Constant-temperature cell culture equipment for research and development of cattle akabane disease inactivated vaccine

Abstract

The invention relates to the technical field of cell culture, in particular to constant-temperature cell culture equipment for cattle akabane disease inactivated vaccine research and development, which comprises a cell culture box, the cell culture box comprises a bottom cover, a plurality of partition plates and a top cover, and a bottom chamber, a middle chamber and a top chamber are arranged in the cell culture box. A plurality of n-shaped frames are further arranged above the bottom cover, a first liquid inlet space is formed between the n-shaped frames and the partition plate, and a second liquid inlet space is formed between the n-shaped frames and the top cover. Liquid pumping assemblies are arranged in the bottom-layer chamber, the middle-layer chamber and the top-layer chamber, each liquid pumping assembly comprises a liquid baffle, the liquid baffle is connected with the first end of a pull rod, and the second end of the pull rod extends to the outer side of the cell culture box. According to the cell culture device, the liquid baffle can block the inlets of all the cavities, only after the liquid baffle moves, a nutrient solution can enter all the cavities, meanwhile, the liquid baffle can exhaust all air in all the cavities, the air cannot occupy the space in all the cavities, and the cell culture process cannot be hindered.

Description

Technical Field

[0001] The invention relates to the field of cell culture technology, and specifically relates to a constant temperature cell culture device for the research and development of an inactivated vaccine for bovine Akabane disease. Background Art

[0002] Akabane disease in cattle is a viral infectious disease caused by Akabane virus. The disease mainly affects ruminants such as cattle and sheep, causing symptoms such as miscarriage, premature birth, and arthrogryposis and hydrocephalus syndrome in newborns. In view of the huge threat of Akabane disease to the breeding industry, vaccine research and development has become an important means of preventing and controlling the disease. The expression of specific proteins, monoclonal antibodies, interferons and viral vaccine products through large-scale in vitro culture of animal cells has become the most common technology at present. The use of cell culture devices can harvest a large number of cells at one time. For example, the Chinese invention patent with patent application number "CN201811182765.6" provides a cell culture device and a cell box thereof, which can realize perfusion culture. The nutrient solution first flows from the liquid inlet into the outermost bottom chamber, and then flows from the bottom to the top through the middle chambers directly above, and finally reaches the top chamber directly above. The nutrient solution flows from the bottom to the top through the odd-numbered columns of chambers, and then flows from the top to the bottom through the even-numbered columns of chambers. After flowing through the last column of chambers, the nutrient solution flows out of the cell box from the liquid outlet.

[0003] The disadvantage of the device is that each chamber in the device is filled with air. Only when the nutrient solution enters each chamber in sequence can the air in several chambers be discharged in sequence, so that the nutrient solution fills several chambers. Since the top surface of each chamber of the device is flat, the air may form bubbles and cannot be discharged smoothly. The bubbles will occupy the space in each chamber, hindering the entire cell culture process. Summary of the invention

[0004] The purpose of the present invention is to provide a constant temperature cell culture device for the research and development of an inactivated vaccine for Akabane disease in cattle. A liquid baffle plate can block the entrance of each chamber. Only after the liquid baffle plate moves can the nutrient solution enter each chamber. At the same time, the liquid baffle plate can also exhaust all the air in each chamber. The air will not occupy the space in each chamber, and will not hinder the cell culture process.

[0005] To achieve the above object, the present invention provides the following technical solution: A constant-temperature cell culture device for the research and development of an inactivated vaccine against bovine Akabane disease, comprising a cell culture box. The cell culture box includes a bottom cover, a plurality of partition plates and a top cover stacked in sequence from bottom to top. A bottom layer chamber is provided between the bottom cover and the lowermost partition plate. A middle layer chamber is provided between two adjacent partition plates. A top layer chamber is provided between the uppermost partition plate and the top cover. The length of the bottom cover is greater than the length of the partition plate. The length of the partition plate is the same as the length of the top cover. A plurality of U-shaped frames stacked in sequence from bottom to top are further provided above the bottom cover. The plurality of U-shaped frames are respectively fixedly connected to the plurality of partition plates and the top cover. A first liquid inlet space is formed between the U-shaped frame and the partition plate. A second liquid inlet space is formed between the U-shaped frame and the top cover. The first liquid inlet space, the second liquid inlet space, the bottom layer chamber, the middle layer chamber and the top layer chamber communicate with each other. The cross-sectional shapes of the bottom layer chamber, the middle layer chamber and the top layer chamber are the same. Liquid pumping assemblies are provided in the bottom layer chamber, the middle layer chamber and the top layer chamber. The liquid pumping assembly includes a liquid blocking plate. The cross-sectional shape of the liquid blocking plate matches the cross-sectional shape of the bottom layer chamber. The liquid blocking plate is connected to the first end of a pull rod. The pull rod is located on the side away from the U-shaped frame. The length of the pull rod is greater than the length of the bottom cover. The second end of the pull rod extends to the outside of the cell culture box.

[0006] Preferably, it further includes a perfusion assembly. The perfusion assembly includes a liquid inlet box. The liquid inlet box is provided above the uppermost U-shaped frame and is fixedly connected to the U-shaped frame. The interior of the liquid inlet box communicates with the second liquid inlet space. A liquid inlet pipe is provided on the side wall of the liquid inlet box. The liquid blocking plate is fixedly connected to the first end of a liquid outlet pipe. The length of the liquid outlet pipe is greater than the width of the liquid blocking plate. The liquid outlet pipe penetrates through the liquid blocking plate. The second end of the liquid outlet pipe is located on the side away from the U-shaped frame. A plug is provided at the second end of the liquid outlet pipe. The first end of the plug extends into the liquid outlet pipe. The diameter of the second end of the plug is the same as the diameter of the liquid outlet pipe. The plug is detachably connected to the liquid outlet pipe. A first avoidance hole is provided on the side wall of the cell culture box. The position of the first avoidance hole corresponds to the position of the liquid outlet pipe. The shape of the first avoidance hole matches the cross-sectional shape of the liquid outlet pipe.

[0007] Preferably, the height of the first end of the liquid inlet box is greater than that of the second end. An exhaust port is provided on the top surface of the first end of the liquid inlet box. A discharge pipe is provided in the exhaust port. The discharge pipe is vertically arranged and communicated with the interior of the liquid inlet box. A pipe cap is provided at the top of the discharge pipe. The pipe cap is detachably connected to the discharge pipe. The perfusion assembly further includes a bubble guiding assembly. The bubble guiding assembly includes: a regulating plate provided in the liquid inlet box, between the exhaust port and the second end of the liquid inlet box. The first end of the regulating plate is hinged to the inner wall of the liquid inlet box. The first end of the regulating plate and the discharge pipe are on the same side. The liquid inlet pipe and the exhaust port are on the same side and below the exhaust port. A stop block is provided in the liquid inlet box and below the second end of the regulating plate. The stop block is fixedly connected to the liquid inlet box. The cross-sectional shape of the stop block is circular. There is a gap between the stop block and the second end of the regulating plate. A first control assembly is provided on the top surface of the liquid inlet box for driving the regulating plate to rotate.

[0008] Preferably, the first control assembly includes: a driving block. A sliding groove is provided on the top surface of the second end of the liquid inlet box. The length extension direction of the sliding groove is the same as that of the regulating plate. The driving block is provided in the sliding groove and can slide along the inner wall of the sliding groove. The top surface of the driving block is provided with anti-slip lines. A metal plate is fixedly provided on the top surface of the middle part of the regulating plate and directly below the first end of the sliding groove. A clamping groove is provided on the side wall of the first end of the sliding groove. A clamping block is fixedly provided on the side wall of the driving block. The shape of the clamping block matches the shape of the clamping groove. A first magnet is fixedly provided on the bottom surface of the driving block. A second magnet is fixedly provided on the top surface of the second end of the regulating plate and directly below the second end of the sliding groove. The same-sex magnetic poles of the first magnet and the second magnet are arranged oppositely.

[0009] Preferably, a connecting seat is fixedly provided on the end surface of the liquid baffle away from the C-shaped frame. The first end of the pull rod is detachably connected to the connecting seat. A second avoidance hole is provided on the side wall of the cell culture box. The pull rod is provided in the second avoidance hole. The diameter of the second avoidance hole is greater than the diameter of the pull rod.

[0010] Preferably, in the vertical direction, the positions of the bottom layer chamber, several middle layer chambers and the top layer chamber correspond to each other. A position adjusting assembly is provided on the side of the cell culture box away from the C-shaped frame. The second ends of several pull rods are all connected to the position adjusting assembly. The position adjusting assembly is used for driving several pull rods to move synchronously.

[0011] Preferably, the positioning assembly includes: a first driving rod, which is arranged above the second avoidance hole at the top layer; a second driving rod, which is arranged below the second avoidance hole at the bottom layer; a third driving rod, which is arranged between the first driving rod and the second driving rod, and a ring is fixedly provided at the second end of each pull rod, and the third driving rod passes through a plurality of the rings, and the inner diameter of the ring is the same as the diameter of the third driving rod, and the third driving rod can slide along the inner wall of the ring, and the top of the third driving rod is detachably connected to the first driving rod, and the bottom of the third driving rod is detachably connected to the second driving rod.

[0012] Preferably, the third driving rod includes a plurality of assembly columns, and the plurality of assembly columns are stacked in sequence from bottom to top, and two adjacent assembly columns are detachably connected, the assembly column on the top layer is detachably connected to the first driving rod, and the assembly column on the bottom layer is detachably connected to the top of the insertion rod, and a socket is provided on the top surface of the second driving rod, and the socket passes through the second driving rod, and the bottom of the insertion rod extends into the socket and can slide along the inner wall of the socket.

[0013] Preferably, the positioning assembly further includes a second control assembly, which includes: two first connecting rods, both of which are arranged on the side of the first driving rod away from the cell culture box, the first connecting rod is in a V-shaped structure, the first end of the first connecting rod is fixedly connected to the first driving rod, the height of the first end of the first connecting rod is higher than the second end, and there is a gap between the two first connecting rods; a second connecting rod, which is arranged on the side of the second driving rod away from the cell culture box, the second connecting rod is in a V-shaped structure, the first end of the second connecting rod is fixedly connected to the second driving rod, the height of the first end of the second connecting rod is lower than the second end, the second connecting rod is located between the two first connecting rods, and the width of the gap between the two first connecting rods is the same as the width of the second connecting rod; an elastic band, which is sleeved on the second ends of the two first connecting rods and the second end of the second connecting rod, the second end of the first connecting rod is provided with a first groove, the second end of the second connecting rod is provided with a second groove, the two ends of the elastic band are respectively arranged in the first groove and the second groove, and the width of the elastic band, the width of the first groove and the width of the second groove are all the same.

[0014] Preferably, a force-bearing rod is provided at each end of the second connecting rod, and the length extension direction of the force-bearing rod is perpendicular to the length extension direction of the second connecting rod. The second connecting rod is fixedly connected to the first end of the force-bearing rod, and a handle is provided on each side of the bottom cover, and the handle is fixedly connected to the bottom cover. A guide groove is provided on the bottom surface of the handle, and the second end of the force-bearing rod extends into the guide groove and can slide along the inner wall of the guide groove. The length of the force-bearing rod is less than the length of the pull rod.

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

[0016] (i) In the present invention, the liquid baffle plate can block the entrance of each chamber. Only after the liquid baffle plate moves can the nutrient solution enter each chamber. At the same time, the liquid baffle plate can also exhaust all the air in each chamber. The air will not occupy the space in each chamber, and will not hinder the cell culture process.

[0017] (ii) The present invention also includes a perfusion component, which includes a bubble guide component. The bubble guide component can drive the bubbles in the nutrient solution in the liquid inlet box to move, so that the bubbles can definitely move to the exhaust port, preventing the bubbles from staying inside the liquid inlet box.

[0018] (III) The present invention further comprises a positioning assembly, which comprises a first driving rod, a second driving rod, a third driving rod and a second control assembly. The positioning assembly is used to drive a plurality of pull rods to move synchronously, the second control assembly can ensure that the positioning assembly has no tendency to rotate, and the third driving rod can be normally away from the cell culture box. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 An axonometric view of the present invention at one angle;

[0020] Figure 2 It is an axonometric view of another angle of the present invention;

[0021] Figure 3 for Figure 2 The enlarged view of point A in the middle;

[0022] Figure 4 is an axonometric view of the bottom cover of the present invention;

[0023] Figure 5 is an axonometric view of the partition in the present invention;

[0024] Figure 6 for Figure 5 The enlarged view of point B in the middle;

[0025] Figure 7 It is an axonometric view of the top cover in the present invention;

[0026] Figure 8 for Figure 7 Enlarged view of point C in the middle;

[0027] Fig. 9 It is an axonometric view of the liquid extraction component in the present invention;

[0028] Fig.10 for Fig. 9 The enlarged view of point D in the middle;

[0029] Fig.11 It is a front cross-sectional view of the perfusion assembly of the present invention when the adjustment plate is away from the stopper;

[0030] Fig.12 for Fig.11 Enlarged view of point E in the middle;

[0031] Fig.13 It is a front cross-sectional view of the perfusion assembly of the present invention after the adjustment plate is close to the stopper;

[0032] Fig.14 is an axonometric exploded view of the first driving rod and the third driving rod in the present invention;

[0033] Fig.15 for Fig.14 The enlarged view of F in the middle;

[0034] Fig.16 for Fig.14 Enlarged view of G in the middle;

[0035] Fig.17 is an axonometric view of the second driving rod in the present invention;

[0036] Fig.18 is an axonometric cross-sectional view of the present invention;

[0037] Fig.19 It is an axonometric diagram of the present invention after the position adjustment component drives the liquid extraction component to move;

[0038] Fig. 20 for Fig.19 The enlarged view of the H in the middle;

[0039] Fig.21 It is an axonometric view of the present invention after the positioning assembly is removed;

[0040] Fig. 22 Axonometric view of the present invention after reducing the number of partitions.

[0041] Reference numerals include:

[0042] 1-cell culture box, 11-bottom cover, 12-partition, 13-top cover, 14-shaped frame, 15-first liquid inlet space, 16-second liquid inlet space, 17-first avoidance hole, 18-second avoidance hole, 19-handle, 191-guide groove, 2-liquid extraction assembly, 21-liquid baffle, 211-connecting seat, 22-pull rod, 23-liquid outlet pipe, 24-plug, 25-ring, 3-perfusion assembly, 31-liquid inlet box, 311-exhaust port, 312-slide, 313-clamping groove, 32-liquid inlet pipe, 33-exhaust pipe, 34-pipe cap, 35-bubble guide assembly, 351-adjustment Plate, 352-block, 353-first control component, 3531-driving block, 35311-anti-slip groove, 35312-clamping block, 3532-metal plate, 3533-first magnet, 3534-second magnet, 4-positioning component, 41-first driving rod, 42-second driving rod, 421-jack, 43-third driving rod, 431-assembly column, 432-insertion rod, 44-second control component, 441-first connecting rod, 4411-first groove, 442-second connecting rod, 4421-second groove, 443-elastic binding band, 45-force rod, 5-drain pipe. DETAILED DESCRIPTION

[0043] 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.

[0044] Embodiment 1

[0045] See also Figure 1-22 The present invention provides a technical solution: a constant temperature cell culture device for the research and development of an inactivated vaccine for Akabane disease in cattle, comprising a cell culture box 1, wherein the cell culture box 1 comprises a bottom cover 11, a plurality of partitions 12 and a top cover 13 stacked in sequence from bottom to top. The top surface of the bottom cover 11 is provided with a first tank, the top and bottom surfaces of the partition 12 are both provided with a second tank, and the bottom surface of the top cover 13 is both provided with a third tank, the first tank and the second tank form a bottom chamber, the two opposite second tanks form a middle chamber, and the second tank and the third tank form a top chamber. The number of partitions 12 can be increased or decreased according to actual needs, and the height and internal volume of the cell culture box 1 can be changed to facilitate the control of the number of cultured cells, so that a large number of cells can be harvested in one culture. The bottom cover 11, the plurality of partitions 12 and the top cover 13 can be fixed together by welding, and the cell culture box 1 has a sealing function, which helps to reduce the risk of contamination during cell culture.

[0046] See also Figure 1-2 , Figure 5-9 and Figure 18-20 , above the bottom cover 11, there are also a number of U-shaped frames 14 stacked in sequence from bottom to top. A first liquid inlet space 15 is formed between the U-shaped frame 14 and the partition plate 12, and a second liquid inlet space 16 is formed between the U-shaped frame 14 and the top cover 13. Both the partition plate 12 and the top cover 13 can be fixed to the U-shaped frame 14 by welding. A liquid pumping assembly 2 is provided in each of the bottom chamber, the middle chamber, and the top chamber. The liquid pumping assembly 2 includes a liquid blocking plate 21. The liquid blocking plate 21 is connected to the first end of the pull rod 22, and the second end of the pull rod 22 extends to the outside of the cell culture box 1.

[0047] Please refer to Figure 1-2 , Figure 5-9 and Figure 18-20 , when the present invention is in use, first, push all the pull rods 22 to drive all the liquid blocking plates 21 to move forward to block the entrances at the front ends of each chamber. Then, inject nutrient solution into the first liquid inlet space 15 and the second liquid inlet space 16. The nutrient solution can gradually fill the first liquid inlet space 15 and the second liquid inlet space 16. At this time, the nutrient solution cannot enter each chamber. And during this process, the bubbles in the nutrient solution in the first liquid inlet space 15 and the second liquid inlet space 16 can float up and quickly leave the first liquid inlet space 15 and the second liquid inlet space 16. When the first liquid inlet space 15 and the second liquid inlet space 16 are filled with the nutrient solution, slowly pull the pull rod 22 to drive the liquid blocking plate 21 to move backward, and the nutrient solution can enter each chamber from the first liquid inlet space 15 and the second liquid inlet space 16. While pulling the pull rod 22, continue to inject nutrient solution into the first liquid inlet space 15 and the second liquid inlet space 16. The liquid blocking plate 21 moves to the limit position and abuts against the rear side walls of each chamber, and the nutrient solution will fill the bottom chamber, the middle chamber, and the top chamber. During the process of the liquid blocking plate 21 moving backward, the air originally existing in each chamber will be discharged to the outside of the cell culture box 1 through the gap between the pull rod 22 and the side wall of the cell culture box 1. Since there will be no bubble residue in the first liquid inlet space 15 and the second liquid inlet space 16, no bubbles will enter the bottom chamber, the middle chamber, and the top chamber. The air will not occupy the space in each chamber and will not hinder the process of cell culture.

[0048] Embodiment 2

[0049] On the basis of Embodiment 1, please refer to Figure 1-2 , the present invention further includes a perfusion assembly 3. The perfusion assembly 3 includes a liquid inlet box 31. The liquid inlet box 31 is arranged above the topmost U-shaped frame 14. A liquid inlet pipe 32 is provided on the side wall of the liquid inlet box 31. Nutrient solution can be injected into the liquid inlet box 31 through the liquid inlet pipe 32, and the nutrient solution flows downward and enters the first liquid inlet space 15 and the second liquid inlet space 16 in the cell culture box 1. Please refer to Figure 1-3 and Figure 9-10The liquid baffle 21 is fixedly connected to the first end of the liquid outlet pipe 23, and the second end of the liquid outlet pipe 23 is provided with a plug 24. The plug 24 and the liquid outlet pipe 23 are detachably connected, and a first avoidance hole 17 is provided on the side wall of the cell culture box 1. When the liquid baffle 21 blocks the entrance of the front end of each chamber, due to the presence of the plug 24, the nutrient solution cannot pass through the liquid outlet pipe 23 and move to the other side of the liquid baffle 21. Please refer to Figure 9-10 A connecting seat 211 is also provided on the liquid baffle plate 21, and the first end of the pull rod 22 is detachably connected to the connecting seat 211. A second avoidance hole 18 is provided on the side wall of the cell culture box 1, and the pull rod 22 is arranged in the second avoidance hole 18. The diameter of the second avoidance hole 18 is larger than the diameter of the pull rod 22, and the diameter of the second avoidance hole 18 is larger than the diameter of the connecting seat 211.

[0050] See also Figure 19-21 , during the backward movement of the liquid baffle plate 21, the air originally existing in each chamber will be discharged to the outside of the cell culture box 1 through the second avoidance hole 18. When the liquid baffle plate 21 moves to the extreme position and abuts against the rear side wall of each chamber, the second end of the liquid outlet pipe 23 extends into the first avoidance hole 17, the plug 24 extends to the outside of the cell culture box 1, the connecting seat 211 extends into the second avoidance hole 18, and the pull rod 22 extends to the outside of the cell culture box 1 as a whole. In this embodiment, the plug 24 is threadedly connected to the liquid outlet pipe 23, and the pull rod 22 is threadedly connected to the connecting seat 211. Next, the pull rod 22 is first twisted to remove the pull rod 22, and then the plug 24 is twisted to remove the plug 24, and then the drain pipe 5 is installed at the second end of each liquid outlet pipe 23. Several drain pipes 5 can be connected to the collection bottle, the collection bottle is connected to the peristaltic pump through a circulation pipeline, and the peristaltic pump is connected to the inlet pipe 32 through the inlet pipeline to form a perfusion system. Therefore, the present invention can realize perfusion culture, which can continuously and uninterruptedly provide nutrient delivery for the cell growth process. At the same time, dynamic culture is closer to the growth environment of cells in the body, which is beneficial to improving cell activity to express more proteins, monoclonal antibodies, cytokines, etc., and is beneficial to obtaining more accurate and effective experimental data in the research and development process of bovine Akabane disease inactivated vaccine.

[0051] Embodiment 3

[0052] Based on Example 2, please refer to Figure 1-2 and Figure 11-13The height of the first end of the liquid inlet box 31 is greater than the height of the second end, and an exhaust port 311 is provided on the top surface of the first end of the liquid inlet box 31. An exhaust pipe 33 is provided in the exhaust port 311. A pipe cap 34 is provided on the top of the exhaust pipe 33. The pipe cap 34 is detachably connected to the exhaust pipe 33. When the present invention is in use, after the liquid baffle 21 blocks the entrance of the front end of each chamber, the pipe cap 34 is screwed off, and the nutrient solution is injected into the liquid inlet box 31 through the liquid inlet pipe 32. The first liquid inlet space 15, the second liquid inlet space 16 and the liquid inlet box 31 will be gradually filled, and the original air in the first liquid inlet space 15, the second liquid inlet space 16 and the liquid inlet box 31, as well as the bubbles in the nutrient solution can float up and move toward the exhaust port 311 along the top inner wall of the liquid inlet box 31, and the air and bubbles can be discharged from the liquid inlet box 31 through the exhaust pipe 33. When the air in the liquid inlet box 31 is exhausted, the tube cap 34 is screwed back on, and the pull rod 22 is slowly pulled to drive the liquid baffle 21 to move backward, and the nutrient solution is continuously injected into the liquid inlet box 31 through the liquid inlet tube 32 .

[0053] See also Figure 1-2 , Figure 11-13 and Fig.18 The perfusion assembly 3 also includes a bubble guide assembly 35, which can drive the bubbles in the nutrient solution in the liquid inlet box 31 to move, so that the bubbles can definitely move to the exhaust port 311, and prevent the bubbles from staying inside the liquid inlet box 31. The bubble guide assembly 35 includes an adjustment plate 351, a stopper 352 and a first control assembly 353. The first end of the adjustment plate 351 is hinged to the inner wall of the liquid inlet box 31, the stopper 352 is located below the second end of the adjustment plate 351, and the first control assembly 353 is arranged on the top surface of the liquid inlet box 31. The cross-sectional shape of the stopper 352 is circular, so when the bubble contacts the bottom of the stopper 352, it can move along the side wall of the stopper 352 to the top of the stopper 352, and the bubble will not be restricted by the stopper 352. When the bubble floats up in the liquid inlet box 31, it will first contact the bottom surface of the adjustment plate 351, and then the first control assembly 353 works to drive the adjustment plate 351 to rotate, and the second end of the adjustment plate 351 swings back and forth. The stopper 352 can limit the second end of the adjustment plate 351. When the second end of the adjustment plate 351 hits the stopper 352, the adjustment plate 351 cannot continue to rotate downward. During the reciprocating swing of the adjustment plate 351, the bubbles below the adjustment plate 351 will gradually be pushed toward the exhaust port 311.

[0054] See also Figure 11-13The first control assembly 353 includes a driving block 3531, a metal plate 3532, a first magnet 3533 and a second magnet 3534. The driving block 3531 is arranged in the slide groove 312 on the top surface of the liquid inlet box 31, and the driving block 3531 is provided with an anti-slip pattern 35311 and a clamping block 35312, and a clamping groove 313 is provided on the side wall of the slide groove 312. The first magnet 3533 is fixedly arranged on the bottom surface of the driving block 3531, the metal plate 3532 is fixedly arranged in the middle of the adjustment plate 351, and the second magnet 3534 is fixedly arranged at the second end of the adjustment plate 351, and the like magnetic poles of the first magnet 3533 and the second magnet 3534 are arranged oppositely.

[0055] See also Figure 11-13 When the present invention is not in use, the clamping block 35312 is located in the clamping groove 313 to prevent the driving block 3531 from moving at will. At this time, the first magnet 3533 can absorb the metal plate 3532, and the adjustment plate 351 is close to the top inner wall of the liquid inlet box 31. After the nutrient solution is injected into the liquid inlet box 31, the operator presses the anti-slip pattern 35311 to drive the driving block 3531 to move back and forth in the slide groove 312. If the driving block 3531 moves to the top of the second magnet 3534, the first magnet 3533 and the second magnet 3534 repel each other, and the second magnet 3534 drives the adjustment plate 351 to swing downward and hit the stopper 352. If the driving block 3531 moves to the top of the metal plate 3532, the first magnet 3533 attracts the metal plate 3532, and the metal plate 3532 drives the adjustment plate 351 to swing upward and again close to the top inner wall of the liquid inlet box 31. The above operation is repeated continuously until the regulating plate 351 gradually pushes the bubbles in the liquid inlet box 31 toward the exhaust port 311 .

[0056] Embodiment 4

[0057] Based on Example 2, please refer to Figure 1-2 The present invention further comprises a position adjustment component 4, which is arranged on the side of the cell culture box 1 away from the shaped frame 14, and the second ends of the plurality of pull rods 22 are connected to the position adjustment component 4, and the position adjustment component 4 is used to drive the plurality of pull rods 22 to move synchronously. Figure 1-2 and Figure 14-17The positioning assembly 4 includes a first driving rod 41, a second driving rod 42 and a third driving rod 43. A collar 25 is fixedly provided at the second end of each pull rod 22. The third driving rod 43 passes through a plurality of collars 25. The top of the third driving rod 43 is detachably connected to the first driving rod 41, and the bottom of the third driving rod 43 is detachably connected to the second driving rod 42. When the first driving rod 41 and the second driving rod 42 move away from the cell culture box 1 at the same time, the third driving rod 43 can be driven to move together, and the third driving rod 43 drives the plurality of pull rods 22 connected thereto to move together, so that the plurality of liquid baffles 21 are away from the entrances of the respective chambers, and the nutrient solution can enter the bottom chamber, the middle chamber and the top chamber at the same time.

[0058] See also Figure 1-2 , Figure 14-17 The third driving rod 43 includes a plurality of assembly columns 431, two adjacent assembly columns 431 are detachably connected, the assembly column 431 at the top is detachably connected to the first driving rod 41, and the assembly column 431 at the bottom is detachably connected to the top of the insertion rod 432. In this embodiment, the assembly column 431, the first driving rod 41 and the insertion rod 432 are all connected by threads. A plug hole 421 is provided on the top surface of the second driving rod 42, and the bottom of the insertion rod 432 extends into the plug hole 421. Please refer to Figure 1-2 and Fig. 22 When the number of partitions 12 is increased or decreased according to actual needs, the length of the third driving rod 43 can also be changed, and it is only necessary to increase or decrease the number of assembly columns 431 so that the size of the adjustment component 4 matches the cell culture box 1.

[0059] In the present invention, the first driving rod 41 and the second driving rod 42 must move simultaneously for the position adjustment assembly 4 to work properly. If the operator only pulls one of the first driving rod 41 or the second driving rod 42, the position adjustment assembly 4 will tend to rotate, causing the cell culture box 1 to apply a reaction force to the pull rod 22, and the third driving rod 43 cannot normally move away from the cell culture box 1. Fig.19 The positioning component 4 further includes a second control component 44 , which can ensure that the positioning component 4 has no tendency to rotate and the third driving rod 43 can be normally away from the cell culture box 1 .

[0060] See also Figure 1-2 , Figure 14-17 and Fig.19The second control assembly 44 includes two first connecting rods 441, a second connecting rod 442 and an elastic band 443. The first connecting rod 441 and the second connecting rod 442 are both V-shaped structures. The first connecting rod 441 is fixedly connected to the first driving rod 41, and there is a gap between the two first connecting rods 441. The second connecting rod 442 is fixedly connected to the second driving rod 42, and the second connecting rod 442 is located between the two first connecting rods 441. The elastic band 443 is sleeved on the two first connecting rods 441 and the second connecting rod 442. When the second control assembly 44 is required to work, the operator pulls the elastic band 443, and the top and bottom of the elastic band 443 can pull the first connecting rod 441 and the second connecting rod 442 to move simultaneously, and the first connecting rod 441 and the second connecting rod 442 can respectively pull the first driving rod 41 and the second driving rod 42 to move simultaneously. During this process, the first driving rod 41 and the second driving rod 42 are evenly stressed, the positioning assembly 4 has no tendency to rotate, and the third driving rod 43 can be normally away from the cell culture box 1 .

[0061] See also Figure 14-17 and Fig.19 The second end of the first connecting rod 441 is provided with a first groove 4411, the second end of the second connecting rod 442 is provided with a second groove 4421, and the two ends of the elastic band 443 are respectively arranged in the first groove 4411 and the second groove 4421, and the width of the elastic band 443, the width of the first groove 4411 and the width of the second groove 4421 are all the same. Therefore, when the operator pulls the elastic band 443, the elastic band 443 will not be separated from the first connecting rod 441 and the second connecting rod 442.

[0062] See also Figure 2 and Fig.19 When the height of the cell culture box 1 is high, the top of the elastic band 443 is connected to the first connecting rod 441, and the bottom of the elastic band 443 is connected to the second connecting rod 442, and the second connecting rod 442 does not enter the inner side of the two first connecting rods 441. Fig.19 and Fig. 22 When the height of the cell culture box 1 is shorter, the top of the elastic band 443 is connected to the second connecting rod 442, and the bottom of the elastic band 443 is connected to the first connecting rod 441, and the second connecting rod 442 enters the inner side of the two first connecting rods 441. In either case, the elastic band 443 can fix the first connecting rod 441 and the second connecting rod 442 together. When it is necessary to change the length of the third driving rod 43, it is only necessary to remove the elastic band 443 from the first connecting rod 441 and the second connecting rod 442, and lift the first driving rod 41 upward, so that the plurality of assembly columns 431 can leave the ring 25, and the insertion rod 432 can leave the insertion hole 421, and finally the number of assembly columns 431 can be increased or decreased as needed.

[0063] See also Figure 19-21 When the third driving rod 43 drives the plurality of pull rods 22 to move to the limit position, the elastic band 443 is removed, the first driving rod 41 is lifted upward to make the third driving rod 43 leave the collar 25, and then each pull rod 22 is twisted to remove the pull rod 22 from the connecting seat 211. Then, the plurality of plugs 24 are removed, and the plurality of drain pipes 5 can be installed.

[0064] See also Figure 1-2 and Figure 4 A handle 19 is provided on each side of the bottom cover 11, and the present invention can be stably picked up by the handle 19, so that the present invention is easy to move. Figure 4 , Fig.17 and Fig.19 , a force-bearing rod 45 is provided at each end of the second connecting rod 442, and a guide groove 191 is provided on the bottom surface of the handle 19. When a plurality of liquid baffles 21 block the entrance at the front end of each chamber, the force-bearing rod 45 extends into the guide groove 191, and the force-bearing rod 45 is received inside the handle 19. When the third driving rod 43 moves away from the cell culture box 1, the second driving rod 42 drives the force-bearing rod 45 to leave the handle 19. The force-bearing rod 45 can increase the stability of the position adjustment component 4, and the position adjustment component 4 itself will not fall over regardless of whether the position adjustment component 4 is connected to the liquid extraction component 2.

[0065] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations falling within the meaning and scope of the equivalent elements of the claims be included in the invention. Any reference numeral in a claim should not be considered as limiting the claim to which it relates.

[0066] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A constant-temperature cell culture device for the research and development of an inactivated vaccine against bovine Akabane disease, comprising a cell culture box. The cell culture box includes a bottom cover, a plurality of partition plates, and a top cover stacked in sequence from bottom to top. A bottom layer chamber is provided between the bottom cover and the lowermost partition plate, a middle layer chamber is provided between adjacent two partition plates, and a top layer chamber is provided between the topmost partition plate and the top cover. It is characterized in that: The length of the bottom cover is greater than the length of the partition plate, the length of the partition plate is the same as the length of the top cover. A plurality of U-shaped frames stacked in sequence from bottom to top are further provided above the bottom cover. The plurality of U-shaped frames are respectively fixedly connected to the plurality of partition plates and the top cover. A first liquid inlet space is formed between the U-shaped frame and the partition plate, and a second liquid inlet space is formed between the U-shaped frame and the top cover. The first liquid inlet space, the second liquid inlet space, the bottom layer chamber, the middle layer chamber, and the top layer chamber are communicated with each other; The cross-sectional shapes of the bottom layer chamber, the middle layer chamber, and the top layer chamber are the same. Liquid extraction components are provided in the bottom layer chamber, the middle layer chamber, and the top layer chamber. The liquid extraction component includes a liquid blocking plate. The cross-sectional shape of the liquid blocking plate matches the cross-sectional shape of the bottom layer chamber. The liquid blocking plate is connected to the first end of a pull rod. The pull rod is located on the side away from the U-shaped frame. The length of the pull rod is greater than the length of the bottom cover. The second end of the pull rod extends to the outside of the cell culture box.

2. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 1, characterized in that: It further includes a perfusion component. The perfusion component includes a liquid inlet box. The liquid inlet box is provided above the topmost U-shaped frame and is fixedly connected to the U-shaped frame. The inside of the liquid inlet box is communicated with the second liquid inlet space. A liquid inlet pipe is provided on the side wall of the liquid inlet box; The liquid blocking plate is fixedly connected to the first end of a liquid outlet pipe. The length of the liquid outlet pipe is greater than the width of the liquid blocking plate. The liquid outlet pipe penetrates through the liquid blocking plate. The second end of the liquid outlet pipe is located on the side away from the U-shaped frame. A plug is provided at the second end of the liquid outlet pipe. The first end of the plug extends into the liquid outlet pipe. The diameter of the second end of the plug is the same as the diameter of the liquid outlet pipe. The plug is detachably connected to the liquid outlet pipe. A first avoidance hole is provided on the side wall of the cell culture box. The position of the first avoidance hole corresponds to the position of the liquid outlet pipe. The shape of the first avoidance hole matches the cross-sectional shape of the liquid outlet pipe.

3. The constant temperature cell culture equipment for the research and development of the inactivated vaccine of bovine Akabane disease according to claim 2, characterized in that: The height of the first end of the liquid inlet box is greater than the height of the second end. An exhaust port is provided on the top surface of the first end of the liquid inlet box. A exhaust pipe is provided in the exhaust port. The exhaust pipe is vertically arranged and is communicated with the inside of the liquid inlet box. A pipe cap is provided at the top of the exhaust pipe. The pipe cap is detachably connected to the exhaust pipe; The perfusion component further includes a bubble guiding component. The bubble guiding component includes: The regulating plate is arranged in the liquid inlet box, between the exhaust port and the second end of the liquid inlet box. The first end of the regulating plate is hinged to the inner wall of the liquid inlet box. The first end of the regulating plate and the exhaust pipe are on the same side. The liquid inlet pipe and the exhaust port are on the same side and below the exhaust port. The stopper is arranged in the liquid inlet box and below the second end of the regulating plate. The stopper is fixedly connected to the liquid inlet box. The cross-sectional shape of the stopper is circular. There is a gap between the stopper and the second end of the regulating plate. The first control assembly is arranged on the top surface of the liquid inlet box and is used to drive the regulating plate to rotate.

4. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 3, characterized in that: The first control assembly includes: The driving block. A sliding groove is arranged on the top surface of the second end of the liquid inlet box. The length extension direction of the sliding groove is the same as the length extension direction of the regulating plate. The driving block is arranged in the sliding groove and can slide along the inner wall of the sliding groove. The top surface of the driving block is provided with anti-slip lines. The metal plate is fixedly arranged on the top surface of the middle part of the regulating plate and directly below the first end of the sliding groove. A clamping groove is arranged on the side wall of the first end of the sliding groove. A clamping block is fixedly arranged on the side wall of the driving block. The shape of the clamping block matches the shape of the clamping groove. The first magnet is fixedly arranged on the bottom surface of the driving block. The second magnet is fixedly arranged on the top surface of the second end of the regulating plate and directly below the second end of the sliding groove. The same-sex magnetic poles of the first magnet and the second magnet are arranged oppositely.

5. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 1, characterized in that: A connecting seat is fixedly arranged on the end surface of the liquid baffle away from the U-shaped frame. The first end of the pull rod is detachably connected to the connecting seat. A second avoidance hole is arranged on the side wall of the cell culture box. The pull rod is arranged in the second avoidance hole. The diameter of the second avoidance hole is larger than the diameter of the pull rod.

6. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 5, characterized in that: In the vertical direction, the positions of the bottom layer chamber, several middle layer chambers and the top layer chamber correspond one by one. A position adjustment assembly is arranged on the side of the cell culture box away from the U-shaped frame. The second ends of several pull rods are all connected to the position adjustment assembly. The position adjustment assembly is used to drive several pull rods to move synchronously.

7. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 6, characterized in that: The position adjustment assembly includes: The first driving rod is arranged above the second avoidance hole at the topmost layer. The second driving rod is arranged below the second avoidance hole at the bottommost layer. The third driving rod is arranged between the first driving rod and the second driving rod. A collar is fixedly arranged at the second end of each pull rod. The third driving rod passes through several collars. The inner diameter of the collar is the same as the diameter of the third driving rod. The third driving rod can slide along the inner wall of the collar. The top of the third driving rod is detachably connected to the first driving rod. The bottom of the third driving rod is detachably connected to the second driving rod.

8. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 7, characterized in that: The third driving rod includes a plurality of assembly columns, which are stacked in sequence from bottom to top, and two adjacent assembly columns are detachably connected. The assembly column on the top layer is detachably connected to the first driving rod, and the assembly column on the bottom layer is detachably connected to the top of the insertion rod. A socket is provided on the top surface of the second driving rod, and the socket passes through the second driving rod. The bottom of the insertion rod extends into the socket and can slide along the inner wall of the socket.

9. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 7, characterized in that: The positioning component further includes a second control component, and the second control component includes: Two first connecting rods are both arranged on a side of the first driving rod away from the cell culture box, the first connecting rod is in a V-shaped structure, the first end of the first connecting rod is fixedly connected to the first driving rod, the first end of the first connecting rod is higher than the second end, and there is a gap between the two first connecting rods; a second connecting rod, arranged on a side of the second driving rod away from the cell culture box, the second connecting rod being in a V-shaped structure, the first end of the second connecting rod being fixedly connected to the second driving rod, the height of the first end of the second connecting rod being lower than the second end, the second connecting rod being located between the two first connecting rods, and the width of the gap between the two first connecting rods being the same as the width of the second connecting rod; An elastic band is sleeved on the second ends of the two first connecting rods and the second ends of the second connecting rods. The second ends of the first connecting rods are provided with a first groove, and the second ends of the second connecting rods are provided with a second groove. The two ends of the elastic band are respectively arranged in the first groove and the second groove. The width of the elastic band, the width of the first groove and the width of the second groove are all the same.

10. The constant temperature cell culture equipment for the research and development of the inactivated vaccine for Akabane disease of cattle according to claim 9, characterized in that: A force-bearing rod is provided at each end of the second connecting rod, and the length extension direction of the force-bearing rod is perpendicular to the length extension direction of the second connecting rod. The second connecting rod is fixedly connected to the first end of the force-bearing rod. A handle is provided on each side of the bottom cover, and the handle is fixedly connected to the bottom cover. A guide groove is provided on the bottom surface of the handle, and the second end of the force-bearing rod extends into the guide groove and can slide along the inner wall of the guide groove. The length of the force-bearing rod is less than the length of the pull rod.

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