Efficient suspension cell culture bin based on dynamic centrifugation

Abstract

The invention relates to an efficient suspension cell culture bin based on dynamic centrifugation, and solves the technical problems that an existing cell culture device is complex to operate and high in pollution risk, and a culture medium cannot be replaced in a cell amplification stage. The device comprises a channel shaft, a first connector, a second connector, a cover plate, a bin body, an upper liquid circulation mechanism and a lower liquid circulation mechanism, the channel shaft is provided with an inner channel and an outer channel, the first connector is connected with the channel shaft, the second connector is connected with the channel shaft, and the lower liquid circulation mechanism and the upper liquid circulation mechanism are located in the bin body. According to the invention, culture medium replacement can be dynamically and continuously carried out in a cell amplification stage.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cell culture, in particular to a high-efficiency culture bin for suspension cells based on dynamic centrifugation. BACKGROUND

[0002] In recent years, with the rapid development of cell therapy technology, individualized immunotherapy represented by CAR-T cells, TCR-T cells and natural killer (NK) cells has shown significant clinical efficacy in the fields of tumors and autoimmune diseases. With the surge in clinical demand, the manufacturing process of cell therapy products is evolving towards high efficiency, safety, standardization and automation.

[0003] Currently, the expansion of T cells, NK cells and other immune cells mainly adopts adherent culture or suspension static culture, which is often completed through open culture bottles (such as T bottles, cell bags) and multi-step manual medium replacement. For cell expansion, please refer to the invention patent application with the application publication number CN120082511A, and for cell culture device, please refer to the invention patent application with the application publication number CN113214970A. However, the above-mentioned methods mainly have the following technical problems: complex operation steps, high risk of contamination; during the cell expansion stage, the medium cannot be replaced, because the expanded cells are suspension cells, and during the cell culture process, the cells are distributed in the medium, and at this time, the replacement will cause the cultured cells to be excluded. SUMMARY

[0004] The present application is to solve the technical problems of the existing cell culture device, such as complex operation, high risk of contamination and inability to replace the medium during the cell expansion stage, and provides a high-efficiency culture bin for suspension cells based on dynamic centrifugation.

[0005] The present application provides a high-efficiency culture bin for suspension cells based on dynamic centrifugation, which comprises a channel shaft, a first joint, a second joint, a cover plate, a bin body, an upper liquid flow mechanism, a lower liquid flow mechanism and a sealing ring. The channel shaft is provided with an inner channel and an outer channel, and the outer channel is located at the periphery of the inner channel. The first joint is connected with the channel shaft, and the first joint is in communication with the inner channel. The second joint is connected with the channel shaft, and the second joint is in communication with the outer channel. The lower liquid flow mechanism and the upper liquid flow mechanism are located in the bin body, and the lower liquid flow mechanism is fixedly connected to the bottom surface of the bin body. The lower liquid flow mechanism comprises a lower flow passage support part and two lower flow passages. The two lower flow passages are respectively connected with the lower flow passage support part, and are symmetrically arranged on both sides of the lower flow passage support part. The lower flow passage support part is provided with an axial center through hole, and the lower flow passage is provided with a radial channel. The radial channel is in communication with the center through hole, and there is a gap between the end of the lower flow passage and the side wall of the bin body. The upper liquid flow mechanism includes an upper flow channel support component and two upper flow channels. The two upper flow channels are respectively connected to the upper flow channel support component and are located on both sides of the upper flow channel support component and arranged symmetrically. The upper flow channels are provided with an open radial channel at the top. The upper flow channel support component is provided with a receiving cavity with an open top. The open radial channel at the top communicates with the receiving cavity with the open top. The upper flow channel support component is provided with a central shaft hole. The upper flow channel support component is fixedly connected to the lower flow channel support component. The channel shaft passes through the cover plate and is rotatably connected to the cover plate via two bearings. The lower end of the channel shaft passes through the central shaft hole of the upper flow channel support component, and a sealing ring is connected between the lower end of the channel shaft and the central shaft hole. The cover plate is fixedly connected to the upper flow channel support component, and a cavity is formed between the inner wall of the cover plate and the chamber. The receiving cavity with the top opening of the upper flow channel support component is connected to the outer channel of the channel shaft. There is a gap between the end of the upper flow channel and the side wall of the chamber, and the top of the upper flow channel is attached to the inner wall of the cover plate to form a channel. The lower end face of the channel shaft abuts against the top of the lower flow channel support component, and the axial center through hole of the lower flow channel support component is connected to the inner channel of the channel shaft. There is a radial gap between the end of the upper flow channel and the end of the lower flow channel.

[0006] Preferably, the radial channel of the lower liquid flow mechanism is a semi-circular channel with an open lower part, and the open lower semi-circular channel forms a channel with the bottom surface of the tank body.

[0007] Preferably, the lower flow channel of the lower liquid flow mechanism is provided with an arc-shaped notch at its end.

[0008] Preferably, the upper open radial channel of the upper flow channel is an upper open semi-circular radial channel.

[0009] Preferably, the bottom end of the open semi-circular radial channel at the top is provided with an arc-shaped notch.

[0010] Preferably, the high-efficiency culture chamber for suspended cells based on dynamic centrifugation further includes a locking connection part, which is fixedly connected to the central part of the outside of the chamber.

[0011] Preferably, the cover plate is provided with a fixing sleeve, which is inserted into the receiving cavity of the top opening of the upper flow channel support component and is fixedly connected to the side wall of the receiving cavity of the top opening; the fixing sleeve is provided with two holes, which are respectively connected to two upper open radial channels.

[0012] Preferably, the fixed sleeve is provided with a step, and the end face of the outer channel of the channel shaft abuts against the step of the fixed sleeve.

[0013] Preferably, a sealing gasket is connected to the step of the fixed sleeve, and the end face of the outer channel abuts against the sealing gasket.

[0014] Preferably, the container is transparent.

[0015] The beneficial effect of this disclosure is that it enables dynamic and continuous culture medium replacement, allowing for culture medium replacement during the cell expansion phase.

[0016] It facilitates the adjustment of amplification density to meet requirements. It also facilitates the dynamic control of culture space and cell growth environment.

[0017] Significantly reduce the risk of pollution.

[0018] Further features and aspects of this disclosure will be clearly described in the following detailed description with reference to the accompanying drawings. Attached Figure Description

[0019] Figure 1 This is an isometric view of a high-efficiency suspension cell culture chamber based on dynamic centrifugation; Figure 2 yes Figure 1 Bottom view of the culture chamber shown; Figure 3 yes Figure 1 The diagram shows the internal structure of the culture chamber. Figure 4 yes Figure 1 Exploded view of the culture chamber shown; Figure 5 yes Figure 3 The diagram shows the assembly relationship between the channel shaft, the upper liquid flow mechanism, and the lower liquid flow mechanism in the structure shown. Figure 6 yes Figure 5 The diagram shows the structure of the upper liquid flow mechanism and the lower liquid flow mechanism. Figure 7 yes Figure 3 The diagram shows the structure in which the channel shaft is connected to the lower flow channel support component. Figure 8 yes Figure 3 The diagram shows the structure in which the channel shaft is connected to the upper liquid flow mechanism. Figure 9 It is an isometric drawing of the lower liquid flow mechanism; Figure 10 yes Figure 9 An isometric view of the lower liquid flow mechanism from another perspective; Figure 11 It is an isometric drawing of the upper liquid flow mechanism; Figure 12 yes Figure 11 A top view of the upper liquid flow mechanism shown; Figure 13 yesFigure 3 Figure 2 is a structural diagram of the fixed connection between the cover plate and the upper flow passage support member; Figure 14 Figure 3 is a schematic diagram of the entire liquid in the chamber being divided into two layers under high-speed rotation of the culture bin; Figure 15 Figure 4 is a diagram of the positional relationship between the end of the upper flow passage and the end of the lower flow passage in the radial direction; Figure 16 Figure 5 is a diagram of the positional relationship between the end of the upper flow passage and the end of the lower flow passage in the radial direction; Figure 3 Figure 6 is an enlarged view of part P in Figure 5.

[0020] Figure 7 is a diagram of the symbols in the figures. 1. Passage shaft, 101. First joint, 102. Second joint, 103. Inner passage, 104. Outer passage; 2. Cover plate, 2-1. Fixed sleeve portion, 2-1-1. Hole; 3. Bin body, 3-1. Bottom surface, 3-2. Annular step, 301. Locking connection portion; 4. Upper liquid flow passage mechanism, 401. Upper flow passage support member, 401-1. Top open accommodating cavity, 401-2. Center shaft hole, 402. Upper flow passage, 402-1. Upper open semicircular radial passage, 402-2. Circular-arc-shaped notch, 403. Lower portion; 5. Lower liquid flow passage mechanism, 501. Lower flow passage support member, 501-1. Axial center through hole, 502. Lower flow passage, 502-1. Radial passage, 502-2. Circular-arc-shaped notch, 503. Upper portion; 6. Bearing; 7. Bearing, 8. Sealing ring; 9. Annular liquid layer, 10. Inner annular layer. DETAILED DESCRIPTION

[0021] The application will be further described in detail below with reference to the accompanying drawings.

[0022] The specific embodiments described below are merely preferred embodiments of the application and the scope of protection of the application is not limited thereto. Based on or according to the principles, concepts, and spirits of the application, some changes or modifications can be made by those skilled in the art, and the technical solutions formed by these changes or modifications should be included in the scope of protection of the application.

[0023] As Figures 1-8As shown, the dynamic centrifugation-based high-efficiency culture bin for suspended cells comprises a channel shaft 1, a first joint 101, a second joint 102, a cover plate 2, a bin body 3, an upper liquid flow mechanism 4, a lower liquid flow mechanism 5, a bearing 6, a bearing 7, and a sealing ring 8. A cavity is formed between the inner wall of the cover plate 2 and the bin body 3, the channel shaft 1 passes through the cover plate 2, and the lower end of the channel shaft 1 extends into the bin body 3. The channel shaft 1 is rotationally connected to the cover plate 2 through the bearings 6 and 7, and is provided with an inner channel 103 and an outer channel 104, with the outer channel 104 located at the periphery of the inner channel 103. The first joint 101 is connected to the channel shaft 1 and communicates with the inner channel 103, and the second joint 102 is connected to the channel shaft 1 and communicates with the outer channel 104. The lower liquid flow mechanism 5 and the upper liquid flow mechanism 4 are located in the bin body 3, the lower liquid flow mechanism 5 is fixedly installed on the bottom surface of the bin body 3, and the upper liquid flow mechanism 4 is fixedly connected to the lower liquid flow mechanism 5. The locking connection part 301 is fixedly connected to the central part outside the bin body 3. The bin body 3 is circular as a whole.

[0024] The lower liquid flow mechanism 5 comprises a lower flow passage support part 501 and two lower flow passages 502, the two lower flow passages 502 are respectively connected to the lower flow passage support part 501, and the two lower flow passages 502 are located on both sides of the lower flow passage support part 501 and are symmetrically arranged. The lower flow passage support part 501 is provided with an axial center through hole 501-1, and the lower flow passage 502 is provided with a radial passage 502-1 which communicates with the center through hole 501-1. There is a certain distance gap between the end of the lower flow passage 502 and the side wall of the bin body 3 (that is, there is a gap between the end of the radial passage 502-1 and the side wall of the bin body 3). As shown in Figure 3 、 Figure 9 and Figure 10 As shown, the shape of the radial passage 502-1 is preferably a lower-opened semicircular channel (the cross section is semicircular), in which case the radial passage 502-1 forms a channel for liquid flow with the bottom surface 3-1 of the bin body 3. Further preferably, a circular-arc-shaped notch 502-2 is arranged at the end of the lower flow passage 502, which communicates with the radial passage 502-1, which is more conducive to liquid flow.

[0025] The end surface of the lower end of the channel shaft 1 abuts against the top of the lower flow passage support part 501, and the axial center through hole 501-1 communicates with the inner channel 103.

[0026] The upper liquid flow passage mechanism 4 comprises an upper flow passage support component 401 and two upper flow passages 402, which are respectively connected with the upper flow passage support component 401 and symmetrically arranged on both sides of the upper flow passage support component 401. The upper flow passage 402 is provided with an upper open semicircular radial passage 402-1 (the cross section is semicircular). The upper flow passage support component 401 is provided with a top open accommodating cavity 401-1, and the upper open semicircular radial passage 402-1 is in communication with the top open accommodating cavity 401-1. The upper flow passage support component 401 is provided with a central shaft hole 401-2 for allowing the lower end of the passage shaft 1 to pass through.

[0027] Referring to Figure 3 , the upper liquid flow passage mechanism 4 is fixedly connected with the lower liquid flow passage mechanism 5, and the specific connection manner can be that the lower part 403 of the upper flow passage support component 401 is sleeved on the upper part 503 of the lower flow passage support component 501 for fixation. The lower end of the passage shaft 1 passes through the central shaft hole 401-2 of the upper flow passage support component 401, and a sealing ring 8 is connected between the lower end of the passage shaft 1 and the central shaft hole 401-2. The cover plate 2 is fixedly connected with the upper flow passage support component 401. The top open accommodating cavity 401-1 is in communication with the outer passage 104 of the passage shaft 1. There is a certain gap between the end of the upper flow passage 402 and the side wall of the cartridge body 3 (that is, there is a gap between the end of the upper open semicircular radial passage 402-1 and the side wall of the cartridge body 3), and the top of the upper flow passage 402 is attached to the inner wall of the cover plate 2 to form a passage for liquid flow. In order to better allow the liquid to flow into the upper open semicircular radial passage 402-1, referring to Figure 12 , a circular arc-shaped notch 402-2 is arranged at the bottom of the end of the upper flow passage 402, and the circular arc-shaped notch 402-2 is in communication with the upper open semicircular radial passage 402-1. As shown in Figure 16 , the lowermost part of the side wall of the cartridge body 3 is provided with an annular step 3-2.

[0028] Referring to Figure 13 , one specific implementation manner that the cover plate 2 is fixedly connected with the upper flow passage support component 401 is that the cover plate 2 is provided with a fixed sleeve part 2-1, the fixed sleeve part 2-1 is inserted into the top open accommodating cavity 401-1 of the upper flow passage support component 401, and the fixed sleeve part 2-1 is fixedly connected with the side wall of the top open accommodating cavity 401-1. In this case, two holes 2-1-1 are arranged on the fixed sleeve part 2-1, the holes 2-1-1 are in communication with the upper open semicircular radial passage 402-1, and the two holes 2-1-1 respectively correspond to the two upper open semicircular radial passages 402-1. The holes 2-1-1 are in communication with the top open accommodating cavity 401-1. In order to further improve the sealing effect, referring to Figure 13, the end face of the outer channel 104 abuts against the step of the fixed sleeve part 2-1; further optimization, a sealing gasket is connected on the step of the fixed sleeve part 2-1, and the end face of the outer channel 104 abuts against the sealing gasket.

[0029] Reference Figure 15 There is a spacing L in the radial direction between the end of the upper flow channel 402 and the end of the lower flow channel 502, and the end of the lower flow channel 502 is inside and the end of the upper flow channel 402 is outside. That is, the end of the upper flow channel 402 and the end of the lower flow channel 502 are not aligned. The specific value of the spacing L is determined according to the cell type, for example, the spacing L is 1 mm for T cells.

[0030] The working process of the above-mentioned high-efficiency culture bin for suspended cells based on dynamic centrifugation is as follows: First, fix and position the channel shaft 1; connect the locking connection part 301 with the output shaft of the rotary drive motor.

[0031] Second, inject the prepared cell suspension from the first joint 101, and the cell suspension flows into the inner channel 103, then sequentially passes through the axial center through hole 501-1 and the radial channel 502-1, and finally flows out from the end of the radial channel 502-1 and enters the bin body 3. It should be noted that the PH value, oxygen content, temperature and the like can be set according to the known conditions of the prior art.

[0032] Third, carry out cell expansion culture work under the condition that the culture bin does not rotate or rotates at a low speed, and culture for a period of time. Regarding the condition of low speed rotation, start the rotary drive motor to make the cover plate 2 and the bin body 3 rotate with the channel shaft 1 as the reference, and the rotating speed is 300 rpm. During the culture process, oxygen, carbon dioxide and other gases are filled into the chamber through the second joint 102 (the gases enter the chamber from the outer channel 104 and then enter the chamber through the upper open semicircular radial channel 402-1), and the PH value, oxygen content and other environmental indicators are adjusted; in addition, the culture temperature is controlled at 37°C by the heat exchange system. It should be noted that the PH value, oxygen content, temperature and the like can be set according to the known conditions of the prior art.

[0033] In order to monitor the environmental parameters in the chamber, conventional methods can be used, such as setting sensors in the chamber to sense temperature, pH, dissolved oxygen, CO2 concentration, humidity and other environmental parameters. In order to better realize monitoring, a parameter sensing patch sensor with high sensitivity and rapid response capability can be selected, and the sensing patch sensor is fixed on the inner wall of the chamber, and the optical signal of the sensing patch sensor is collected in a non-contact manner through an optical fiber collection module, and the optical signal is transmitted to a data processing instrument.

[0034] In the fourth step, during the culture process, culture medium is added through the first connector 101, and the culture medium is output into the chamber through the radial channel 502-1 of the lower flow channel 502. After the culture medium has been in action for a period of time, the speed of the rotary drive motor is adjusted to make the chamber 3 rotate at high speed (e.g., 1000 rpm), forming a stable cell density gradient field. At this time, the entire liquid in the chamber is divided into two layers (the reason for the formation of two layers in cell density gradient centrifugation is the combined effect of the density difference of cells or particles in the sample and the distribution of the density gradient medium. Components with different densities migrate to the medium region that matches their own density under the action of centrifugal force, thus forming two layers. This can be understood as the cells being pressed against the side wall of the chamber due to the difference in weight between the light and heavy culture medium). The suspension containing cells forms an annular liquid layer 9 against the side wall of the chamber 3, and the used culture medium also forms an inner annular layer 10, that is, the used culture medium is located in the inner layer, such as... Figure 14 As shown, the central area of ​​the chamber is empty and without liquid, and the annular liquid layer 9 is located above the annular step 3-2. Next, the used culture medium (inner annular layer 10) is drawn outward through the first connector 101 and flows in from the opening at the end of the radial channel 502-1. After passing through the radial channel 502-1, it enters the axial central through hole 501-1 and finally exits from the first connector 101 through the inner channel 103 (generally, most of the used culture medium is drawn away). Since the annular liquid layer 9 is attached to the side wall of the chamber 3 and is located above the annular step 3-2, and the bottom of the annular liquid layer 9 is far away from the opening at the end of the radial channel 502-1, the annular liquid layer 9 (containing cell suspension) will not be drawn away and will remain in the chamber.

[0035] After the used culture medium is discharged, new culture medium can be added as needed. First, adjust the speed of the rotary drive motor to make the chamber 3 rotate at a low speed. Then, add the new culture medium through the first connector 101. After the new culture medium has been in effect for a period of time, adjust the speed of the rotary drive motor to make the chamber 3 rotate at a high speed. Discharge the culture medium in the same way as described above.

[0036] New culture medium can be added multiple times as needed.

[0037] The fifth step, after the amplification culture is completed, is to drain and collect the suspension containing cells. There are two specific methods. The first specific process is to separate the culture medium and cells by rotating the chamber 3 at high speed. Then, the used culture medium is first aspirated outward through the first connector 101 to drain, and then the liquid containing cells is aspirated outward through the second connector 102 to drain.

[0038] The second specific process is to stop the rotation driving motor and make the whole culture chamber static; then add culture medium (elution to achieve the purpose of purification) through the first joint 101; then add buffer solution through the first joint 101, start the rotation driving motor to make the chamber body 3 rotate at low speed for mixing; then, while keeping the chamber body 3 rotating at low speed, the liquid containing cells in the chamber is sucked out through the second joint 102, flows into the opening at the end of the upper open semicircular radial channel 402-1, then enters the top open containing cavity 401-1 through the upper open semicircular radial channel 402-1, and finally is discharged from the second joint 102 through the outer channel 104.

[0039] It can be seen that the above process realizes dynamic medium replacement, and the medium is replaced during the cell expansion stage, so that the cell culture is better. In addition, the cell density can also be adjusted conveniently.

[0040] In order to verify the effect of layered extraction, the core evaluation index is the density of cells (cells / mL) taken out by the extracted medium. By observing the used medium sample discharged from the first joint 101 under a microscope and counting the cells, it is quantitatively evaluated whether the cells are mistakenly extracted under the condition that the stable layered state is formed by high-speed centrifugation. If the cell density ratio in the discharged used medium liquid should be much lower than 5%, and the ideal situation should be <1%, it indicates that the layered structure is reliable, and the liquid extraction operation will not cause substantial interference to the cells.

[0041] It should be noted that the chamber body 3 can be made of polystyrene to make the chamber body 3 transparent, so as to facilitate observation or visual monitoring through a camera or convenient signal transmission of a sensor. When visual monitoring is performed through a camera, the camera collects images from the bottom of the chamber body 3, and then analyzes the cell density according to the images (the specific analysis method can be performed by using a conventional method), adjusts the whole culture process, and then adjusts the cell density to meet the requirements.

[0042] It should be noted that for the second collection process of the fifth step, the liquid can be sucked out through the first joint 101 instead of the second joint 102.

Claims

1. A high-efficiency suspension cell culture chamber based on dynamic centrifugation, characterized in that, The system includes a channel shaft, a first connector, a second connector, a cover plate, a chamber body, an upper liquid flow mechanism, a lower liquid flow mechanism, and a sealing ring. The channel shaft has an inner channel and an outer channel, with the outer channel located around the inner channel. The first connector is connected to the channel shaft and communicates with the inner channel. The second connector is also connected to the channel shaft and communicates with the outer channel. The lower and upper liquid flow mechanisms are located within the chamber body. The lower liquid flow mechanism is fixedly connected to the bottom surface of the chamber body. The lower liquid flow mechanism includes a lower flow channel support component and two lower flow channels. The two lower flow channels are respectively connected to the lower flow channel support component and are located on both sides of the lower flow channel support component and arranged symmetrically. The lower flow channel support component has an axial central through hole, and the lower flow channels have radial channels that communicate with the central through hole. There is a gap between the end of the lower flow channel and the side wall of the chamber body. The upper liquid flow mechanism includes an upper flow channel support component and two upper flow channels. The two upper flow channels are respectively connected to the upper flow channel support component and are located on both sides of the upper flow channel support component and arranged symmetrically. Each upper flow channel has an open radial channel at the top. The upper flow channel support component has a receiving cavity with an open top. The open radial channel at the top communicates with the receiving cavity with the open top. The upper flow channel support component has a central shaft hole. The upper flow channel support component is fixedly connected to the lower flow channel support component. The channel shaft passes through the cover plate and is rotatably connected to the cover plate via two bearings. The lower end of the channel shaft passes through the central shaft hole of the upper flow channel support component, and a sealing ring is connected between the lower end of the channel shaft and the central shaft hole. The cover plate is fixedly connected to the upper flow channel support component, and a cavity is formed between the inner wall of the cover plate and the hopper body. The receiving cavity with the top opening of the upper flow channel support component is connected to the outer channel of the channel shaft. There is a gap between the end of the upper flow channel and the side wall of the hopper body, and the top of the upper flow channel is attached to the inner wall of the cover plate to form a channel. An annular step is provided at the bottom of the side wall of the hopper body. The lower end face of the channel shaft abuts against the top of the lower flow channel support component, and the axial central through hole of the lower flow channel support component communicates with the inner channel of the channel shaft. There is a radial distance between the end of the upper flow channel and the end of the lower flow channel.

2. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 1, characterized in that, The radial channel of the lower liquid flow mechanism is a semi-circular channel with an open lower part, which forms a channel with the bottom surface of the tank body.

3. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 2, characterized in that, The lower flow channel of the lower liquid flow mechanism has an arc-shaped notch at its end.

4. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 1, 2, or 3, characterized in that, The upper open radial channel of the upper flow channel is an upper open semi-circular radial channel.

5. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 4, characterized in that, The bottom end of the open semi-circular radial channel at the top is provided with an arc-shaped notch.

6. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 1, 2, or 3, characterized in that, The high-efficiency suspension cell culture chamber based on dynamic centrifugation also includes a locking connection part, which is fixedly connected to the central part of the outside of the chamber.

7. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 1, characterized in that, The cover plate is provided with a fixing sleeve, which is inserted into the receiving cavity of the top opening of the upper flow channel support component and is fixedly connected to the side wall of the receiving cavity of the top opening; the fixing sleeve is provided with two holes, which are respectively connected to two upper open radial channels.

8. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 7, characterized in that, The fixed sleeve is provided with a step, and the end face of the outer channel of the channel shaft abuts against the step of the fixed sleeve.

9. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 8, characterized in that, A sealing gasket is connected to the step of the fixed sleeve, and the end face of the outer channel abuts against the sealing gasket.

10. The high-efficiency suspension cell culture chamber based on dynamic centrifugation according to claim 1, characterized in that, The container is transparent.

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