073] Preferred embodiments of the present invention are described below in separate sections.

[0074] Embodiment 1 A cell culture vessel comprising a cell culture chamber for culturing cells, an inlet configured for the flow of liquid to fill the cell culture vessel, and an outlet configured for the discharge of fluid from the cell culture vessel, A filling sensor is provided to detect when the liquid inside the cell culture vessel reaches a predetermined filling level at a specific location in the cell culture vessel during the filling of the cell culture vessel, and the filling sensor is configured to generate a detection signal when the liquid inside the cell culture vessel reaches the filling level. An actuator configured to change the orientation of the cell culture vessel in response to the detection signal, A cell culture system equipped with the following features.

[0075] Embodiment 2 The at least one filling sensor includes a first filling sensor, which is configured to detect when the liquid in the cell culture vessel reaches a first filling level and to generate a first detection signal when the liquid in the cell culture vessel reaches the first filling level. The cell culture system according to Embodiment 1, wherein the actuator is configured to change the orientation of the cell culture vessel from a first orientation to a second orientation in response to the first detection signal.

[0076] Embodiment 3 The at least one filling sensor includes a second filling sensor, which is configured to detect when the liquid in the cell culture vessel reaches a second filling level and to generate a second detection signal when the liquid in the cell culture vessel reaches the second filling level. The cell culture system is configured to stop filling the cell culture vessel in response to the second detection signal, The cell culture system according to Embodiment 1 or 2, wherein the second filling water level is different from the first filling water level.

[0077] Embodiment 4 The cell culture system according to any one of embodiments 1 to 3, wherein the cell culture vessel is a multilayer cell culture vessel.

[0078] Embodiment 5 A cell culture system according to any one of embodiments 1 to 4, further comprising a control device configured to receive the detection signal from at least one of the sensors and to control the actuator in response to the detection signal.

[0079] Embodiment 6 The cell culture system according to any one of embodiments 1 to 5, further comprising a pump fluidly connected to the inlet of the cell culture vessel.

[0080] Embodiment 7 The cell culture system according to embodiment 6, wherein the control device is configured to control the flow rate of the pump.

[0081] Embodiment 8 The cell culture vessel comprises a filling side where the inlet and outlet are located, and a support-facing side adjacent to the filling side, and when in the first position, the support-facing side faces a substantially horizontal support member below the cell culture vessel. When in the first position, the side surface of the support facing the support forms a first angle with respect to the support member, the outlet is at a first distance from the support member, and the inlet is closer to the support member than the outlet. When in the second position described above, the side surface of the support facing the support forms a composite angle with respect to the support member, and the composite angle includes (i) a second angle formed between the length of the side surface of the support facing the support and the support member, and (ii) a third angle formed between the width of the side surface of the support facing the support and the support member. A cell culture system according to any one of embodiments 2 to 7, wherein, when in the second posture, the outlet is at a second distance from the support member, and the second distance is greater than the first distance.

[0082] Embodiment 9 The cell culture system according to any one of embodiments 1 to 8, wherein at least one filling sensor is attached to the outside of the cell culture vessel and detects the filling water level through the wall of the cell culture vessel.

[0083] Embodiment 10 The cell culture vessel comprises a plurality of cell culture chambers and a manifold connecting the plurality of cell culture chambers. The cell culture system according to embodiment 9, wherein the at least one packing sensor is attached to the manifold.

[0084] Embodiment 11 The cell culture system according to any one of embodiments 1 to 10, wherein the at least one packing sensor is removable and reusable from the cell culture vessel.

[0085] Embodiment 12 The cell culture system further comprises a flow controller arranged to detect the fluid pressure inside the piping of the cell culture system, The cell culture system according to any one of embodiments 1 to 11, wherein the flow rate controller is configured to reduce the flow rate of the cell culture system in response to the fluid pressure in the piping being above a predetermined pressure value.

[0086] Embodiment 13 The cell culture vessel is further provided with a valve at its outlet. The cell culture system according to any one of embodiments 1 to 12, wherein the valve is configured to close the fluid path of the outlet or nearby of the cell culture vessel, thereby stopping the filling of the cell culture vessel.

[0087] Embodiment 14 The cell culture system according to embodiment 13, wherein the valve is a pinch valve provided outside the tube connected to the outlet.

[0088] Embodiment 15 The cell culture system according to embodiment 13 or 14, wherein the valve is configured to close the fluid path in response to at least one of the detection signal from one of the at least one filling sensors and the fluid pressure in the piping.

[0089] Embodiment 16 The cell culture system according to any one of embodiments 1 to 15, wherein the at least one packing sensor includes at least one of an optical sensor, a through-beam sensor, or a photoelectric sensor.

[0090] Embodiment 17 The cell culture system according to any one of embodiments 1 to 16, further comprising a multiposition support configured to support the cell culture vessel.

[0091] Embodiment 18 The cell culture system according to embodiment 17, wherein the actuator is attached to the multiposition support and configured to change the orientation of the multiposition support with respect to a horizontal support surface on which the multiposition support is positioned.

[0092] Embodiment 19 The actuator has an adjustable length including a first length and a second length, A cell culture system according to any one of embodiments 2 to 18, wherein the cell culture vessel is in a first position when it is at the first length, and the cell culture vessel is in a second position when it is at the second length.

[0093] Embodiment 20 The cell culture system according to Embodiment 19, wherein the first length is greater than the second length.

[0094] Embodiment 21 The cell culture system according to any one of embodiments 1 to 20, wherein the actuator comprises at least one of a solenoid, a piezoelectric material, a pneumatic piston, and a hydraulic piston.

[0095] Embodiment 22 The system further comprises a third filling sensor, which is positioned to detect when the liquid in the cell culture vessel reaches a third filling level, and is configured to generate a third detection signal when the liquid in the cell culture vessel reaches the third filling level. The cell culture system according to any one of Embodiments 3 to 21, wherein the third filling water level is different from the first filling water level and the second filling water level.

[0096] Embodiment 23 The cell culture system according to embodiment 22, wherein the control device is configured to slow down the filling rate of the cell culture vessel based on a third detection signal from the third filling sensor.

[0097] Embodiment 24 A cell culture system according to any one of embodiments 1 to 23, wherein when the liquid is filled into the cell culture vessel, the liquid reaches the first filling level before the second filling level.

[0098] Embodiment 25 A cell culture system according to any one of embodiments 22 to 24, wherein when filling the cell culture vessel with liquid, the liquid reaches the third filling level after the first filling level and before the second filling level.

[0099] Embodiment 26 The cell culture system according to any one of embodiments 1 to 25, wherein the control device is configured to stop filling the cell culture vessel based on the second detection signal from the second filling sensor.

[0100] Embodiment 27 The multi-position support is The main base portion is placed on the support member when the upright position is set. A support surface provided in the upright configuration setting at a position offset vertically from the main base, which supports the cell culture vessel when the cell culture vessel is on the support surface, and It has an intermediate surface that extends from the main base to the support surface, and has a boundary portion that extends at an angle inclined with respect to the side surface of the main base and connects to the main base, The cell culture system according to any one of embodiments 17 to 26, wherein the multi-position support has a tilted configuration in which the multi-position support is rotated around the boundary while the multi-position support supports the cell culture vessel on the support surface, so that the support surface is closer to the support member than in the upright configuration.

[0101] Embodiment 28 The cell culture system according to Embodiment 27, wherein the multi-position support further has a sub-base that is placed on the support member when the upright position is set.

[0102] Embodiment 29 The aforementioned support surface is the first support surface, The multi-position support further has a second support surface located between the main base and the sub-base, The cell culture system according to Embodiment 28, wherein the second support surface supports the cell culture vessel when the cell culture vessel is on the second support surface.

[0103] Embodiment 30 The cell culture system according to Embodiment 29, wherein the first support surface and the second support surface are substantially on the same plane, and the same plane is inclined with respect to the main base.

[0104] Embodiment 31 The cell culture system according to embodiment 29 or 30, wherein the multi-position support further has a support flange that engages with the filled side of the cell culture vessel to restrain the cell culture vessel on the first support surface and the second support surface.

[0105] Embodiment 32 The cell culture system according to any one of embodiments 29 to 31, wherein the multi-position support further comprises other support flanges extending from the first support surface that engage with the rear side of the cell culture vessel opposite to the filled side.

[0106] Embodiment 33 The cell culture system according to any one of embodiments 29 to 32, wherein the second support surface is spaced apart from the support member.

[0107] Embodiment 34 A cell culture system according to any one of embodiments 27 to 33, wherein the support surface is connected to the intermediate surface at another boundary portion which is at an angle inclined with respect to the side surface of the main base.

[0108] Embodiment 35 The cell culture system according to Embodiment 34, wherein the angles of inclination of the two boundary portions with respect to the side surface of the main base portion are substantially the same.

[0109] Embodiment 36 A method for changing the filling angle of a cell culture apparatus comprising a cell culture vessel including a plurality of cell culture modules fluidly connected to each other by a fluid manifold and an air manifold, The steps include connecting the cell culture device to a multi-position support having tilted and upright position settings, The step of filling the cell culture device with the multi-position support, which is set to either the upright position or the tilted position, The steps include detecting that the cell culture apparatus has been filled to a first filling level using a first filling sensor located outside the cell culture module, Based on the step detected by the first filling sensor, the multi-position support is changed from one of the tilted positioning setting and the upright positioning setting to the other of the tilted positioning setting and the upright positioning setting, The steps include detecting, using a second filling sensor located outside the cell culture module, that the cell culture apparatus has been filled to a second filling level different from the first filling level, Based on the step detected by the second filling sensor, the filling process is stopped. A method that includes this.

[0110] Embodiment 37 The method according to embodiment 36, wherein each of the plurality of cell culture modules includes a plurality of cell culture chambers.

[0111] Embodiment 38 The multi-position support is The main base portion that is placed on the support member when the upright arrangement is set as described above, A support surface provided in the upright configuration setting at a position offset vertically from the main base, which supports the cell culture apparatus when the cell culture apparatus is on the support surface, and It has an intermediate surface that extends from the main base to the support surface, and has a boundary portion that extends at an angle inclined with respect to the side surface of the main base and connects to the main base, The method according to Embodiment 36 or 37, wherein in the tilted configuration, the multi-position support is rotated around the boundary while supporting the cell culture apparatus on the support surface, so that the support surface is closer to the support member than in the upright configuration.

[0112] Embodiment 39 The method according to Embodiment 36, wherein the filling step includes filling the cell culture device while the cell culture device is supported by the multi-position support configured to be upright.

[0113] Embodiment 40 The method according to Embodiment 37, further comprising the step of tilting the cell culture apparatus using the multiposition support by rotating the multiposition support about the boundary portion.

[0114] Embodiment 41 The method according to any one of embodiments 36 to 40, wherein the step of changing the multi-position support from one of the tilted positioning setting and the upright positioning setting to the other includes the step of moving an actuator attached to the multi-position support. [Explanation of Symbols]

[0115] 10, 210 Cell culture equipment 12, 14, 16, 212, 214, 216 Cell Culture Modules 18, 1030 Cell Culture Chamber 20, 220 First manifold (fluid manifold) 22,222 Second manifold (air manifold) 24 stacked layers 25 Airway space 26 Cell culture surface 28 membrane 30, 30' sidewall base structure 31, 33, 35 Spacers 32, 32' column structure 34, 34' struct 40 Side view (of cell culture apparatus) 42 Support member 50, 250 Multi-Position Supports 52 Bottom (of the multi-position support) 54 (Top of multi-position support) 56, 58 (ends of the multi-position support) 60, 62 (Sides of the multi-position support) 64, 66 Position tabs 68 (Bottom edge of cell culture apparatus) 70 Main base 71, 73 recesses 72 Main support surface 74, 80, 84 (intermediate surface of multi-position support) 75. Flexed section 76, 77, 82, 86 (boundary of multi-position support) 78 Secondary support surface 79 Subbase 88 Handle 90, 94 Support flange 100 (Rear of cell culture apparatus) 102 Front (of the cell culture apparatus) 110 (Rear corner of the cell culture apparatus) 112 (Corner of cell culture apparatus) 114 Bottom (of the cell culture device) 116 (Top of cell culture device) 200 Cell Culture Systems 300 Actuators 301 Piston 302 First filling sensor 303 First filling water level 304 Second filling sensor 305 Second filling water level 307 Third filling water level 310 (Filling port for cell culture system) 312 (Exhaust port of the cell culture system) 314 Inlet valve 316 Outlet valve 318 Flow control sensor 400 planes 1000 cell culture vessels (cell culture equipment) 1010 Bottom (of the cell culture vessel) 1011, 1013, 1015, 1017 Side walls (of cell culture vessels) 1020 Top surface (of cell culture vessel) 1035 Culture bottom (chamber bottom) 1040 Filling Column 1045 (Cell culture vessel) filling port (access port) 1050 Exhaust column 1055 (Exhaust port / access port of cell culture vessel) 1060 First filling position 1070 Second filling position 1080 Incubation posture (incubation position) 1090 work surface

Claims

1. A cell culture vessel comprising a cell culture chamber for culturing cells, an inlet configured for the flow of liquid to fill the cell culture vessel, and an outlet configured for the discharge of fluid from the cell culture vessel, A filling sensor comprising a first filling sensor, wherein the first filling sensor is positioned to detect when the liquid in the cell culture vessel reaches a first filling level at a predetermined position in the cell culture vessel during filling of the cell culture vessel, and is configured to generate a first detection signal when the liquid in the cell culture vessel reaches the first filling level, A multi-position support configured to support the cell culture vessel, The multi-position support is The main base that is placed on the support member when set upright, A support surface provided in the upright configuration setting at a position offset vertically from the main base, which supports the cell culture vessel when the cell culture vessel is on the support surface, and It has an intermediate surface that extends from the main base to the support surface, and has a boundary portion that extends at an angle inclined with respect to the side surface of the main base and connects to the main base, The multi-position support has a tilted configuration in which, while supporting the cell culture vessel on the support surface, the multi-position support is rotated around the boundary so that the support surface is closer to the support member than in the upright configuration. An actuator attached to the multi-position support and configured to change the orientation of the cell culture vessel from a first orientation to a second orientation in response to the first detection signal, A cell culture system equipped with the following features.

2. The at least one filling sensor further includes a second filling sensor, the second filling sensor being configured to detect when the liquid in the cell culture vessel reaches a second filling level, and to generate a second detection signal when the liquid in the cell culture vessel reaches the second filling level. The cell culture system is configured to stop filling the cell culture vessel in response to the second detection signal. The cell culture system according to claim 1, wherein the second filling water level is different from the first filling water level.

3. The cell culture vessel comprises a plurality of cell culture chambers and a manifold connecting the plurality of cell culture chambers. The cell culture system according to claim 1 or 2, wherein the at least one filling sensor is attached to the manifold.

4. The cell culture system according to claim 2, further comprising a control device configured to receive the first detection signal or the second detection signal and to control the actuator in response to the first detection signal or the second detection signal.

5. The cell culture vessel further comprises a pump connected to the inlet of the cell culture vessel, The cell culture system according to claim 4, wherein the control device is configured to control the flow rate of the pump.

6. The cell culture vessel comprises a filling side where the inlet and outlet are located, and a support-facing side adjacent to the filling side, and when in the first position, the support-facing side faces a substantially horizontal support member below the cell culture vessel. When in the first position, the side surface of the support facing the support forms a first angle with respect to the support member, the outlet is at a first distance from the support member, and the inlet is closer to the support member than the outlet. When in the second position described above, the side surface of the support facing the support forms a composite angle with respect to the support member, the composite angle includes (i) a second angle formed between the length of the side surface of the support facing the support and the support member, and (ii) a third angle formed between the width of the side surface of the support facing the support and the support member. The cell culture system according to any one of claims 1 to 5, wherein when in the second position, the outlet is at a second distance from the support member, and the second distance is greater than the first distance.

7. The cell culture system according to any one of claims 1 to 6, wherein the at least one filling sensor is attached to the outside of the cell culture vessel and detects the filling water level through the wall of the cell culture vessel.

8. The cell culture system further comprises a flow controller arranged to detect the fluid pressure inside the piping of the cell culture system, The cell culture system according to any one of claims 1 to 7, wherein the flow rate controller is configured to reduce the flow rate of the cell culture system in response to the fluid pressure in the piping being above a predetermined pressure value.

9. The cell culture vessel is further provided with a valve at its outlet. The cell culture system according to claim 2 or claim 3 as a reference to claim 2, wherein the valve is configured to close the fluid path at or near the outlet of the cell culture vessel in response to at least one of the first detection signal, the second detection signal, and the fluid pressure in the piping, thereby stopping the filling of the cell culture vessel.

10. The cell culture system according to any one of claims 1 to 9, wherein the at least one packing sensor includes at least one of an optical sensor, a through-beam sensor, or a photoelectric sensor.

11. The actuator has an adjustable length including a first length and a second length, The first length is greater than or equal to the second length, The cell culture system according to any one of claims 1 to 10, wherein the cell culture vessel is in a first position when it is at the first length, and the cell culture vessel is in a second position when it is at the second length.

12. The cell culture system according to any one of claims 1 to 11, wherein the actuator comprises at least one of a solenoid, a piezoelectric material, a pneumatic piston, and a hydraulic piston.

13. The system further includes a third filling sensor, which is positioned to detect when the liquid in the cell culture vessel reaches a third filling level, and is configured to generate a third detection signal when the liquid in the cell culture vessel reaches the third filling level. The third filling water level is different from the first filling water level and the second filling water level. The cell culture system according to claim 4, wherein the control device is configured to slow down the filling rate of the cell culture vessel based on a third detection signal from the third filling sensor.

14. The multi-position support further includes a sub-base portion which is placed on the support member when the upright configuration is set, The support surface is the first support surface, The multi-position support further has a second support surface located between the main base and the sub-base, The second support surface supports the cell culture vessel when the cell culture vessel is on the second support surface. The cell culture system according to claim 1, wherein the first support surface and the second support surface are substantially on the same plane, and the same plane is inclined with respect to the main base.

15. A method for changing the filling angle of a cell culture apparatus comprising a cell culture vessel including a plurality of cell culture modules fluidly connected to each other by a fluid manifold and an air manifold, A step of connecting the cell culture device to a multi-position support having tilted and upright position settings, The multi-position support is The main base portion that is placed on the support member when the upright arrangement is set as described above, A support surface provided in the upright configuration setting at a position offset vertically from the main base, which supports the cell culture apparatus when the cell culture apparatus is on the support surface, and The intermediate surface extends from the main base to the support surface, and further comprises an intermediate surface that connects to the main base at a boundary portion that extends at an angle inclined with respect to the side surface of the main base, In the tilted configuration, the multi-position support is rotated around the boundary while supporting the cell culture apparatus on the support surface, so that the support surface is closer to the support member than in the upright configuration; The step of filling the cell culture device with the multi-position support, which is set to either the upright position or the tilted position, The steps include detecting that the cell culture apparatus has been filled to a first filling level using a first filling sensor located outside the cell culture module, Based on the step detected by the first filling sensor, the multi-position support is changed from one of the tilted positioning setting and the upright positioning setting to the other of the tilted positioning setting and the upright positioning setting, The steps include detecting, using a second filling sensor installed outside the cell culture module, that the cell culture apparatus has been filled to a second filling level different from the first filling level, Based on the step detected by the second filling sensor, the filling process is stopped. A method that includes this.

16. The method according to claim 15, wherein each of the plurality of cell culture modules includes a plurality of cell culture chambers.

17. The method according to claim 15 or 16, further comprising the step of tilting the cell culture apparatus using the multiposition support by rotating the multiposition support about the boundary portion.

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