Cell culture external circulation ultrafiltration device

Abstract

The utility model discloses a cell culture external circulation ultrafiltration device which is characterized in that a backflow pipeline is provided with a second valve, a second peristaltic pump and a waste liquid ultrafiltration fiber column, the liquid inlet end of a filtering pipeline is connected with a reaction bottle, the liquid outlet end of the filtering pipeline is connected with a circulation bottle, the liquid inlet end of the backflow pipeline is connected with the circulation bottle, and the liquid outlet end of the backflow pipeline is connected with the circulation bottle. The first valve is arranged close to the circulating bottle, the second valve is arranged close to the reaction bottle, and the first peristaltic pump and the second peristaltic pump are both bidirectional peristaltic pumps. Liquid in the reaction bottle flows to the circulation bottle through the first peristaltic pump and the harvesting ultrafiltration fiber column, cells are intercepted by the harvesting ultrafiltration fiber column, then the first valve is closed, the first peristaltic pump is reversely started, the cells flow back into the reaction bottle, and the operation is repeated, so that products can be efficiently and continuously harvested, and the device is particularly suitable for perfusion culture; reutilization of the culture medium is realized, and biological material cost is reduced.

Description

Technical Field

[0001] The utility model relates to the technical field of cell culture, in particular to an external circulation ultrafiltration device for cell culture. Background Art

[0002] Fixed-bed bioreactors have been successfully used for many years in high-throughput perfusion processes for vaccines and gene therapy, facilitating scale-up and commercial production. Perfusion culture systems require large volumes of culture medium to be continuously input and output, resulting in a low degree of automation throughout the entire culture process. Even the slightest operational error during the extensive manual operation can easily lead to contamination, necessitating termination and re-initiation of the culture. This is not only difficult and labor-intensive, but also leads to low production efficiency and limits the ability to increase culture volume. Therefore, designing an efficient external circulation perfusion device is crucial for the continuous collection of harvested products, removal of metabolic waste, recycling of culture medium, replenishment, circulation, and harvesting. Utility Model Content

[0003] Aiming at the technical problem of low automation level of existing perfusion culture systems, the utility model proposes a cell culture external circulation ultrafiltration device, which realizes the reuse of culture medium, reduces the cost of biological materials, avoids the risk of contamination in manual operation, and has high efficiency.

[0004] The technical solution adopted by the present invention is as follows: A cell culture external circulation ultrafiltration device, comprising a circulation bottle, a reaction bottle, a filtration pipeline and a reflux pipeline, the filtration pipeline is provided with a first valve, a first peristaltic pump and a harvest liquid ultrafiltration fiber column, the reflux pipeline is provided with a second valve, a second peristaltic pump and a waste liquid ultrafiltration fiber column, the liquid inlet end of the filtration pipeline is connected to the reaction bottle, the liquid outlet end of the filtration pipeline is connected to the circulation bottle, the liquid inlet end of the reflux pipeline is connected to the circulation bottle, the liquid outlet end of the reflux pipeline is connected to the circulation bottle, the first valve is arranged close to the circulation bottle, the second valve is arranged close to the reaction bottle, and the first peristaltic pump and the second peristaltic pump are respectively bidirectional peristaltic pumps.

[0005] Optionally, the pore size range of the harvested liquid ultrafiltration fiber column is 0.1 to 0.5 μm, and the pore size range of the waste liquid ultrafiltration fiber column is 10 to 30 nm.

[0006] Optionally, the filtering pipeline is provided with a first pressure controller, and the reflux pipeline is provided with a second pressure controller, the first pressure controller is connected to the first peristaltic pump, and the second pressure controller is connected to the second peristaltic pump.

[0007] Optionally, the reaction bottle is provided with an air inlet pipeline, an air outlet pipeline, a DO electrode, a humidity detector, an acid and alkali agent pipeline and a pH electrode.

[0008] Optionally, the reaction bottle is provided with a stirring shaft, and the stirring shaft is provided with stirring blades.

[0009] Optionally, a partition is provided inside the reaction bottle, which divides the inside of the reaction bottle into a reaction zone and a stirring zone. The stirring shaft and the stirring blade are located in the stirring zone. The partition is provided with multiple through holes that connect the stirring zone and the reaction zone.

[0010] Optionally, the circulation bottle is provided with a filter duct, a sampling duct and a reflux duct, the filter duct is connected to the filter pipeline, the reflux duct is connected to the reflux pipeline, the liquid outlet of the filter duct is against the inner wall of the circulation bottle, and the liquid inlet of the sampling duct and the liquid inlet of the reflux duct are located at the bottom of the circulation bottle.

[0011] Optionally, the bottle cap is provided with a through hole for the filter duct and the return duct to pass through, and an air filter element is installed in the through hole.

[0012] Optionally, a magnetic stirrer is provided at the bottom of the circulation bottle.

[0013] Optionally, the outer wall of the reaction bottle is covered with a thermostat.

[0014] The beneficial effects of the present invention are as follows: the liquid in the reaction bottle flows to the circulation bottle through the first peristaltic pump and the harvesting ultrafiltration fiber column, wherein the cells are retained by the harvesting ultrafiltration fiber column, and then the first valve is closed, the first peristaltic pump is started in reverse, and the cells flow back into the reaction bottle, and such operation is repeated, so that the product can be harvested efficiently and continuously, and it is particularly suitable for perfusion culture; when the reaction bottle needs to be replenished with fluid, the second peristaltic pump is turned on, and the liquid in the circulation bottle flows to the reaction bottle through the waste liquid ultrafiltration fiber column, the virus in the waste liquid is retained by the waste liquid ultrafiltration fiber column, and the liquid enters the reaction bottle for replenishment, thereby realizing the reuse of the culture medium, reducing the cost of biological materials, and avoiding the risk of contamination in manual operation, with high efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This is a schematic diagram of the structure of the cell culture external circulation ultrafiltration device proposed in an embodiment of the present utility model;

[0016] The markings in each accompanying drawing are: 1. circulation bottle; 2. reaction bottle; 3. filtration pipeline; 4. reflux pipeline; 5. first valve; 6. first peristaltic pump; 7. harvest liquid ultrafiltration fiber column; 8. second valve; 9. second peristaltic pump; 10. waste liquid ultrafiltration fiber column; 11. first pressure controller; 12. second pressure controller; 13. air inlet pipeline; 14. air outlet pipeline; 15. DO electrode; 16. humidity detector; 17. pH electrode; 18. acid and alkali agent pipeline; 19. stirring shaft; 20. stirring blade; 21. partition; 22. reaction zone; 23. stirring zone; 24. through hole; 25. filtration duct; 26. reflux duct; 27. bottle cap; 28. air filter element; 29. ​​magnetic stirrer; 30. sampling duct; 31. thermostat; 32. first collection port; 33. second collection port. DETAILED DESCRIPTION

[0017] The present application will be further described in detail below with reference to the accompanying drawings and examples.

[0018] like Figure 1As shown, this embodiment discloses a cell culture external circulation ultrafiltration device, including a circulation bottle 1, a reaction bottle 2, a filtration pipeline 3 and a reflux pipeline 4, the filtration pipeline 3 is provided with a first valve 5, a first peristaltic pump 6 and a harvest liquid ultrafiltration fiber column 7, the reflux pipeline 4 is provided with a second valve 8, a second peristaltic pump 9 and a waste liquid ultrafiltration fiber column 10, the liquid inlet end of the filtration pipeline 3 is connected to the reaction bottle 2, the liquid outlet end of the filtration pipeline 3 is connected to the circulation bottle 1, the liquid inlet end of the reflux pipeline 4 is connected to the circulation bottle 1, the liquid outlet end of the reflux pipeline 4 is connected to the circulation bottle 1, the first valve 5 is arranged close to the circulation bottle 1, the second valve 8 is arranged close to the reaction bottle 2, the first peristaltic pump 6 and the second peristaltic pump 9 are bidirectional peristaltic pumps respectively. The liquid in the reaction bottle 2 flows to the circulation bottle 1 through the first peristaltic pump 6 and the harvesting ultrafiltration fiber column, wherein the cells are retained by the harvesting ultrafiltration fiber column, and the harvesting liquid ultrafiltration fiber column 7 is provided with a first collecting port 32. The viruses and the waste ultrafiltered by the harvesting liquid ultrafiltration fiber column 7 are collected by the first collecting port 32, and then the first valve 5 is closed, and the first peristaltic pump 6 is started in reverse, and the cells flow back into the reaction bottle 2. Such repeated operations can efficiently and continuously harvest the product, which is particularly suitable for perfusion culture; when the reaction bottle 2 needs to be replenished with fluid, the second peristaltic pump 9 is turned on, and the liquid in the circulation bottle 1 flows to the reaction bottle 2 through the waste liquid ultrafiltration fiber column 10, and the viruses in the waste liquid are retained by the waste liquid ultrafiltration fiber column 10. The waste liquid ultrafiltration fiber column 10 is provided with a second collecting port 33. The viruses and the waste ultrafiltered by the waste liquid ultrafiltration fiber column 10 are collected by the second collecting port 33, and the liquid enters the reaction bottle 2 for replenishment, thereby realizing the reuse of the culture medium, reducing the cost of biological materials, and avoiding the risk of contamination in manual operation, with high efficiency. The pore size of the harvested liquid ultrafiltration fiber column 7 is in the range of 0.1 to 0.5 μm, and the pore size of the waste liquid ultrafiltration fiber column 10 is in the range of 10 to 30 nm. The harvested liquid ultrafiltration fiber column 7 allows viruses to pass through but does not allow cells to pass through. The waste liquid ultrafiltration fiber column 10 does not allow viruses to pass through.

[0019] like Figure 1As shown, the filtration line 3 is equipped with a first pressure controller 11, and the return line 4 is equipped with a second pressure controller 12. The first pressure controller 11 is connected to the first peristaltic pump 6, and the second pressure controller 12 is connected to the second peristaltic pump 9. The first pressure controller 11 controls the speed of the first peristaltic pump 6 by sensing changes in the hydraulic pressure within the filtration line 3, and the second pressure controller 12 controls the speed of the second peristaltic pump 9 by sensing changes in the hydraulic pressure within the return line 4. The reaction flask is equipped with an air inlet line 13, an air outlet line 14, a DO electrode 15, a humidity detector 16, an acid and alkali reagent line 18, and a pH electrode 17. The air inlet line 13 and the air outlet line 14 are used to balance the air pressure inside the reaction flask. The DO electrode 15 is used to detect the oxygen concentration in the reaction solution. The acid and alkali reagent line 18 is used to add acid or alkali reagents. The pH electrode is used to detect the pH value of the reaction solution. The humidity detector 16 is used to detect the solution humidity.

[0020] like Figure 1 As shown, the reaction flask is provided with a stirring shaft 19, which is provided with stirring blades 20. The stirring shaft 19 is connected to a stirring motor. A partition 21 is provided inside the reaction flask, which divides the interior of the reaction flask into a reaction zone 22 and a stirring zone 23. The stirring shaft 19 and stirring blades 20 are located in the stirring zone 23. The partition 21 is provided with a plurality of through holes 24 that connect the stirring zone 23 with the reaction zone 22. This prevents the stirring shaft and stirring blades from damaging the cells in the reaction zone 22 during high-speed rotation.

[0021] like Figure 1 As shown, the circulating bottle 1 is equipped with a filter conduit 25, a sampling conduit 30, and a return conduit 26. The filter conduit 25 is connected to the filter line 3, and the return conduit 26 is connected to the return line 4. The outlet of the filter conduit 25 abuts the inner wall of the circulating bottle 1, while the inlets of the sampling conduit and the return conduit 26 are located at the bottom of the circulating bottle 1. The outlet of the filter conduit 25 is close to the bottle wall, which prevents bubbles from forming when the liquid flows out. This reduces the risk of bubbles damaging the product. The circulating bottle 1 is equipped with a bottle cap 27, which has a through hole 24 for the filter conduit 25 and return conduit 26 to pass through. An air filter 28 is installed in the through hole 24. The air filter 28 is a disc-shaped element that passes through the filter conduit 25 and return conduit 26. The air filter 28 prevents external air from contaminating the culture solution in the bottle. A magnetic stirrer 29 is installed at the bottom of the circulating bottle 1. The magnetic rotor of the magnetic stirrer 29 is in the shape of an elliptical rugby ball. During the rotation process, the shear force on the cells is small and bubbles are not easily formed.

[0022] like Figure 1As shown, the outer wall of the reaction bottle is covered with a thermostat 31, which is a U-shaped container that controls the temperature of the reaction bottle and provides a stable growth temperature for the cell culture in the reaction bottle. The thermostat 31 is a prior art.

[0023] It is understandable that the specific embodiments described above are only used to explain the relevant utility model, rather than to limit the utility model. It should also be noted that, for the convenience of description, only the parts related to the utility model are shown in the accompanying drawings. Multiple technical solutions in the same embodiment, as well as multiple technical solutions in different embodiments, can be arranged and combined to form new technical solutions that do not have contradictions or conflicts. Any equivalent structural transformation made using the contents of the description and drawings of this utility model, directly or indirectly used in other related technical fields, are also included in the protection scope of this utility model.

Claims

1. A cell culture external circulation ultrafiltration device, characterized in that: It includes a circulation bottle, a reaction bottle, a filtration pipeline and a reflux pipeline. The filtration pipeline is provided with a first valve, a first peristaltic pump and a harvest liquid ultrafiltration fiber column. The reflux pipeline is provided with a second valve, a second peristaltic pump and a waste liquid ultrafiltration fiber column. The liquid inlet end of the filtration pipeline is connected to the reaction bottle, the liquid outlet end of the filtration pipeline is connected to the circulation bottle, the liquid inlet end of the reflux pipeline is connected to the circulation bottle, and the liquid outlet end of the reflux pipeline is connected to the circulation bottle. The first valve is arranged close to the circulation bottle, the second valve is arranged close to the reaction bottle, and the first peristaltic pump and the second peristaltic pump are respectively bidirectional peristaltic pumps.

2. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: The pore size of the harvested liquid ultrafiltration fiber column is in the range of 0.1 to 0.5 μm, and the pore size of the waste liquid ultrafiltration fiber column is in the range of 10 to 30 nm.

3. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: The filtering pipeline is provided with a first pressure controller, and the reflux pipeline is provided with a second pressure controller. The first pressure controller is connected to the first peristaltic pump, and the second pressure controller is connected to the second peristaltic pump.

4. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: The reaction bottle is provided with an air inlet pipeline, an air outlet pipeline, a DO electrode, a humidity detector, an acid and alkali agent pipeline and a pH electrode.

5. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: The reaction bottle is provided with a stirring shaft, and the stirring shaft is provided with stirring blades.

6. The cell culture external circulation ultrafiltration device according to claim 5, characterized in that: A partition is provided inside the reaction flask, which divides the inside of the reaction flask into a reaction zone and a stirring zone. The stirring shaft and the stirring blade are located in the stirring zone. The partition is provided with a plurality of through holes that connect the stirring zone and the reaction zone.

7. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: The circulation bottle is provided with a filter duct, a sampling duct and a reflux duct. The filter duct is connected to the filter pipeline, the reflux duct is connected to the reflux pipeline, the liquid outlet of the filter duct is against the inner wall of the circulation bottle, and the liquid inlet of the sampling duct and the liquid inlet of the reflux duct are located at the bottom of the circulation bottle.

8. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: The circulation bottle is provided with a bottle cap, the bottle cap is provided with a through hole for the filter conduit and the return conduit to pass through, and an air filter element is installed in the through hole.

9. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: A magnetic stirrer is provided at the bottom of the circulation bottle.

10. The cell culture external circulation ultrafiltration device according to claim 1, characterized in that: The outer wall of the reaction bottle is covered with a thermostat.

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