Biological reaction device and stirring method

Abstract

The invention discloses a biological reaction device and a stirring method, and relates to the technical field of bioreactors, the biological reaction device comprises a container, an execution device, a sensor and a control device, the execution device comprises a base and a plurality of stirring paddles arranged on the base, the base is detachably installed on the container, the stirring paddles can be inserted into the container, and the sensor is arranged on the container. Each stirring paddle comprises a rotating shaft and a plurality of paddles, the paddles are inclined to the horizontal plane, the rotating directions of every two adjacent stirring paddles are opposite, liquid in the containing cavity can flow along the rotating axis of the stirring paddles in the rotating process of the stirring paddles, and the mixing effect of the cells and the culture solution is improved. The sensor is inserted into the channel of the rotating shaft and extends out of the stirring paddle, parameters in a culture solution can be detected in real time, and the control device can receive a detection signal of the sensor and control the rotating speed of the stirring paddle based on the detection signal, so that full contact and reaction of cells and the culture solution are guaranteed, and the biological culture effect is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of bioreactors, and in particular to a bioreactor and a stirring method. Background Art

[0002] A bioreactor is a highly engineered device used to promote the growth, reproduction or metabolism of biologically active substances (such as microorganisms, plant cells, animal cells or enzymes) under controlled conditions to produce specific biological products or carry out biotransformation processes. Current bioreactors are equipped with a stirring device to increase the mixing degree of cells and culture fluids through stirring to achieve the effect of increasing the reaction rate. However, the stirring function is single and the stirring effect is poor. Summary of the invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention provides a bioreactor and a stirring method, which can improve the stirring effect and increase the bioreaction rate.

[0004] A bioreactor according to a first aspect of the present invention comprises: A container having a cavity; The actuator comprises a base and a plurality of stirring paddles arranged on the base, the base is detachably mounted on the container, the stirring paddles can be inserted into the cavity, the rotation directions of two adjacent stirring paddles are opposite, the stirring paddles comprise a rotating shaft and a plurality of paddles arranged along the circumference of the rotating shaft, the paddles are inclined to a horizontal plane, and the rotating shaft has channels running through both ends; A sensor is inserted into the channel and extends out of the stirring paddle, and the sensor is used to detect the solution in the cavity and generate a detection signal; The control device is electrically connected to the execution device and the sensor. The control device is provided with a switch. The control device can receive a detection signal from the sensor and can control the rotation speed of the stirring paddle based on the detection signal.

[0005] According to a biological reaction device of the first aspect of the present invention, there are at least the following beneficial effects: the embodiment is provided with a container, an execution device, a sensor and a control device, the container is provided with a cavity, the execution device includes a base and a plurality of stirring paddles provided on the base, the base is detachably mounted on the container, the stirring paddle can be inserted into the cavity, the stirring paddle includes a rotating shaft and a plurality of paddles arranged along the circumference of the rotating shaft, the paddles are inclined to the horizontal plane, the rotating shaft has a channel running through both ends, during the rotation of the stirring paddle, it helps to make the liquid in the cavity flow along the direction of the rotation axis of the stirring paddle, improve the mixing effect of cells and culture fluid, and help to promote the reaction rate. In addition, the rotation directions of two adjacent stirring paddles are opposite, which helps to improve the stirring efficiency of the liquid in the cavity, improve the mixing efficiency of cells and culture fluid, and thus improve the reaction rate. The sensor is inserted into the channel and extends out of the stirring paddle, which is beneficial to improving the compactness of the space while being able to detect the parameters in the culture solution in real time. The sensor is used to detect the solution in the cavity and generate a detection signal. The control device can receive the detection signal from the sensor, and the control device can control the rotation speed of the stirring paddle based on the detection signal. The control device can automatically adjust the rotation speed of the stirring paddle according to the state parameters of the culture solution in real time to ensure that the cells are fully in contact and react with the culture solution, which is beneficial to improving the biological culture effect.

[0006] According to an embodiment of the first aspect of the present invention, a driving gear is provided on the end of the rotating shaft away from the stirring paddle, and the driving gears on two adjacent rotating shafts are meshed with each other.

[0007] According to an embodiment of the first aspect of the present invention, the execution device is provided with a motor, which is mounted on the base and is connected to any one of the stirring paddles.

[0008] According to an embodiment of the first aspect of the present invention, the driving gear includes a driving wheel and a driven wheel, and the stirring paddle connected to the motor power is provided with a driving wheel, and the driving wheel is meshed with the driven wheel.

[0009] According to an embodiment of the first aspect of the present invention, the stirring paddle includes a first stirring paddle and a second stirring paddle that are adjacent to each other, and the inclination directions of the blades between the first stirring paddle and the second stirring paddle are opposite.

[0010] According to an embodiment of the first aspect of the present invention, the base is provided with a positioning edge, and a limiting groove is provided on the outer edge of the container. When the actuator is installed on the container, the positioning edge is inserted into the limiting groove.

[0011] According to an embodiment of the first aspect of the present invention, a plurality of unit cavities are provided in the chamber, and a stirring paddle is inserted into each unit cavity.

[0012] According to an embodiment of the first aspect of the present invention, the blade includes a connecting end close to the rotating shaft and a free end away from the rotating shaft, the inclination angle between the outer edge of the free end and the axis of the rotating shaft is a first angle, the inclination angle between the edge of the connecting end and the axis of the rotating shaft is a second angle, and the first angle and the second angle are not equal.

[0013] According to an embodiment of the second aspect of the present invention, there is provided a stirring method, which is applied to the above-mentioned bioreactor, and comprises the following steps: Step 1: Place the culture medium and cells in the container, place the actuator on the container, and insert the positioning edge into the limiting groove; Step 2: Start the switch, the motor drives any stirring paddle to rotate, the driving wheel drives several driven wheels to rotate, and the mixed liquid in the chamber is stirred. At this time, the motor reaches the first speed; Step 3: Place the actuator and the container in a carbon dioxide culture chamber for cell culture. During this process, the sensor measures parameter A of the pH value and parameter B of the dissolved oxygen content in the solution, and transmits parameter A and parameter B to the control device.

[0014] A bioreactor according to the second aspect of the present invention has at least the following beneficial effects: In this embodiment, the execution device can be conveniently installed by cooperating with the positioning edge and the limiting groove, and can be quickly adapted to the existing container for cell culture, with good flexibility of use; the speed of the motor can be adjusted in time by real-time detection of parameter indicators in the culture medium through the sensor, and the speed of the stirring paddle can be automatically adjusted according to the state parameters of the culture medium in real time to ensure that the cells are in full contact and reaction with the culture medium, which is beneficial to improving the biological culture effect.

[0015] According to an embodiment of the second aspect of the present invention, in step 3, when A is less than 6.8, the control device changes the speed of the motor to a second speed; when B is less than 10000 ppm, the control device changes the speed of the motor to a second speed, and the second speed is greater than the first speed.

[0016] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 An isometric view of a bioreactor in an embodiment of the first aspect of the present invention; Figure 2 A bottom view of an execution device in an embodiment of the first aspect of the present invention; Figure 3An isometric view of an execution device in an embodiment of the first aspect of the present invention; Figure 4 An axonometric view of a first container in an embodiment of the first aspect of the present invention; Figure 5 An axonometric view of a second container in an embodiment of the first aspect of the present invention; Figure 6 A first cross-sectional view of an execution device in an embodiment of the first aspect of the present invention; Figure 7 for Figure 6 A magnified view of center A; Figure 8 A second cross-sectional view of the execution device in the embodiment of the first aspect of the present invention; Fig. 9 for Figure 8 Magnified view of B.

[0018] Reference numerals: Actuator 100; base 101; upper cover 102; motor 103; mounting slot 104; driving wheel 105; driven wheel 106; switch 107; positioning edge 108; limiting slot 109; First stirring paddle 110; paddle 111; rotating shaft 112; channel 113; first axis 114; connecting end 115; free end 116; first angle 117; second angle 118; second axis 119; The second stirring paddle 120 ; the containing cavity 121 ; the unit cavity 122 ; the sensor 123 ; and the container 124 . DETAILED DESCRIPTION

[0019] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0020] In the description of the present invention, it should be understood that descriptions involving orientations, such as up, down, front, back, left, right, etc., and orientations or positional relationships indicated are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0021] In the description of the present invention, "several" means one or more, "more" means more than two, "greater than", "less than", "exceed" etc. are understood as not including the number itself, and "above", "below", "within" etc. are understood as including the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing the technical features, and cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0022] In the description of the present invention, unless otherwise clearly defined, terms such as setting, installing, connecting, etc. should be understood in a broad sense, and technicians in the relevant technical field can reasonably determine the specific meanings of the above terms in the present invention based on the specific content of the technical solution.

[0023] Reference Figure 1 and Figure 2 In the first aspect of the present invention, a bioreactor includes a container 124 and an execution device 100, the execution device 100 is detachably mounted on the container 124, the container 124 is provided with a cavity 121, the execution device 100 includes a base 101 and a plurality of stirring paddles arranged on the base 101, the base 101 is detachably mounted on the container 124, it can be understood that, with reference to Figure 3 and Figure 4 The base 101 is provided with a positioning edge 108 extending toward the bottom, and a limiting groove 109 is provided on the outer edge of the container 124. When the actuator 100 is installed on the container 124, the positioning edge 108 can be inserted into the limiting groove 109, thereby realizing the installation and positioning of the actuator 100 and the container 124 and improving the convenience of use.

[0024] It can be understood that in this embodiment, 6 stirring paddles are provided, and the stirring paddles can be inserted into the cavity 121. The stirring paddles are used to stir the culture solution in the cavity 121 so that the culture solution and the cells are fully in contact. The stirring paddles include a rotating shaft 112 and a plurality of blades 111 arranged circumferentially along the rotating shaft 112. The blades 111 are inclined to the horizontal plane. During the rotation of the stirring paddles, it helps to make the liquid in the cavity 121 flow along the direction of the rotating axis of the stirring paddles, improve the mixing effect of the cells and the culture solution, and help to promote the reaction rate. At the same time, the actuator 100 is provided with a motor 103, and the base 101 is provided with a covering upper cover 102. The motor 103 is installed on the upper cover 102 and is connected to any stirring paddle. A driving gear is provided at one end of the rotating shaft 112 away from the stirring paddle. The driving gears on the two adjacent rotating shafts 112 are meshed with each other, so that the rotation directions of the two adjacent stirring paddles are opposite, which helps to improve the stirring efficiency of the liquid in the cavity 121, improve the mixing efficiency of the cells and the culture solution, and thus improve the reaction rate. Furthermore, the driving gear includes a driving wheel 105 and a driven wheel 106. The stirring paddle connected to the motor 103 is provided with the driving wheel 105. The driving wheel 105 is meshed with the driven wheel 106 to achieve power transmission.

[0025] Refer to 2 and Figure 3, the stirring paddle includes a first stirring paddle 110 and a second stirring paddle 120 which are arranged adjacent to each other, and the inclination direction of the paddle 111 between the first stirring paddle 110 and the second stirring paddle 120 is opposite. It can be understood that during the cell culture process, the cells are deposited at the bottom of the chamber 121, and the cells consume the dissolved oxygen and nutrients of the culture solution in the process of continuous growth and metabolism, thereby resulting in a low concentration of nutrients and a low pH value at the bottom, which is not conducive to the continuous growth of the cells. Therefore, when the motor 103 drives the driving wheel 105 to rotate, the driving wheel 105 and the driven wheel 106 are externally meshed, so the driving wheel 105 rotates in opposite directions to the driven wheel, and the stirring paddle installed on the driving wheel 105 and the stirring paddle installed on the driven wheel 106 rotate in opposite directions, and can create vortices of different directions in the chamber 121 during rotation, which can enhance the mixing effect of the culture solution and cells, and improve the utilization rate of nutrients by cells, thereby facilitating the growth of cells. At the same time, since the inclination directions of the blades 111 between the adjacent first stirring paddle 110 and the second stirring paddle 120 are opposite, that is, the first stirring paddle 110 and the second stirring paddle 120 are mirror images of each other, that is, the inclination directions of the blades 111 between the stirring paddle installed by the driving wheel 105 and the stirring paddle installed by the driven wheel 106 are opposite, and the rotation directions of the first stirring paddle 110 and the second stirring paddle 120 are opposite, when rotating, the first stirring paddle 110 and the second stirring paddle 120 can make the culture solution move in the same direction along the rotation axis in the culture solution, so that convection is generated between the upper and lower layers of the culture solution, and the cells located at the bottom of the cavity 121 float upward, which is beneficial to maintaining the uniform distribution of nutrients and dissolved oxygen in the culture solution, which is beneficial to cell growth.

[0026] Further, refer to Figure 2 The blade 111 includes a connecting end 115 close to the rotating shaft 112 and a free end 116 far from the rotating shaft 112. The outer edge of the free end 116 is inclined at a first angle 117 with the axis of the rotating shaft 112. The edge of the connecting end 115 is inclined at a second angle 118 with the axis of the rotating shaft 112. The first angle 117 is not equal to the second angle 118. Figure 6 and Figure 7, along the width direction of the blade 111, a straight line passing through the blade 111 and parallel to the outer edge of the free end 116 is defined as the first axis 114, a straight line passing through the blade 111 and parallel to the edge of the connecting end 115 is defined as the second axis 119, a first angle 117 is an angle between the first axis 114 and the rotation axis, and a second angle 118 is an angle between the second axis 119 and the rotation axis. In this embodiment, the first angle 117 is greater than the second angle 118, that is, the inclination angle of the free end 116 relative to the rotation axis is greater than that of the connecting end 115. 115 is an inclination angle relative to the rotation axis. In the present embodiment, at least three blades 111 are arranged. Since the connection end 115 of the blade 111 is closer to the rotation axis, when the inclination angle is too large, the circumferential width of the blade 111 is large, which is not conducive to the spatial arrangement of multiple blades 111. The free end 116 is located at the periphery of the stirring paddle and has ample arrangement space. Therefore, when the first angle 117 is greater than the second angle 118, the surface of the blade 111 can have a curved surface, and when the stirring paddle rotates, it can effectively reduce the resistance while improving the stirring efficiency of the liquid.

[0027] Further, refer to Figure 5 , the chamber 121 is also provided with a plurality of unit chambers 122, each of which is plugged with a stirring paddle, and each unit chamber 122 can be used to place different types of microorganisms for cultivation, so the execution device 100 can stir different culture dishes at the same time, which can improve the flexibility of use. Furthermore, multiple motors 103 can be provided, and each stirring paddle is driven by a separate motor 103, and the stirring speed can be adjusted according to different culture dishes to meet the use requirements.

[0028] Reference Figure 8 and Fig. 9 , the rotating shaft 112 has a channel 113 running through both ends, and the actuator 100 is provided with a sensor 123, which is used to detect the pH value and dissolved oxygen concentration in the culture solution. The sensor 123 is installed on the base 101 and inserted into the channel 113 and extends out of the stirring paddle. In addition, there is a gap between the radial outer peripheral wall of the sensor 123 and the inner wall of the channel 113 to prevent the sensor 123 from rotating with the stirring paddle, and can detect the parameters in the culture solution in real time while improving the compactness of the space. The sensor 123 can detect the solution in the cavity 121 and generate a detection signal. The control device can receive the detection signal from the sensor 123, and the control device can control the rotation speed of the stirring paddle based on the detection signal, and can automatically adjust the rotation speed of the stirring paddle according to the state parameters of the culture solution in real time to ensure that the cells are fully in contact and react with the culture solution, which is conducive to improving the biological culture effect. Further, a mounting groove 104 is provided in the upper cover 102, and the end of the sensor 123 away from the stirring paddle is fixed in the mounting groove 104. It can be understood that the control device is provided with a switch 107.

[0029] According to an embodiment of the second aspect of the present invention, there is provided a stirring method, which is applied to the above-mentioned bioreactor, and comprises the following steps: Step 1: Place the culture medium and cells in the cavity 121, place the actuator 100 on the container 124, and insert the positioning edge 108 into the limiting groove 109. Through the cooperation between the positioning edge 108 and the limiting groove 109, quick installation can be achieved, which is conducive to improving the convenience of use.

[0030] Step 2: Start the switch 107, the motor 103 starts and drives any stirring paddle to rotate, the driving wheel 105 drives a plurality of driven wheels 106 to rotate, so that the first stirring paddle 110 and the second stirring paddle 120 rotate synchronously in opposite directions, and stir the mixed liquid in the cavity 121, so that the culture liquid in the cavity 121 convects from top to bottom or from bottom to top, thereby improving the mixing degree of cells and culture liquid. At this time, the motor 103 reaches the first speed and maintains the first speed for uniform stirring.

[0031] Step 3: Place the actuator 100 and the container 124 in the carbon dioxide culture chamber for cell culture. During this process, the sensor 123 measures the parameter A of the pH value and the parameter B of the dissolved oxygen content in the solution, and transmits the parameter A and the parameter B to the control device. Furthermore, the distance between the detection part of the sensor 123 and the bottom of the chamber 121 is less than half of the depth of the culture solution, that is, the detection part of the sensor 123 can at least detect the parameters of the bottom layer of the culture solution, so as to more accurately detect the parameters of the culture solution near the cells, so as to adjust the rotation speed in time to ensure that the cells are in a good growth environment.

[0032] It can be understood that in this embodiment, it is necessary to control the pH value to be between 6.8-7.8, and the dissolved oxygen content to be between 10000-50000ppm, so when A is less than 6.8, the control device changes the speed of the motor 103 to reach the second speed; when B is less than 10000ppm, the control device changes the speed of the motor 103 to reach the second speed, and the second speed is greater than the first speed. It can be understood that due to the metabolism of the cells, the pH value of the culture solution near the cells will decrease. When the pH value parameter decreases, the speed of the motor 103 is increased, so that the stirring paddle increases the disturbance of the culture solution, so that the upper and lower layers of the culture solution produce more convection, so that the nutrients and dissolved oxygen tend to be uniform.

[0033] It is understandable that a high pH value (i.e., alkaline) or high dissolved oxygen content in the culture dish environment will also have an adverse effect on the cells. When A>7.8, the control device changes the speed of the motor 103 to the third speed; when B>50000ppm, the control device changes the speed of the motor 103 to the third speed, which is less than the first speed and greater than zero. It is understandable that during the cell culture process, the actuator 100 and the container 124 are placed in the carbon dioxide culture chamber, and the nutrient solution can dissolve part of the carbon dioxide to meet the needs of cell growth. When the pH value is high or the dissolved oxygen content is too high, the carbon dioxide has a certain regulating ability and can properly maintain the pH value and dissolved oxygen concentration. Therefore, by reducing the speed of the motor 103 to the third speed, waiting for the cells to consume the dissolved oxygen and other nutrients in the culture solution and the dissolution of carbon dioxide in the culture solution, the parameters of the culture solution tend to appropriate values ​​to create a good cell growth environment.

[0034] The embodiments of the present invention are described in detail above with reference to the accompanying drawings, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge scope of ordinary technicians in the relevant technical field without departing from the purpose of the present invention.

Claims

1. A bioreactor, characterized in that: include: A container having a cavity; An actuator, comprising a base and a plurality of stirring paddles arranged on the base, wherein the base is detachably mounted on the container, the stirring paddles can be inserted into the cavity, the rotation directions of two adjacent stirring paddles are opposite, the stirring paddles comprise a rotating shaft and a plurality of paddles arranged along the circumference of the rotating shaft, the paddles are inclined to a horizontal plane, and the rotating shaft has channels running through both ends; A sensor, inserted into the channel and extending out of the stirring paddle, the sensor being used to detect parameters of the solution in the cavity and generate a detection signal; The control device is electrically connected to the execution device and the sensor. The control device is provided with a switch. The control device can receive the detection signal from the sensor and can control the rotation speed of the stirring paddle based on the detection signal.

2. A bioreactor according to claim 1, characterized in that: A driving gear is provided on one end of the rotating shaft away from the stirring paddle, and the driving gears on two adjacent rotating shafts are meshed with each other.

3. A bioreactor according to claim 2, characterized in that: The actuator is provided with a motor, which is mounted on the base and is connected to any one of the stirring paddles.

4. A bioreactor according to claim 3, characterized in that: The driving gear comprises a driving wheel and a driven wheel. The stirring paddle connected to the motor power is provided with a driving wheel, and the driving wheel is meshed with the driven wheel.

5. A bioreactor according to claim 1, characterized in that: The stirring paddle comprises a first stirring paddle and a second stirring paddle which are adjacent to each other, and the inclination directions of the blades between the first stirring paddle and the second stirring paddle are opposite to each other.

6. A bioreactor according to claim 1, characterized in that: The base is provided with a positioning edge, and the outer edge of the container is provided with a limiting groove. When the execution device is installed on the container, the positioning edge is inserted into the limiting groove.

7. A bioreactor according to claim 1, characterized in that: A plurality of unit cavities are arranged in the chamber, and a stirring paddle is inserted into each of the unit cavities.

8. A bioreactor according to claim 7, characterized in that: The blade includes a connecting end close to the rotating shaft and a free end away from the rotating shaft, the outer edge of the free end is inclined at a first angle to the axis of the rotating shaft, the edge of the connecting end is inclined at a second angle to the axis of the rotating shaft, and the first angle is not equal to the second angle.

9. A stirring method, characterized in that: A bioreactor according to any one of claims 1 to 8, comprising a container, an actuator, a sensor and a control device, wherein the actuator comprises a motor, a driving wheel, a driven wheel and a stirring paddle, the control device is provided with a switch, and the container has a cavity, comprising the following steps: Step 1: placing the culture fluid and cells in the cavity, placing the actuator on the container, and inserting the positioning edge into the limiting groove; Step 2: Start the switch, the motor drives any one of the stirring paddles to rotate, the driving wheel drives a plurality of the driven wheels to rotate, and the mixed liquid in the chamber is stirred, and at this time the motor reaches a first speed; Step 3: Place the actuator and the container in a carbon dioxide culture chamber for cell culture. During this process, the sensor measures parameter A of the pH value and parameter B of the dissolved oxygen content in the solution, and transmits parameter A and parameter B to the control device.

10. A bioreactor according to claim 9, characterized in that: In step 3, when A is less than 6.8, the control device changes the rotation speed of the motor to a second rotation speed; when B is less than 10000 ppm, the control device changes the rotation speed of the motor to a second rotation speed, and the second rotation speed is greater than the first rotation speed.

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