Perfusion module and perfusion culture system

Abstract

The present invention provides a perfusion module and a perfusion culture system, which include: a bioreactor module, including at least one bioreactor; a perfusion module, including at least one perfusion chamber, each perfusion chamber is connected to a pressure regulating pipe for air flow in or out, and the pressure regulating pipe is provided with a pump assembly that can open and close the pressure regulating pipe and change the air flow delivery direction; when there are multiple perfusion chambers, two adjacent perfusion chambers are connected through the pressure regulating pipe; the bioreactor, the perfusion module and the filtration module are connected in sequence, and if there are more than two perfusion chambers in the perfusion module, all the perfusion chambers are arranged in parallel; the cell culture fluid in the bioreactor flows into the perfusion chamber in the perfusion module, and then flows out of the perfusion chamber into the filter for filtration, and the permeate in the filter flows into the harvesting device through the permeate outlet. The present invention solves the problem of high shear force in the perfusion culture system.

Description

Technical Field

[0001] The invention relates to a perfusion culture device, in particular to a perfusion module and a perfusion culture system. Background Art

[0002] The mainstream direction of the current large-scale fermentation process of animal cells is the stirred suspension continuous fermentation process, especially the large-scale production of recombinant proteins such as antibodies. In industrial production, the amplification principle and process control of suspension culture process parameters are easier to understand and master than other fermentation systems. Therefore, this process is often selected in large-scale animal cell fermentation production. Suspension fermentation technology (Suspension Culture) refers to a fermentation method in which cells are freely suspended in the culture medium to grow and proliferate. At present, this technology is developing rapidly internationally and has gradually matured.

[0003] The perfusion culture process is a commonly used fermentation process, which mostly uses a stirred cell fermentation system, and a tubular system can also be used. During the fermentation process, the culture medium is continuously perfused into the fermenter and flows out at the same flow rate. The advantage of the perfusion process is that it can continuously remove cell debris and by-products during the fermentation process, and reduce the possible effects of various enzymes released after cell death on the product. At the same time, due to the continuous perfusion of fresh culture medium to provide a more constant culture environment, a higher unit volume yield can be obtained. Since the perfusion system mostly uses a cell retention and recovery system, high-density fermentation of cells can be achieved.

[0004] At present, the selection of animal cell culture methods is mainly determined by the characteristics of cell growth and the properties of the target protein. The significant advantage of perfusion culture is that the protein can be separated out in time with the culture medium, the protein stays in the reactor for a short time, and is less degraded by various hydrolases in the culture system, which is conducive to the improvement of protein quality. This feature is extremely beneficial for the production of chemically unstable proteins, such as enzymes and coagulation factors. When cell growth and protein quality are not limited by the culture method, for example, for the production of drugs such as antibodies with relatively stable chemical properties, whether to choose cell perfusion culture or fed-batch culture requires a comprehensive consideration of factors such as cost, benefit, scale, risk, and operational flexibility.

[0005] In the current diversified production environment, biotechnology companies are increasingly inclined to develop highly flexible and efficient production and manufacturing processes. Perfusion culture has been widely used in the bioengineering industry as an effective means to increase the yield of recombinant proteins with low stability. Perfusion culture can achieve the same amount of protein obtained by large-scale production of fed-batch culture with small bioreactors, realizing the miniaturization of culture scale and increasing the flexibility of operation.

[0006] With the large-scale application of suspended cells, the perfusion culture technology of suspended cells is also developing day by day. Cell retention is one of the process elements of suspended cell perfusion culture. How to effectively retain cells without damaging the cells has become the focus and challenge of the process. The current cell retention equipment is mainly designed based on the principles of filtration, sedimentation, and centrifugation, including rotary filters, vortex filters, inclined sedimentators, hydrocyclones, etc. The cell retention equipment designed based on the filtration principle can separate cells 100%, but the filter membrane is easily blocked by cell debris, defoaming agents, etc., which eventually leads to the termination of perfusion culture. Centrifugal separation of cells may cause cell damage. Neither sedimentation nor centrifugation can completely retain cells, and the cell separation effect will also be affected by the perfusion rate. In addition, whether it can be simply and effectively scaled up has also become a limiting condition for the application of most cell retention equipment.

[0007] The Alternating Tangential Flow (ATF) system designed based on the hollow fiber column retention principle has the advantages of effectively alleviating filter membrane clogging and low shear force, and is currently the best cell retention equipment. The hollow fiber column is connected to the reactor through a pipe, and the other side is driven by the repeated alternation of high-pressure air pumping and vacuum pumping negative pressure to drive the silicone diaphragm pump to move, so that the animal cell culture fluid passes through the hollow fiber column in and out of the reactor. This process also achieves full flushing of the hollow fiber column under low shear force, effectively reducing the blockage caused by the accumulation effect of the filter cake of the hollow fiber column. Using the ATF system, higher cell density and higher protein yield can be achieved at a higher perfusion rate, and it can also be effectively scaled up to large-scale production through the ratio of culture volume to hollow fiber column membrane area.

[0008] However, the above-mentioned alternating tangential flow system (i.e., ATF system) also has the following defects: the processing capacity of the current largest model of the ATF perfusion system (such as ATF10) is 500-800L / set (or each batch), and multiple ATF systems need to be connected in parallel for ton-level or larger-scale reactors, which requires users to invest a lot of money; due to design defects, the ATF system has a certain dead volume, and the reciprocating motion of the liquid in the dead volume is not conducive to cell culture.

[0009] Therefore, a low-cost, large-scale (1000-30000 L) perfusion culture system is needed. Summary of the invention

[0010] In view of the above-mentioned shortcomings of the prior art, an object of the present invention is to provide a perfusion module and a perfusion culture system, which are used to solve the problems of high cost and high shear force of large-scale reactors in the prior art.

[0011] To achieve the above-mentioned purpose and other related purposes, the present invention provides a perfusion module, which includes at least one perfusion chamber, each of which is connected to a pressure regulating tube for airflow in or out, and the pressure regulating tube is provided with a pump assembly that can open and close the pressure regulating tube and change the airflow delivery direction; when there are multiple perfusion chambers, two adjacent perfusion chambers are connected through the pressure regulating tube. By connecting each perfusion chamber to a pressure regulating tube, the airflow out or in from the perfusion chamber can be adjusted as needed, so that the cell culture fluid can flow better, the problem of high shear force of the perfusion culture system is solved, and large-scale cell culture is realized.

[0012] The present invention also provides a perfusion culture system, comprising:

[0013] A bioreactor module, comprising at least one bioreactor;

[0014] The perfusion module comprises at least one perfusion chamber, each of which is connected to a pressure regulating tube for air flow in or out, and the pressure regulating tube is provided with a pump assembly for switching the pressure regulating tube on and off and changing the air flow delivery direction; when there are multiple perfusion chambers, two adjacent perfusion chambers are connected through the pressure regulating tube;

[0015] The filtration module comprises at least one filter, wherein the filter has an inlet end and an outlet end which are arranged opposite to each other, wherein the inlet end is provided with a cell culture fluid inlet, and the outlet end is provided with a permeate outlet and a V cell culture fluid outlet;

[0016] The bioreactor, the perfusion module and the filtration module are sequentially connected. If there are more than two perfusion chambers in the perfusion module, all the perfusion chambers are arranged in parallel; the cell culture fluid in the bioreactor flows into the perfusion chamber in the perfusion module, and then flows out of the perfusion chamber into the filter for filtration, and the permeate in the filter flows into the harvesting device through the permeate outlet.

[0017] Preferably, the inlet end of the filter is also provided with a culture medium inlet, and the culture medium inlet is connected to a culture medium supply system.

[0018] Preferably, the cell culture fluid outlet of the filter is connected to the bioreactor through a reflux pipe, and the cell culture fluid filtered by the filter flows back into the bioreactor through the reflux pipe.

[0019] Preferably, the culture medium supply system is connected to the bioreactor via a culture medium delivery pipe.

[0020] Preferably, when the perfusion module has one perfusion chamber, the top of the perfusion chamber is connected to the top of the bioreactor via a pressure regulating pipeline.

[0021] Preferably, the pump assembly is a diaphragm pump assembly.

[0022] Preferably, the perfusion chamber is connected to an airflow mass flow controller via an airflow delivery tube.

[0023] Preferably, the air flow conveying pipe is provided with a gas filter and / or a switch valve.

[0024] Preferably, the bioreactor is connected to the perfusion chamber via an infusion tube, and an on-off valve for opening and closing the infusion tube is provided on the infusion tube.

[0025] Preferably, the filtering module is provided with two filters arranged in parallel, and the two filters work alternately.

[0026] As described above, the perfusion culture system of the present invention has the following beneficial effects: by connecting the bioreactor, the perfusion chamber and the filter through pipelines, the arrangement of multiple perfusion chambers can realize large-scale (1000-30000L) perfusion culture; and by connecting each perfusion chamber to a pressure regulating tube, the airflow out of or in the perfusion chamber can be adjusted as needed, that is, the positive pressure regulation of the perfusion chamber is realized, the cell culture fluid is better flowed, and the problem of high shear force in the perfusion culture system is solved. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Shown is a schematic diagram of an embodiment of the perfusion culture system of the present invention.

[0028] Figure 2 FIG. 4 is a schematic diagram showing another embodiment of the perfusion culture system of the present invention.

[0029] Component number description

[0030] 1 Bioreactor

[0031] 2.2a, 2b Perfusion chamber

[0032] 3.3a,3b Filters

[0033] 4 Culture medium supply system

[0034] 5 Harvesting device

[0035] 6 Mass Flow Controller

[0036] 7 Harvest Pump

[0037] 9 Delivery Pump

[0038] 10 Electromagnetic flowmeter

[0039] 8, 12, 13, 17, 18, 16, valve 23, 22

[0041] 14. Switch valve

[0042] 15 Gas filter

[0043] 16 Diaphragm Pump Assembly

[0044] 11, 20 On / Off Valve

[0045] 19 Normally open valve

[0046] 21 Protection valve

[0047] 101 Infusion tube

[0048] 102 Reflux pipe

[0049] 103 Culture medium delivery tube

[0050] 104 Delivery pipe

[0051] 105 Regulator DETAILED DESCRIPTION

[0052] The following is a description of the implementation of the present invention by means of specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0053] See also Figure 1 to Figure 2 . It should be noted that the structures, proportions, sizes, etc. illustrated in the drawings of this specification are only used to match the contents disclosed in the specification for people familiar with this technology to understand and read, and are not used to limit the limiting conditions for the implementation of the present invention, so they have no substantial technical significance. Any modification of the structure, change in the proportional relationship, or adjustment of the size should still fall within the scope of the technical content disclosed by the present invention without affecting the effects and purposes that can be achieved by the present invention. At the same time, the terms such as "upper", "lower", "left", "right", "middle" and "one" quoted in this specification are only for the convenience of description, and are not used to limit the scope of the implementation of the present invention. Changes or adjustments in their relative relationships should also be regarded as the scope of the implementation of the present invention without substantially changing the technical content.

[0054] A core of the present invention is to provide a perfusion module capable of realizing positive pressure regulation. Figure 1 and Figure 2 As shown, the perfusion module includes at least one perfusion chamber 2, each perfusion chamber 2 is connected to a pressure regulating tube 105 for air flow in or out, and the pressure regulating tube 105 is provided with a pump assembly for opening and closing the pressure regulating tube 105 and changing the air flow direction; when there are multiple perfusion chambers 2, two adjacent perfusion chambers 2 are connected through the pressure regulating tube 105. By connecting each perfusion chamber 2 with a pressure regulating tube 105, the air flow out or in from the perfusion chamber can be adjusted as needed, so that the cell culture fluid can flow better, the problem of high shear force of the perfusion culture system is solved, and large-scale cell culture is realized.

[0055] If there is only one perfusion chamber 2, the pressure regulating tube connected to it can be used independently; when the scale of cell culture fluid is large, multiple perfusion chambers 2 can be arranged in parallel and used alternately, and two adjacent perfusion chambers 2 are connected by a pressure regulating tube 105, so that the gas in the perfusion chamber 2 can flow out when the cell culture fluid is injected, which is convenient for the entry of the cell culture fluid, and the gas can flow in on the side of discharging the cell culture fluid, thereby increasing the pressure in the perfusion chamber and facilitating the discharge of the cell fluid.

[0056] like Figure 1 and Figure 2 As shown, this embodiment provides a perfusion culture system, which includes:

[0057] A bioreactor module, comprising at least one bioreactor 1;

[0058] The perfusion module is the above-mentioned perfusion module, specifically: comprising at least one perfusion chamber 2, each perfusion chamber 2 is connected to a pressure regulating tube 105 for air flow in or out, and the pressure regulating tube 105 is provided with a pump assembly that can turn on and off the pressure regulating tube 105 and change the air flow delivery direction. In this embodiment, the pump assembly can be a diaphragm pump assembly; when there are multiple perfusion chambers 2, two adjacent perfusion chambers 2 are connected through the pressure regulating tube 105;

[0059] The filtration module comprises at least one filter 3, wherein the filter 3 has an inlet end and an outlet end which are arranged opposite to each other, wherein the inlet end is provided with a cell culture fluid inlet, and the outlet end is provided with a permeate outlet and a cell culture fluid outlet;

[0060] The bioreactor 1, the perfusion module and the filtration module are connected in sequence. If there are more than two perfusion chambers 2 in the perfusion module, all the perfusion chambers 2 are arranged in parallel; the cell culture fluid in the bioreactor 1 flows into the perfusion chamber 2 in the perfusion module, and at the same time, the gas in the perfusion chamber 2 is discharged through the pressure regulating tube 105, and the cell culture fluid then flows out of the perfusion chamber 2 and enters the filter 3 for filtration, and the permeate in the filter 3 flows into the harvesting device 5 through the permeate outlet.

[0061] The present invention connects the bioreactor 1, the perfusion chamber 2, and the filter 3 through pipelines, and the arrangement of multiple perfusion chambers can realize large-scale (1000-30000L) perfusion culture. Moreover, each perfusion chamber is connected to a pressure regulating tube, and the airflow out of or in the perfusion chamber can be adjusted according to needs, that is, the positive pressure regulation of the perfusion chamber is realized, the cell culture fluid is better flowed, and the problem of high shear force of the perfusion culture system is solved.

[0062] In order to better recycle the culture medium, in this embodiment, a culture medium inlet is also provided at the inlet end of the filter 3, and the culture medium inlet is connected to the culture medium supply system 4. The culture medium supply system 4 is directly connected to the filter 3, and can flush the filter, thereby improving the utilization efficiency of the filter 3. The culture medium supply system 4 includes a culture medium storage container and a delivery pump 9, and a valve 18 is provided on the pipeline connected to the filter, and the valve 18 is controlled to open and close as needed.

[0063] Furthermore, the culture medium supply system 4 is connected to the bioreactor 1 via a culture medium delivery pipe 103 , on which a valve 12 is disposed. The culture medium is delivered to the bioreactor 1 by opening and closing the valve 12 , so as to better supplement nutrients for the cell culture fluid in the bioreactor 1 .

[0064] To facilitate the circulation of the cell culture fluid, the cell culture fluid outlet of the filter 3 in this embodiment is connected to the bioreactor 1 through the reflux pipe 102, that is, the cell culture fluid filtered by the filter 3 can be refluxed into the bioreactor 1 through the reflux pipe 102, which also increases the scale of the cell culture fluid.

[0065] In this embodiment, the perfusion module has only one perfusion chamber 2. Figure 1 As shown, the top of the perfusion chamber 2 is connected to the top of the bioreactor 1 through a pressure regulating tube 105, and a diaphragm pump assembly 16 is provided on the pressure regulating tube 105 to enable and disable the pressure regulating tube 105 and change the direction of air flow. In this embodiment, the perfusion chamber 2 and the bioreactor 1 are connected through the pressure regulating tube 105. When the cell culture fluid in the perfusion chamber 2 is emptied, the valve on the delivery tube 104 connecting the perfusion chamber 2 and the filter 3 is closed, and at the same time, the on-off valve 11 on the infusion tube 101 connected to the perfusion chamber 2 is opened, and the diaphragm pump assembly 16 on the pressure regulating tube 105 is opened, so that the gas in the perfusion chamber 2 can enter the bioreactor 1, and the cell culture fluid flows into the perfusion chamber 2. The arrangement of the pressure regulating tube 105 and the diaphragm pump assembly 16 can positively adjust the air pressure in the perfusion chamber 2, facilitate the circulation of the cell culture fluid, and solve the problem of high shear force of the perfusion culture system.

[0066] In order to better adapt to large-scale cell culture fluid, the perfusion module in this embodiment has more than two perfusion chambers 2, see Figure 2 As shown, adjacent perfusion chambers 2 are connected through a pressure regulating tube 105, and a diaphragm pump assembly is provided on the pressure regulating tube 105 to open and close the pressure regulating tube 105 and change the direction of air flow. In this embodiment, the diaphragm pump assembly is a pipeline diaphragm pump composed of valves 17, 8, and 16. The three valves are opened and closed in sequence to realize the pumping of air above the liquid surface in the perfusion chamber to the left or right, so as to facilitate the cell culture fluid to enter the corresponding perfusion chamber and also facilitate the output of the cell culture fluid to another perfusion chamber.

[0067] The bioreactor 1 is connected to the perfusion chamber 2 through a liquid infusion tube 101. The liquid infusion tube 101 is provided with an on-off valve 11 for opening and closing the liquid infusion tube. Figure 2 As shown, each branch connected to the perfusion chamber can be equipped with an on-off valve 11, 20, and a normally open valve 19 is configured on the main pipeline. In order to facilitate the perfusion chamber 2a, 2b to transport the cell culture fluid to the flow module, in this embodiment, valves 23, 22 are provided at the delivery port of each perfusion chamber 2a, 2b. The opening and closing of the valves 23, 22 can realize the opening or closing of the perfusion chamber 2a, 2b, and when opened, the cell culture fluid in the perfusion chamber 2a, 2b can be output. For better control, a protective valve 21 and an electromagnetic flowmeter 10 are also provided on the delivery pipe 104 connecting the perfusion chamber 2a, 2b to the filtration module, so as to more accurately control the delivery of the cell culture fluid; to ensure safety, a pressure gauge is provided on each perfusion chamber 2 to detect the pressure inside it.

[0068] The above filter 3 contains a hollow fiber membrane column, which can be called a hollow fiber type filter. Hollow fiber (HF) type filters give a longer working life, and they can obtain many sizes, configurations, materials, pore sizes and porosities. In addition, the filter 3 does not need to be limited to the use of hollow fiber filters, and it can also be inserted into other separation devices in the hollow fiber housing.

[0069] In this embodiment, the filter module is provided with two filters 3a and 3b arranged in parallel, and the two filters 3a and 3b work alternately to ensure normal use of the entire system.

[0070] As an embodiment of two perfusion chambers and two filters, the specific working process is as follows: Figure 2As shown, in this embodiment, the two perfusion chambers 2a and 2b are used alternately, the normally open valve 19 is normally open, the on-off valve 11 is opened, and the cell culture fluid in the bioreactor 1 is transported to the perfusion chamber 2a through the infusion tube 101. In this embodiment, a liquid level gauge is provided in each perfusion chamber 2 to detect whether the cell culture fluid therein has reached the highest liquid level and the lowest liquid level, thereby controlling whether the perfusion chamber should input or output the cell culture fluid. When the cell culture fluid in the perfusion chamber 2a reaches the maximum working volume, the on-off valve 11 is closed, and the on-off valve 20, the valve 23 and the valve 14 are opened. At this time, the bioreactor 1 is transported to the perfusion chamber 2b through the infusion tube 101. During the transportation, the valves in the diaphragm pump assembly are opened in sequence according to the valves 16-8-17 to pump the air above the liquid level in the perfusion chamber 2b into the perfusion chamber 2a, that is, the air flow direction in the pressure regulating tube 105 is from left to right. At the same time, compressed air is introduced into the perfusion chamber 2a by the mass flow controller 6 to ensure that the perfusion chamber 2a has sufficient perfusion pressure, so that the cell culture fluid in the perfusion chamber 2a flows into the filter 3 through the delivery tube 104 for filtration. The two filters 3a and 3b are used alternately to ensure that there is always one filter in working condition; the permeate after the filter is transported to the harvesting device 5 by the harvesting pump 7 through the permeate outlet, and the cell culture fluid flows back to the bioreactor 1 through the reflux tube 102.

[0071] When the cell culture fluid in the perfusion chamber 2b reaches the maximum volume and the cell culture fluid in the perfusion chamber 2a reaches the minimum liquid level, the on-off valve 11 is controlled to open, the on-off valve 20 is closed, the valve 22 is opened, and the valve 23 is closed. The cell culture fluid in the bioreactor 1 is transported to the perfusion chamber 2b through the infusion tube 101. At this time, the valves in the diaphragm pump assembly are opened in sequence according to valves 17-8-16, so that the air above the liquid level in the perfusion chamber 2a is pumped into the perfusion chamber 2b, that is, the air flow direction in the pressure regulating tube 105 is from right to left. At the same time, compressed air is introduced into the perfusion chamber 2b by the mass flow controller 6 to ensure that the perfusion chamber 2b has sufficient perfusion pressure, so that the cell culture fluid in the perfusion chamber 2b flows into the filter 3 through the delivery tube 104 for filtration.

[0072] The perfusion chambers 2a and 2b are used reciprocally in sequence to ensure the normal delivery of the cell culture fluid, thereby increasing the utilization scale of the entire system.

[0073] The culture medium supply system 4 is used to flush the filter 3 on the one hand, and is connected to the bioreactor 1 through the culture medium delivery pipe 103 on the other hand to replenish the culture medium for the bioreactor.

[0074] In order to better transport the cell culture fluid from the perfusion chamber to the filter, each perfusion chamber 2a, 2b in this embodiment is connected to a mass flow controller 6 through an air flow delivery tube. A gas filter 15 and / or a switch valve 14 are provided on the air flow delivery tube, and the gas filter 15 ensures the sterility of the gas supplied by the mass flow controller 6 to the perfusion chamber.

[0075] The mass flow controller 6 and other actuators such as valves or pumps are sent action instructions by the controller and need to be set according to the process parameters when used. The mass flow controller 6 is known and can be used according to methods known to those skilled in the art.

[0076] In this embodiment, multiple perfusion chambers can be provided as needed, such as three or more, and adjacent perfusion chambers can be connected by a pressure regulating tube, or the perfusion chamber can be connected to the bioreactor through a pressure regulating tube to achieve positive regulation of the air pressure in the perfusion chamber. In this embodiment, multiple perfusion chambers 2 are used alternately and / or in parallel, so that a large volume of cell culture fluid can be flowed, the permeate harvest is increased, and the entire system is easy to control and implement. The material of the perfusion chamber is usually 316L stainless steel plus heat-resistant borosilicate glass.

[0077] In summary, the perfusion culture system of the present invention can realize large-scale (1000-30000L) perfusion culture, and the high shear force problem of the perfusion culture system is solved by setting up a gas positive pressure perfusion chamber. At the same time, the system can be sterilized online, which is convenient for operation. Therefore, the present invention effectively overcomes various shortcomings in the prior art and has high industrial utilization value.

[0078] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A perfusion culture system, characterized in that: include: A bioreactor module, comprising at least one bioreactor; The perfusion module comprises at least one perfusion chamber, each of which is connected to a pressure regulating tube for air flow in or out, and the pressure regulating tube is provided with a pump assembly for switching the pressure regulating tube on and off and changing the air flow delivery direction; when there are multiple perfusion chambers, two adjacent perfusion chambers are connected through the pressure regulating tube; The filtration module comprises at least one filter, wherein the filter has an inlet end and an outlet end which are arranged opposite to each other, wherein the inlet end is provided with a cell culture fluid inlet, and the outlet end is provided with a permeate outlet and a cell culture fluid outlet; The bioreactor, the perfusion module and the filtration module are sequentially connected. If there are more than two perfusion chambers in the perfusion module, all the perfusion chambers are arranged in parallel; the cell culture fluid in the bioreactor flows into the perfusion chamber in the perfusion module, and then flows out of the perfusion chamber into the filter for filtration, and the permeate in the filter flows into the harvesting device through the permeate outlet.

2. The perfusion culture system according to claim 1, characterized in that: The inlet end of the filter is also provided with a culture medium inlet, and the culture medium inlet is connected to a culture medium supply system.

3. The perfusion culture system according to claim 1, characterized in that: The cell culture fluid outlet of the filter is connected to the bioreactor through a reflux pipe, and the cell culture fluid filtered by the filter flows back into the bioreactor through the reflux pipe.

4. The perfusion culture system according to claim 2, characterized in that: The culture medium supply system is connected to the bioreactor via a culture medium delivery pipe.

5. The perfusion culture system according to claim 1, characterized in that: When the perfusion module has one perfusion chamber, the top of the perfusion chamber is connected to the top of the bioreactor through the pressure regulating tube.

6. The perfusion culture system according to claim 1, characterized in that: The pump assembly is a diaphragm pump assembly.

7. The perfusion culture system according to claim 1, characterized in that: The perfusion chamber is connected to an airflow mass flow controller via an airflow delivery tube.

8. The perfusion culture system according to claim 7, characterized in that: The air flow conveying pipe is provided with a gas filter and / or a switch valve.

9. The perfusion culture system according to claim 7, characterized in that: The bioreactor is connected to the perfusion chamber through an infusion tube, and an on-off valve for opening and closing the infusion tube is arranged on the infusion tube.

10. The perfusion culture system according to claim 1, characterized in that: The filter module is provided with two filters arranged in parallel, and the two filters work alternately.

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