A reactor for culturing animal cells in suspension based on collagen microcarriers

Abstract

The invention discloses a reactor for culturing animal cells in suspension based on collagen microcarriers, comprising an outer shell, a base being arranged at the lower end of the outer shell, a support seat being arranged at the upper end of the outer shell, an upper cover being arranged on the upper side of the support seat, a magnetic coupler being fixedly connected to the upper side of the upper cover, a motor being fixedly connected to the upper end of the magnetic coupler, an inner reaction tank being arranged inside the outer shell, a stirring shaft being arranged inside the inner reaction tank, a cooling coil being arranged in the middle of the stirring shaft, and the upper end of the stirring shaft being fixedly connected to the output shaft of the magnetic coupler; through the designed first push blade and the pushing stirring mechanism, when in use, the first push blade and the second push blade on the pushing stirring mechanism cooperate with the anti-shear chamfer and the inner stirring groove during stirring, thereby reducing the shear force of the first push blade and the second push blade on the cells during stirring, thereby reducing the phenomenon of cell damage.

Description

Technical Field

[0001] The invention belongs to the technical field of animal cell culture, and in particular relates to a reactor based on collagen microcarrier suspension culture of animal cells. Background Art

[0002] Animal cell culture is to remove the relevant tissue from the animal body, disperse it into single cells (using trypsin or collagenase), and then place it in a suitable culture medium to allow these cells to grow and proliferate. Cell culture refers to the growth of cells under in vitro conditions. Animal cells no longer form individuals during the process of individual cell culture. The nutrients required for in vitro cell culture are basically the same as those in vivo, for example, sugars, amino acids, inorganic salts, growth factors, trace elements, etc. The culture medium that strictly formulates the above substances required by the cells according to their types and required quantities is called a synthetic culture medium. Since some components of the internal environment in which animal cells live have not been studied clearly, animal serum needs to be added to provide an environment similar to that in the body. Therefore, when using synthetic culture media, some natural ingredients such as serum and plasma are usually added.

[0003] When animal cells are cultured, stirring is usually used to mix the prepared raw materials. If the shear force is too large during mixing, the cells will be cut and crushed. In addition, stirring and mixing during use will cause uneven mixing from top to bottom due to the height. For this reason, we propose a reactor for suspending animal cells based on collagen microcarriers. Summary of the invention

[0004] The object of the present invention is to provide a reactor for suspending and culturing animal cells based on collagen microcarriers to solve the problems raised in the above background technology.

[0005] To achieve the above-mentioned purpose, the present invention provides the following technical solutions: A reactor for culturing animal cells based on collagen microcarrier suspension, comprising an outer shell, a base is provided at the lower end of the outer shell, a support seat is provided at the upper end of the outer shell, an upper cover is provided on the upper side of the support seat, a magnetic coupler is fixedly connected to the upper side of the upper cover, a motor is fixedly connected to the upper end of the magnetic coupler, an inner reaction tank is provided inside the outer shell, a stirring shaft is provided inside the inner reaction tank, a cooling coil is provided in the middle of the stirring shaft, the upper end of the stirring shaft is fixedly connected to the output shaft of the magnetic coupler, a first push blade is provided on the outer surface of the stirring shaft, a pushing and stirring mechanism is provided at the inner bottom of the inner reaction tank, the pushing and stirring mechanism comprises a support frame fixedly mounted on the inner bottom of the inner reaction tank and a rotating disk rotatably mounted on the support frame, a second push blade is provided on the outer surface of the rotating disk, the second push blade comprises a blade body, one side of the blade body is provided with an anti-shear chamfer, an inner stirring tank is provided on the inner side of the blade body, and the first push blade and the second push blade have the same structure.

[0006] Preferably, the pushing stirring mechanism also includes a bevel gear, which is fixedly mounted on the rotating shaft of the rotating disk and the lower end of the stirring shaft, and the stirring shaft and the bevel gear are meshed and driven by the rotating disk, and the stirring shaft is rotatably connected to the support frame through a bearing.

[0007] Preferably, the rotating disk and the second push blade are centrally symmetrically mounted on a support frame, and the shape of the support frame is set to be cylindrical.

[0008] Preferably, a support pad is provided at the bottom of the base, a moving mechanism is provided inside the shell, and the shape of the base is set to be a truncated cone.

[0009] Preferably, the moving mechanism includes a pushing plate slidably installed inside the base, a spring is provided on the outer side of the pushing plate, a moving wheel is provided at the lower end of the pushing plate, an eccentric rotating wheel is rotatably connected to the inner upper end of the base, and a handle is provided at one end of the eccentric rotating wheel.

[0010] Preferably, the position of the eccentric wheel corresponds to the position of the push plate, and a slot is provided on the upper surface of the push plate, and the shape of the slot matches the shape of one end of the eccentric wheel.

[0011] Preferably, the outer shell comprises an outer protective shell, an inner supporting layer is arranged inside the outer protective shell, and a thermal insulation filling is arranged on the inner side of the inner supporting layer.

[0012] Preferably, the cross-sectional shape of the inner supporting layer is set to be a honeycomb shape, and the thermal insulation filler is installed in the honeycomb holes of the inner supporting layer.

[0013] Preferably, a constant temperature furnace is arranged inside the outer shell, and the inner reaction tank is located inside the constant temperature furnace.

[0014] Preferably, a pressing block is provided on the inner side of the upper cover, an inlet and outlet pipe is provided on the lower side of the pressing block, the pressing block is pressed tightly against the upper side of the inner reaction tank, a pressure explosion-proof gauge and inlet and outlet valves are provided on both sides of the upper cover, and the inlet and outlet valves are connected to the interior of the inner reaction tank through the inlet and outlet pipes, and the pressure explosion-proof gauge is connected to the interior of the inner reaction tank through a built-in pipeline.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] 1. Through the designed first push blade and push stirring mechanism, when in use, the first push blade and the second push blade on the push stirring mechanism cooperate with the anti-shear chamfer and the inner stirring groove during stirring, thereby reducing the shear force of the first push blade and the second push blade on the cells during stirring, thereby reducing the phenomenon of cell damage.

[0017] 2. Through the designed pushing stirring mechanism and the first pushing blade, when in use, the transmission of the bevel gear drives the rotating disk and the second pushing blade to rotate when the stirring shaft rotates, so that the second pushing blade and the rotating disk push the culture solution at the bottom outward for stirring. At the same time, the rotation of the stirring shaft drives the first pushing blade to rotate, pushing the culture solution downward, so that the culture solution rolls and stirs in the inner reaction tank, thereby achieving a better mixing effect.

[0018] 3. Through the designed moving mechanism, when in use, the eccentric wheel is driven to rotate by turning the handle, so that the eccentric wheel pushes the push plate to slide, and the moving wheel is pushed out. Then the reactor can be pushed to move by the moving wheel. When in use, the eccentric wheel can be rotated in the opposite direction, and the spring pushes the push plate and the moving wheel to reset and retract them into the base. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a structural schematic diagram of the present invention;

[0020] Figure 2 It is a schematic diagram of the internal structure of the present invention;

[0021] Figure 3 For the present invention Figure 2 The enlarged structural diagram at A in the middle;

[0022] Figure 4 It is a schematic diagram of the push blade structure of the present invention;

[0023] Figure 5 It is a schematic diagram of the structure of the mobile mechanism of the present invention;

[0024] Figure 6 It is a schematic diagram of the shell structure of the present invention;

[0025] In the figure: 1. outer shell; 2. base; 3. upper cover; 4. support seat; 5. inlet and outlet valve; 6. pressure explosion-proof gauge; 7. magnetic coupler; 8. motor; 9. moving mechanism; 10. support pad; 11. pressing block; 12. stirring shaft; 13. first push blade; 14. cooling coil; 15. pushing stirring mechanism; 16. inner reaction tank; 17. inlet and outlet pipe; 18. constant temperature furnace; 19. rotating disk; 20. second push blade; 21. support frame; 22. bevel gear; 23. blade body; 24. anti-shear chamfer; 25. inner stirring tank; 26. handle; 27. eccentric wheel; 28. push plate; 29. ​​slot; 30. spring; 31. moving wheel; 32. outer protective shell; 33. inner support layer; 34. thermal insulation filling. DETAILED DESCRIPTION

[0026] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] See also Figures 1 to 6The present invention provides a technical solution: a reactor for culturing animal cells based on collagen microcarrier suspension, comprising a shell 1, a base 2 is arranged at the lower end of the shell 1, a support seat 4 is arranged at the upper end of the shell 1, an upper cover 3 is arranged on the upper side of the support seat 4, a magnetic coupler 7 is fixedly connected to the upper side of the upper cover 3, a motor 8 is fixedly connected to the upper end of the magnetic coupler 7, an inner reaction tank 16 is arranged inside the shell 1, a stirring shaft 12 is arranged inside the inner reaction tank 16, a cooling coil 14 is arranged in the middle of the stirring shaft 12, the upper end of the stirring shaft 12 is fixedly connected to the output shaft of the magnetic coupler 7, a first push blade 13 is arranged on the outer surface of the stirring shaft 12, a pushing and stirring mechanism 15 is arranged on the inner bottom of the inner reaction tank 16, the pushing and stirring mechanism 15 comprises a support frame 21 fixedly mounted on the inner bottom of the inner reaction tank 16 and a rotating disk 19 rotatably mounted on the support frame 21, and a second push blade 20 is arranged on the outer surface of the rotating disk 19, The second push blade 20 includes a blade body 23, one side of the blade body 23 is provided with an anti-shear chamfer 24, the inner side of the blade body 23 is provided with an inner stirring groove 25, and the first push blade 13 and the second push blade 20 have the same structure, and the pushing and stirring mechanism 15 also includes a bevel gear 22, which is fixedly mounted on the rotating shaft of the rotating disk 19 and the lower end of the stirring shaft 12, and the stirring shaft 12 and the bevel gear 22 are meshed and transmitted through the rotating disk 19, and the stirring shaft 12 is rotatably connected to the support frame 21 through a bearing, and the stirring shaft 12 drives the rotating disk 19 and the second push blade 20 to rotate through the meshing of the bevel gear 22, and at the same time, the stirring shaft 12 drives the first push blade 13 to rotate, and the first push blade 13 pushes the liquid to move downward, and the rotating disk 19 drives the second push blade 20 to rotate and push the liquid from the bottom to both sides, so that the liquid is tumbled and stirred in the inner reaction tank 16, and the anti-shear chamfer 24 is used to reduce the shear force during stirring, thereby reducing the cell breakage.

[0028] For further information, see Figure 2 and Figure 3 The rotating disk 19 and the second push blade 20 are centrally symmetrically mounted on a support frame 21, and the shape of the support frame 21 is set to be cylindrical.

[0029] For further information, see Figure 1 and Figure 5A support pad 10 is provided at the bottom of the base 2, and a moving mechanism 9 is provided inside the shell 1. The shape of the base 2 is set to be a truncated cone. The moving mechanism 9 includes a pushing plate 28 slidably installed inside the base 2, and a spring 30 is provided on the outer side of the pushing plate 28. A moving wheel 31 is provided at the lower end of the pushing plate 28. An eccentric wheel 27 is rotatably connected to the inner upper end of the base 2, and a handle 26 is provided at one end of the eccentric wheel 27. The position of the eccentric wheel 27 corresponds to the position of the pushing plate 28. A card slot 29 is provided on the upper surface of the pushing plate 28, and the shape of the card slot 29 matches the shape of one end of the eccentric wheel 27. Lifting the handle 26 drives the eccentric wheel 27 to rotate, so that the eccentric wheel 27 slides down against the pushing plate 28, and pushes the moving wheel 31 to slide out of the base 2, and the reactor is pushed to move by the moving wheel 31.

[0030] For further information, see Figure 6 The outer shell 1 includes an outer protective shell 32, an inner supporting layer 33 is arranged inside the outer protective shell 32, and a thermal insulation filling 34 is arranged on the inner side of the inner supporting layer 33. The cross-sectional shape of the inner supporting layer 33 is set to a honeycomb shape, and the thermal insulation filling 34 is installed in the honeycomb holes of the inner supporting layer 33.

[0031] For further information, see Figure 2 A constant temperature furnace 18 is arranged inside the outer shell 1 , and the inner reaction tank 16 is located inside the constant temperature furnace 18 .

[0032] For further information, see Figure 2 A pressing block 11 is arranged on the inner side of the upper cover 3, and an inlet and outlet pipe 17 is arranged on the lower side of the pressing block 11. The pressing block 11 is pressed tightly on the upper side of the inner reaction tank 16. A pressure explosion-proof gauge 6 and an inlet and outlet valve 5 are arranged on both sides of the upper cover 3. The inlet and outlet valve 5 is connected to the interior of the inner reaction tank 16 through the inlet and outlet pipe 17, and the pressure explosion-proof gauge 6 is connected to the interior of the inner reaction tank 16 through a built-in pipeline.

[0033] The cultured collagen microcarriers have the characteristic of combining with substances such as fibronectin in the culture medium containing bovine serum. Therefore, collagen is a kind of excellent matrix with a wide source that can be used as a preparation of cell culture medium.

[0034] The constant temperature furnace 18 adopts far infrared heating element as heater, and uses constant temperature controller to keep the temperature of digestion process constant.

[0035] The working principle and use process of the present invention are as follows: when in use, the collagen microcarrier culture solution is added into the inner reaction tank 16, and the cells to be cultured are added into the inner reaction tank 16, the stirring shaft 12 is driven to rotate by the motor 8 and the magnetic coupler 7, the stirring shaft 12 drives the rotating disk 19 and the second push leaf 20 to rotate through the meshing of the bevel gear 22, and at the same time, the stirring shaft 12 drives the first push leaf 13 to rotate, the first push leaf 13 pushes the liquid downward, the rotating disk 19 drives the second push leaf 20 to rotate and push the liquid from the bottom to both sides, so that the liquid rolls and stirs in the inner reaction tank 16, so that the stirring is more uniform, and the anti-shear chamfer 24 is used to reduce the shearing during stirring. force, thereby reducing cell breakage, and the inner stirring tank 25 forms a vortex during stirring, so that the stirring is more sufficient. At the same time, the constant temperature furnace 18 maintains a constant temperature, so that the cells are cultured at a suitable temperature. When the reactor needs to be moved, it is only necessary to lift the handle 26 to drive the eccentric wheel 27 to rotate, so that the eccentric wheel 27 pushes the push plate 28 to slide down, and push the moving wheel 31 to slide out of the base 2, and the moving wheel 31 is used to push the reactor to move. After moving to the use position, the handle 26 is retracted to lift the eccentric wheel 27, and the push plate 28 is lifted by the elastic force of the spring 30, so that the moving wheel 31 is retracted and the support pad 10 is supported on the ground for use, which is more stable during use.

[0036] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0037] The above description is only used to illustrate the technical solution of the present invention rather than to limit it. Other modifications or equivalent substitutions made to the technical solution of the present invention by ordinary technicians in this field should be included in the scope of the claims of the present invention as long as they do not depart from the spirit and scope of the technical solution of the present invention.

Claims

1. A reactor for culturing animal cells in suspension based on collagen microcarriers, comprising a housing (1), characterized in that: The lower end of the shell (1) is provided with a base (2), the upper end of the shell (1) is provided with a support seat (4), the upper side of the support seat (4) is provided with an upper cover (3), the upper side of the upper cover (3) is fixedly connected to a magnetic coupler (7), the upper end of the magnetic coupler (7) is fixedly connected to a motor (8), an inner reaction tank (16) is provided inside the shell (1), a stirring shaft (12) is provided inside the inner reaction tank (16), a cooling coil (14) is provided in the middle of the stirring shaft (12), the upper end of the stirring shaft (12) is fixedly connected to the output shaft of the magnetic coupler (7), and the outer surface of the stirring shaft (12) is provided with a a push blade (13), wherein a push stirring mechanism (15) is arranged at the inner bottom of the inner reaction tank (16), wherein the push stirring mechanism (15) comprises a support frame (21) fixedly mounted at the inner bottom of the inner reaction tank (16) and a rotating disk (19) rotatably mounted on the support frame (21), wherein a second push blade (20) is arranged on the outer surface of the rotating disk (19), wherein the second push blade (20) comprises a blade body (23), wherein one side of the blade body (23) is provided with an anti-shear chamfer (24), and an inner stirring groove (25) is arranged on the inner side of the blade body (23), and the first push blade (13) and the second push blade (20) have the same structure; The pushing and stirring mechanism (15) further comprises a bevel gear (22), wherein the bevel gear (22) is fixedly mounted on the rotating shaft of the rotating disk (19) and the lower end of the stirring shaft (12), and the stirring shaft (12) and the bevel gear (22) are meshed and driven by the rotating disk (19), and the stirring shaft (12) is rotatably connected to the support frame (21) via a bearing; The rotating disk (19) and the second push blade (20) are mounted on a support frame (21) in a centrally symmetrical manner, and the support frame (21) is configured to be cylindrical in shape.

2. A reactor based on collagen microcarrier suspension culture of animal cells according to claim 1, characterized in that: A support pad (10) is provided at the bottom of the base (2), a moving mechanism (9) is provided inside the housing (1), and the shape of the base (2) is set to be a truncated cone.

3. A reactor based on collagen microcarrier suspension culture of animal cells according to claim 2, characterized in that: The moving mechanism (9) comprises a push plate (28) slidably mounted inside the base (2), a spring (30) being arranged on the outer side of the push plate (28), a moving wheel (31) being arranged at the lower end of the push plate (28), an eccentric rotating wheel (27) being rotatably connected to the inner upper end of the base (2), and a handle (26) being arranged at one end of the eccentric rotating wheel (27).

4. A reactor based on collagen microcarrier suspension culture of animal cells according to claim 3, characterized in that: The position of the eccentric rotating wheel (27) corresponds to the position of the pushing plate (28); a slot (29) is provided on the upper surface of the pushing plate (28); and the shape of the slot (29) matches the shape of one end of the eccentric rotating wheel (27).

5. A reactor based on collagen microcarrier suspension culture of animal cells according to claim 1, characterized in that: The housing (1) comprises an outer protective shell (32), an inner supporting layer (33) is arranged inside the outer protective shell (32), and a thermal insulation filler (34) is arranged inside the inner supporting layer (33).

6. A reactor based on collagen microcarrier suspension culture of animal cells according to claim 5, characterized in that: The cross-sectional shape of the inner support layer (33) is set to be a honeycomb shape, and the thermal insulation filler (34) is installed in the honeycomb holes of the inner support layer (33).

7. The reactor based on collagen microcarrier suspension culture of animal cells according to claim 1, characterized in that: A constant temperature furnace (18) is arranged inside the outer shell (1), and the inner reaction tank (16) is located inside the constant temperature furnace (18).

8. The reactor based on collagen microcarrier suspension culture of animal cells according to claim 1, characterized in that: A pressing block (11) is arranged on the inner side of the upper cover (3), and an inlet and outlet pipe (17) is arranged on the lower side of the pressing block (11). The pressing block (11) is pressed tightly against the upper side of the inner reaction tank (16). A pressure explosion-proof gauge (6) and an inlet and outlet valve (5) are arranged on both sides of the upper cover (3), and the inlet and outlet valve (5) is connected to the interior of the inner reaction tank (16) through the inlet and outlet pipe (17), and the pressure explosion-proof gauge (6) is connected to the interior of the inner reaction tank (16) through a built-in pipeline.

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