Hollow fiber bioreactor beneficial to nutrition and gas transmission

Abstract

The invention provides a hollow fiber bioreactor beneficial to nutrition and gas transmission, and particularly relates to the technical field of cell culture, the hollow fiber bioreactor beneficial to nutrition and gas transmission comprises a first hollow fiber membrane, a second hollow fiber membrane and a sealing glue outer layer, wherein the plurality of first hollow fiber membranes and the plurality of second hollow fiber membranes are arranged at intervals, the first arrangement module comprises a plurality of first hollow fiber structures which are arranged in parallel, the second arrangement module comprises a plurality of second hollow fiber structures which are arranged in parallel, and nutrient substances enter the culture cavity through porous structures on the membrane walls of the first hollow fiber structures; gas enters the culture cavity through the gas permeation structure of the second hollow fiber structure, nutrient substances can be evenly conveyed to cells through the first hollow fiber membrane and the second hollow fiber membrane which are arranged in a crossed mode by means of the porous structure and the gas permeation structure, and meanwhile the gas can be introduced to regulate and control the gas microenvironment.

Description

Technical Field

[0001] This invention relates to the field of cell culture technology, and in particular to a hollow fiber bioreactor that facilitates nutrient and gas transport. Background Technology

[0002] Traditional two-dimensional cell culture is a monolayer culture carried out at the bottom of a culture dish. This culture method lacks three-dimensional contact between cells and physiological support from the ECM, making it difficult to truly simulate the in vivo microenvironment, resulting in significant differences between cell behavior and physiological state.

[0003] Currently, three-dimensional culture technology is being utilized, such as three-dimensional cell culture through hollow fiber membrane bioreactors. Hollow fiber membrane bioreactors (HFMBRs), as an advanced cell culture system, provide a highly biomimetic microenvironment for cell function and metabolic activities, which can improve the efficient diffusion of nutrients and create favorable conditions for cell growth and tissue construction.

[0004] However, existing hollow fiber bioreactors still have several technical bottlenecks. For example, the random distribution structure of the fiber membrane leads to spatial heterogeneity in nutrient transport, which not only affects the efficiency of material transfer but also significantly reduces the quality uniformity of cultured cells or tissues. The device system lacks precise gas microenvironment control capabilities, which increases the necrosis rate of cells or tissues during large-scale culture. Summary of the Invention

[0005] To address the issues of existing hollow fiber bioreactors lacking gas microenvironment regulation and failing to achieve uniformity in mass transfer, this invention proposes a hollow fiber bioreactor that facilitates nutrient and gas transport.

[0006] This invention is achieved through the following technical solution: This invention proposes a hollow fiber bioreactor that facilitates nutrient and gas transport, comprising a first hollow fiber membrane, a second hollow fiber membrane, and a sealing outer layer, wherein: Multiple first hollow fiber membranes and second hollow fiber membranes are arranged alternately. The first arrangement module includes multiple parallel first hollow fiber structures, and the second arrangement group includes multiple parallel second hollow fiber structures. The first hollow fiber structures and second hollow fiber structures have a certain arrangement angle. The sealing outer layer encapsulates the first hollow fiber structures and second hollow fiber structures inside, and forms a culture chamber in the space between the outer surfaces of the first hollow fiber structures and second hollow fiber structures and the sealing outer layer. The membrane wall of the first hollow fiber structure is provided with a porous structure, and the membrane wall of the second hollow fiber structure is provided with a gas permeation structure. Nutrients enter the culture chamber through the porous structure on the membrane wall of the first hollow fiber structure, and gas enters the culture chamber through the gas permeation structure of the second hollow fiber structure.

[0007] Furthermore, the first hollow fiber structure is made of one of polyvinylidene fluoride, cellulose diacetate, cellulose triacetate, polyethersulfone, polysulfone, or polyacrylonitrile.

[0008] Furthermore, the second hollow fiber structure is made of one of polypropylene, polyimide, poly4-methyl-1-pentene, polyvinylidene fluoride, polytetrafluoroethylene, or polydimethoxysilane.

[0009] Furthermore, the outer layer of the sealant is provided with a first liquid inlet and a first liquid outlet. The first liquid inlet is connected to one end of the plurality of first hollow fiber structures, and the first liquid outlet is connected to the other end of the plurality of first hollow fiber structures.

[0010] Furthermore, the outer layer of the sealant is also provided with a first air inlet and a first air outlet. The first air inlet is connected to one end of a plurality of second hollow fiber structures, and the first air outlet is connected to the other end of a plurality of second hollow fiber structures.

[0011] Furthermore, the outer layer of the sealant is also provided with a second liquid inlet and a second liquid outlet, which are connected to the culture chamber.

[0012] Furthermore, the inner diameter of the first hollow fiber structure is 200-1500 μm, and the wall thickness of the first fiber membrane is 30-300 μm.

[0013] Furthermore, the inner diameter of the second hollow fiber structure is 200-1500 μm, and the wall thickness of the second fiber membrane is 70-500 μm.

[0014] Furthermore, the arrangement angle between the first hollow fiber structure and the second hollow fiber structure is 30-90°.

[0015] Furthermore, the vertical distance between the first hollow fiber membrane and the second hollow fiber membrane is 0-5000μm, the arrangement spacing between adjacent first hollow fiber structures is 0-10000μm, and the arrangement spacing between adjacent second hollow fiber structures is 0-10000μm.

[0016] The beneficial effects of this invention are: (1) The hollow fiber bioreactor proposed in this invention, which is beneficial to nutrient and gas transport, uses a first hollow fiber membrane and a second hollow fiber membrane arranged in a cross pattern, and utilizes a porous structure and a gas permeation structure to uniformly transport nutrients to cells while permeating and transporting gases to regulate the gas microenvironment.

[0017] (2) The hollow fiber bioreactor proposed in this invention facilitates the transport of nutrients and gases by uniformly transporting nutrients and gases through the cross-arranged first and second hollow fiber membranes, which has higher integration and portability. Attached Figure Description

[0018] Figure 1 The diagram shows the structure of the first and second hollow fiber membranes of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 2 This is a structural diagram of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 3 This is a cross-sectional structural diagram of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 4 This is a diagram of the first hollow fiber membrane structure of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 5 This is a cross-sectional view of the first hollow fiber structure of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 6 This is a diagram of the porous structure of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 7 This is a cross-sectional view of the second hollow fiber structure of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 8 This is a gas permeation structure diagram of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. Figure 9 This is a cell growth diagram of the hollow fiber bioreactor of the present invention, which is beneficial for nutrient and gas transport. In the figure: 1. First hollow fiber structure; 2. Second hollow fiber structure; 3. Sealing outer layer; 4. First liquid inlet; 5. First liquid outlet; 6. First air inlet; 7. First air outlet; 8. Second liquid inlet; 9. Second liquid outlet. The realization of the purpose, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0019] To more clearly and completely illustrate the technical solution of the present invention, the present invention will be further described below with reference to the accompanying drawings.

[0020] Please refer to Figures 1-9 This invention proposes a hollow fiber bioreactor that facilitates nutrient and gas transport, comprising a first hollow fiber membrane, a second hollow fiber membrane, and a sealing outer layer 3, wherein: Multiple first hollow fiber membranes and second hollow fiber membranes are arranged alternately. The first arrangement module includes multiple parallel first hollow fiber structures 1, and the second arrangement group includes multiple parallel second hollow fiber structures 2. The first hollow fiber structures 1 and the second hollow fiber structures 2 have a certain arrangement angle. The sealing outer layer 3 encapsulates the first hollow fiber structures 1 and the second hollow fiber structures 2 inside, and forms a culture chamber in the space between the outer surfaces of the first hollow fiber structures 1 and the second hollow fiber structures 2 and the sealing outer layer 3. The membrane wall of the first hollow fiber structure 1 is provided with a porous structure, and the membrane wall of the second hollow fiber structure 2 is provided with a gas permeation structure. Nutrients enter the culture chamber through the porous structure on the membrane wall of the first hollow fiber structure 1, and gas enters the culture chamber through the gas permeation structure of the second hollow fiber structure 2.

[0021] In a specific embodiment, the first hollow fiber structure 1 and the second hollow fiber structure 2 are similar to tubes. Gas or liquid can be introduced into the interior of the first hollow fiber structure 1 and the second hollow fiber structure 2. Multiple first hollow fiber structures 1 and multiple second hollow fiber structures 2 are arranged in parallel to form a first hollow fiber membrane and a second hollow fiber membrane, respectively. Multiple first hollow fiber membranes are arranged laterally, and multiple second hollow fiber membranes are arranged vertically between adjacent first hollow fiber membranes to form a cross structure. Then, the sealing outer layer 3 covers the entire cross structure. The space between the sealing outer layer 3 and the outer surface of the first hollow fiber structure 1 and the second hollow fiber structure 2 is a culture chamber. Cells are attached to the outer surface of the first hollow fiber structure 1 and the second hollow fiber structure 2. Nutrients and gases flow inside the first hollow fiber structure 1 and the second hollow fiber structure 2. Nutrients and gases are uniformly introduced into the culture chamber through the porous structure of the first hollow fiber structure 1 and the gas permeation structure of the second hollow fiber structure 2 and absorbed by the cells, thereby realizing 3-day cell culture.

[0022] In one embodiment, the morphology of the second hollow fiber structure 2 is preferably a woven reference. Figure 4 The gas permeation structure of the second hollow fiber structure 2 is preferably a dense skin type.

[0023] Furthermore, the first hollow fiber structure 1 is made of one of polyvinylidene fluoride, cellulose diacetate, cellulose triacetate, polyethersulfone, polysulfone, or polyacrylonitrile.

[0024] In a specific embodiment, the material of the first hollow fiber structure 1 is preferably polyethersulfone, and secondarily polyethersulfone.

[0025] Furthermore, the second hollow fiber structure 2 is made of one of polypropylene, polyimide, poly4-methyl-1-pentene, polyvinylidene fluoride, polytetrafluoroethylene, or polydimethoxysilane.

[0026] In a specific embodiment, the material of the second hollow fiber structure 2 is preferably poly4-methyl-1-pentene, and secondarily polypropylene.

[0027] Furthermore, the outer sealing layer 3 is provided with a first liquid inlet 4 and a first liquid outlet 5. The first liquid inlet 4 is connected to one end of a plurality of first hollow fiber structures 1, and the first liquid outlet 5 is connected to the other end of a plurality of first hollow fiber structures 1.

[0028] In a specific embodiment, the first liquid inlet 4 delivers nutrients into the interior of each first hollow fiber structure 1, and the first hollow fiber structure 1 then uniformly transmits the nutrients into the cell culture chamber through diffusion, dispersion or convection via the porous structure on its surface.

[0029] Furthermore, the outer sealing layer 3 is also provided with a first air inlet 6 and a first air outlet 7. The first air inlet 6 is connected to one end of a plurality of second hollow fiber structures 2, and the first air outlet 7 is connected to the other end of a plurality of second hollow fiber structures 2.

[0030] In a specific implementation, the first air inlet 6 allows the mixture of carbon dioxide and air to be permeated into the cell culture chamber through the second hollow fiber structure 2, thereby achieving efficient and uniform gas transmission.

[0031] Furthermore, the outer layer 3 of the sealant is also provided with a second liquid inlet 8 and a second liquid outlet 9, which are connected to the culture chamber.

[0032] In a specific embodiment, the second liquid inlet 8 is used to introduce the cell suspension, and the second liquid outlet 9 is used to discharge the cell suspension, thereby injecting the cell suspension onto the surfaces of the first hollow fiber membrane and the second hollow fiber membrane.

[0033] Furthermore, the inner diameter of the first hollow fiber structure 1 is 200-1500 μm, and the wall thickness of the first fiber membrane is 30-300 μm.

[0034] In a specific embodiment, the preferred inner diameter of the first hollow fiber structure 1 is 200-400 μm, the wall thickness is 40-70 μm, the molecular weight cutoff of the first hollow fiber structure 1 is 5-1000 kD, and the porosity is 40-80%, preferably 10-30 kD, and secondly 10-500 kD.

[0035] Furthermore, the inner diameter of the second hollow fiber structure 2 is 200-1500 μm, and the wall thickness of the second fiber membrane is 70-500 μm.

[0036] In a specific embodiment, the preferred inner diameter of the second hollow fiber structure 2 is 200-400 μm, the wall thickness is 80-150 μm, and the nitrogen flux of the second hollow fiber structure 2 is 0.2-20 mL·cm. -2 ·min -1 ·bar -1 Oxygen flux is 0.2-30 mL·cm⁻¹ -2 ·min -1 ·bar -1 The carbon dioxide flux is 0.3-40 mL·cm⁻¹. -2 ·min -1 ·bar -1 The preferred nitrogen flux is 2-8 mL·cm⁻¹. -2 ·min -1 ·bar -1 Oxygen flux is 3-15 mL·cm⁻¹ -2 ·min -1 ·bar -1 The carbon dioxide flux is 6-25 mL·cm⁻¹ -2 ·min -1 ·bar -1 .

[0037] Furthermore, the arrangement angle between the first hollow fiber structure 1 and the second hollow fiber structure 2 is 30-90°.

[0038] In a specific embodiment, the arrangement angle between the first hollow fiber membrane and the second hollow fiber membrane is preferably 90°.

[0039] Furthermore, the vertical distance between the first hollow fiber membrane and the second hollow fiber membrane is 0-5000μm, the arrangement spacing between adjacent first hollow fiber structures 1 is 0-10000μm, and the arrangement spacing between adjacent second hollow fiber structures 2 is 0-10000μm.

[0040] In a specific embodiment, the arrangement spacing between adjacent first hollow fiber structures 1 and the arrangement spacing between adjacent second hollow fiber structures 2 are preferably 300-1000 μm, while the vertical distance between the first hollow fiber membrane and the second hollow fiber membrane is preferably 0-500 μm.

[0041] Example 1: The reactor consists of a first hollow fiber membrane made of polyethersulfone and a second hollow fiber membrane made of poly4-methyl-1-pentene arranged in a cross-arrangement at an angle of 90°, with no vertical distance between the two membranes. The first hollow fiber membrane has an inner diameter of 200 μm, a wall thickness of 50 μm, a porosity of 70%, a molecular weight cutoff of 20 kD, and a minimum distance of 0.7 mm between adjacent first hollow fiber structures 1. The second hollow fiber structure 2 is a woven structure with dense inner and outer surfaces, an inner diameter of 200 μm, a wall thickness of 100 μm, and a minimum distance of 0.3 mm between adjacent second hollow fiber structures 2. During reactor operation, the cell suspension is circulated and infused into the outer surfaces of the first hollow fiber structure 1 and the second hollow fiber structure 2 through the second liquid inlet 8 and the second liquid outlet 9. During stable operation, the cells adhere to the outer surfaces and grow. The culture medium circulates within the first hollow fiber structure 1 through the first liquid inlet 4 and the first liquid outlet 5, and the nutrients are uniformly transported into the cell culture chamber through diffusion, dispersion, or convection via the porous structure, thus achieving nutrient supply. The first air inlet 6 continuously introduces a mixture of 5% carbon dioxide and 95% air at a certain pressure. The gas slowly and uniformly diffuses into the cell culture chamber through the inner cavity of the second hollow fiber membrane, achieving efficient and uniform gas transport. This ensures a uniform supply of gas and nutrients during 3D cell culture, and the cell growth status is determined by detecting cell metabolites.

[0042] Example 2: The reactor consists of a first hollow fiber membrane made of polyethersulfone and a second hollow fiber membrane made of poly4-methyl-1-pentene arranged in a cross-arrangement at an angle of 90°, with a vertical distance of 0 between the two membranes. The first hollow fiber membrane has an inner diameter of 200 μm, a wall thickness of 50 μm, a porosity of 70%, a molecular weight cutoff of 20 kD, and a minimum distance of 1 mm between adjacent first hollow fiber structures 1; the second hollow fiber structure 2 is a woven structure with dense inner and outer surfaces, an inner diameter of 200 μm, a wall thickness of 100 μm, and a minimum distance of 0.7 mm between adjacent second hollow fiber structures 2; During reactor operation, the cell suspension is circulated and infused into the outer surfaces of the first hollow fiber structure 1 and the second hollow fiber structure 2 through the second liquid inlet 8 and the second liquid outlet 9. During stable operation, the cells adhere to the outer surfaces and grow. The culture medium circulates within the first hollow fiber structure 1 through the first liquid inlet 4 and the first liquid outlet 5, and the nutrients are uniformly transported into the cell culture chamber through diffusion, dispersion, or convection via the porous structure, thus achieving nutrient supply. The first air inlet 6 continuously introduces a mixture of 5% carbon dioxide and 95% air at a certain pressure. The gas slowly and uniformly diffuses into the cell culture chamber through the inner cavity of the second hollow fiber membrane, achieving efficient and uniform gas transport. This ensures a uniform supply of gas and nutrients during 3D cell culture, and the cell growth status is determined by detecting cell metabolites.

[0043] Of course, the present invention may have many other embodiments. Based on this embodiment, other embodiments obtained by those skilled in the art without any creative effort are all within the scope of protection of the present invention.

Claims

1. A hollow fiber bioreactor that facilitates nutrient and gas transfer, characterized in that, The first hollow fiber membrane, the second hollow fiber membrane and the glue outer layer are included. The first hollow fiber structure and the second hollow fiber structure are arranged at a certain arrangement angle, the glue outer layer encapsulates the first hollow fiber structure and the second hollow fiber structure inside, and forms a culture cavity between the outer surface of the first hollow fiber structure and the second hollow fiber structure and the glue outer layer, the membrane wall of the first hollow fiber structure is provided with a porous structure, and the membrane wall of the second hollow fiber structure is provided with a gas permeation structure, nutrients enter the culture cavity through the porous structure on the membrane wall of the first hollow fiber structure, and gas enters the culture cavity through the gas permeation structure of the second hollow fiber structure.

2. The hollow fiber bioreactor to facilitate nutrient and gas transfer of claim 1, wherein, The first hollow fiber structure is made of one of polyvinylidene fluoride, diacetic cellulose, triacetic cellulose, polyether sulfone, polysulfone and polyacrylonitrile.

3. The hollow fiber bioreactor to facilitate nutrient and gas transfer of claim 1, wherein, The second hollow fiber structure is made of one of polypropylene, polyimide, poly-4-methyl-1-pentene, polyvinylidene fluoride, polytetrafluoroethylene and polydimethylsiloxane.

4. The hollow fiber bioreactor to facilitate nutrient and gas transfer of claim 3, wherein, The glue outer layer is provided with a first liquid inlet and a first liquid outlet, one end of the first liquid inlet is communicated with the first hollow fiber structure, and the other end of the first liquid outlet is communicated with the first hollow fiber structure.

5. The hollow fiber bioreactor for facilitating nutrient and gas transfer of claim 2, wherein, The glue outer layer is further provided with a first gas inlet and a first gas outlet, one end of the first gas inlet is connected with the second hollow fiber structure, and the other end of the first gas outlet is connected with the second hollow fiber structure.

6. The hollow fiber bioreactor for facilitating nutrient and gas transfer of claim 5, wherein, The glue outer layer is further provided with a second liquid inlet and a second liquid outlet, and the second liquid inlet and the second liquid outlet are communicated with the culture cavity.

7. The hollow fiber bioreactor for nutrient and gas transfer of claim 1, wherein, The inner diameter of the first hollow fiber structure is 200-1500 μm, and the wall thickness of the first fiber membrane is 30-300 μm.

8. The hollow fiber bioreactor for nutrient and gas transfer of claim 1, wherein, The inner diameter of the second hollow fiber structure is 200-1500 μm, and the wall thickness of the second fiber membrane is 70-500 μm.

9. The hollow fiber bioreactor for nutrient and gas transfer of claim 1, wherein, The arrangement angle between the first hollow fiber structure and the second hollow fiber structure is 30-90°.

10. The hollow fiber bioreactor for nutrient and gas transfer of claim 1, wherein, The vertical distance between the first hollow fiber membrane and the second hollow fiber membrane is 0-5000 μm, the arrangement distance between adjacent first hollow fiber structures is 0-10000 μm, and the arrangement distance between adjacent second hollow fiber structures is 0-10000 μm.

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