A bioreactor

Abstract

This invention relates to the field of biological technology, specifically providing a bioreactor, including a container, a gas exchange tube, and a fixator. The gas exchange tube is at least partially disposed within the container. The gas exchange tube includes a first section and a second section connected together, each with multiple exchange holes on its wall. The fixator secures the gas exchange tube to the container. The fixator has fixing slots, the area of ​​which is not less than a preset value. The second section is disposed within the fixing slots, with at least some of the exchange holes located on the side of the second section opposite to the fixing slots. This invention, through the cooperation of the gas exchange tube and the fixator, effectively increases the number of exchange holes for gas exchange, meeting the gas consumption requirements of the target culture medium and resulting in good culture effects.

Description

Technical Field

[0001] This utility model relates to the field of biology, and in particular to a bioreactor. Background Technology

[0002] Cell culture refers to the technique of simulating the in vivo environment in vitro to enable cells to survive, grow, and maintain specific functions. It is widely used in cell biology, molecular biology, and other fields. Among these, the gaseous environment is a crucial factor that cannot be ignored in cell culture. The concentrations and interactions of different gases can affect cell metabolism, proliferation, differentiation, and functional status.

[0003] In existing technologies, an aeration plate is typically installed at the bottom of the cell culture container to generate bubbles and provide the necessary gas for cell growth. However, the number of aeration holes on the aeration plate for generating bubbles is limited. With other parameters remaining constant, it is impossible to generate enough bubbles for gas exchange, thus failing to meet the gas consumption requirements during cell culture and affecting the cell culture effect. Summary of the Invention

[0004] The bioreactor provided in this embodiment of the invention at least solves the problem that the number of aeration holes in conventional aeration plates is limited and cannot meet the gas consumption requirements during cell culture. By combining gas exchange tubes and fixation devices, the number of exchange holes is effectively increased to carry out gas exchange, thereby meeting the gas consumption requirements of the target culture object and achieving good culture results.

[0005] This invention provides a bioreactor, comprising a container; a gas exchange tube, at least partially disposed within the container; the gas exchange tube comprising a first section and a second section connected to each other, both the first and second sections having multiple exchange holes on their walls; a fixture for fixing the gas exchange tube to the container; the fixture having a fixing groove, the area of ​​the fixing groove on the fixture being not less than a preset value; the second section being disposed within the fixing groove, at least a portion of the exchange holes being disposed on the side of the second section opposite to the fixing groove.

[0006] In one embodiment of the present invention, the fixing slot includes a first spiral groove segment and a second spiral groove segment. The first spiral groove segment and the second spiral groove segment are both spirally arranged from the middle of the fixing device to the outside. The ends of the first spiral groove segment and the second spiral groove segment located in the middle of the fixing device are interconnected. The ends of the first spiral groove segment and the second spiral groove segment located on the outside of the fixing device are both located on the side wall of the fixing device on the same side.

[0007] In one embodiment of the present invention, the inner wall of the container is provided with a plurality of fixing members arranged at intervals in sequence, and at least a portion of the first pipe segment is connected to the fixing members.

[0008] In one embodiment of this utility model, the first pipe section includes: an air inlet section connected to the fixing member; an air outlet section connected to the fixing member; the air outlet section and the air inlet section are arranged side by side; a first bending section is disposed between the air inlet section and the second pipe section, and respectively connects the air inlet section and the second pipe section; and a second bending section is disposed between the air outlet section and the second pipe section, and respectively connects the air outlet section and the second pipe section; wherein, both the first bending section and the second bending section are bent toward the inner wall of the container.

[0009] In one embodiment of the present invention, the fixture includes a first fixing surface and a second fixing surface disposed opposite to each other. The first fixing surface is provided with the fixing slot, and the second fixing surface is provided with a first snap-fit ​​member. The bottom of the container is provided with a liquid outlet connector, and a second snap-fit ​​member is provided on the side of the liquid outlet connector near the fixture. The second snap-fit ​​member is snap-fitted to the first snap-fit ​​member.

[0010] In one embodiment of this utility model, a clearance hole is further provided on the second fixing surface, the clearance hole extending through to the first fixing surface; the first locking member includes a plurality of first locking blocks, which are sequentially spaced along the circumference of the clearance hole, and each first locking block is provided with a first locking hook; the liquid outlet connector is further provided with a third locking member, at least a portion of which is disposed within the clearance hole, the third locking member having a receiving hole and a second locking hook disposed outside the receiving hole; the second locking member is circumferentially disposed on the first fixing surface. On the outside of the three-clamp connector, the second clamp connector is provided with a first clamp slot and a plurality of second clamp blocks. The first clamp slot is clamped and connected to the first clamp hook. Each second clamp block is respectively disposed between two adjacent first clamp blocks and is clamped and connected to the first clamp block. The container is also provided with a stirrer. The stirrer includes a stirring shaft and a stirring paddle disposed on the stirring shaft. The stirring shaft includes a first end, which is disposed in the receiving hole. A second clamp slot is provided on the side wall of the first end, and the second clamp slot is clamped and connected to the second clamp hook.

[0011] In one embodiment of this utility model, an air inlet connector and an air outlet connector are provided on the top surface of the container, and both the air inlet connector and the air outlet connector are sealed to the container; the air inlet section of the gas exchange pipe is connected to the air inlet connector, and the air outlet section of the gas exchange pipe is connected to the air outlet connector.

[0012] In one embodiment of the present invention, the device further includes a gas supply component connected to the gas inlet connector, the gas supply component being used to introduce the target gas into the gas exchange pipe; and a pressure regulating component connected to the gas outlet connector, the pressure regulating component being used to adjust the gas pressure of the target gas.

[0013] In one embodiment of this utility model, the container is used to hold a biological culture medium; at least a portion of the gas exchange tube is disposed within the biological culture medium, and the gas exchange tube is configured to circulate a target gas; wherein the hydraulic pressure of the biological culture medium is less than the gas pressure of the target gas, and a target pressure difference is satisfied between the gas pressure of the target gas and the hydraulic pressure of the biological culture medium, so that the target gas passes through the exchange hole and forms attached bubbles on the outer wall of the orifice of the exchange hole, and the attached bubbles are used to exchange gases with the biological culture medium.

[0014] In one embodiment of this utility model, at least one of the following features is included: the wall thickness of the gas exchange tube. Satisfy the relation, The diameter of the exchange hole Satisfy the relation, The porosity of the exchange holes on the gas exchange pipe Satisfy the relation, The target pressure difference Satisfy the relation, .

[0015] Compared with the prior art, the above-mentioned technical solution of this utility model has the following beneficial effects:

[0016] The bioreactor of this invention achieves gas exchange with the biological culture medium by replacing the aeration plate in the prior art with a gas exchange tube. The wall area of ​​the gas exchange tube is larger than that of a conventional aeration plate, thus allowing for a greater number of exchange holes to be set on the gas exchange tube to meet the gas consumption requirements of the target culture medium and ensure good biological culture results.

[0017] Meanwhile, a retainer with a fixing slot secures the gas exchange tube to the container. During biological culture, the fixing slot keeps the gas exchange tube in place, preventing it from shaking or moving freely during aeration and stirring. This also prevents interference between the gas exchange tube and other components within the container, extending its lifespan. Furthermore, the fixing slot allows for the installation of longer gas exchange tubes within the container, enabling the creation of more exchange pores for gas exchange. This not only fully meets the gas consumption requirements of the target culture medium but also facilitates a rapid increase in the gas content of the biological culture medium. Attached Figure Description

[0018] The accompanying drawings, which are included to provide a further understanding of the present invention and constitute a part of this invention, illustrate exemplary embodiments of the present invention and, together with the description thereof, serve to explain the present invention and do not constitute an undue limitation thereof. In the drawings:

[0019] Figure 1 This is a schematic diagram of the structure of the bioreactor in a preferred embodiment of the present invention.

[0020] Figure 2 This is a schematic diagram of the internal structure of the container in a preferred embodiment of the present invention.

[0021] Figure 3 This is one of the structural schematic diagrams of the fixator in a preferred embodiment of this utility model.

[0022] Figure 4 This is a partial cross-sectional view of the bioreactor in a preferred embodiment of the present invention.

[0023] Figure 5 This is a schematic diagram of the gas exchange tube in a preferred embodiment of the present invention.

[0024] Figure 6 This is the second structural schematic diagram of the fixator in a preferred embodiment of this utility model.

[0025] Figure 7 This is a schematic diagram of the liquid outlet connector and stirring shaft in a preferred embodiment of the present invention.

[0026] Figure 8 This is a schematic diagram of the liquid outlet connector and the fixing device in a preferred embodiment of the present invention.

[0027] Figure 9 This is a cross-sectional view of the liquid outlet connector in a preferred embodiment of the present invention.

[0028] Figure 10 This is a cross-sectional view of the gas exchange tube in a preferred embodiment of the present invention.

[0029] Figure 11 This is a schematic diagram of the structure of the bioreactor in a preferred embodiment of the present invention.

[0030] The above figures include the following reference numerals:

[0031] 10. Container; 11. Top surface; 111. Air inlet connector; 112. Air outlet connector; 113. Liquid inlet connector; 12. Fixing component; 13. Liquid outlet connector; 131. Second snap-fit ​​component; 1311. First snap-fit ​​groove; 1312. Second snap-fit ​​block; 132. Third snap-fit ​​component; 1321. Receiving hole; 1322. Second snap-fit ​​hook; 14. Test connector;

[0032] 20. Biological culture medium; 21. Target culture object;

[0033] 30. Gas exchange pipe; 31. Gas exchange section; 311. Exchange port; 3121. Second pipe section; 3122. Inlet section; 3123. Outlet section; 3124. First bend section; 3125. Second bend section; 32. First connecting part; 33. Second connecting part;

[0034] 40. Target gas; 41. Adhering bubbles;

[0035] 51. Gas supply components; 52. Pressure regulating components; 53. Outer pipe body;

[0036] 60. Fixing device; 61. First fixing surface; 611. Fixing slot; 6111. First spiral groove segment; 6112. Second spiral groove segment; 62. Second fixing surface; 621. First snap-fit ​​component; 6211. First snap-fit ​​block; 6212. First snap-fit ​​hook; 63. Clearance hole;

[0037] 70. Stirrer; 71. Stirring shaft; 711. First end; 7111. Second locking groove; 712. Second end; 72. Stirring paddle; 73. Connecting component. Detailed Implementation

[0038] It should be noted that, where there is no conflict, the embodiments and features in the embodiments of this utility model can be combined with each other. The present utility model will now be described in detail with reference to the accompanying drawings and embodiments.

[0039] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the exemplary embodiments according to the present invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0040] Unless otherwise specifically stated, the relative arrangement, numerical expressions, and values ​​of the components and steps described in these embodiments do not limit the scope of this invention. It should also be understood that, for ease of description, the dimensions of the various parts shown in the drawings are not drawn to actual scale. Techniques, methods, and devices known to those skilled in the art may not be discussed in detail, but where appropriate, such techniques, methods, and devices should be considered part of the specification. In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limitations. Therefore, other examples of exemplary embodiments may have different values. It should be noted that similar reference numerals and letters in the following drawings denote similar items; therefore, once an item is defined in one drawing, it need not be further discussed in subsequent drawings.

[0041] It is important to note that the gaseous environment is a crucial factor that cannot be ignored during cell culture. The concentrations and interactions of different gases can affect cell metabolism, proliferation, differentiation, and functional status. Taking the culture of conventional stem cells as an example, oxygen needs to be introduced into the culture medium to allow for gas exchange between the oxygen and the stem cells in the culture medium, thus promoting cell growth.

[0042] In existing technologies, culture bags are typically used to culture stem cells. Besides the culture medium and stem cells, the culture bag also contains a stirrer and an aeration plate. The aeration plate is connected to an external air supply device and has multiple aeration holes. During stem cell culture, oxygen is supplied to the aeration plate via the external air supply device. The oxygen passes through the aeration holes in the aeration plate, forming numerous bubbles. These bubbles enter the culture medium, suspend, and exchange gases with the stem cells.

[0043] However, these aeration holes are usually located on the same surface of the aeration plate, and the number of aeration holes is limited by the size of the aeration plate. When the size of the aeration plate is limited, the number of aeration holes is also limited. When dealing with stem cells that have high gas consumption requirements, conventional aeration plates often cannot meet their gas requirements, affecting cell culture results.

[0044] To solve the above problems, refer to Figure 1 and Figure 2 As shown, this embodiment of the present invention provides a bioreactor, including a container 10, a gas exchange tube 30, and a fixator 60.

[0045] Container 10 is used to house the corresponding components and biological culture medium 20, providing a simulated in vivo environment for the target culture subject 21 for biological culture. Those skilled in the art can design different containers 10 according to actual needs, such as rigid containers or flexible containers. Preferably, in this embodiment of the invention, disposable containers are selected as containers 10. After use, sterilization and cleaning are not required; instead, new containers are replaced to effectively ensure the sterility of the biological culture and reduce the risk of cross-contamination.

[0046] The biological culture medium 20 is used to provide basic nutrition for the target culture object 21 and maintain its living environment. For example, it includes glucose, amino acids, vitamins, etc., to meet the energy requirements and material synthesis of the target culture object 21. The biological culture medium 20 is prior art. During biological culture, a container 10 pre-filled with the biological culture medium 20 can be selected, or a container 10 without the biological culture medium 20 can be selected, with the biological culture medium 20 added to the container 10 when needed. Alternatively, the biological culture medium 20 can be added to the container 10 during the culture process via a corresponding connector.

[0047] The target culture object 21 refers to the organism to be cultured, including cells, probiotics, etc. In this embodiment of the invention, the target culture object 21 is mainly cells, such as adherent cells (e.g., HEK293 cells), HeLa cells, MSC mesenchymal stem cells, etc. Different target culture objects 21 have different gas requirements. For example, oxygen needs to be introduced when culturing stem cells, while some other cells require gases such as carbon dioxide.

[0048] The gas exchange tube 30 is the core component of the bioreactor described in this invention, used to replace the aeration disc in the prior art, and to provide the target gas 40 required for the target culture medium 21. Those skilled in the art can configure the gas exchange tube 30 with the appropriate target gas 40 according to the actual target culture medium 21.

[0049] In order to provide target gas 40 to the target culture object 21 inside the container 10 to meet the gas requirements of the target culture object 21, at least a portion of the gas exchange tube 30 is disposed inside the container 10 to introduce the external target gas 40 into the interior of the container 10.

[0050] Those skilled in the art can configure the arrangement of the gas exchange pipe 30 and the container 10 according to actual needs. For example, only a portion of the gas exchange pipe 30 can be placed inside the container 10, and the portion of the gas exchange pipe 30 outside the container 10 can be connected to a corresponding component to achieve ventilation. Alternatively, the entire gas exchange pipe 30 can be placed inside the container 10, and a corresponding connector can be provided on the container 10 to connect the gas exchange pipe 30 inside the container 10 with an external pipeline to achieve ventilation.

[0051] The gas exchange tube 30 includes a gas exchange section 31 for immersion in the biological culture medium 20. Those skilled in the art can configure different gas exchange sections 31 on different gas exchange tubes 30 according to actual needs to meet different biological culture requirements. For example, if it is required that the container 10 be filled with the biological culture medium 20, then the portion of the gas exchange tube 30 located inside the container 10 can be configured as a gas exchange section 31. If there is a liquid level requirement for the biological culture medium 20 inside the container 10, then only the portion of the gas exchange tube 30 located inside the container 10 and immersed in the biological culture medium 20 can be configured as a gas exchange section 31.

[0052] The gas exchange unit 31 includes a first pipe section and a second pipe section 3121 that are connected to each other. Multiple exchange holes 311 are provided on the walls of both the first pipe section and the second pipe section 3121. Those skilled in the art can set the specific shape of the exchange holes 311 according to actual needs, such as square through holes, triangular through holes, etc. Preferably, the exchange holes 311 are set as circular through holes.

[0053] The gas exchange tube 30 is configured to allow the flow of target gas 40, which can pass through the exchange hole 311 and form bubbles, and exchange gases with the target culture object 21 in the biological culture medium 20 through the bubbles.

[0054] Understandably, when an aeration tray is set in container 10, the size of the aeration tray is often limited by the volume of container 10, resulting in a limited area of ​​the tray surface where the aeration holes are located. The limited area of ​​the tray surface can only accommodate a limited number of aeration holes. When facing a target culture object 21 with high gas consumption requirements, conventional aeration trays often cannot meet its gas consumption requirements, affecting the cell culture effect.

[0055] The gas exchange tube 30, being a tubular structure, differs from a conventional aeration plate. Firstly, the wall area of ​​the gas exchange tube 30 is larger than that of a conventional aeration plate, meaning that more exchange holes 311 can be set on the gas exchange tube 30 for gas exchange, thereby meeting the gas consumption requirements of the target culture medium 21 and ensuring good biological culture results. Secondly, thanks to the elongated structure of the gas exchange tube 30, the internal space of the container 10 can be fully utilized to accommodate it, allowing the biological culture medium 20 in different parts of the container 10 to exchange gases with the air bubbles, thus meeting the gas requirements of the target culture medium 21.

[0056] The gas exchange tube 30 can be a hollow tube. When the target gas 40 is flowing through it for gas exchange, the target gas 40 is introduced into one axial end of the gas exchange tube 30 and discharged from the other end, thus achieving gas exchange. Alternatively, a blind hole can be set along the axial direction on a shaft to obtain a gas exchanger, in which the target gas 40 is introduced only through the opening of the blind hole.

[0057] It is worth noting that waste gas, such as carbon dioxide, is generated during the biological culture process. The waste gas discharge function can be achieved by setting other corresponding components on the container 10, rather than through the gas exchange pipe 30.

[0058] When the gas exchange tube 30 is configured as a tubular structure, round tubes, square tubes, etc., can be selected. Among them, round tubes have many ready-made products in the existing technology, which is conducive to cost control. Under the same inner diameter, the outer wall contact area of ​​the square tube is larger, which is conducive to setting more exchange holes 311, forming more bubbles, thereby increasing the gas content in the biological culture medium 20.

[0059] The flow of the target gas 40 is prior art. Those skilled in the art can integrate corresponding components on the container 10 to achieve gas supply according to actual needs, or they can install corresponding components on the outside of the container 10 to achieve gas supply. For example, an air pump or air compressor, pressure gauge, etc. can be configured to provide the target gas 40.

[0060] Considering that the gas exchange tube 30 is not integrally formed with the container 10, and other components, such as a stirrer 70, are often installed in the container 10, the gas exchange tube 30 may interfere with these components during biological culture. Therefore, it is necessary to fix the gas exchange tube 30. Those skilled in the art can devise appropriate methods to fix the gas exchange tube 30 according to actual needs, such as welding, snap-fitting, etc.

[0061] In this embodiment of the invention, the gas exchange tube 30 is fixed to the container 10 by a fixture 60. The fixture 60 is connected to the container 10, and those skilled in the art can determine the connection method between the fixture 60 and the container 10 according to actual needs, such as welding or snap-fitting. Those skilled in the art can determine the shape of the fixture 60 according to actual needs, such as square or disc-shaped. Those skilled in the art can determine the specific installation position of the fixture 60 according to actual needs, such as placing it on the side wall or bottom of the container 10.

[0062] The fixture 60 is provided with a fixing slot 611, and the second pipe section 3121 is set in the fixing slot 611. By using the fixture 60 with the fixing slot 611, the gas exchange pipe 30 can be effectively fixed, preventing it from shaking during ventilation and stirring.

[0063] The second pipe section 3121 is located in the fixing slot 611 of the fixture 60. The wall of the fixing slot 611 will block the exchange hole 311 on the second pipe section 3121. Therefore, during assembly, care should be taken to place at least part of the exchange hole 311 on the side of the second pipe section 3121 away from the fixing slot 611 to ensure that the second pipe section 3121 located in the fixing slot 611 can carry out normal gas exchange through these exchange holes 311.

[0064] Meanwhile, the area of ​​the fixed slots 611 on the fixture 60 is not less than a preset value. To ensure that the area of ​​the fixed slots 611 meets the requirements, those skilled in the art can set the specific position, number and arrangement of the fixed slots 611 on the fixture 60 according to actual needs.

[0065] For example, the fixture 60 is configured as a cuboid structure with six faces, and a fixing slot 611 is provided on each face of the fixture 60. The second pipe segment 3121 of the gas exchange pipe 30 is respectively disposed in the corresponding fixing slot 611. Alternatively, the fixing slot 611 is provided on only one face of the fixture 60, and the fixing slot 611 is configured as a coiled structure or a snake-shaped structure to improve space utilization and ensure that the arrangement area of ​​the fixing slots 611 on the fixture 60 meets the preset value requirement.

[0066] The preset value is an empirical value that can be flexibly adjusted according to actual conditions. For example, it can be set to no less than 20% of the total surface area of ​​the fastener 60; for example, it can be set to 20%, 30%, 40%, etc. of the total surface area of ​​the fastener 60.

[0067] With the fixture 60 in place, the gas exchange tube 30 can be secured, preventing it from shaking or moving freely during aeration or stirring, thus preventing interference between the gas exchange tube 30 and other components within the container 10 and extending its service life. Simultaneously, the fixture 60 allows for the installation of a longer gas exchange tube 30 within the container 10 using the fixing slots 611, enabling the installation of more exchange holes 311 for gas exchange. This not only fully meets the gas consumption requirements of the target culture medium 21 but also facilitates a rapid increase in the gas content of the biological culture medium 20.

[0068] The bioreactor of this invention achieves gas exchange with the biological culture medium 20 by replacing the aeration plate in the prior art with a gas exchange pipe 30. The wall area of ​​the gas exchange pipe 30 is larger than that of a conventional aeration plate, so a greater number of exchange holes 311 are provided on the gas exchange pipe 30 for gas exchange, thereby meeting the gas consumption requirements of the target culture object 21 and ensuring good biological culture results.

[0069] Meanwhile, the gas exchange tube 30 is fixed to the container 10 by the fixture 60 with the fixing slot 611. During the biological culture process, the fixing slot 611 secures the gas exchange tube 30, preventing it from shaking or moving randomly during aeration and stirring, thus preventing interference between the gas exchange tube 30 and other components inside the container 10 and extending its service life. Furthermore, the fixing slot 611 allows for the installation of a longer gas exchange tube 30 within the container 10, enabling the installation of more exchange holes 311 for gas exchange. This not only fully meets the gas consumption requirements of the target culture medium 21 but also facilitates a rapid increase in the gas content of the biological culture medium 20.

[0070] Reference Figure 3 As shown, in some embodiments of the bioreactor described in this utility model, the fixing slot 611 is configured as a coiled structure. Specifically, the fixing slot 611 includes a first spiral groove segment 6111 and a second spiral groove segment 6112. Both the first spiral groove segment 6111 and the second spiral groove segment 6112 are spirally arranged from the middle of the fixture 60 outwards. The ends of the first spiral groove segment 6111 and the second spiral groove segment 6112 located in the middle of the fixture 60 are interconnected, and the ends of the first spiral groove segment 6111 and the second spiral groove segment 6112 located on the outer side of the fixture 60 are both located on the sidewall of the fixture 60 on the same side.

[0071] To fully utilize the area of ​​the fixture 60 and improve space utilization, the first spiral groove segment 6111 and the second spiral groove segment 6112 are at least partially arranged side by side. When both the first spiral groove segment 6111 and the second spiral groove segment 6112 are located on the same surface of the fixture 60, the middle part of the fixture 60 can be a circular area with the center of the two groove segments as its center and a radius set to a preset size. When the fixture 60 is a cuboid structure, the sidewalls on the same side of the fixture 60 refer to the same rectangular surface; when the fixture 60 is a cylindrical structure, the sidewalls on the same side of the fixture 60 refer to the semi-cylindrical curved surfaces obtained by dividing the cylinder along its axis.

[0072] By setting the fixing slot 611 of this coiled structure, the area of ​​the fixture 60 can be fully utilized to set a longer fixing slot 611. By fixing the longer second tube segment 3121 with the extended fixing slot 611, the contact area between the outer wall of the gas exchange tube 30 and the biological culture medium 20 is effectively increased, which facilitates more gas bubbles to exchange gas and thus rapidly increases the gas content.

[0073] In some other embodiments, when the fixing slot 611 is configured as a snake-shaped structure, the fixing slot 611 includes a plurality of first U-shaped bending slot segments and second U-shaped bending slot segments connected in sequence, with the bending directions of the two types of U-shaped bending slot segments being opposite.

[0074] Reference Figure 1 , Figure 2 and Figure 5 As shown, in some embodiments of the bioreactor described in this utility model, an air inlet connector 111 and an air outlet connector 112 are provided on the top surface 11 of the container 10.

[0075] In existing bioreactors, connectors are mature existing technologies, including Luer connectors, threaded connectors, and flange connectors. Those skilled in the art can configure specific models of the two types of connectors according to actual needs. Positioning the inlet connector 111 and the outlet connector 112 on the top surface 11 of the container 10 facilitates installation work by personnel.

[0076] It is worth noting that both the air inlet connector 111 and the air outlet connector 112 are sealed to the container 10 to avoid cross-contamination or leakage. Preferably, the two connectors are sealed to the container 10 by welding.

[0077] With both an inlet connector 111 and an outlet connector 112 installed on the container 10, the gas exchange pipe 30 does not require the integration of additional components. It can be connected to corresponding components outside the container 10 via the inlet connector 111 and the outlet connector 112 to achieve pressure control of the target gas 40. This effectively improves the space utilization within the container 10, reduces the volume of the bioreactor, and enhances portability. Furthermore, the gas inlet connector 111, in conjunction with corresponding components, enables gas introduction, while the outlet connector 112, in conjunction with corresponding components, enables precise pressure control. This spatial division of labor effectively decouples functions, contributing to improved stability, safety, and reliability.

[0078] The first section of the gas exchange pipe 30 includes an inlet section 3122 and an outlet section 3123. The inlet section 3122 is connected to the inlet connector 111 and the second section 3121, respectively. The outlet section 3123 is connected to the outlet connector 112 and the second section 3121, respectively.

[0079] A first connecting part 32 is provided at the end of the air intake section 3122 opposite to the second pipe section 3121, and the air intake section 3122 is connected to the air intake connector 111 through the first connecting part 32. A second connecting part 33 is provided at the end of the air outlet section 3123 opposite to the second pipe section 3121, and the air outlet section 3123 is connected to the air outlet connector 112 through the second connecting part 33.

[0080] Technicians in the field can configure two types of connection parts and their specific connection methods according to actual needs. For example, the intake section 3122 is Luer-connected to the intake connector 111 via the first connection part 32 to achieve communication; the exhaust section 3123 is Luer-connected to the exhaust connector 112 via the second connection part 33 to achieve communication. This can effectively ensure the sealing and stability of the connection.

[0081] Reference Figure 4 As shown, in some embodiments, the bioreactor of this invention further includes a gas supply component 51, which is connected to an inlet connector 111 and is used to introduce the target gas 40 into the gas exchange pipe 30; or / and, it also includes a pressure regulating component 52, which is connected to an outlet connector 112 and is used to control the pressure of the target gas 40. Thus, there are three possible scenarios.

[0082] In the first type, the bioreactor only includes a gas supply unit 51, which is connected to the gas inlet connector 111 on the container 10 to introduce the target gas 40 into the gas exchange pipe 30. The component responsible for controlling the gas pressure of the target gas 40 can be configured independently.

[0083] The second type of bioreactor only includes a pressure regulating component 52, which is connected to the gas outlet connector 112 on the container 10 to control the pressure of the target gas 40 during the biological culture process. The component responsible for introducing the target gas 40 into the gas exchange pipe 30 can be configured independently.

[0084] The third type of bioreactor is equipped with a gas supply component 51 and a pressure regulating component 52. The gas supply component 51 is directly connected to the gas inlet connector 111 on the container 10 to introduce the target gas 40 into the gas exchange pipe 30. The pressure regulating component 52 is connected to the gas outlet connector 112 on the container 10 to control the gas pressure of the target gas 40 during the biological culture process.

[0085] Both the air supply component 51 and the pressure regulating component 52 are existing technologies, and those skilled in the art can configure them according to actual needs. They are illustrated in the accompanying drawings as block diagrams. For example, the air supply component 51 can be configured as a peristaltic pump, compressor, etc., and the pressure regulating component 52 can be configured as a pressure reducing valve, safety valve, pressure regulating valve, etc.

[0086] When an air supply component 51 and / or a pressure regulating component 52 are provided, an outer tube 53 can be provided to achieve communication between the corresponding connectors and components. Preferably, the outer tube 53 is a silicone tube, which can effectively adapt to the peristaltic pump and prevent liquid from overflowing and causing contamination.

[0087] This structure allows for precise control of the intake volume and aeration rate of the target gas 40 by working well with the inlet connector 111 and outlet connector 112 on container 10. Specifically, the intake volume of the bioreactor can be estimated by adjusting the supply rate of the gas supply component 51; how this estimation is done is existing technology and will not be elaborated further. With a fixed opening size for the exchange port 311, the pressure difference between the target gas 40 and the hydraulic pressure of the biological culture medium 20 can be adjusted by using the pressure regulating component 52, thereby controlling the bubble size and achieving different aeration rates to meet the needs of different biological culture stages, offering greater flexibility.

[0088] Preferably, the retainer 60 is located at the bottom of the container 10. With an air inlet 111 and an air outlet 112 provided on the top surface 11 of the container 10, the retainer 60 effectively secures the gas exchange tube 30, preventing it from shaking during aeration and stirring. Furthermore, it ensures that the biological culture medium 20 in different parts of the container 10 can fully exchange gases with the air bubbles to meet the gas requirements of the target culture medium 21.

[0089] Reference Figure 4 and Figure 5As shown, in some embodiments of the bioreactor described in this utility model, the inner wall of the container 10 is provided with a plurality of sequentially spaced fasteners 12, and at least a portion of the first pipe section is connected to the fasteners 12. Those skilled in the art can configure the fasteners 12 according to actual needs; preferably, the fasteners 12 are configured as double-ended pipe clamps.

[0090] Both the inlet section 3122 and the outlet section 3123 of the first pipe section are connected to the fixing member 12, and the outlet section 3123 is arranged in parallel with the inlet section 3122. By setting this structure, the contact interference between the gas exchange pipe 30 and the agitator 72 can be further reduced, ensuring the stability of the bioreactor and extending the service life of the gas exchange pipe 30 and the agitator 72.

[0091] Furthermore, refer to Figure 5 As shown, in some embodiments of the bioreactor of this invention, the first pipe section further includes a first bend section 3124 and a second bend section 3125. The first bend section 3124 is disposed between the air inlet section 3122 and the second pipe section 3121, and is connected to both the air inlet section 3122 and the second pipe section 3121. The second bend section 3125 is disposed between the air outlet section 3123 and the second pipe section 3121, and is connected to both the air outlet section 3123 and the second pipe section 3121. Both the first bend section 3124 and the second bend section 3125 are bent towards the inner wall of the container 10.

[0092] During biological culture, a stirrer 70 is typically installed inside the container 10. The stirrer 70 includes a stirring shaft 71 and stirring paddles 72 mounted on the stirring shaft 71. To achieve thorough mixing, multiple stirring paddles 72 are often arranged sequentially at intervals on the stirring shaft 71 to stir simultaneously. Although a retainer 60 and a fixing member 12 are provided inside the container 10 to fix the gas exchange tube 30, the portion of the gas exchange tube 30 located between the retainer 60 and the fixing member 12 will inevitably interfere with the stirring paddles 72.

[0093] Based on this, the corresponding part of the gas exchange tube 30 is bent toward the inner wall of the container 10 to obtain the first bent section 3124 and the second bent section 3125, which can effectively reduce the possibility of contact interference between the gas exchange tube 30 and the stirring paddle 72 and extend the service life of the gas exchange tube 30 and the stirring paddle 72.

[0094] Reference Figure 6 , Figure 7 and Figure 8As shown, in some embodiments of the bioreactor described in this utility model, the fixator 60 includes a first fixing surface 61 and a second fixing surface 62 disposed opposite to each other. Preferably, the fixator 60 is configured as a cylindrical structure to better fit the bag-shaped container 10. The first fixing surface 61 is provided with a fixing groove 611, and the second fixing surface 62 is provided with a first snap-fit ​​member 621, which is used to connect with the container 10.

[0095] Correspondingly, a liquid outlet connector 13 is provided at the bottom of the container 10, which enables the container 10 to drain liquid. The draining structure of the liquid outlet connector 13 is prior art and will not be described in detail. Preferably, the liquid outlet connector 13 is welded and fixed to the bottom of the container 10. A second snap-fit ​​member 131 is provided on the side of the liquid outlet connector 13 near the fixture 60, and the second snap-fit ​​member 131 is snap-fitted to the first snap-fit ​​member 621. Those skilled in the art can design the specific structures of the two snap-fit ​​members according to actual needs to achieve snap-fit ​​fixation. By providing the first snap-fit ​​member 621 and the second snap-fit ​​member 131, the separate container 10 and the fixture 60 can be easily assembled and fixed.

[0096] Furthermore, refer to Figure 7 , Figure 8 and Figure 9 As shown, in some embodiments of the bioreactor described in this utility model, the second fixed surface 62 is also provided with a clearance hole 63, which extends through to the first fixed surface 61. The clearance hole 63 is mainly used to avoid the stirring shaft 71 of the stirrer 70.

[0097] Specifically, the stirring shaft 71 includes a first end 711 and a second end 712. The first end 711 is located at the bottom of the container 10, while the second end 712 is located at the top of the container 10. A connecting member 73 is provided on the top surface 11 of the container 10, and the connecting member 73 is rotatably connected to the container 10. The second end 712 of the stirring shaft 71 is connected to the connecting member 73. In actual use, the connecting member 73 is driven to rotate relative to the container 10 by a corresponding driving component, thereby driving the stirring shaft 71 and the stirring paddle 72 to rotate, thus achieving stirring. A second locking groove 7111 is provided on the side wall of the first end 711.

[0098] The fixing groove 611 is coiled around the outside of the opening of the avoidance hole 63 to avoid contact interference between the second pipe section 3121 inside the fixing groove 611 and the stirring shaft 71.

[0099] The first latching component 621 includes a plurality of first latching blocks 6211, which are arranged sequentially at intervals along the circumference of the clearance hole 63. Each first latching block 6211 is provided with a first latching hook 6212.

[0100] Correspondingly, the liquid outlet connector 13 is also provided with a third locking member 132, at least part of which is located within the clearance hole 63. The third locking member 132 is provided with a receiving hole 1321 and a second locking hook 1322 located outside the receiving hole 1321. The first end 711 of the stirring shaft 71 is located within the receiving hole 1321.

[0101] The second snap-fit ​​member 131 of the liquid outlet connector 13 is arranged around the outside of the third snap-fit ​​member 132. The second snap-fit ​​member 131 is provided with a first snap-fit ​​groove 1311 and a plurality of second snap-fit ​​blocks 1312. The first snap-fit ​​groove 1311 is snap-fitted to the first snap-fit ​​hook 6212; each second snap-fit ​​block 1312 is respectively disposed between two adjacent first snap-fit ​​blocks 6211 and snap-fitted to the first snap-fit ​​block 6211. The second snap-fit ​​groove 7111 on the first end 711 is snap-fitted to the second snap-fit ​​hook 1322.

[0102] This structure ensures that the liquid outlet connector 13 and the stirring shaft 71, as well as the liquid outlet connector 13 and the retainer 60, are fixed together. For the liquid outlet connector 13 and the stirring shaft 71, the engagement of the receiving hole 1321 and the first end 711, as well as the second locking groove 7111 and the second locking hook 1322, prevents the stirring shaft 71 from moving radially or axially while still allowing it to rotate. For the liquid outlet connector 13 and the retainer 60, the engagement of the first locking block 6211 and the second locking block 1312, as well as the engagement of the first locking groove 1311 and the first locking hook 6212, allows for easy assembly of the retainer 60 while effectively preventing the retainer 60 from rotating or moving axially along the stirring shaft 71 during its rotation.

[0103] Preferably, the stirring shaft 71 can move axially relative to the connecting component 73, enabling the stirring shaft 71 to be compatible with containers 10 of different sizes and models. For example, a pin is provided on the second end 712 of the stirring shaft 71, while a socket is provided on the connecting component 73. A slot corresponding to the pin is provided on the wall of the socket. Through the engagement of the socket and the pin, axial movement is achieved while preventing relative rotation between the stirring shaft 71 and the connecting component 73.

[0104] As is well known, stem cells lack cell walls, making them extremely sensitive to mechanical stress. During stem cell culture, the mechanical stress experienced by stem cells mainly comes from the shear force generated by the rotation of the agitator and the shear force generated when suspended air bubbles burst. The shear force generated by the agitator 72 at low speeds is negligible; therefore, the greatest source of shear force during stem cell culture is the shear force generated when suspended air bubbles burst.

[0105] When oxygen is directly introduced through the gas exchange tube 30 or the existing aeration plate, due to the low density of the gas, the oxygen enters the culture medium and is first encapsulated and forms bubbles due to the surface tension of the culture medium. Then, it rises due to the buoyancy of the culture medium. During the rising process, due to the action of the agitator 72, the bubbles continuously aggregate from small bubbles into large bubbles and break down from large bubbles into small bubbles. The shear force generated in this process can cause stem cells to be damaged and die, resulting in a low cell survival rate.

[0106] Meanwhile, because the bubbles rise to the surface due to buoyancy in the culture medium, their residence time in the medium is relatively short. This short residence time makes it difficult to meet the requirements for increasing the oxygen content of the culture medium, and thus, to meet the oxygen consumption of stem cells. In existing technologies, the conventional solution is to increase the aeration rate to increase the number of bubbles in the culture medium. However, an increased number of bubbles means more bubble bursts, generating more shear forces, which in turn leads to an increased stem cell mortality rate.

[0107] Furthermore, depending on the actual operating conditions, the aperture of the aeration holes on different aeration plates varies, in order to control the size of the introduced air bubbles. However, during actual culture, opening the culture bag to replace the aeration plate can easily lead to cross-contamination, affecting the purity and consistency of stem cells. Therefore, once an aeration plate is selected, it is often difficult to change, the aeration rate cannot be adjusted, and flexibility is low.

[0108] To solve the above problems, in some embodiments of the bioreactor described in this utility model, the gas pressure of the target gas 40 is controlled so that the target gas 40 passes through the exchange hole 311 and forms attached bubbles 41 on the outer wall of the orifice of the exchange hole 311. The attached bubbles 41 are used to exchange gases with the biological culture medium 20.

[0109] With the target gas 40 flowing through the gas exchange tube 30, the pressure of the target gas 40 is controlled to ensure a target pressure difference is achieved inside and outside the gas exchange tube 30. Specifically, in this embodiment, the internal pressure of the gas exchange tube 30, i.e., the pressure of the target gas 40, can be controlled and adjusted; the external pressure of the gas exchange tube 30 is the hydraulic pressure of the biological culture medium 20. The hydraulic pressure of the biological culture medium 20 is less than the pressure of the target gas 40.

[0110] Reference Figure 2 , Figure 10 and Figure 11As shown, under the condition of a target pressure difference, the biological culture medium 20 located outside the gas exchange tube 30 cannot overcome the target pressure difference and enter the gas exchange tube 30. It can only rely on surface tension to encapsulate part of the target gas 40, forming attached bubbles 41. However, the target gas 40 flowing inside the gas exchange tube 30 can pass through the exchange hole 311 and enter the biological culture medium 20, where it is encapsulated and forms attached bubbles 41 on the outer wall of the pore of the exchange hole 311, maintaining its attachment. The size of the attached bubbles 41 may change due to gas exchange or pressure control.

[0111] It is worth noting that when the target gas 40 enters the biological culture medium 20, it does not become a suspended bubble as in the prior art, but rather forms an attached bubble 41 on the outer wall of the orifice of the exchange port 311. That is, the attached bubble 41 adheres to the outer wall of the gas exchange tube 30. Of course, in cases where the attached bubble 41 is large, exceeding the orifice diameter of the exchange port 311, the attached bubble 41 may also partially contact the outer wall of the exchange tube near the orifice. Those skilled in the art can set a corresponding target pressure difference according to actual needs, so that the target gas 40 forms attached bubbles 41, rather than detaching from the gas exchange tube 30 and suspending in the biological culture medium 20.

[0112] Based on this, firstly, the attached bubbles 41 do not frequently aggregate and disperse, generating shear forces, like conventional suspended bubbles in the prior art. This effectively reduces the number of target culture objects 21 lost or killed due to shear forces, thus improving the survival rate of the target culture objects 21.

[0113] Secondly, since the attached bubbles 41 are attached to the gas exchange tube 30, they will not rise due to liquid buoyancy like suspended bubbles in the prior art. This ensures more complete gas exchange, effectively increasing the gas content of the biological culture medium 20 and meeting the gas consumption of the target culture object 21.

[0114] Finally, although the size of the exchange holes 311 on the gas exchange tube 30 is fixed, the size of the attached bubbles 41 can still be adjusted by controlling the gas pressure of the target gas 40. With the size of the attached bubbles 41 adjustable, the bioreactor can achieve different aeration rates to meet the needs of different biological cultivation stages, thus offering greater flexibility.

[0115] In actual biological culture, taking stem cell culture as an example, the corresponding stem cells and biological culture medium 20 are first placed in container 10, such that the biological culture medium 20 submerges at least part of the gas exchange tube 30. Then, oxygen is introduced into the gas exchange tube 30, and the oxygen pressure is controlled so that oxygen molecules can pass through the exchange pores 311 of the gas exchange tube 30 into the biological culture medium 20. Under the action of the surface tension of the biological culture medium 20, they are enveloped by the biological culture medium 20, forming attached bubbles 41 on the outer wall of the pore opening 311. The oxygen molecules in the attached bubbles 41 can dissolve into the biological culture medium 20 and achieve gas exchange with the stem cells through the transfer of the biological culture medium 20. It is worth noting that the formation of attached bubbles 41 and gas exchange may occur simultaneously.

[0116] Furthermore, refer to Figure 10 and Figure 11 As shown, the bioreactor of this invention, in some embodiments, includes at least one of the following features:

[0117] Wall thickness of gas exchange tube Satisfy the relation, .

[0118] Orifice diameter of the exchange port Satisfy the relation, .

[0119] The porosity of the exchange pores on the gas exchange tube Satisfy the relation, .

[0120] Target pressure difference Satisfy the relation, .

[0121] The wall thickness of gas exchange tube 30 The aperture of the exchange hole 311 and the porosity of the exchange hole 311 on the gas exchange tube 30 These are key parameters of the gas exchange tube 30. These parameters can affect the target pressure difference between the gas pressure of the target gas 40 and the hydraulic pressure of the biological culture medium 20. This determines whether the target gas 40 can form attached bubbles 41 on the outer wall of the orifice of the exchange hole 311, as well as the size and distribution of the attached bubbles 41.

[0122] Among them, the target pressure difference and the wall thickness of gas exchange tube 30 It is positively correlated with the target pressure difference. and the aperture of the exchange hole 311 Opening ratio They are all negatively correlated.

[0123] Specifically, regarding the wall thickness of the gas exchange tube 30... If the thickness is too small, less than 0.2mm, the gas exchange tube 30 is prone to damage during installation, making it difficult to meet usage requirements. The wall thickness of the gas exchange tube 30... When the thickness is too large, exceeding 0.5mm, while the tube strength increases, the difficulty of drilling holes in the tube also increases significantly. The exchange hole 311 is more likely to be machined as a blind hole instead of a through hole. With an increased proportion of blind holes, the air permeability will decrease for the same parameters. Furthermore, it also increases installation difficulty and cost. And the wall thickness of the gas exchange tube 30... While meeting the requirements of 0.2mm to 0.5mm, the gas exchange tube 30 combines flexibility with pressure resistance and is easy to install.

[0124] The diameter of the exchange hole 311 If the pore size is too small, less than 0.1 μm, the air permeability of the exchange orifice 311 will decrease significantly, and it will be prone to clogging, making manufacturing difficult. Actual measurements show that the current limiting pore size for the exchange orifice 311 is 0.05 μm. [The last sentence appears to be incomplete and possibly refers to a different topic: "Within the pore size of the exchange orifice 311..."] If the pore size is too large, exceeding 3 μm, even slight pressure fluctuations can cause the attached air bubbles 41 to detach from the gas exchange tube 30 and become suspended in the biological culture medium 20. Furthermore, if the target gas 40 is not adequately filtered, the large-aperture exchange pores 311 significantly increase the likelihood of bacterial entry and contamination. For reference, the diameter of bacteria is approximately 0.5 to 5 μm, the diameter of stem cells is approximately 8 μm, the diameter of water molecules is approximately 0.4 nm, and the diameter of oxygen molecules is approximately 0.346 nm.

[0125] The orifice ratio refers to the ratio of the total area of ​​the holes in a given area of ​​an exchange pipe to the surface area of ​​that area. If the opening ratio is too low, less than 30%, the air permeability will be greatly reduced, often requiring increased air supply pressure to maintain the air permeability, thus increasing air consumption. (Regarding the opening ratio...) When the value is too large, exceeding 80%, the measured open area ratio... For every 10% increase, the effective bearing area of ​​the material decreases by about 15%, which will cause permanent deformation of the gas exchange tube 30 when the pressure exceeds the standard value, and will easily cause internal blockage of the gas exchange tube 30.

[0126] Determine the wall thickness of gas exchange tube 30 The aperture of the exchange hole 311 and porosity In this case, it is also necessary to determine the gas viscosity of target gas 40. and gas flow rate The gas viscosity of target gas 40 and gas flow rate How to measure this is existing technology and will not be elaborated further. After determining the relevant parameters, the target pressure difference can be calculated based on these parameters. .

[0127] Specifically, it is expressed as:

[0128] .

[0129] In the formula, This is the Kozeny-Kaman constant, typically with a value between 3 and 5.

[0130] Below, we will take MSC mesenchymal stem cells as an example and demonstrate their culture using bioreactors with different parameters.

[0131] First, MSC mesenchymal stem cells were expanded and cultured in culture dishes. After 4 days, the cell density reached 8.54 × 10⁻⁶ cells / day. 6 The cell count was 94.81% (cells per mL).

[0132] Next, these MSC mesenchymal stem cells were seeded into bioreactors with different parameters and cultured in existing devices. During culture, Stem-Ex hMSC expansion medium (manufacturer: Yishengke) was used as the basal medium, and the cells were cultured at a constant temperature of 36.5℃. The gas flow rate was determined according to different cell growth stages. After 5 days of culture, cells were sampled and trypsin digested. The viable cell density and cell viability were calculated using a cell counter.

[0133] Example 1: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0134] Example 2: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0135] Example 3: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0136] Example 4: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 30%.

[0137] Example 5: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set it to 80%.

[0138] Example 6: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0139] Example 7: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0140] Comparative Example 1: The wall thickness of gas exchange tube 30 in a bioreactor. Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0141] Comparative Example 2: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0142] Comparative Example 3: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set it to 20%.

[0143] Comparative Example 4: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 90%.

[0144] Comparative Example 5: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0145] Comparative Example 6: Wall thickness of gas exchange tube 30 in a bioreactor Set as The aperture of the exchange hole 311 Set as Open area ratio Set to 55%.

[0146] Comparative Example 7: Using a conventional device with an aeration plate in the prior art, the aeration hole diameter on the aeration plate is... .

[0147] It is worth noting that in the actual culture process, the target pressure difference needs to be adjusted according to the different gas consumption of cells in the early and middle stages of stem cell culture. Adjustments will be made.

[0148] Among them, for the target pressure difference of part of the gas exchange tube 30 The calculated result will exceed 0.65 bar. In this case, it is not advisable to use it according to the calculated value. It is necessary to increase the oxygen content of the introduced gas to control it and prevent the target pressure difference from being too large and damaging the gas exchange tube 30.

[0149] If the calculated target pressure difference is too small, the actual pipe resistance and whether pressure control can actually be achieved need to be considered. If not, the gas exchange pipe 30 needs to be replaced to adjust the relevant parameters and the target pressure difference needs to be recalculated.

[0150] Table 1. Culture results of different samples

[0151]

[0152] Based on the comparison of Comparative Examples 1, 2, 1, 2, and 3 in Table 1, it can be seen that in Comparative Example 1, the wall thickness of the gas exchange tube 30 was too small, causing the tube to break after pressurization, thus preventing the experiment from continuing. In Comparative Example 2, the wall thickness of the gas exchange tube 30 was too large, increasing the blind hole rate and the target pressure difference, resulting in poor gas permeability and a low final cell survival rate. In actual use, it is necessary to increase the oxygen content of the gas to improve the survival rate.

[0153] Based on the comparison of Comparative Examples 3, 4, 2, 4, and 5 in Table 1, it can be seen that in Comparative Example 3, the opening ratio of the gas exchange tube 30 is too small, resulting in poor air permeability and a low final cell survival rate. In Comparative Example 4, the opening ratio of the gas exchange tube 30 is too large. In actual use, after air is introduced, under the action of the stirring paddle 72, the attached air bubbles 41 detach from the outer wall of the pores of the gas exchange tube 30 and enter the liquid suspension, resulting in a low final cell survival rate.

[0154] Based on the comparison of Comparative Examples 5, 6, 2, 6, and 7 in Table 1, it can be seen that in Comparative Example 5, the pore size of the exchange hole 311 in the gas exchange tube 30 is too small, resulting in the gas source pressure not meeting the usage requirements and virtually no attached bubbles 41 being generated. In Comparative Example 6, the pore size of the exchange hole 311 is too large. In actual use, after aeration, under the action of the stirring paddle 72, the attached bubbles 41 detach from the outer wall of the pore of the gas exchange tube 30 and enter the liquid suspension, resulting in a low final cell survival rate.

[0155] Based on the comparison of Examples 1, 2, 3, 4, 5, 6, 7 and Comparative Example 7 in Table 1, it can be seen that the cell density and cell survival rate of stem cell culture using the bioreactor described in the embodiments of this utility model are superior to the conventional device using an aeration plate in the prior art.

[0156] In some embodiments of the bioreactor described in this invention, the gas exchange tube 30 is configured as a polytetrafluoroethylene (PTFE) tube. Polytetrafluoroethylene (PTFE), also known as Teflon, has excellent heat resistance, cold resistance, and corrosion resistance, a low coefficient of friction, and good biocompatibility, making it well-suited for use in bioreactors to achieve biological culture.

[0157] In the production process, a polytetrafluoroethylene (PTFE) tube can be prepared first by using a PTFE film, and then the PTFE tube can be stretched along its own axis with a preset tension to form an exchange hole 311 on the tube wall, thus obtaining a gas exchange tube 30.

[0158] Reference Figure 1As shown, in some embodiments of the bioreactor described in this utility model, the top surface 11 of the container 10 is further provided with multiple liquid inlet connectors 113, through which various nutrients are supplemented to the biological culture medium 20 during the biological culture process. Those skilled in the art can set the number and model of the liquid inlet connectors 113 according to actual needs to adapt to different instruments.

[0159] A test connector 14 is provided on the side wall of container 10. Exemplarily, the test connector 14 includes a DO value measuring connector, a pH value measuring connector, a temperature measuring connector, and a test drain connector.

[0160] DO value refers to dissolved oxygen, representing the concentration of dissolved oxygen in the water. During biological culture, stem cells and attached air bubbles 41 are extremely small, making gas exchange difficult to observe directly with the naked eye. In this situation, a DO value measuring instrument can be used with a DO value measuring connector. DO value measuring instruments are existing technology, and the measurement method will not be elaborated further. Real-time DO values ​​are used to determine the gas exchange status and whether the oxygen content in container 10 meets the requirements for stem cell culture. Generally, after oxygen is introduced for a certain period, the DO value will increase due to gas exchange; after a further period, as the oxygen is consumed, the DO value will decrease.

[0161] The pH measurement connector can be used with a corresponding pH measuring instrument to measure the pH value and ensure that the acidity or alkalinity within container 10 meets the requirements for biological culture. The pH measuring instrument is existing technology, and the method for measuring pH will not be elaborated further.

[0162] Preferably, each container 10 has two different models of DO and pH measurement connectors to accommodate two different testing schemes in the prior art.

[0163] The temperature measuring connector is used in conjunction with a temperature measuring instrument to measure the internal temperature of container 10, ensuring that the internal temperature of container 10 meets the requirements for biological culture. The temperature measuring instrument is existing technology, and its measurement method will not be described in detail.

[0164] The test drain connector is used to sample the liquid in container 10 at any time during the biological culture process.

[0165] Preferably, each test connector 14 on the side wall of container 10 is located at approximately 20% of the height of container 10 to facilitate operation by staff.

[0166] Working principle: First, a polytetrafluoroethylene (PTFE) film is selected to prepare the PTFE tube. The thickness of the PTFE film is the final wall thickness of the gas exchange tube 30. Satisfy the relation, .

[0167] Next, the polytetrafluoroethylene (PTFE) tube is stretched along its own axis under a preset tension to form exchange holes 311 on the tube wall, thus obtaining a gas exchange tube 30. The diameter of the exchange holes 311 is... Satisfy the relation, The porosity of the exchange hole 311 on the gas exchange tube 30 Satisfy the relation, .

[0168] Next, the components are assembled, including connecting the two ends of the gas exchange pipe 30 to the inlet connector 111 and the outlet connector 112 respectively, setting the second section 3121 of the gas exchange pipe 30 in the fixing slot 611 of the fixture 60, and fixing the inlet section 3122 and the outlet section 3123 to the double-ended pipe clamps on the inner wall of the container 10.

[0169] After assembly, the corresponding sterilization process is carried out to obtain the finished bioreactor.

[0170] During stem cell culture, the container 10, gas supply unit 51, and pressure regulating unit 52 are first connected via silicone tubing to complete the assembly of each component. Then, biological culture medium 20 and stem cells are introduced into the container 10. When the DO value is lower than the preset value, oxygen is introduced into the gas exchange tube 30 through the cooperation of gas supply unit 51 and pressure regulating unit 52, so that oxygen passes through the exchange hole 311 and forms attached bubbles 41 on the outer wall of the orifice of the exchange hole 311. The attached bubbles 41 exchange gases with the biological culture medium 20.

[0171] The oxygen pressure is calculated in advance based on the wall thickness of the gas exchange tube 30, the pore size of the exchange hole 311, the porosity of the exchange hole 311 on the gas exchange tube 30, the oxygen gas viscosity, and the gas flow rate. This ensures that attached bubbles 41 can be formed, reducing the shear force generated by the collapse of suspended bubbles, improving the survival rate of stem cells, and preventing the gas exchange tube 30 from being damaged by excessively high gas pressure.

[0172] For ease of description, spatial relative terms such as "above," "on top of," "on the upper surface of," "above," etc., are used herein to describe the spatial positional relationship of a device or feature as shown in the figures to other devices or features. It should be understood that spatial relative terms are intended to encompass different orientations in use or operation beyond the orientation of the device as described in the figures. For example, if the device in the figures were inverted, a device described as "above" or "on top of" other devices or structures would subsequently be positioned as "below" or "under" other devices or structures. Thus, the exemplary term "above" can include both "above" and "below." The device may also be positioned in other different ways (rotated 90 degrees or in other orientations), and the spatial relative descriptions used herein will be interpreted accordingly.

[0173] Furthermore, it should be noted that the use of terms such as "first" and "second" to define components is merely for the purpose of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore cannot be construed as limiting the scope of protection of this utility model.

[0174] The above are merely preferred embodiments of this utility model and are not intended to limit the scope of this utility model. Various modifications and variations can be made to this utility model by those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of this utility model should be included within the protection scope of this utility model.

Claims

1. A bioreactor, characterized in that, include: container; A gas exchange tube is at least partially disposed within the container; The gas exchange pipe includes a first pipe section and a second pipe section that are connected to each other, and multiple exchange holes are provided on the pipe walls of both the first pipe section and the second pipe section. A fixture is used to fix the gas exchange tube to the container; the fixture is provided with a fixing slot, and the area of ​​the fixing slot on the fixture is not less than a preset value; the second tube segment is disposed in the fixing slot, and at least part of the exchange hole is disposed on the side of the second tube segment away from the fixing slot.

2. The bioreactor according to claim 1, characterized in that: The fixing slot includes a first spiral groove segment and a second spiral groove segment. The first spiral groove segment and the second spiral groove segment are both spirally arranged from the middle of the fixing device to the outside. The ends of the first spiral groove segment and the second spiral groove segment located in the middle of the fixing device are connected to each other. The ends of the first spiral groove segment and the second spiral groove segment located on the outside of the fixing device are both located on the side wall of the fixing device on the same side.

3. The bioreactor according to claim 1 or 2, characterized in that: The inner wall of the container is provided with a plurality of fasteners arranged at intervals in sequence, and at least a portion of the first pipe section is connected to the fasteners.

4. The bioreactor of claim 3, wherein, The first pipe section includes: The air intake section is connected to the aforementioned fixing component; An exhaust section is connected to the fixing member; the exhaust section and the intake section are arranged side by side. A first bend is disposed between the intake section and the second pipe section, and connects the intake section and the second pipe section respectively; and, The second bend section is disposed between the air outlet section and the second pipe section, and is connected to the air outlet section and the second pipe section respectively; Both the first and second bending sections are bent toward the inner wall of the container.

5. The bioreactor according to claim 1 or 2, characterized in that: The fastener includes a first fixing surface and a second fixing surface disposed opposite to each other. The first fixing surface is provided with the fixing slot, and the second fixing surface is provided with a first snap-fit ​​component. The container is provided with a liquid outlet connector at the bottom, and a second snap-fit ​​component is provided on the side of the liquid outlet connector near the fixture. The second snap-fit ​​component is snap-fitted to the first snap-fit ​​component.

6. The bioreactor according to claim 5, characterized in that: The second fixing surface is also provided with a clearance hole, which extends through the first fixing surface; the first snap-fit ​​component includes a plurality of first snap-fit ​​blocks, which are arranged sequentially at intervals along the circumference of the clearance hole, and each first snap-fit ​​block is provided with a first snap-fit ​​hook; The liquid outlet connector is also provided with a third snap-fit ​​component, at least a portion of which is disposed within the clearance hole. The third snap-fit ​​component has a receiving hole and a second snap-fit ​​hook disposed outside the receiving hole. The second snap-fit ​​component is arranged around the outside of the third snap-fit ​​component. The second snap-fit ​​component has a first snap-fit ​​groove and a plurality of second snap-fit ​​blocks. The first snap-fit ​​groove is snap-fitted to the first snap-fit ​​hook. Each second snap-fit ​​block is disposed between two adjacent first snap-fit ​​blocks and snap-fitted to the first snap-fit ​​block. The container is also equipped with a stirrer, which includes a stirring shaft and a stirring paddle mounted on the stirring shaft. The stirring shaft includes a first end, which is disposed in the receiving hole. A second snap-fit ​​groove is provided on the side wall of the first end, and the second snap-fit ​​groove is snap-fitted to a second snap-fit ​​hook.

7. The bioreactor according to claim 1, characterized in that: An air inlet and an air outlet are provided on the top surface of the container, and both the air inlet and the air outlet are sealed to the container. The inlet section of the gas exchange pipe is connected to the inlet connector, and the outlet section of the gas exchange pipe is connected to the outlet connector.

8. The bioreactor of claim 7, wherein, Also includes: A gas supply component, which is connected to the gas inlet connector, is used to introduce target gas into the gas exchange pipe; as well as, A pressure regulating component is connected to the gas outlet connector and is used to adjust the gas pressure of the target gas.

9. The bioreactor according to claim 1, 7, or 8, characterized in that: The container is used to hold the biological culture medium; At least a portion of the gas exchange tube is disposed within the biological culture medium, and the gas exchange tube is configured to circulate the target gas. Wherein, the hydraulic pressure of the biological culture medium is less than the gas pressure of the target gas, and the gas pressure of the target gas and the hydraulic pressure of the biological culture medium satisfy a target pressure difference, so that the target gas passes through the exchange hole and forms attached bubbles on the outer wall of the orifice of the exchange hole, and the attached bubbles are used to exchange gases with the biological culture medium.

10. The bioreactor of claim 9, wherein, Includes at least one of the following features: The wall thickness of the gas exchange tube satisfies the relationship, ; The aperture of the exchange hole satisfies the relationship, ; The opening ratio of the exchange holes on the gas exchange tube satisfies the relationship, ; the target pressure difference satisfies the relationship, .

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