Culture device

Abstract

A culture device is provided having a base having a first surface and a second surface fixedly disposed relative to the first surface, and a flexible sidewall extending from the base and defining an interior volume of the device, the first surface and the second surface being non-coplanar.

Description

Technical Field

[0001] The present invention relates to a device, in particular a culture device for culturing cells, a method for culturing cells and a system for culturing cells. Background Art

[0002] As is well known, in the field of cell culture, cell growth proceeds from the so-called initial "lag phase" to the so-called "logarithmic phase", in which the cells adapt to the culture environment and adopt a slow growth rate, and in the "logarithmic phase", the cells proliferate exponentially and consume the nutrients in their growth medium. When the cells consume the nutrients in their growth medium, it is necessary to remove the spent medium and introduce fresh growth medium to ensure that the cells continue to proliferate. In addition, as the cells proliferate, the density of the cell culture increases, and it is necessary to transfer the cell culture from one culture device to another larger culture device, which has fresh growth medium. This process is called the passage of cell culture. This is also required for adherent cell culture, in which cells are attached or attached to an appropriate surface. In adherent cell culture, adherent cells will proliferate and eventually occupy the entire surface area available in the culture device. Therefore, it is necessary to transfer the adherent cell culture from one surface to another larger surface to maintain the desired proliferation rate.

[0003] Typically, the process of passaging involves transferring cells from one culture vessel to another, usually larger, vessel. For adherent cells, the process is more complex in that cells that are attached or attached to a surface must be dissociated or detached from the surface before being transferred to another culture vessel. Many dissociation procedures are known in the art, such as shaking or agitating the culture vessel, vigorous pipetting, scraping the cells from the surface, or using reagents such as trypsin, dispase, TrypLE TM (ThermoFisher Scientific), collagenase, EDTA, or Accutase TM (Innovative Cell Technologies)) for enzymatic dissociation.

[0004] There are many disadvantages in the current method of transferring cells between various culture containers. First, by using interconnected pipes, additional containers or other devices that may not be fully sterilized before use, there is a risk that cells are introduced into or introduced through a non-sterile environment. Second, the process of transferring cells requires manual intervention, especially in the case of adherent cell culture, which therefore introduces the risk of operator error and may damage sterility during use. Third, cell yield is usually adversely affected by such a transfer process because it is difficult to ensure that most or all of the cells have been taken out of the first container and then transferred to the second container. Specifically, during the transfer process, cells are usually lost through pipette ends, pipelines, etc. Fourth, in currently known culture containers, it is usually complicated to remove the final cell product from the container at the end of the process. Fifth, using several containers is not an effective use of space, thereby affecting the scalability of the process. Typically, cells are removed by pipetting or by a pumping mechanism, which may introduce additional complexity to the system, have the associated risk of damaging sterility, and have the risk of operator error. In addition, such a pipetting for transferring materials requires a laminar flow hood, which requires the highest level of personal protective equipment and a particle-free environment. As such, typical material transfer via pipetting results in a more costly, time-consuming and labor-intensive process that requires highly skilled operators.

[0005] Therefore, an object of the present invention is to solve or at least alleviate the above-mentioned shortcomings. Specifically, an object of the present invention is to provide a culture device and method, which avoids the need for transferring cells or "going down" of cells between two or more culture devices (such as two culture devices, three culture devices, four culture devices or more culture devices). More specifically, an object of the present invention is to provide a culture device and method, wherein cells can be gone down or otherwise moved in a single culture device, thereby ensuring a sterile environment and higher cell yield. Such a culture device and method can also be more suitable for automation. Another object of the present invention is to provide a culture device, in which material (such as cells) is easier to take out. Another object of the present invention is to provide a culture device, in which material (such as cells) can be mixed, resuspended or otherwise agitated in a culture medium. Summary of the invention

[0006] According to one aspect of the present invention, a device is provided, which includes a base and a side wall, wherein the base includes a first surface and a second surface fixedly arranged relative to the first surface, and the side wall extends from the base and defines an internal volume of the device, wherein the first surface and the second surface are not coplanar.

[0007] That is, the present invention provides a device, for example, a culture device or a device that can be used for culture materials (such as cell materials). The device includes a base and side walls upright from the base. The base includes more than one surface, i.e., a plurality of surfaces. At least one of these surfaces forms an angle with respect to a horizontal plane (i.e., a transverse plane) extending perpendicular to the central longitudinal axis of the device, specifically a predetermined, pre-set or fixed angle. That is, the device may include a plurality of surfaces, each of which is fixedly arranged relative to each other (i.e., cannot move). The transverse plane can be at least partially defined by the base, such as by a specific point of the base set in the transverse plane, or by a side of the base extending in the transverse plane, as further described below.

[0008] In other words, without reference to a specific plane (such as a horizontal plane or a transverse plane), the base is non-planar. That is, the base includes a plurality of surfaces, at least one of which is not coplanar with respect to another one or more surfaces. That is, at least one of the surfaces (such as the first surface) can form an angle between 1 degree and 179 degrees relative to another surface (such as the second surface). The angle can be positive or negative relative to the transverse plane. That is, a culture surface can form an angle from +1 degree to +179 degrees or from -1 degree to -179 degrees relative to another culture surface. In a specific example, a surface can form an angle from 90 degrees to 179 degrees relative to another surface.

[0009] In other words, the base includes a plurality of surfaces, such as a first surface and a second surface, each of which extends in a plane. The planes of the first surface and the second surface are not coplanar. In other words, the planes of the first surface and the second surface form an angle. The angle may be the angle described above or an angle described in more detail below.

[0010] In some embodiments, a culture device is provided, comprising a base and a sidewall extending from the base in an axial direction along a central longitudinal axis, the sidewall defining an internal volume of the device, wherein the base comprises a plurality of surfaces, wherein at least one of the surfaces is non-coplanar with respect to one or more other surfaces.

[0011] Preferably, the sidewall is a flexible sidewall. Optionally, the flexible sidewall is a compressible sidewall. Alternatively, the sidewall is a rigid sidewall.

[0012] This provides the advantage that each surface can be independently arranged to a horizontal position, i.e., arranged in a horizontal plane or a transverse plane. In this way, when the device is redirected from one horizontal surface to another horizontal surface, materials (such as cell cultures) can be transferred or "passed down" between the surfaces without having to leave the device. Therefore, the need for multiple devices is avoided. In this way, the device ensures a continuous aseptic environment, reduces operator manipulation, and therefore reduces relevant errors, and is more suitable for automated processing. In addition, this provides the advantage that small volume cell cultures (such as suspension cell cultures) can be mixed, stirred or resuspended before adding additional cell culture media to the container. For example, before adding additional culture media to the container, a volume of approximately 10mL to approximately 200mL of sedimentation suspension cells can be resuspended in its culture media, which is generally referred to as "expansion" of volume.

[0013] It should be noted that any suitable material can be cultured in the culture device described herein, such as but not limited to biological materials, bacteria, algae, synthetic meat, human tissue or animal tissue, or human cells or animal cells. In particular, the culture device described herein is suitable for culturing adherent cells and / or suspended cells.

[0014] Each of the plurality of surfaces may be arranged fixedly relative to each other. That is, each of the plurality of surfaces is provided in a manner fixed relative to each other. In this respect, the surfaces are arranged statically, i.e. cannot move relative to each other. This can be achieved by means of a rigid surface and / or a rigid base.

[0015] Alternatively, in the case of a substantially circular base, each surface is formed as an angled or sloped sector. That is, they may each be formed as a sloped wedge. Alternatively, in the case of a substantially circular base, each surface is formed as an angled or sloped cylindrical sector. Alternatively, each sector may slope downwardly towards a point or axis of the base, such as a central origin or axis of the base.

[0016] Optionally, the plurality of surfaces include a first surface and a second surface. Optionally, the second surface is separated from the first surface. The first surface and the second surface may have any suitable arrangement, such as side by side or adjacent arrangement or concentric arrangement.

[0017] Optionally, the first surface or the plane in which the first culture surface is arranged or located forms a first angle with respect to a horizontal plane, ie a transverse plane extending substantially perpendicularly to a central longitudinal axis of the device.

[0018] Optionally, the first angle is from approximately 1 degree to approximately 179 degrees.

[0019] Optionally, the first angle is from approximately 1 degree to approximately 90 degrees.

[0020] Optionally, the first angle is from approximately 1 degree to approximately 45 degrees.

[0021] Optionally, the first angle is from approximately 1 degree to approximately 30 degrees.

[0022] Optionally, the first angle is from approximately 5 degrees to approximately 30 degrees.

[0023] Optionally, the first angle is from approximately 5 degrees to approximately 25 degrees.

[0024] Optionally, the second surface or a plane in which the second surface is arranged or located forms a second angle with respect to a horizontal plane (ie a transverse plane extending substantially perpendicularly to a central longitudinal axis of the device).

[0025] Optionally, the second angle is from approximately 1 degree to approximately 179 degrees.

[0026] Optionally, the second angle is from approximately 1 degree to approximately 90 degrees.

[0027] Optionally, the second angle is from approximately 1 degree to approximately 45 degrees.

[0028] Optionally, the second angle is from approximately 1 degree to approximately 30 degrees.

[0029] Optionally, the second angle is from approximately 5 degrees to approximately 30 degrees.

[0030] Optionally, the second angle is from approximately 5 degrees to approximately 25 degrees.

[0031] Optionally, the first angle and the second angle are different or unequal.

[0032] Optionally, each first angle is from approximately 1 degree to approximately 45 degrees, optionally from approximately 5 degrees to approximately 25 degrees, wherein each of the first angle and the second angle are different or unequal.Any of the above-described desired ranges may be used in combination.

[0033] Optionally, the surface area of ​​the first surface is equal to the surface area of ​​the second surface. Alternatively, the surface area of ​​the first surface is not equal to the surface area of ​​the second surface.

[0034] Optionally, a ratio between a surface area of ​​the first surface and a surface area of ​​the second surface is approximately 1:9 to approximately 1:1.

[0035] Optionally, the ratio is approximately 1:9, or approximately 1:8, or approximately 1:7, or approximately 1:6, or approximately 1:5, or approximately 1:4, or approximately 1:3, or approximately 1:2, or approximately 1:1.

[0036] It should be understood that such a ratio may represent the surface area of ​​the first surface relative to the second surface or the surface area of ​​the second surface relative to the first surface.

[0037] Optionally, the device and / or the base (i.e., the device, the base, or the device and the base) can be moved about an axis so that each of the first surface and the second surface can be positioned in a horizontal plane. This can be the case where the first surface, the second surface, or the first surface and the second surface form an angle relative to the horizontal plane. The axis can be coaxial with the adjacent area between the first surface and the second surface. In some examples, the axis can be formed non-coaxially with the adjacent area between the first surface and the second surface.

[0038] Optionally, the base further includes a third surface that is fixedly arranged relative to each of the first surface and the second surface and is not coplanar with each of the first surface and the second surface. That is, the third surface can be fixedly arranged relative to the first surface and the second surface. That is, the third surface can be non-coplanar with the first surface and the second surface.

[0039] Optionally, the device and / or the base (i.e., the device, the base, or the device and the base) can be moved around two axes so that each of the first surface, the second surface, and the third surface can be positioned in a horizontal plane. That is, the base can be moved around the first axis so that one or more of the surfaces can be positioned in a horizontal plane. That is, the base can be moved around the second axis so that one or more of the surfaces can be positioned in a horizontal plane. In some examples, the axis can be coaxial with the adjacent area between the surfaces. In some examples, the axes can be formed perpendicular to each other. Optionally, the base can be moved around more than two axes so that each of the first surface, the second surface, and the third surface can be positioned in a horizontal plane. Any number of axes can be envisioned.

[0040] Optionally, the third surface is separate from the first surface and the second surface.The first surface, the second surface and the third surface may have any suitable arrangement, such as side by side or adjacent arrangement, concentric arrangement or arrangement in a triangle (ie a circular sector or a cylindrical sector).

[0041] Optionally, the third surface or a plane in which the third surface is arranged or located forms a third angle with respect to a horizontal plane (ie a transverse plane extending substantially perpendicularly to a central longitudinal axis of the device).

[0042] Optionally, the third angle is from approximately 1 degree to approximately 179 degrees.

[0043] Optionally, the third angle is from approximately 1 degree to approximately 90 degrees.

[0044] Optionally, the third angle is from approximately 1 degree to approximately 45 degrees.

[0045] Optionally, the third angle is from approximately 1 degree to approximately 30 degrees.

[0046] Optionally, the third angle is from approximately 5 degrees to approximately 30 degrees.

[0047] Optionally, the third angle is from approximately 5 degrees to approximately 25 degrees.

[0048] Optionally, the first angle, the second angle and the third angle are different or unequal.

[0049] Optionally, each of the first angle, the second angle, and the third angle is selected from approximately 1 degree to approximately 45 degrees, optionally selected from approximately 1 degree to approximately 30 degrees, optionally selected from approximately 5 degrees to approximately 25 degrees, wherein each of the first angle, the second angle, and the third angle is different or unequal. Any of the above-described desired ranges may be used in combination.

[0050] Optionally, the first angle is approximately 20 degrees, the second angle is approximately 30 degrees, and the third angle is approximately 45 degrees.

[0051] Although three surfaces have been described above, it is easy to understand that other surfaces such as the fourth, fifth, sixth, etc. surfaces can also be formed as part of the device. Optionally, each surface forms a different angle relative to the horizontal plane (i.e., a transverse plane extending substantially perpendicular to the central longitudinal axis of the device). Optionally, in the case of any number of surfaces, the device and / or the base (i.e., the device, the base, or the device and the base) can be moved around two (or more) axes so that each of the surfaces can be positioned in a horizontal plane. That is, the base can be moved around a first axis so that one or more of the surfaces can be positioned in a horizontal plane. That is, the base can be moved around a second axis so that one or more of the surfaces can be positioned in a horizontal plane. In some examples, the axis can be coaxial with the adjacent areas between the surfaces. That is, the number of axes can be equal to the number of adjacent areas between the surfaces. In some examples, the axes can be formed perpendicular to each other. Optionally, the base can be moved around more than two axes so that each of the surfaces can be positioned in a horizontal plane. Any number of axes can be envisioned.

[0052] Optionally, the surface area of ​​the first surface, the surface area of ​​the second surface and the surface area of ​​the third surface are equal. Alternatively, the surface area of ​​the first surface, the surface area of ​​the second surface and the surface area of ​​the third surface are not equal.

[0053] Optionally, a ratio between the surface area of ​​the first surface, the surface area of ​​the second surface and the surface area of ​​the third surface is approximately 1:2:3.

[0054] Optionally, a ratio between the surface area of ​​the first surface, the surface area of ​​the second surface, and the surface area of ​​the third surface is approximately 1:3:6.

[0055] Optionally, the surface area of ​​the first surface is 25 cm 2 , the surface area of ​​the second surface is 75cm 2 , and the surface area of ​​the third surface is 150cm 2 .

[0056] This offers the advantage that cells can be transferred to larger and larger surface areas as they proliferate.

[0057] Optionally, the base further comprises at least one port. Optionally, the base further comprises at least one outlet.

[0058] This provides the advantage that material, such as cells, can be removed from the device.

[0059] Optionally, one surface or each surface includes a port or outlet. The port or outlet can be arranged on the first surface, the second surface and / or the third surface. The port or outlet can be arranged on any surface.

[0060] Optionally, one or more ports are provided at or adjacent to an abutment region between adjacent surfaces. Optionally, one or more outlets are provided at or adjacent to an abutment region between adjacent surfaces.

[0061] This provides the advantage that a maximum amount of material is harvested from the device. In particular, the first surface and / or the second surface may be pivoted so as to direct the material towards the port, thereby ensuring maximum harvesting efficiency.

[0062] Optionally, the flexible side wall comprises a corrugated wall.

[0063] Optionally, the flexible sidewall comprises a bellows sidewall. The bellows sidewall may comprise a series of deformable regions alternating with rigid sections. The flexible sidewall (such as a bellows sidewall) may be held within an external supporting (optionally surrounding) frame.

[0064] This provides the advantage that the compressible wall element is movable in response to pivoting of the base (ie the first and second surfaces). This also provides the advantage of providing a compressible device that can be stretched and / or compressed by an external force to promote mixing of the contents.

[0065] Optionally, at least one of the surfaces comprises an adherent cell culture substrate or coating.

[0066] This provides the advantage that the device is suitable for use with adherent cells, and thus adherent cells can be "passaged" between surfaces within the device.

[0067] Optionally, the adherent cell culture substrate or coating material comprises an extracellular matrix.

[0068] Optionally, the adherent cell culture substrate or coating material is selected from the group consisting of collagen, fibronectin, vitronectin, laminin, gelatin, poly-lysine, cellulose or a combination thereof.

[0069] Optionally, the base comprises polystyrene, polycarbonate, low density polyethylene, high density polyethylene, silicone or a thermoplastic elastomer. In a particular embodiment, the base comprises silicone, in particular substantially breathable silicone.

[0070] Optionally, at least one of the surfaces or each surface comprises polystyrene, polycarbonate, low density polyethylene, high density polyethylene, silicone or a thermoplastic elastomer.In a particular embodiment, the base comprises silicone, in particular a substantially breathable silicone.

[0071] Optionally, the base comprises a composite material, such as a silicone inner wall and a substantially air impermeable outer wall.

[0072] Optionally, the base comprises a substantially air impermeable thermoplastic elastomer.

[0073] The above materials provide a variety of advantages. For example, a base or surface composed of polystyrene allows adherent cells to attach directly to the surface, thereby reducing the need for additional substrates or coating materials. In addition, a base composed of silicone provides air permeability at the base of the culture device, thereby avoiding hypoxic conditions during use of the culture device. In addition, a base composed of thermoplastic elastomer provides the necessary biocompatibility and mechanical integrity for the base of the device, but can provide an airtight base, which may be advantageous for certain materials to be cultured within the device.

[0074] Optionally, the base is rigid. That is, the base comprises a rigid or non-flexible material.

[0075] Optionally, each of the surfaces is rigid or inflexible. That is, each surface comprises a rigid or inflexible material. Each surface can be rigidly arranged, i.e. cannot be flexed or bent relative to each other.

[0076] Optionally, the base includes a first side and an opposite second side. The first side may include each culture surface. The second side may be substantially planar. The second side may include a material block, such as a wedge, forming the property of one or more surfaces being angled. The second side may include a substantially horizontal surface having a material block extending axially therefrom. The material block may be integrally formed with the second side. Optionally, the second side includes a base and at least one wall erected from the base in an axial direction (i.e., along a longitudinal axis). The at least one wall may form an angled surface.

[0077] Optionally, at least one of the surfaces forms an angle relative to the second side of the base. That is, one or more of the surfaces located on the first side of the base forms an angle relative to the second side of the base. Optionally, the second side of the base defines a transverse plane, i.e. is arranged in the transverse plane.

[0078] Optionally, the device includes a top spaced apart from the base by a side wall. Thus, the top is disposed at the distal end of the side wall and the base is disposed at the proximal end of the side wall. The top may include an opening. The opening may be threaded internally or externally.

[0079] Preferably, the device is a culture device. Optionally, the culture device is a biological culture device, such as a cell culture device. In the case of a culture device, the first surface can be a first culture surface and / or the second surface can be a second culture surface. Additional surfaces (e.g., a third surface) can also form a culture surface (e.g., a third culture surface, etc.).

[0080] According to another aspect of the present invention, a culture device is provided, which includes a base and a flexible side wall extending from the base in an axial direction along a central longitudinal axis, the flexible side wall defining an internal volume of the device, wherein the base includes a plurality of culture surfaces, at least one of the culture surfaces forming an angle relative to a transverse plane defined by at least a portion of the base and extending substantially perpendicular to the central longitudinal axis.

[0081] According to another aspect of the present invention, there is provided a system for culturing cells, the system comprising:

[0082] A culture device as described herein; and

[0083] An interface member is operatively coupled to the top of the device.

[0084] Optionally, the interface member is threadably coupled to a threaded opening of the device, which threaded opening is optionally formed in a top wall of the device.

[0085] Optionally, the interface member comprises one or more ports for material to enter / leave the device. Optionally, one or more ports comprise a septum seal.

[0086] According to another aspect of the present invention, there is provided a system for culturing cells, the system comprising:

[0087] An apparatus as described herein; and

[0088] An actuator is configured to engage at least a portion of the device and move the device between a first configuration in which the first surface is horizontal and a second configuration in which the second surface is horizontal.

[0089] That is, a "horizontal plane" refers to a horizontal plane, ie, a transverse plane extending substantially perpendicular to a central longitudinal axis of the device.

[0090] Optionally, the actuator is configured to move the device and / or the base (ie the device, the base, or both) about at least one axis, or two axes, or two or more axes.

[0091] Optionally, the actuator is configured to move the device and / or the base (i.e., the device, the base, or the device and the base) around two axes so that each of the surfaces can be positioned in a horizontal plane. That is, the base can move around a first axis so that one or more of the surfaces can be positioned in a horizontal plane. That is, the base can move around a second axis so that one or more of the surfaces can be positioned in a horizontal plane. In some examples, the axis can be coaxial with the adjacent area between the surfaces. In some examples, the axes can be formed perpendicular to each other. Optionally, the base can move around more than two axes so that each of the surfaces can be positioned in a horizontal plane. Any number of axes can be envisioned.

[0092] Optionally, the actuator is configured to engage at least a portion of the base and move the base about at least one axis in a transverse plane. In this way, the actuator causes movement between the first configuration and the second configuration.

[0093] Optionally, the actuator is configured to engage at least a portion of the base and move the base around two axes so that each of the surfaces can be positioned in a horizontal plane. That is, the base can move around a first axis so that one or more of the surfaces can be positioned in a horizontal plane. That is, the base can move around a second axis so that one or more of the surfaces can be positioned in a horizontal plane. In some examples, the axis can be coaxial with the adjoining area between the surfaces. In some examples, the axes can be formed perpendicular to each other. Optionally, the base can move around more than two axes so that each of the surfaces can be positioned in a horizontal plane. Any number of axes can be envisioned.

[0094] This offers the advantage that the orientation of each culture surface can be controlled by the actuator, either by manual or automatic manipulation of the actuator.

[0095] Optionally, the actuator is electrically coupled to the processor.

[0096] Optionally, the processor is configured to actuate the actuator according to a set of predefined parameters.

[0097] Optionally, the processor may be electrically coupled to one or more sensors of the culture device.

[0098] Optionally, the processor may be configured to receive signals from one or more sensors and actuate the actuator in response to the received signals.

[0099] Optionally, the actuator is further configured to engage at least a portion of the base and move the base along the longitudinal axis. The longitudinal axis may be a central longitudinal axis of the culture device.

[0100] This provides the advantage that the entire base is translated, allowing mixing of the contents of the culture device by compressing and / or stretching the flexible side walls of the culture device.

[0101] Optionally, the system further comprises an interface member as discussed in the above aspects.

[0102] According to another aspect of the present invention, there is provided a kit of parts comprising:

[0103] A culture device as described herein; and

[0104] An interface plate as described herein, arranged to be operatively coupled to a culture device, and / or an actuator as described herein, configured to engage at least a portion of a base and move the base about at least one axis within a transverse plane.

[0105] According to another aspect of the present invention, there is provided a method for culturing cells, the method comprising the following steps:

[0106] providing a culture device having a first culture surface and a second culture surface, wherein at least one of the first culture surface and the second culture surface forms an angle relative to a transverse plane defined by at least a portion of the base and extending substantially perpendicular to the central longitudinal axis;

[0107] orienting the first culture surface in a plane substantially parallel or coplanar with the transverse plane;

[0108] culturing cells on a first culture surface;

[0109] orienting the second culture surface in a plane substantially parallel or coplanar with the transverse plane, thereby passage cells from the first culture surface to the second culture surface; and

[0110] The cells are cultured on the second culture surface.

[0111] According to another aspect of the present invention, there is provided a method for culturing cells, the method comprising the following steps:

[0112] providing a culture device as described herein;

[0113] orienting the first culture surface in a plane substantially parallel or coplanar with the transverse plane;

[0114] culturing cells on a first culture surface;

[0115] orienting the second culture surface in a plane substantially parallel or coplanar with the transverse plane, thereby passage cells from the first culture surface to the second culture surface; and

[0116] The cells are cultured on the second culture surface.

[0117] According to another aspect of the present invention, there is provided a method for culturing a biological material, the method comprising the following steps:

[0118] providing a first surface and a second surface fixedly disposed relative to the first surface, wherein the first surface and the second surface are not coplanar;

[0119] orienting the first surface into a substantially horizontal plane;

[0120] culturing biological material on a first surface;

[0121] orienting the second surface into a substantially horizontal plane, thereby transferring the biological material from the first surface to the second surface; and

[0122] The biological material is cultured on the second surface.

[0123] This provides the advantage that biological material (such as cell cultures) can be transferred or passaged between the two surfaces without the need to transfer the material to another container. Therefore, this method ensures a continuous sterile environment, reduces operator manipulation and therefore reduces related errors, and is more suitable for automated processing.

[0124] Optionally, the step of orienting the first surface into a substantially horizontal plane comprises rotating the device and / or the base (i.e., the device, the base, or the device and the base) about a first axis. Optionally, the step of orienting the second surface into a substantially horizontal plane comprises rotating the device and / or the base (i.e., the device, the base, or the device and the base) about the first axis or a second (i.e., different) axis.

[0125] Optionally, the biological material comprises tissue or cells.

[0126] Optionally, the cells are adherent cells, and each of the first surface and the second surface comprises an adherent cell culture substrate or coating material.

[0127] Optionally, the method further comprises the step of introducing culture medium into the device.

[0128] Optionally, the method further comprises the step of removing the culture medium from the device.

[0129] Optionally, the method further comprises the step of genetically modifying the cellular material within the device.

[0130] Optionally, the method further comprises the step of introducing viruses and / or magnetic beads into the device.

[0131] Optionally, the method further comprises the step of introducing growth factors, cytokines, etc. into the device.

[0132] Optionally, the method further comprises the step of removing biological material (such as cells) from the device.

[0133] According to another aspect of the present invention, there is provided a method for culturing adherent cells, the method comprising the following steps:

[0134] providing a device as described herein;

[0135] orienting the first surface in a plane substantially parallel or coplanar with the transverse plane;

[0136] culturing adherent cells on a first surface;

[0137] detaching the adherent cells from the first surface;

[0138] orienting the second surface into a plane substantially parallel or coplanar with the transverse plane, thereby passage the adherent cells from the first surface to the second surface; and

[0139] Adherent cells are cultured on the second surface.

[0140] This offers the advantage that adherent cell cultures can be transferred or passaged between two culture surfaces without the need to transfer the cells to another container. Thus, this approach ensures a continuous sterile environment, reduces operator manipulation and, therefore, associated errors, and is more amenable to automated processing.

[0141] Optionally, prior to the step of detaching the adherent cells from the first culture surface, a step of washing the adherent cell culture is provided.

[0142] Optionally, the step of washing may include introducing a washing solution (such as a buffered saline solution) into the device. The step of washing may also include removing the washing solution from the device.

[0143] Optionally, the step of separating the adherent cells from the first surface comprises introducing a dissociation reagent into the device. Optionally, the dissociation reagent comprises trypsin, dispase, TrypLE TM , collagenase, EDTA or Accutase TM Etc. Generally, any suitable dissociation reagent or method can be used to isolate adherent cells.

[0144] Alternatively, any adherent cells can be cultured in the device. The adherent cells can include one or more of mesenchymal stem cells, induced pluripotent stem cells (IPS) cells, embryonic stem cells (ESC), cancer cell lines, human embryonic kidney (HEK) cells, HeLa cells, fibroblasts or epithelial cells, etc.

[0145] Optionally, the method further comprises:

[0146] Culturing adherent cells on a first substrate disposed on a first surface; and / or

[0147] Adherent cells are cultured on a second substrate disposed on the second surface.

[0148] Optionally, the method further comprises the step of introducing culture medium into the device. Optionally, the method further comprises the step of removing culture medium from the device.

[0149] Optionally, the method further comprises the step of genetically modifying the cellular material within the device.

[0150] Optionally, the method further comprises the step of introducing viruses and / or magnetic beads into the device.

[0151] Optionally, the method further comprises the step of introducing growth factors, cytokines, etc. into the device.

[0152] Optionally, the method further comprises a step of removing material (such as cells) from the device. In such a step, the cells may be separated from their respective culture surfaces before being removed.

[0153] Optionally, the methods discussed herein are performed in an apparatus discussed herein.

[0154] Typically, the method discussed herein can be carried out in any appropriate order. In addition, as required, the method discussed herein can include the further step of passage cells to another culture surface (such as the third culture surface or the fourth culture surface). In those cases, the method can further include the step of directing the third surface, the fourth surface, etc., to a substantially horizontal plane. In some examples, the step of directing the third surface, the fourth surface, etc., to a substantially horizontal plane includes rotating the device and / or the base around the first axis or the second axis (i.e., the axis that is different from the first axis). In some examples, the axis can be coaxial with the adjacent area between the surfaces. In some examples, the axis can be formed perpendicular to each other. Any number of axes can be envisioned.

[0155] The optional features noted with respect to the methods herein apply to each of the method "aspects" described herein.

[0156] According to yet another aspect, there is provided use of the device described herein or the system described herein in the cultivation of adherent cells.

[0157] According to yet another aspect, there is provided use of the device described herein or the system described herein in the cultivation of cells in suspension.

[0158] According to yet another aspect, a kit of parts is provided, comprising a device as described herein and an interface member configured to be operatively coupled to a top portion of the device.

[0159] It will be apparent to those skilled in the art that the various elements, features and functions are applicable to all aspects of the invention, whether described as a single aspect or otherwise. BRIEF DESCRIPTION OF THE DRAWINGS

[0160] Example embodiments of the present invention will now be described hereinafter, by way of example only, with reference to the accompanying drawings, in which:

[0161] Figure 1 shows a side view of a culture device according to the present invention;

[0162] Figure 2Shown with interface board attached Figure 1 A side view of a culture device;

[0163] Figure 3 A side view of another culture device according to the present invention is shown;

[0164] Figure 4 Shown with interface board attached Figure 3 A cross-sectional view of a culture device;

[0165] Figure 5 Shows Figures 1 to 4 A top view of a culture device;

[0166] Figure 6 A side view of another culture device with an interface plate attached according to the present invention is shown;

[0167] Figure 7 Shows Figure 6 A top view of a culture device;

[0168] Figure 8 A top view of another culture device according to the present invention is shown;

[0169] Fig. 9 Shown along Figure 8 A view of the culture device in section AA;

[0170] Fig.10 Shown along Figure 8 A cross-sectional view of the culture device BB;

[0171] Fig.11 Shown along Figure 8 Cross-sectional view of the CC culture device;

[0172] Fig.12 (a) and Fig.12 (b) shows a top view of another culture device according to the present invention;

[0173] Fig.13 (a) and Fig.13 (b) shows a top view of another culture device according to the present invention;

[0174] Fig.14 Showing top views of (a) a square base, (b) another square base, (c) a triangular base, and (d) an octagonal base of various culture devices according to the present invention;

[0175] Fig.15 Shows Figure 3 and Figure 4 a first configuration of the culture device during use; and

[0176] Fig.16 Shows Figure 3 and Figure 4 The culture device is in a second configuration during use. DETAILED DESCRIPTION

[0177] The described example embodiments relate to a device and a method. The described example embodiments mainly relate to devices and methods in cell processing (particularly cell therapy and / or gene therapy), but are not limited thereto. As will be appreciated by those skilled in the art, the present device and method can be easily applied to adherent cells, suspended cells, human cells, animal cells, etc. The present device and method can be used for cell therapy and / or gene therapy, tissue engineering, cell agriculture, including laboratory or cell cultured meat products, water treatment and other similar technical fields. In addition, the present device can be applied to other fields, such as chemical processing, agriculture, etc. The present device and method are not limited to any specific use described herein. The use of "culture device" and "culture surface" is not intended to be limiting in any way, but is merely an example of a device and surface that can be used.

[0178] Certain terms used in the following description are for convenience only and are not limiting. The words "upper", "lower", "upwardly" and "downwardly" refer to directions referenced in the drawings and are directions relative to the components when assembled and installed. The words "interior", "inwardly" and "exteriorly", "outwardly" refer to directions toward and away from a specified centerline or geometric center (e.g., central axis) of the element, respectively, and the specific meanings are obvious from the context of the specification. In addition, the terms "proximally" (i.e., closer) and "distally" (i.e., away from) refer to positions relative to a body or attachment point.

[0179] In addition, as used herein, the terms "connected", "fixedly coupled" and the like are intended to include a direct connection between two components without any other components interposed therebetween, and an indirect connection between the components with one or more other components interposed therebetween. The terms include the words specifically mentioned above, their derivatives and words of similar meanings.

[0180] In addition, unless otherwise indicated, the use of ordinal adjectives (such as "first", "second", "third", etc.) merely refers to different instances of similar objects and does not mean that the objects so described must be in a given order in time, space, order, or any other aspect. The same reference numerals are used to describe the same features, and different examples are distinguished by increasing the base number of 100.

[0181] Figure 1 and Figure 2A culture device 10 is shown, which includes a base 12 and a flexible side wall 14. The flexible side wall 14 extends from the base 12 along a central longitudinal axis L of the culture device (i.e., in an axial direction). The flexible side wall 14 extends from a proximal end adjacent to and connected to the base 12 toward a distal end. The distal end includes an opening 16, the inner circumference of which can be threaded. The flexible side wall 14 is formed as a bellows side wall and includes a series of deformable regions 18, which are interlaced with rigid regions 20 located therebetween. The culture device 10 also includes an optional outlet port 22, and an interface plate 24 coupled to the culture device includes a plurality of optional inlet ports 26, such as Figure 2 As shown, it is formed as a septum seal and enables material to enter the culture device 10 and / or leave the culture device.

[0182] Typically, the base 12 comprises a rigid material and the flexible sidewall 14 comprises an elastic material. The base 12 may be composed of polystyrene, polypropylene, polycarbonate, low-density polyethylene or high-density polyethylene (LDPE or HDPE), a thermoplastic elastomer, silicone, etc. The flexible sidewall 14 may be composed of silicone, a substantially air-permeable or substantially air-impermeable thermoplastic elastomer, a flexible polymer film, etc. Any combination of materials for the base 12 and the flexible sidewall 14 is contemplated. Likewise, in embodiments not shown, the sidewall may be composed of a rigid material.

[0183] Specific reference Figure 2 , the culture device 10 includes an interface plate 24 coupled to its opening 16. The interface plate 24 can be threadedly coupled to the threaded interior of the opening 16. Likewise, other coupling methods can be envisioned, such as push-fit or clip-fit. The interface plate 24 includes a plurality of ports 26 formed as septum seals for material to enter and exit the culture device 10 through the interface plate 24. The plurality of ports 26 can allow access to the culture device 10 by using a pin connector (not shown).

[0184] like Figure 2 As shown, the base 12 of the culture device 10 includes a first side 12a and an opposite second side 12b. The first side 12a forms a first culture surface 28 and a second culture surface 30, which are separated by an abutment area 32. The first culture surface 28 and the second culture surface 30 are positioned at a fixed angle relative to each other, which will be further described herein. Figure 1As shown, the first culture surface 28 is angled relative to the second side 12b of the base 12, forming a fixed angle α relative to the second side 12b. In this case, the second side 12b extends in a transverse (i.e., horizontal) plane extending perpendicular to the central longitudinal axis L and is arranged in this transverse plane. That is, the second side 12b is planar. Again, with specific reference to Figure 2 , the angle α between the first culture surface 28 and the second side 12b is fixed and is formed by means of the material block 12c. The cross section of the material block 12c is substantially triangular. In this way, the second side 12b of the base 12 is provided as a continuous (i.e., flat) surface, and the material block 12c provides the angled first culture surface 28. In other words, the material block 12c is provided as a so-called wedge, thereby providing a surface 28 at a fixed angle.

[0185] The first culture surface 28 and the second culture surface 30 can be the same or different materials. In some examples, particularly those related to culturing adherent cell types, the first culture surface 28 and / or the second culture surface 30 include a coating material or substrate that allows cell attachment, or are composed of a suitable material that allows cell attachment, such as polystyrene. The adjacent area 32 generally separates the first culture surface 28 and the second culture surface 30. The culture device 10 includes a plurality of optional outlet ports 22, which are disposed on the second culture surface 30, adjacent to the adjacent area 32, although the plurality of optional outlet ports can alternatively be formed at the adjacent area 32 or on the first culture surface 28. In this example, one port 22 is shown, but more ports or no ports are also contemplated.

[0186] Figure 3 and Figure 4 Another culture device 100 is shown, which is respectively Figure 1 and Figure 2 The culture device 10 is substantially similar, and therefore the same elements will not be described in detail. Figure 3 and Figure 4 In the example, no egress port is provided ( Figure 1 and Figure 2 22 in the figure). In addition, Figure 3 and Figure 4 In the example, no material block is provided ( Figure 2 Thus, the base 112 is provided with a fixed angle β extending between the first culture surface 128 and a transverse plane T extending perpendicularly to the central longitudinal axis L. The transverse plane T is partially defined by the base 112, in particular by the second culture surface 130 and the adjacent area 132.

[0187] Figure 5 Shows Figures 1 to 41. A top view of a culture device 10 is shown. As shown, a base 12, 112 is provided, wherein a first culture surface 28, 128 and a second culture surface 30, 130 are separated by an adjacent area 32, 132. An optional outlet port 22 (present in Figure 1 and Figure 2 ) is shown in dashed lines to illustrate its optional nature. The optional outlet port 22 can be located on any of the culture surfaces 28, 128, 30, 130. Likewise, there can be multiple outlet ports (not shown).

[0188] Figure 6 A side view of another culture device 200 is shown. The culture device 200 is substantially similar to Figure 3 and Figure 4 Specifically, the culture device 200 includes a base 212, flexible sidewalls 214, an interface plate 224, and an optional port 226, as described above.

[0189] The culture device 200 includes a first culture surface 228 separated from a second culture surface 230 by an adjacent region 232. The first culture surface 228 and the second culture surface 230 are positioned relative to each other at a fixed angle, which will be further described herein. In this example, the first culture surface 228 forms a fixed angle α relative to the transverse plane T, and the second culture surface 230 forms a fixed angle β relative to the transverse plane T. The transverse plane T is partially defined by the base 212 because the adjacent region 232 (that is, the axis defined by the adjacent region 232) is arranged in the transverse plane T. In this example, angle α and angle β are shown as being different. In other examples, they can be the same. In this particular example, α is 30 degrees, and β is 45 degrees.

[0190] Figure 7 Shown in top view Figure 6 The culture device 200. As shown in the figure, the first culture surface 228 and the second culture surface 230 of the base 212 are separated by an adjacent area 232.

[0191] It should be noted that although Figure 6 and Figure 7 The culture device is basically similar to Figure 3 and Figure 4 The culture device 100 may also be introduced Figure 1 and Figure 2The base 212 may include other features of the culture device 10. For example, the first culture surface 228 and / or the second culture surface 230, or even the adjacent region 232, may include an optional outlet port. In addition, the base 212 may be provided with blocks of material disposed on either side of the adjacent region 232 to provide an angled first culture surface 228 and second culture surface 230 while ensuring a continuous and planar second side (i.e., bottom side) of the base 212.

[0192] Figures 8 to 11 Another culture device 300 is shown. The culture device 300 is substantially similar to the culture device 300 of Figure 6 and Figure 7 The culture device 200 is described above, except that the culture device has additional (i.e., three in total) culture surfaces. Figures 9 to 11 As shown, culture device 300 includes features of base 312 , flexible sidewalls 314 , interface plate 324 , and port 326 .

[0193] Specifically, Figure 8 As shown, the culture device 300 includes a base 312 having a first culture surface 328, a second culture surface 330 and a third culture surface 334. The first culture surface 328, the second culture surface 330 and the third culture surface 334 are each positioned at a fixed angle relative to each other, which will be further described herein. The first culture surface 328 is separated from the second culture surface 330 by a first adjacent area 332. The second culture surface 330 is separated from the third culture surface 334 by a second adjacent area 336. The third culture surface 334 is separated from the first culture surface 328 by a second adjacent area 336. The first culture surface 328, the second culture surface 330 and the third culture surface 334 are each formed into a sector, i.e. a circular sector, in which the base 312 is circular according to this example. That is, each of the first culture surface 328, the second culture surface 330 and the third culture surface 334 is formed by two radii and an arc.

[0194] Fig. 9 Shown by Figure 8 , and shows the relationship between the first culture surface 328 and the third culture surface 334 separated by the second abutment area 336. As shown, the first culture surface 328 forms a fixed angle α with respect to the transverse plane T. In this example, α is 20 degrees. As shown, the third culture surface 334 forms a fixed angle γ with respect to the transverse plane T. In this example, γ is 45 degrees.

[0195] Fig.10 Shown by Figure 8BB, and shows the relationship between the second culture surface 330 and the third culture surface 334 separated by the second adjacent area 336. As shown, the second culture surface 330 forms a fixed angle β with respect to the transverse plane T. In this example, β is 30 degrees. As shown, and as also described above with respect to Fig. 9 As described, the third culture surface 334 forms a fixed angle γ with respect to the transverse plane T. In the example shown, γ is 45 degrees.

[0196] Fig.11 Shown by Figure 8 336) and shows the relationship between the first culture surface 328 and the second culture surface 330 separated by the second adjacent area 332. As shown in the figure, and also as described above with respect to Fig. 9 As described above, the first culture surface 328 forms a fixed angle α relative to the transverse plane T. In the example shown, α is 20 degrees. Fig.10 As shown and described, the second culture surface 330 forms a fixed angle β with respect to the transverse plane T. In the example shown, β is 30 degrees.

[0197] As described above, the base 312 of the culture device 300 is thus formed of three angled or inclined cylindrical sectors, thereby forming each of the culture surfaces 328, 330, 334. The surface area of ​​each culture surface 328, 330, 334 gradually increases from the first culture surface 328 to the third culture surface 334, and forms a surface area ratio of 1:3:6 (i.e., the surface area of ​​the first culture surface: the surface area of ​​the second culture surface: the surface area of ​​the third culture surface). In a specific example, the first culture surface 328 has a surface area of ​​25 cm 2 The second culture surface 330 has a surface area of ​​75 cm 2 The third culture surface 334 has a surface area of ​​150 cm 2 "Surface area" refers to a surface that forms corresponding fixed angles α, β, γ with respect to a transverse plane T. The area defining the "surface area" (i.e., the area of ​​the culture surface that extends in the plane of the culture surface) defines the area on which cells, tissues or other materials can be cultured.

[0198] Fig.12 (a) and Fig.12 (b) shows another culture device 400, which has a base 412, which includes three culture surfaces: a first culture surface 428, a second culture surface 430 and a third culture surface 434. The first culture surface 428 is separated from the second culture surface 430 by a first adjacent area 432. The second culture surface 430 is separated from the third culture surface 434 by a second adjacent area 436. Fig.12 As shown in (a), the first abutment region 432 and the second abutment region 436 each extend across a chord of the substantially circular base 412 and are parallel to each other. The first culture surface 428, the second culture surface 430, and the third culture surface 434 each have approximately equal surface areas, such that the ratio between the first culture surface 428, the second culture surface 430, and the third culture surface 434 is approximately 1:1:1. Fig.12 (b), in another example, the first abutment region 432 extends across a chord of the substantially circular base 412, and the second abutment region 436 extends across a diameter of the substantially circular base 412. Thus, the first abutment region 432 and the second abutment region 436 extend parallel to each other. The first culture surface 428, the second culture surface 430, and the third culture surface 434 each have a gradually increasing surface area, such that the ratio between the first culture surface 428, the second culture surface 430, and the third culture surface 434 is approximately 1:2:3, or in another example 1:3:6.

[0199] In any example, at least two of the culture surfaces 428, 430, 434 form a fixed angle relative to the transverse plane, similar to that described in the previous example. In a specific example, the first culture surface 428 and the third culture surface 434 form a fixed angle relative to the transverse plane, while the second culture surface 430 is substantially parallel or coplanar with the transverse plane. The first culture surface 428 and the third culture surface 434 can be at equal or different fixed angles relative to the transverse plane.

[0200] Fig.13 (a) and Fig.13 (b) shows a culture device 500 having a base 512 comprising four culture surfaces: a first culture surface 528, a second culture surface 530, a third culture surface 534 and a fourth culture surface 538. Four adjoining regions are also provided: a first adjoining region 540, a second adjoining region 542, a third adjoining region 544 and a fourth adjoining region 546. The first culture surface 528 is separated from the second culture surface 530 by the first adjoining region 540. The second culture surface 530 is separated from the third culture surface 534 by the second adjoining region 542. The third culture surface 534 is separated from the fourth culture surface 538 by the third adjoining region 544. The fourth culture surface 538 is separated from the first culture surface 528 by the fourth adjoining region 546.

[0201] Fig.13 (a) and Fig.13 The difference between (b) is the point where adjacent regions 540, 542, 544, 546 meet. Fig.13In (a), each of the adjacent regions 540, 542, 544, 546 intersects at a central origin O, which is located along the central longitudinal axis of the device. Fig.13 In the example of (a), the surface area of ​​each culture surface 528, 530, 534, 538 is equal. Alternatively, each adjacent region 540, 542, 544, 546 intersects at a non-central point P, which is arranged along an axis other than the central longitudinal axis of the device. Fig.13 In the example of (b), the surface area of ​​each culture surface 528, 530, 534, 538 is different.

[0202] In any example, at least three of the culture surfaces 528, 530, 534, 538 form a fixed angle relative to the transverse plane, similar to that described in the previous example. In a specific example, the first culture surface 528, the second culture surface 530, and the third culture surface 534 each form a fixed angle relative to the transverse plane, while the fourth culture surface 538 is substantially parallel or coplanar with the transverse plane. The first culture surface 528, the second culture surface 530, and the third culture surface 534 can be at equal or different fixed angles relative to the transverse plane.

[0203] Fig.14 (a) to Fig.14 (d) shows different cross-sectional shapes of a base 612, 712, 812, 912 in a culture device, each having an adjacent region 632, 732, 832, 932 separating a first culture surface 628, 728, 828, 929 from a second culture surface 630, 730, 830, 930. Fig.14 (a) shows a generally square base 612 including an abutment region 632 extending between opposing sides of the square base 612 . Fig.14 (b) shows a generally square base 712 including an abutment region 732 extending between adjacent sides of the square base 712 . Fig.14 (c) shows a generally triangular base 812 including an abutment region 832 extending between adjacent sides of the triangular base 812 . Fig.14 (d) shows a generally octagonal base 912 including an abutment region 932 extending between generally opposing corners of the octagonal base 912. As will be appreciated by those skilled in the art, the base 912 may take on any geometric shape or cross-sectional profile, and the abutment region 932 may extend between any suitable edges, sides, corners, etc. of such a base 912.

[0204] exist Fig.14 (a) to Fig.14In the example of (d), at least one or both of the first culture surface 628, 728, 828, 928 and the second culture surface 630, 730, 830, 930 form a fixed angle with respect to the transverse plane T as described herein. The angle formed can be any angle contemplated herein.

[0205] Currently described Figures 1 to 14 The culture device can be used to perform cell passage in the following manner. Specifically, the fixed nature of the angles between the culture surfaces (or, in fact, between a culture surface and the transverse plane T) enables cells to be passaged between the surfaces by moving the base (i.e., moving the transverse plane T at a predetermined angle) so that each culture surface is positioned in a horizontal plane in a predetermined order. In the specific exemplary embodiments discussed below, reference is made to Figure 3 and Figure 4 The use of culture device 100 is described herein, although such use is equally applicable to any other culture device described herein.

[0206] Example 1: Suspension cell culture

[0207] In one example, a suspension cell culture (i.e., a suspension of cells of the desired type in an appropriate culture medium) is introduced into Figure 3 and Figure 4 100. Specifically, the cell culture can be loaded from an external receptacle having a pin connector (not shown) into the volume of the culture device 100. The pin connector can cause piercing of a septum seal within the external receptacle and can also cause piercing of one of the ports 126 of the interface plate 124, thereby fluidly connecting the external receptacle and the culture device 100. Thus, the cell culture is introduced into the culture device 100.

[0208] Prior to or after the introduction of the suspension cell culture, an external actuator (not shown) can rotate the entire base 112 so that the first culture surface 128 is substantially horizontal (i.e., flush or coplanar with the transverse plane T). Fig.15 An external actuator (not shown) can cause the base to pivot or rotate about an axis substantially aligned with the abutment region 132. It should be noted that the flexible sidewalls 114 allow the base 112 to pivot in this manner. Fig.15 Position shown.

[0209] The cells within the cell culture are allowed to settle onto the base wall 112, i.e., onto the first culture surface 128. Optionally, the base 112 may be agitated by external actuation, such as pivoting the base around an axis defined by the abutment area 132, so that all cells move to, and thus reside on, the first culture surface 128. Alternatively, the culture device 100 may be tilted in its entirety so that the cells settle and reside on the first culture surface 128.

[0210] Allow the cells in the culture to proliferate, thereby achieving the exponential growth of the cells. During the proliferation period, suitable materials such as growth factors, fresh culture media, proteins, magnetic beads, antibodies, viruses, etc. can be added to the cell culture. Such materials can be introduced from an external receiver with a pin connector, similar to the introduction of the above-mentioned cell culture. During the proliferation period, the base 112 (i.e., the first culture surface 128 and the second culture surface 130) can be pivoted around the axis defined by the adjacent area 132 in a cyclic manner for mixing and / or resuspending the cell culture. Typically, the cyclic pivoting around the axis defined by the adjacent area 132 can cause turbulence in the cell culture device 100 to allow the contents therein to mix and / or resuspend. The cyclic pivoting around the axis defined by the adjacent area 132 can be provided between any appropriate angle (such as +10 degrees and -10 degrees). Allow the cell culture to proliferate until the desired cell density is reached.

[0211] Optionally, a culture medium replacement step is provided. Specifically, a portion of the cell culture medium (i.e., the exhausted or used culture medium) is removed from the culture device 100 and replaced with fresh culture medium. The culture medium can be removed by the port 126 of the interface plate 124. By using an external receiver with a pin connector, fresh culture medium is introduced through the port 126 of the interface plate 124, similar to the introduction of the above-mentioned cell culture. Similarly, as will be appreciated by those skilled in the art, a culture medium replacement step can be provided after cell passage as described below. In some examples, a culture medium replacement step can be provided before and after cell passage as described below.

[0212] The culture device 100, and in particular the base 112 thereof, is pivoted about an axis defined by the abutment region 132. Specifically, an external actuator (not shown) causes the entire base 112 to pivot so that the second culture surface 130 is substantially horizontal, i.e., flush or coplanar with the transverse plane T. This Fig.16. In this way, cells are transferred or passaged from the first culture surface 128 to the second culture surface 130 by gravity so that all cells reside on the second culture surface 130. The cell culture can then be allowed to proliferate until the desired cell density is reached. During proliferation, the base 112 can be pivoted in a cyclic manner around the axis defined by the adjacent area 132 for mixing and / or resuspending the cell culture. Typically, the cyclic pivoting around the axis defined by the adjacent area 132 can cause turbulence within the cell culture device 100 to allow the contents therein to be mixed and / or resuspended. The cyclic pivoting around the axis defined by the adjacent area 132 can be provided at angles between +10 degrees and -10 degrees.

[0213] Optionally, further transfer or passage of cells may be performed, i.e., from the second culture surface 130 back to the first culture surface 128. In this case, the above process is reversed, even if the entire base 112 is removed from the second culture surface 130. Fig.16 Move back to Fig.15 In this way, further mixing or passage is achieved, which can promote oxygenation of the cell culture. The cell culture is allowed to proliferate until the desired cell density is reached.

[0214] Furthermore, pivoting of the base 112 about the axis defined by the abutment area 132 may be performed for the purpose of mixing and / or resuspending the cell culture. Thus, after mixing and / or resuspending, the cells may be allowed to settle on one of the first culture surface 128 and the second culture surface 130, depending on which culture surface is placed in the horizontal plane.

[0215] Finally, once the cell culture reaches maturity and a desired density is achieved, the cell culture is harvested through port 126 of interface plate 124. Alternatively, where there are optional ports in the base of the culture device, the cells may be harvested through such ports.

[0216] Example 2: Adherent cell culture

[0217] In another example, adherent cell cultures (i.e., adherent cells in an appropriate culture medium) are introduced into Figure 3 and Figure 4 1. The cell culture device 100 includes a coating material or substrate on each of the first culture surface 128 and the second culture surface 130 to allow cells to attach thereto. Specifically, the cell culture can be loaded into the volume of the culture device 100 from an external receiver having a pin connector (not shown). The pin connector can cause puncture of a septum seal in the external receiver and can also cause puncture of one of the ports 126 of the interface plate 124, thereby fluidly connecting the external receiver and the culture device 100.

[0218] Prior to or after introduction of the adherent cell culture, an external actuator (not shown) can rotate the entire base 112 so that the first culture surface 128 is substantially horizontal (i.e., flush or coplanar with the transverse plane T). Fig.15 An external actuator (not shown) can cause the base to pivot or rotate about an axis substantially aligned with the abutment region 132. It should be noted that the flexible sidewalls 114 allow the base 112 to pivot in this manner. Fig.15 Position shown.

[0219] The cells in the cell culture are allowed to settle onto the base wall 112, ie onto the first culture surface 128, in particular onto its coating material or substrate.

[0220] The cell culture is allowed to proliferate, i.e., by attaching to the first culture surface 128, thereby achieving exponential growth of the cells. The cells can be allowed to attach to the first culture surface 128 and proliferate on the first culture surface within a specific time period, for example, within a time period of 10 minutes to 24 hours, preferably within a time period of 6 hours to 12 hours. During the proliferation period, appropriate materials such as growth factors, fresh culture medium, proteins, magnetic beads, antibodies, viruses, etc. can be added to the cell culture. Such materials can be introduced from an external receiver with a pin connector, similar to the introduction of the above-mentioned cell culture. The cell culture is allowed to proliferate until the desired cell density is reached.

[0221] Optionally, before the cells are passaged from the first culture surface 128 to the second culture surface 130, a culture medium replacement step is provided. Specifically, a portion of the cell culture medium (i.e., the exhausted or used culture medium) is removed from the culture device 100 and replaced with a fresh culture medium. The culture medium can be removed through the port 126 of the interface plate 124. By using an external receiver with a pin connector, fresh culture medium is introduced through the port 126 of the interface plate 124, similar to the introduction of the above-mentioned cell culture. Similarly, as will be appreciated by those skilled in the art, a culture medium replacement step can be provided after cell passage as described below. In some examples, a culture medium replacement step can be provided before and after cell passage as described below.

[0222] Before the adherent cells are transferred or passaged from the first culture surface 128 to the second culture surface 130, the cell culture is washed. Specifically, all or a portion of the cell culture medium is removed from the culture device 100 through one of the ports 126 of the interface plate 124. Subsequently, a washing solution is introduced into the culture device 100. The washing solution can be a buffered saline solution, for example, phosphate buffered saline. Generally, any suitable washing solution can be used to ensure the removal of any reagents, such as serum, calcium or magnesium, which inhibit the effect of the dissociation reagent discussed below. Gently agitate the culture device 100, for example by shaking, so as to wash the cell culture. Agitation can be achieved by means of an external actuator (not shown) moving the base 112. The washing solution is then removed from the culture device 100 through one of the ports 126 of the interface plate 124.

[0223] After washing, and before passaging adherent cells, a dissociation reagent is added to the culture device 100. The dissociation reagent may be preheated to a temperature of 37 degrees Celsius prior to addition. The dissociation reagent may be trypsin or TrypLE TM Or other reagents to allow adherent cells to dissociate from the coating material or substrate on the first culture surface 128. Gently agitate the culture device 10, such as by shaking, to ensure that the cells are completely dissociated from the first culture surface 128. Agitation can also be achieved by moving the base 112 with an external actuator (not shown). The culture device 100 can also be incubated at room temperature for a period of time.

[0224] After the adherent cells are detached from the first culture surface 128, the base 112 is pivoted about the axis defined by the abutment region 132. Specifically, an external actuator (not shown) causes the entire base 112 to pivot so that the second culture surface 130 is substantially horizontal, i.e., flush or coplanar with the transverse plane T. This Fig.16. In this way, cells are transferred or passaged from the first culture surface 128 to the second culture surface 130 by gravity, so that all cells reside on the second culture surface 130. At this stage, fresh cell culture medium can also be added to the culture device through one of the ports 126 in the interface plate 124. Adherent cells are then allowed to attach to the coating material or substrate on the second culture surface 130. The cells are allowed to attach and proliferate within a specific time period, for example, within a time period of 10 minutes to 24 hours, preferably within a time period of 6 hours to 12 hours. Once the cells have been allowed to attach within a specific time period, the cell culture medium containing the dissociation reagent is removed through one of the ports 126 of the interface plate 124. Then, similar to the introduction of the above-mentioned cell culture, fresh cell culture medium is introduced through the port 126 of the interface plate 124 using an external receiver with a pin connector, and the cells are allowed to proliferate on the coating material or substrate on the second culture surface 130. During proliferation, appropriate materials such as growth factors, fresh culture medium, proteins, magnetic beads, antibodies, viruses, etc. can be added to the cell culture. Such materials can be introduced from an external receiver having a pin connector, similar to the introduction of the cell culture described above. The cell culture is allowed to proliferate until the desired cell density is reached.

[0225] Finally, once the cell culture reaches maturity and achieves the desired density, the cell culture is harvested through the port 122 of the interface plate 124. Alternatively, in the case of an optional port in the base of the culture device, the cells can be harvested through such a port. Specifically, the above-mentioned washing steps and dissociation steps can be applied to the cell culture residing on the coating material or substrate on the second culture surface 130 to ensure that the adherent cells are properly dissociated therefrom. Subsequently, the base 112 can be moved or pivoted around the axis defined by the adjacent area 132 by an actuator (not shown) to ensure that all cells are separated from the coating material or substrate. The cells can then be harvested in the manner described above.

[0226] Although the above example scheme describes two culture surfaces, the skilled person will equally apply the above guidance to any number of culture surfaces. Specifically, in the case of three, four or more culture surfaces, the base can also be moved so that cells are passaged from the first culture surface to the second culture surface, to the third surface, etc. For adherent cells, a dissociation step as described above is required between each passage.

[0227] Furthermore, the above example protocols can be performed in any of the devices described herein. Furthermore, the example protocols can be followed in any order, for example, suspension or adherent cells can be first cultured on the second culture surface 130 and then passaged to the first culture surface 128.

[0228] Furthermore, while the use of the apparatus mentioned above is described with respect to movement of the base to allow transfer or "passaging" of material, it is also contemplated that the apparatus as a whole may be moved, tilted, etc. to allow the ordered surface to be positioned in a horizontal manner. This is particularly true for examples where the side walls are rigid.

[0229] It will be appreciated by those skilled in the art that the above embodiments are described by way of example only and are not intended to be limiting, and that various changes and modifications are possible without departing from the scope of the invention as defined by the appended claims. Various modifications to the detailed design as described above are possible, for example, there may be changes in shape, size, material, arrangement, assembly, etc.

[0230] In certain embodiments, the invention may be defined by one or more of the following numbered clauses:

[0231] Clause 1. An apparatus (10, 100, 200, 300) comprising:

[0232] A base (12, 112, 212, 312, 412, 512, 612, 712, 812, 912) comprising a first surface (28, 128, 228, 328, 428, 528, 638, 728, 828, 928) and a second surface (30, 130, 230, 330, 430, 530, 630, 730, 830, 930) fixedly arranged relative to the first surface; and

[0233] a side wall (14, 114, 214, 314) extending from the base and defining an interior volume of the device,

[0234] Wherein, the first surface and the second surface are not coplanar.

[0235] Clause 2. A device according to clause 1, wherein the side wall is a flexible side wall, such as a compressible side wall.

[0236] Clause 3. A device according to clause 1 or clause 2, wherein the first surface forms a first angle (α) relative to a transverse plane (T), the transverse plane extending substantially perpendicular to a central longitudinal axis (L) of the device.

[0237] Clause 4. The device of clause 3, wherein the first angle is from approximately 1 degree to approximately 45 degrees, preferably from approximately 5 degrees to approximately 30 degrees.

[0238] Clause 5. A device according to any preceding clause, wherein the second surface forms a second angle (β) relative to a transverse plane (T), the transverse plane extending substantially perpendicularly to a central longitudinal axis (L) of the device.

[0239] Clause 6. The device of clause 5, wherein the second angle is from approximately 1 degree to approximately 45 degrees, preferably from approximately 5 degrees to approximately 30 degrees.

[0240] Clause 7. An apparatus according to any preceding clause, wherein the ratio between the surface area of ​​the first surface and the surface area of ​​the second surface is approximately 1:9 to approximately 1:1.

[0241] Clause 8. An apparatus according to clause 7, wherein the ratio is approximately 1:3.

[0242] Item 9. An apparatus according to any of the preceding items, wherein the base further includes a third surface (334, 434, 534) fixedly arranged relative to each of the first surface and the second surface, wherein the first surface, the second surface and the third surface are not coplanar.

[0243] Item 10. A culture device according to item 9, wherein the third surface forms a third angle (γ) relative to a transverse plane (T), and the transverse plane extends substantially perpendicular to the central longitudinal axis (L) of the device.

[0244] Item 11. The culture device according to Item 10, wherein the third angle is from approximately 1 degree to approximately 45 degrees, preferably from approximately 5 degrees to approximately 30 degrees.

[0245] Clause 12. The device of any one of clauses 9 to 11, wherein the ratio between the surface areas of the first surface, the second surface and the third surface is approximately 1:2:3 or approximately 1:3:6.

[0246] Clause 13. The device according to any preceding clause, wherein the base further comprises at least one port (22).

[0247] Clause 14. The device of any preceding clause, wherein at least one of the surfaces comprises an adherent cell culture substrate or coating material.

[0248] Item 15. A device according to item 14, wherein each surface comprises an adherent cell culture substrate or coating material.

[0249] Item 16. A device according to any one of Item 14 or Item 15, wherein the adherent cell culture substrate or coating material is selected from a group consisting of collagen, fibronectin, vitronectin, laminin, gelatin, poly-lysine, cellulose or a combination thereof.

[0250] Clause 17. The device of any preceding clause, wherein at least one of the base and / or the surface comprises polystyrene, polycarbonate, low density polyethylene, high density polyethylene, silicone, or a thermoplastic elastomer.

[0251] Clause 18. A device according to any preceding clause, wherein the device is a culture device, preferably a cell culture device.

[0252] Clause 19. A system comprising:

[0253] An apparatus as described in any of clauses 1 to 18; and

[0254] An actuator is configured to engage at least a portion of the device and move the device between a first configuration in which the first surface is horizontal and a second configuration in which the second surface is horizontal.

[0255] Clause 20. A method of culturing a biological material, the method comprising the steps of:

[0256] Providing a first surface (28, 128, 228, 328, 428, 528, 638, 728, 828, 928) and a second surface (30, 130, 230, 330, 430, 530, 630, 730, 830, 930) fixedly disposed relative to the first surface, wherein the first surface and the second surface are not coplanar;

[0257] orienting the first surface into a substantially horizontal plane;

[0258] culturing biological material on the first surface;

[0259] orienting the second surface into a substantially horizontal plane to transfer biological material from the first surface to the second surface; and

[0260] A biological material is cultured on the second surface.

[0261] Clause 21. The method of clause 20, wherein the biological material comprises tissue or cells.

[0262] Item 22. A method according to Item 21, wherein the cells are adherent cells and each of the first surface and the second surface comprises an adherent cell culture substrate or coating material.

[0263] Clause 23. Use of the device according to any one of clauses 1 to 18 or the system according to clause 19 for the cultivation of adherent cells.

[0264] Clause 24. Use of the device according to any one of clauses 1 to 18 or the system according to clause 19 for the cultivation of suspended cells.

[0265] Item 25. A set of parts, comprising:

[0266] An apparatus as described in any of clauses 1 to 18; and

[0267] An interface member is configured to be coupled to the top of the device.

Claims

1. A culture device (10, 100, 200, 300), comprising: A base (12, 112, 212, 312, 412, 512, 612, 712, 812, 912) comprising a first surface (28, 128, 228, 328, 428, 528, 638, 728, 828, 928) and a second surface (30, 130, 230, 330, 430, 530, 630, 730, 830, 930) fixedly arranged relative to the first surface; as well as a flexible side wall (14, 114, 214, 314) extending from the base and defining an interior volume of the device, Wherein, the first surface and the second surface are not coplanar.

2. The culture device according to claim 1, wherein: The sidewall is a compressible sidewall.

3. The culture device according to claim 1 or claim 2, wherein: The first surface forms a first angle (α) relative to a transverse plane (T), the transverse plane extending substantially perpendicularly to a central longitudinal axis (L) of the device.

4. The culture device according to claim 3, wherein: The first angle is from approximately 1 degree to approximately 45 degrees, preferably from approximately 5 degrees to approximately 30 degrees.

5. The culture device according to the preceding claim, wherein: The second surface forms a second angle (β) relative to a transverse plane (T), the transverse plane extending substantially perpendicularly to a central longitudinal axis (L) of the device.

6. The culture device according to claim 5, wherein: The second angle is from approximately 1 degree to approximately 45 degrees, preferably from approximately 5 degrees to approximately 30 degrees.

7. A culture device according to any preceding claim, wherein: A ratio between a surface area of ​​the first surface and a surface area of ​​the second surface is approximately 1:9 to approximately 1:

1.

8. The culture device according to claim 7, wherein: The ratio is approximately 1:

3.

9. A culture device according to any preceding claim, wherein: The base further includes a third surface (334, 434, 534) fixedly disposed relative to each of the first surface and the second surface, wherein the first surface, the second surface, and the third surface are not coplanar.

10. The culture device according to claim 9, wherein: The third surface forms a third angle (γ) relative to a transverse plane (T), the transverse plane extending substantially perpendicularly to a central longitudinal axis (L) of the device.

11. The culture device according to claim 10, wherein: The third angle is from approximately 1 degree to approximately 45 degrees, preferably from approximately 5 degrees to approximately 30 degrees.

12. The culture device according to any one of claims 9 to 11, wherein: A ratio among a surface area of ​​the first surface, a surface area of ​​the second surface, and a surface area of ​​the third surface is approximately 1:2:3 or approximately 1:3:

6.

13. The culture device according to any preceding claim, the base further comprising at least one port (22).

14. A culture device according to any preceding claim, wherein: At least one of the surfaces comprises an adherent cell culture substrate or a coating material.

15. The culture device according to claim 14, wherein: Each surface includes an adherent cell culture substrate or coating material.

16. The culture device according to any one of claims 14 or 15, wherein: The adherent cell culture substrate or coating material is selected from the group consisting of collagen, fibronectin, vitronectin, laminin, gelatin, poly-lysine, cellulose or a combination thereof.

17. A culture device according to any preceding claim, wherein: At least one of the base and / or the surface comprises polystyrene, polycarbonate, low density polyethylene, high density polyethylene, silicone, or a thermoplastic elastomer.

18. A culture device according to any preceding claim, wherein: The culture device is a cell culture device.

19. A system comprising: The culture device according to any one of claims 1 to 18; as well as An actuator is configured to engage at least a portion of the device and move the device between a first configuration in which the first surface is horizontal and a second configuration in which the second surface is horizontal.

20. A method for culturing a biological material, the method comprising the following steps: Providing a first surface (28, 128, 228, 328, 428, 528, 638, 728, 828, 928) and a second surface (30, 130, 230, 330, 430, 530, 630, 730, 830, 930) fixedly disposed relative to the first surface, wherein the first surface and the second surface are not coplanar; orienting the first surface into a substantially horizontal plane; culturing biological material on the first surface; orienting the second surface into a substantially horizontal plane to transfer biological material from the first surface to the second surface; and A biological material is cultured on the second surface.

21. The method according to claim 20, wherein: The biological material includes tissues or cells.

22. The method according to claim 21, wherein: The cells are adherent cells, and each of the first surface and the second surface comprises an adherent cell culture substrate or coating material.

23. Use of the culture device according to any one of claims 1 to 18 or the system according to claim 19 in the culture of adherent cells.

24. Use of the culture device according to any one of claims 1 to 18 or the system according to claim 19 in the culture of suspension cells.

25. A set of parts, including: The culture device according to any one of claims 1 to 18; as well as An interface member is configured to be coupled to the top of the device.

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