Multi-modal controllable fluctuation reactor system for cell culture and applications thereof

Abstract

The application belongs to the technical field of cell culture system, and discloses a multi-mode controllable fluctuation reactor system for cell culture and application thereof. The multi-mode controllable fluctuation reactor system for cell culture comprises a multi-mode controllable fluctuation reactor motion unit and a cell culture unit connected to a control unit respectively, the control unit can control multi-mode controllable fluctuation motion of the multi-mode controllable fluctuation reactor motion unit and monitor cell culture parameters, the control unit can control any one of Z-axis electric lifting platform, X-axis electric stage, Y-axis electric stage and R-axis electric swing table to move individually or in combination, and form multi-mode controllable fluctuation motion according to interpolation algorithm. The application can realize the culture requirement of different cells in different culture stages through different modes of controllable fluctuation motion, make the cell culture process more suitable for the implementation of mixing and separation processes, be suitable for in vitro cell culture, and improve cell culture efficiency and quality.

Description

Technical Field

[0001] This invention relates to the field of cell culture system technology, and in particular to a multi-mode controllable fluctuation reactor system for cell culture and its application. Background Technology

[0002] In the biopharmaceutical industry, large-scale in vitro cell culture is used for the mass production of biological products such as enzymes, hormones, antibodies, proteins, viruses, vaccines, and cells. An increasing number of bioreactor systems are being used for the synthesis and production of these biological products. The production scale of influenza vaccines and rabies vaccines using bioreactors has reached 25,000L, with stirred reactors and airlift reactors being commonly used types. In recent years, with the development of cell therapy and regenerative medicine, stem cells have shown promising applications in the treatment of critical and refractory major diseases. However, due to the limited availability of stem cells and the high demand, large-scale expansion to obtain sufficient quantities of high-quality stem cells is necessary to meet the ever-growing clinical needs.

[0003] Stem cell culture is divided into suspension culture and adherent culture. Currently, commonly used culture reactors are container-type, such as T-flasks or culture dishes. This relies on increasing the number of containers to increase yield, which is difficult to meet the needs of large-scale production. It results in significant waste of human and material resources, difficulty in standardizing operations, susceptibility to bacterial contamination, inability to monitor product performance in real time, and large batch-to-batch quality variations. Therefore, using bioreactors for large-scale expansion of stem cells is an inevitable trend.

[0004] However, stem cells pose new challenges to bioreactors:

[0005] First, stem cells are more sensitive to the culture environment. Unsuitable fluid environments and shear forces can cause stem cells to lose their stemness and activity, or lead to differentiation.

[0006] II. In vitro expansion of stem cells involves stages such as cell adhesion, spreading, proliferation, and detachment. Different stages have different requirements for the culture environment, especially the fluid environment. Existing bioreactors can only provide a single movement mode, which is difficult to meet the mixing requirements of different cells at different culture stages, easily leading to uneven cell mixing and heterogeneous cell growth.

[0007] Third, for adherent cells / stem cells, the addition of microcarriers introduces new factors to consider in the in vitro culture process, including microcarrier particle size, density, and sedimentation rate. Microcarrier culture is complex and places high demands on cell bioreactors; therefore, developing bioreactors suitable for in vitro cell culture, especially stem cell culture, is of great significance. Summary of the Invention

[0008] The purpose of this invention is to provide a multi-mode controllable fluctuation reactor system for cell culture and its application, in order to solve the problem that the bioreactor for in vitro cell culture has a single movement mode, which makes it difficult to meet the culture and mixing needs of different cells at different culture stages.

[0009] To achieve this objective, the present invention adopts the following technical solution:

[0010] A multimodal controlled fluctuation reactor system for cell culture includes:

[0011] The multi-mode controllable wave reactor motion unit is equipped with a Z-axis electric lifting platform, an X-axis electric carrying platform, a Y-axis electric carrying platform, and an R-axis electric swing platform, arranged from bottom to top.

[0012] A cell culture unit includes a tray and a cell culture bag. The tray is mounted on the R-axis motorized swing stage and is detachably connected to the R-axis motorized swing stage. The cell culture bag is placed on the tray.

[0013] The control unit is connected to the multi-mode controllable undulating reactor motion unit to control the multi-mode controllable undulating motion of the multi-mode controllable undulating reactor motion unit. The control unit is also connected to the cell culture unit to monitor cell culture parameters. The control unit can control the individual or combined motion of any one of the Z-axis electric lifting platform, the X-axis electric stage, the Y-axis electric stage, and the R-axis electric swing platform, and form multi-mode controllable undulating motion according to the interpolation algorithm.

[0014] Optionally, the multi-mode controllable wave motion includes any one or more combinations of one-dimensional linear motion, two-dimensional linear motion, two-dimensional curvilinear motion, three-dimensional linear motion, three-dimensional spiral motion, and oscillation.

[0015] Optionally, the interpolation algorithm includes the following motion equations:

[0016] One-dimensional linear motion equations:

[0017] X = V x *t or Y = V y *t or Z = V z *t;

[0018] Two-dimensional linear motion equations:

[0019] X = V x *t

[0020] Y = V y *t

[0021] Two-dimensional curvilinear motion equations:

[0022] X = Vx *t

[0023] Y = Asin(t)

[0024] or,

[0025] X = Asin(t)

[0026] Y = Acos(t)

[0027] Three-dimensional linear motion equations:

[0028] X = V x *t

[0029] Y = V y *t

[0030] Z = V z *t

[0031] Three-dimensional curvilinear motion equations:

[0032] X = Asin(t)

[0033] Y = Acos(t)

[0034] Z = V z *t

[0035] In the formula, X, Y, and Z represent the movement distances of the X-axis electric platform, the Y-axis electric platform, and the Z-axis electric lifting platform along the X, Y, and Z axes, respectively; t represents time; and V represents the distance traveled. x V y and V z These are the component velocities of the X-axis electric platform, the Y-axis electric platform, and the Z-axis electric lifting platform on the X, Y, and Z axes, respectively. A is the maximum movement distance of the X-axis electric platform, the Y-axis electric platform, or the Z-axis electric lifting platform on the X, Y, or Z axes, respectively. They start simultaneously and reach the endpoint simultaneously during movement.

[0036] Optionally, the travel range of the Z-axis electric lifting platform, the X-axis electric carrying platform, and the Y-axis electric carrying platform is (-1000mm) to (+1000mm), and the movement speed does not exceed 1000mm / s. The swing angle of the R-axis electric swing platform is between (-180°) and (+180°), and the swing speed does not exceed 180° / s.

[0037] Optionally, the travel range of the Z-axis electric lifting platform, the X-axis electric carrying platform, and the Y-axis electric carrying platform is (-100mm) to (+100mm), and the movement speed does not exceed 100mm / s. The swing angle of the R-axis electric swing platform is between (-15°) and (+15°), and the swing speed does not exceed 7.5° / s.

[0038] Optionally, the tray is provided with one or more culture positions, which are configured one-to-one with the cell culture bags to meet the needs of laboratory, pilot-scale, intermediate-scale, or large-scale culture.

[0039] Optionally, an electric heating pad is provided on the tray, the cell culture bag is located on the electric heating pad, and the electric heating pad is connected to the control unit.

[0040] Optionally, the cell culture unit further includes a monitoring component and a circulation pump component. The control unit is connected to the monitoring component to receive and display the monitoring parameters of the monitoring component in real time. The control unit is connected to the circulation pump component to provide liquid circulation or gas circulation control for the cell culture bag.

[0041] The present invention also provides an application of a multi-mode controllable undulation reactor system for cell culture, wherein the multi-mode controllable undulation reactor system for cell culture is used to provide multi-mode controllable undulation motion during the adhesion and / or amplification and / or collection phases of cell culture.

[0042] Optionally, during the adhesion and / or amplification and / or collection phases of cell culture, the control unit controls the multi-mode controllable wave reactor motion unit to form multi-mode controllable wave motion. The controllable wave motion during the adhesion phase is: a combination of dynamic and static culture, with one or more controllable wave motion modes set for five minutes every 1-2 hours. The controllable wave motion mode during the amplification phase is: XY plane circular motion combined with R-axis oscillation motion, with the X-axis motion equation X = Asin(t) and the Y-axis motion equation Y = Asin(t), 150 ≤ A ≤ 200, a motion speed of 5-100 mm / s, and an R-axis oscillation range of (-15°)-(+15°), with oscillation speeds of 2.5° / s-7.5° / s. The controllable wave motion mode during the collection phase is: an R-axis oscillation range of (-180°)-(+180°).

[0043] The beneficial effects of this invention are:

[0044] The multi-mode controllable undulation reactor system for cell culture of the present invention, by setting a control unit to control the multi-mode controllable undulation reactor motion unit, facilitates the multi-mode controllable undulation motion of the multi-mode controllable undulation reactor motion unit to provide multi-mode controllable undulation motion. Thus, through controllable undulation motion in different modes, the culture requirements of different cells at different culture stages can be met, providing a gentle and uniform fluid environment, making the cell culture process easier to implement processes such as mixing and separation, suitable for in vitro cell culture, and improving cell culture efficiency and quality.

[0045] The application of the multi-mode controllable wave reactor system for cell culture of the present invention, using the aforementioned multi-mode controllable wave reactor system for cell culture in the adhesion stage and / or amplification stage and / or collection stage of cell culture, can provide corresponding modes of controllable wave motion for different stages of cell culture, making the cell culture process easier to implement processes such as mixing and separation, and improving cell culture efficiency and quality. Attached Figure Description

[0046] Figure 1 This is a schematic diagram of the structure of the multi-mode controllable fluctuation reactor system for cell culture of the present invention;

[0047] Figure 2 This is a schematic diagram of the two-dimensional sine and cosine motion trajectory of the motion unit of the multi-mode controllable wave reactor for cell culture of the present invention.

[0048] Figure 3 This is a schematic diagram of the two-dimensional circular motion trajectory of the motion unit of the multi-mode controllable wave reactor for cell culture of the present invention;

[0049] Figure 4 This is a schematic diagram of the three-dimensional linear motion trajectory of the motion unit of the multi-mode controllable wave reactor for cell culture of the present invention.

[0050] Figure 5 This is a schematic diagram of the three-dimensional spiral curve motion trajectory of the motion unit of the multi-mode controllable wave reactor for cell culture of the present invention.

[0051] In the picture:

[0052] 1. Multi-mode controllable wave reactor motion unit; 11. Z-axis electric lifting platform; 12. X-axis electric stage; 13. Y-axis electric stage; 14. R-axis electric swing stage;

[0053] 2. Cell culture unit; 21. Tray; 211. Heating element; 22. Cell culture bag; 23. Monitoring assembly; 24. Circulation pump assembly;

[0054] 3. Control unit. Detailed Implementation

[0055] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and not intended to limit it. Furthermore, it should be noted that, for ease of description, the accompanying drawings show only the parts relevant to the present invention, and not all of the structures.

[0056] In the description of this invention, unless otherwise explicitly specified and limited, the terms "connected," "linked," and "fixed" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0057] In this invention, unless otherwise explicitly specified and limited, "above" or "below" the second feature can include direct contact between the first and second features, or contact between the first and second features through another feature between them. Furthermore, "above," "over," and "on top" of the second feature includes the first feature directly above or diagonally above the second feature, or simply indicates that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature includes the first feature directly below or diagonally below the second feature, or simply indicates that the first feature is at a lower horizontal level than the second feature.

[0058] In the description of this embodiment, the terms "upper," "lower," "left," and "right," etc., refer to the orientation or positional relationship shown in the accompanying drawings. They are used only for ease of description and simplification of operation, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the present invention. In addition, the terms "first" and "second" are used only for distinction in description and have no special meaning.

[0059] This invention provides a multi-mode controllable fluctuation reactor system for cell culture, such as... Figure 1 As shown, the device includes a multi-mode controllable wave reactor motion unit 1, a cell culture unit 2, and a control unit 3. The multi-mode controllable wave reactor motion unit 1 includes a Z-axis electric lifting platform 11, an X-axis electric stage 12, a Y-axis electric stage 13, and an R-axis electric swing platform 14 arranged sequentially from bottom to top. The cell culture unit 2 includes a tray 21 and cell culture bags 22. The tray 21 is placed on the R-axis electric swing platform 14 and is detachably connected to the R-axis electric swing platform 14. The cell culture bags 22 are placed on the tray 21. The control unit 3 is connected to the multi-mode controllable wave reactor motion unit 1 to control the multi-mode controllable wave motion of the multi-mode controllable wave reactor motion unit 1. The control unit 3 is also connected to the cell culture unit 2 to monitor cell culture parameters. The control unit 3 can control the individual or combined motion of any one of the Z-axis electric lifting platform 11, the X-axis electric stage 12, the Y-axis electric stage 13, and the R-axis electric swing platform 14, and form multi-mode controllable wave motion according to the interpolation algorithm.

[0060] As Figure 1 shown, the motion unit 1 of the multi-mode controllable wave reactor is an XYZR four-axis motion platform. The Z-axis electric lifting platform 11 is fixed on the base, and a Z-axis drive motor is provided to control the lifting motion. The control unit 3 is connected to the Z-axis drive motor to realize the Z-direction height adjustment and motion mode control of the tray 21 or the cell culture bag 22; the X-axis electric stage 12 and the Y-axis electric stage 13 are arranged on the Z-axis electric lifting platform 11 to form an XY plane stage. The displacement of the X-axis electric stage 12 and the Y-axis electric stage 13 is controlled by the X-axis drive motor and the Y-axis drive motor respectively, and both the X-axis drive motor and the Y-axis drive motor are electrically connected to the control unit 3. Both the X-axis drive motor and the Y-axis drive motor are linear motors, and the Z-axis drive motor is a linear motor or a rotary motor plus a worm gear drive unit. The R-axis electric swing table 14 is detachably mounted on the XY plane stage, which can provide R-axis swing motion for cell culture. The R-axis is parallel to the X-axis or the Y-axis.

[0061] In the multi-mode controllable wave reactor system for cell culture of the present invention, the control unit 3 is electrically connected to and controls the motion unit 1 of the multi-mode controllable wave reactor, so that the motion unit 1 of the multi-mode controllable wave reactor can conveniently provide multi-mode controllable wave motion. Thus, through the controllable wave motion, the culture requirements of different cells at different culture stages can be met, making processes such as mixing and separation easier to implement in the cell culture process, which is suitable for in vitro cell culture and improves the efficiency and quality of cell culture.

[0062] Optionally, the multi-mode controllable wave motion includes any one or a combination of more of one-dimensional linear motion, two-dimensional linear motion, two-dimensional curvilinear motion, three-dimensional linear motion, three-dimensional spiral motion and swing motion.

[0063] Among them, for the one-dimensional linear motion, the one-dimensional controllable wave motion can be realized by controlling the motion speed, distance and reciprocation times in the X-axis, Y-axis or Z-axis direction. The combination of the one-dimensional linear motion and the R-axis swing motion can form a linkage, so as to meet the operation requirements of processes such as stirring, mixing and separation in the cell culture process. Similarly, other modes of controllable wave motion can also perform single-mode or multi-mode controllable wave motion by controlling parameters such as speed, distance and reciprocation form.

[0064] Optionally, the motion equations included in the interpolation algorithm are as follows:

[0065] One-dimensional linear motion equation:

[0066] X=V x *t or Y=V y *t or Z=V z *t;

[0067] Two-dimensional linear motion equation:

[0068] X = V x *t

[0069] Y = V y *t

[0070] Two-dimensional curvilinear motion equations:

[0071] X = V x *t

[0072] Y = Asin(t)

[0073] or,

[0074] X = Asin(t)

[0075] Y = Acos(t)

[0076] Three-dimensional linear motion equations:

[0077] X = V x *t

[0078] Y = V y *t

[0079] Z = V z *t

[0080] Three-dimensional curvilinear motion equations:

[0081] X = Asin(t)

[0082] Y = Acos(t)

[0083] Z = V z *t

[0084] In the formula, X, Y, and Z represent the movement distances of the X-axis electric platform 12, Y-axis electric platform 13, and Z-axis electric lifting platform 11 along the X, Y, and Z axes, respectively; t represents time; and V represents the distance traveled. x V y and V z These are the component velocities of the X-axis electric platform 12, Y-axis electric platform 13, and Z-axis electric lifting platform 11 on the X, Y, and Z axes, respectively. A is the maximum movement distance of the X-axis electric platform 12, Y-axis electric platform 13, or Z-axis electric lifting platform 11 on the X, Y, or Z axes, respectively. X, Y, and Z can be interchanged, and they start and reach the endpoint simultaneously during movement.

[0085] The motion equations for the above interpolation algorithms are explained in supplementary terms as follows:

[0086] The XYZR multi-axis platform of the present invention can move independently in the X-axis, Y-axis, Z-axis and R-axis directions, or it can combine movements in the X, Y, Z and R axes to form multi-mode controllable wave motion and realize trajectory motion operation.

[0087] For one-dimensional linear motion, the equation of motion is X = V. x *t or Y = V y *t or Z = V z *t; One-dimensional linear motion refers to the ability of the tray 21 with cell culture bag 22 to move in the X-axis, Y-axis and Z-axis directions respectively. The motion mode is controlled by speed and distance, and combined with reciprocating cyclic motion, controllable wave motion is achieved.

[0088] For two-dimensional linear motion, taking the linear interpolation motion of a mass (in this embodiment, a tray 21 with a cell culture bag 22) in the XY plane in two dimensions as an example, the X-axis and Y-axis start simultaneously and reach their endpoints simultaneously. Let the motion distance in the X-axis direction be X, and the motion distance in the Y-axis direction be Y. If the motion speed of the interpolation main axis is V (the actual speed of each axis is the component speed of the main axis, and the motion component speed of the X-axis is V), then... x The velocity component along the Y-axis is V. y At this point, the trajectory is X = V. x *t and Y = V y *t. It should be noted that the equations for other two-dimensional linear interpolation motions, such as those in the XZ and YZ planes, are the same as those for the XY plane motion.

[0089] Two-dimensional curvilinear motion includes two-dimensional sine and cosine motion and two-dimensional circular motion. Taking the realization of sine and cosine motion in the two-dimensional direction of the XY plane as an example, linear motion is used in the X-axis direction and sine motion is used in the Y-axis direction. The interpolation motion equation is as follows:

[0090] Actual distance traveled on axis X: X = V x *t

[0091] Actual distance traveled along axis Y: Y = Asin(t)

[0092] like Figure 2 As shown, the linear interpolation motion in the XY plane moves from point 0 (X0, Y0) to point 1 (X1, Y1) on the plane. The X-axis and Y-axis start simultaneously and reach their endpoints simultaneously. Let the motion distance in the X-axis direction be X, and the motion distance in the Y-axis direction be Y. If the motion speed of the interpolation master axis is V (the actual speed of each axis is the component speed of the master axis, and the motion speed component speed of the X-axis is V0), then... x The velocity component along the Y-axis is V. y At this time, the trajectory of motion is as follows: Figure 2This yields sinusoidal motion. It's important to note that the equations for other two-dimensional curve interpolation motions, such as those in the XZ and YZ planes, are the same as those for the XY plane motion.

[0093] The circular motion in the two-dimensional XY plane, with sinusoidal motion in the X-axis direction and cosine motion in the Y-axis direction, is interpolated by the following equations:

[0094] Actual trajectory of axis X: X = Asin(t)

[0095] Actual trajectory of motion along axis Y: Y = Acos(t)

[0096] The XY-plane circular interpolation motion moves from point 0 (X0, Y0) to point 1 (X1, Y1) on the plane. The X and Y axes start simultaneously and reach their endpoints simultaneously. Let the distance of motion along the X-axis be X, and the distance along the Y-axis be Y. If the speed of the interpolation master axis is V (the actual speed of each axis is the component of the master axis's speed, and the component of the speed of axis X is V0), then... x The velocity component of the motion along the Y axis is V. y At this time, the trajectory of motion is as follows: Figure 3 As shown, circular motion is formed. It should be noted that other two-dimensional circular curve interpolation motions, such as those in the XZ and YZ planes, have the same equations as those for the XY plane motion. Simply substitute X, Y, and Z to obtain the corresponding motion equations.

[0097] When spatial linear motion is achieved in the XYZ three-dimensional directions, the X-axis, Y-axis, and Z-axis all participate in the interpolation motion. The interpolation motion equations are as follows:

[0098] Actual trajectory of axis X: X = V x *t

[0099] Actual trajectory of axis Y: Y = V y *t

[0100] Actual trajectory of axis Z: Z = V z *t

[0101] The three-axis linear interpolation motion moves from point 0 (X0, Y0, Z0) to point 1 (X1, Y1, Z1) on the plane. The X, Y, and Z axes start simultaneously and reach their endpoints at the same time. Let the motion distance along the X-axis be X, the motion distance along the Y-axis be Y, and the motion distance along the Z-axis be Z. Let the motion speed of the interpolation master axis be V (the actual speed of each axis is the component of the master axis's speed, and the motion component of the X-axis is V). x The velocity component along the Y-axis is V. y The velocity component along the Z-axis is Vz), and the trajectory at this time is as follows: Figure 4 As shown, it forms a linear motion trajectory in three-dimensional space.

[0102] When spatial helical curve motion can be achieved in the XYZ three-dimensional direction, the three axes X, Y and Z participate in the interpolation motion, and the interpolation motion equation is as follows:

[0103] Actual trajectory of axis X: X = Asin(t)

[0104] Actual trajectory of motion along axis Y: Y = Acos(t)

[0105] Actual trajectory of axis Z: Z = V z *t

[0106] The three-axis linear interpolation motion moves from point 0 (X0, Y0, Z0) to point 1 (X1, Y1, Z1) on the plane. The X, Y, and Z axes start simultaneously and reach their endpoints at the same time. Let X be the distance of motion in the X direction, Y be the distance of motion in the Y direction, and Z be the distance of motion in the Z direction. Let the speed of the interpolation master axis be V (the actual speed of each axis is the component of the master axis's speed, and the component of the speed of axis X is V). x The velocity component of the motion along the Y axis is V. y The velocity component of the motion along axis Z is V z At this time, the trajectory of motion is as follows: Figure 5 As shown, this is a three-dimensional spiral curve trajectory.

[0107] It should be noted that, for example, circular motion can be achieved in two-dimensional directions such as the XZ plane and YZ plane, while linear motion can be achieved along the corresponding Y-axis or X-axis. According to the interpolation motion equation, spiral curve motion in different directions can be achieved.

[0108] Any one or more of the above-mentioned curvilinear motions can be combined with the swing motion of the R-axis electric stage 14 to form a richer range of motion modes, meeting the operational needs of different cells and different culture conditions.

[0109] Optionally, the travel range of the Z-axis electric lifting platform 11, the X-axis electric carrying platform 12, and the Y-axis electric carrying platform 13 is (-1000mm) to (+1000mm), and the movement speed does not exceed 1000mm / s. The swing angle of the R-axis electric swing platform 14 is between (-180°) and (+180°), and the swing speed does not exceed 180° / s.

[0110] More preferably, the preferred travel range of the Z-axis electric lifting platform 11, the X-axis electric carrying platform 12, and the Y-axis electric carrying platform 13 is (-100mm) to (+100mm), and the movement speed does not exceed 100mm / s. The swing angle of the R-axis electric swing platform 14 is between (-15°) and (+15°), and the swing speed does not exceed 7.5° / s.

[0111] In this embodiment, the vertical line where the origin of the Z-axis, X-axis, Y-axis and R-axis are located is taken as the center line. When the center of the tray 21 is located on the center line, it is defined as the initial state. The XYZR multi-axis platform can realize continuous motion at a set speed. The control unit 3 outputs control commands to realize multi-mode controllable undulating motion control.

[0112] Optionally, the tray 21 is provided with one or more culture positions, which are set one-to-one with the cell culture bag 22 to meet the needs of laboratory, pilot, intermediate or large-scale culture.

[0113] Typically, cell culture unit 2 uses disposable cell culture bags 22 of various sizes, which can be quickly replaced without washing to increase the frequency of experiments. The cell culture bags 22 can be selected arbitrarily within the size range of 0.05L-5000L according to different culture needs, with a preferred size range of 0.05-500L, and more preferably 0.05-50L. It should be noted that when the cell culture bag 22 is larger, the weight increases. The load-bearing capacity of the tray 21 can be adjusted, its size and model can be adjusted, or a counterweight system can be added to maintain a stable and continuous movement. The tray 21 can bear a maximum load of 10,000 kg, with a preferred load-bearing range of less than 500 kg, more preferably 0-200 kg, more preferably 0-100 kg, and even more preferably 0-50 kg, meeting the culture needs of laboratory, small-scale, pilot-scale, and large-scale production respectively within the selectable range. By setting the power of the Z-axis, X-axis, and Y-axis drive motors, the dimensions and weighing of each axis can be adjusted according to load-bearing requirements, range of motion, and speed. This allows for the addition of a counterweight system and upgrading of the weighing device for large-scale culture, maintaining a stable and continuous motion pattern. The tray 21 is used in conjunction with the cell culture bag 22. The tray 21 is detachably connected to the R-axis motorized stage 14, facilitating tray 21 replacement. The tray 21 can have one or more culture positions; for example, one tray 21 can accommodate 2-100 culture positions.

[0114] Optionally, an electric heating element 211 is provided on the tray 21, and the cell culture bag 22 is located on the electric heating element 211. The electric heating element 211 is connected to the control unit 3.

[0115] like Figure 1 As shown, the electric heating element 211 is an electric heating element or heating wire, which is set at the bottom of the tray 21 and is powered on by the control unit 3 to provide a suitable temperature for the cell culture process.

[0116] Optionally, the cell culture unit 2 also includes a monitoring component 23 and a circulation pump component 24. The control unit 3 is connected to the monitoring component 23 to receive and display the monitoring parameters of the monitoring component 23 in real time. The control unit 3 is connected to the circulation pump component 24 to provide liquid circulation or gas circulation control for the cell culture bag 22.

[0117] In some embodiments, the cell culture process requires parameter testing such as temperature, dissolved oxygen, pH, and carbon dioxide content. Therefore, the monitoring component 23 includes a temperature sensor, a pH meter, a dissolved oxygen detector, and a carbon dioxide concentration detector. The control unit 3 collects the monitoring parameters and, based on these parameters, can control the circulation pump component 24 to start or stop, thereby controlling the cell culture process. The circulation pump component 24 may include a stepper motor, a transmission unit, and a circulation pump, used to circulate and replace the culture medium and gas within the cell culture bag 22. One or more circulation pump components 24 are provided and configured in conjunction with the cell culture bag 22.

[0118] The present invention also provides an application of a multi-mode controllable fluctuation reactor system for cell culture, wherein the multi-mode controllable fluctuation reactor system for cell culture provided by the present invention is used in the adhesion stage and / or amplification stage and / or collection stage of cell culture.

[0119] The multi-mode controllable undulation reactor system for cell culture provided by this invention can be used for suspension culture and adherent cell culture (such as amniotic epithelial stem cells), and is particularly suitable for stem cell culture. It provides gentle, low-shear stress and adapts to appropriate controllable undulation motion modes according to the requirements of different cell culture stages, supporting good in vitro expansion of stem cells and maintenance of cell viability. The multi-mode controllable undulation bioreactor can meet the requirements of different culture scales, supporting needs at different stages from laboratory, pilot-scale, and pilot-scale production.

[0120] Optionally, during the adhesion and / or amplification and / or collection phases of cell culture, the control unit 3 controls the multi-mode controllable wave reactor motion unit 1 to form controllable wave motion. The controllable wave motion mode during the adhesion phase is: dynamic and static combined culture, with controllable wave motion set for five minutes every 1-2 hours; the controllable wave motion mode during the amplification phase is: XY plane circular motion combined with R-axis oscillation motion, X-axis motion equation X=Asin(t), Y-axis motion equation Y=Asin(t), 150≤A≤200, motion speed of 5-100mm / s, R-axis oscillation range (-15°)-(+15°), oscillation speed of 2.5° / s-7.5° / s; the controllable wave motion mode during the collection phase is: R-axis oscillation range (-180°)-(+180°).

[0121] The application of the multi-mode controllable wave reactor system for cell culture of the present invention provides corresponding controllable wave motion for different stages of cell culture, including the adhesion, amplification, and collection stages. This makes the cell culture process easier to implement processes such as mixing and separation, thereby improving cell culture efficiency and quality.

[0122] Specifically, when adherent stem cells are cultured on a scalable support matrix at a three-dimensional scale, the process includes an adhesion phase, a rapid expansion phase, and a collection phase. During the adhesion phase, a combination of static and dynamic methods is required. Static conditions help maintain cell adhesion, while short-term dynamic stirring helps increase the uniformity of cell-carrier contact. After the cell-carrier mixture has been left to stand for 2 hours, the control unit 3 drives the motion unit 1 of the multi-mode controllable wave reactor system to perform controllable wave motion. The motion mode is XY-plane circular motion, with the X-axis moving at X = 100sin(t) and the Y-axis moving at Y = 100sin(t) for 10 minutes to achieve uniform mixing, followed by continued static culture. This cycle is repeated for 48 hours. In some embodiments, the motion conditions can also be that the R-axis electric swing stage 14 is swayed by 12° or 10° at a speed of 3° / s or 4° / s, swaying for 10 minutes to achieve uniform mixing, followed by continued static culture. This cycle is repeated for 48 hours.

[0123] During the expansion phase, fresh culture medium is replenished to the culture system via the circulation pump assembly 24 of cell culture unit 2. Control unit 3 controls the motion unit 1 of the multi-mode controllable undulating reactor system to perform combined motion: the X-axis follows X = 150sin(t), the Y-axis follows Y = 150sin(t), and the R-axis oscillates by 10° at a speed of 4° / s. This motion mode is maintained for 7 days, providing a gentle undulating pattern to thoroughly mix the microcarrier cells. Simultaneously, during the culture process, culture medium is injected into the cell culture bag 22 via the circulation pump assembly 24 every other day. While providing nutrients to the cells, it also removes cellular metabolic products. In some embodiments, the motion equations of the combined motion can also be: X-axis according to X = 200sin(t), Y-axis according to Y = 200sin(t), R-axis electric pendulum 14 oscillates 15°, and the speed is 4° / s; or: XYZ three-dimensional spiral curve motion combined with R oscillation motion mode, the motion conditions are X-axis according to X = 150sin(t), Y-axis according to Y = 150sin(t), Z-axis according to Z = 5t, R-axis oscillates 10°, and the speed is 3° / s.

[0124] During the collection phase, after the culture is completed, the control unit 3 controls the motion unit 1 of the multi-mode controllable wave reactor system to return to zero, and at the same time adjusts the R-axis electric swing stage 14 to swing 180°, tilting the carrier tray 21 to quickly drain the adherent cell culture medium in one go. After adding an appropriate amount of PBS buffer for washing, a certain amount of digestion solution is added. After standing for a period of time, the digestion solution and cells are discharged through the circulation pump assembly 24, the microcarrier is retained, the cells are centrifuged and recovered, and then resuspended for collection and preservation.

[0125] As can be seen from the above process, the control mode of the multi-mode controllable wave reactor system motion unit 1 by the control unit 3 also includes zero-return control, that is, the multi-mode controllable wave reactor system motion unit 1 can directly return to the zero position from any current position, thus improving motion efficiency.

[0126] The multi-mode controllable fluctuation reactor system for cell culture provided by this invention can provide mild shear stress culture conditions for cell culture, thoroughly mix microcarrier-cell cultures, and provide suitable temperature, pH, dissolved oxygen, etc., enabling adjustable and customized culture condition configurations for large-scale cell expansion. Three-dimensional microcarrier-cell co-culture can reduce cell seeding density to 3-4 times lower than two-dimensional seeding density, significantly increasing cell expansion and further enhancing cell proliferation capacity.

[0127] The multi-mode controllable wave reactor system for cell culture provided by this invention can provide multi-mode controllable wave motion. Combined with the R-axis electric swing stage 14, it is designed with multiple tilt angles for convenient sampling. At the same time, the motion unit 1 of the multi-mode controllable wave reactor is equipped with photoelectric limit, direct drive structure, dustproof, splashproof, frictionless, suitable for long-term operation without wear. The multi-mode controllable wave can combine dynamic and static, providing gentle shear force to meet the culture needs of different cells at different culture stages, and is particularly suitable for stem cell culture.

[0128] This invention utilizes disposable cell culture bags 22, which can be customized in multiple sizes to meet the needs of different scales of culture, supporting the needs of different stages from laboratory, pilot-scale, and intermediate-scale production. The control unit 3 controls the multi-mode controllable wave reactor motion unit 1 and the cell culture unit 2 respectively, meeting the mixing requirements of different cell needs. The cell culture system allows for rapid replacement of disposable cell culture bags 22, increasing the frequency of experiments. Simultaneous operation of multiple cell culture bags 22 facilitates monitoring and simulation of relevant experimental needs. The multi-mode controllable wave bioreactor system for cell culture effectively supports good in vitro cell culture, while also enabling rapid cell separation and expandable support matrix. It is particularly effective in supporting the in vitro expansion of stem cells, resulting in high cell numbers, good activity, and better maintenance of stem cell characteristics.

[0129] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art will be able to make various obvious changes, readjustments, and substitutions without departing from the scope of protection of the present invention. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A multi-mode controllable fluctuation reactor system for cell culture, characterized in that, include: The multi-mode controllable wave reactor motion unit (1) is provided with a Z-axis electric lifting platform (11), an X-axis electric carrying platform (12), a Y-axis electric carrying platform (13) and an R-axis electric swing platform (14) from bottom to top. The cell culture unit (2) includes a tray (21) and a cell culture bag (22). The tray (21) is set on the R-axis electric swing stage (14). The tray (21) is detachably connected to the R-axis electric swing stage (14). The cell culture bag (22) is placed on the tray (21). Control unit (3), the control unit (3) is connected to the multi-mode controllable wave reactor motion unit (1) to control the multi-mode controllable wave motion of the multi-mode controllable wave reactor motion unit (1), the control unit (3) is connected to the cell culture unit (2) to monitor cell culture parameters, the control unit (3) controls the combined motion of the Z-axis electric lifting platform (11), the X-axis electric stage (12), the Y-axis electric stage (13) and the R-axis electric swing platform (14), and forms multi-mode controllable wave motion according to the interpolation algorithm; The multi-mode controllable wave motion includes a combination of one-dimensional linear motion, two-dimensional linear motion, two-dimensional curvilinear motion, three-dimensional linear motion, three-dimensional spiral motion, and oscillating motion.

2. The multi-mode controllable fluctuation reactor system for cell culture according to claim 1, characterized in that, The interpolation algorithm includes the following motion equations: One-dimensional linear motion equations: X=V x *t or Y=V y *t or Z=V z *t; Two-dimensional linear motion equations: X=V x *t Y=V y *t Two-dimensional curvilinear motion equations: X=V x *t Y=Asin(t) or, X=Asin(t) Y=Acos(t) Three-dimensional linear motion equations: X=V x *t Y=V y *t Z=V z *t Three-dimensional curvilinear motion equations: X=Asin(t) Y=Acos(t) Z=V z *t In the formula, X, Y, and Z represent the movement distances of the X-axis electric platform (12), the Y-axis electric platform (13), and the Z-axis electric lifting platform (11) along the X, Y, and Z axes, respectively; t represents time; and V represents the movement distance of the X-axis electric platform (12), the Y-axis electric platform (13), and the Z-axis electric lifting platform (11). x V y and V z The values ​​are the component velocities of the X-axis electric platform (12), the Y-axis electric platform (13), and the Z-axis electric lifting platform (11) on the X-axis, Y-axis, and Z-axis, respectively. A is the maximum movement distance of the X-axis electric platform (12), the Y-axis electric platform (13), or the Z-axis electric lifting platform (11) on the X-axis, Y-axis, or Z-axis, respectively. The movement of each axis starts simultaneously and reaches the endpoint simultaneously.

3. The multi-mode controllable fluctuation reactor system for cell culture according to claim 1, characterized in that, The travel range of the Z-axis electric lifting platform (11), the X-axis electric carrying platform (12), and the Y-axis electric carrying platform (13) is (-1000mm) to (+1000mm), and the movement speed does not exceed 1000mm / s. The swing angle of the R-axis electric swing platform (14) is between (-180°) and (+180°), and the swing speed does not exceed 180° / s.

4. The multi-mode controllable fluctuation reactor system for cell culture according to claim 3, characterized in that, The travel range of the Z-axis electric lifting platform (11), the X-axis electric platform (12), and the Y-axis electric platform (13) is (-100mm) to (+100mm), and the movement speed does not exceed 100mm / s. The swing angle of the R-axis electric swing platform (14) is between (-15°) and (+15°), and the swing speed does not exceed 7.5° / s.

5. The multi-mode controllable fluctuation reactor system for cell culture according to claim 1, characterized in that, The tray (21) is provided with one or more culture positions, and the culture positions are set one-to-one with the cell culture bags (22).

6. The multi-mode controllable fluctuation reactor system for cell culture according to claim 5, characterized in that, An electric heating element (211) is provided on the tray (21), the cell culture bag (22) is located on the electric heating element (211), and the electric heating element (211) is connected to the control unit (3).

7. The multi-mode controllable fluctuation reactor system for cell culture according to claim 1, characterized in that, The cell culture unit (2) further includes a monitoring component (23) and a circulation pump component (24). The control unit (3) is connected to the monitoring component (23) to receive the monitoring parameters of the monitoring component (23) and display them in real time. The control unit (3) is connected to the circulation pump component (24) to provide liquid circulation or gas circulation control for the cell culture bag (22).

8. Application of a multi-mode controllable fluctuation reactor system for cell culture, characterized in that, The multimodal controlled undulation reactor system for cell culture according to any one of claims 1-7 is used to provide multimodal controlled undulation motion during the adhesion and / or amplification phases of cell culture.

9. The application of the multi-mode controllable fluctuation reactor system for cell culture according to claim 8, characterized in that, During the adhesion and / or amplification phases of cell culture, the control unit (3) controls the multi-mode controllable wave reactor motion unit (1) to form multi-mode controllable wave motion. The controllable wave motion mode during the adhesion phase is: dynamic and static combined culture, with controllable wave motion set for five minutes every 1-2 hours. The controllable wave motion mode during the amplification phase is: XY plane circular motion combined with R-axis swing motion, X-axis motion equation X=Asin(t), Y-axis motion equation Y=Asin(t), 150≤A≤200, motion speed is 5-100mm / s, R-axis swing range (-15°)-(+15°), swing speed is 2.5° / s-7.5° / s.

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