Cell culture system and cell culture method
The cell culture system addresses the challenge of stable, cost-effective large-scale cell culture by using a culture tank and storage tanks with a separation membrane to adjust and recycle medium components, ensuring prolonged cell culture and improved product quality.
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- ASAHI KASEI KOGYO KABUSHIKI KAISHA
- Filing Date
- 2024-11-05
- Publication Date
- 2026-05-19
AI Technical Summary
Existing cell culture systems struggle to culture cells in large quantities at low cost while maintaining a stable culture environment and product quality, particularly for biopharmaceuticals like antibodies, which are expensive and prone to quality variations.
A cell culture system and method utilizing a culture tank, first and second storage tanks, and a separation membrane to adjust and recycle culture medium components, with fluid machines and detectors to maintain optimal conditions for prolonged cell culture.
The system enables long-term cell culture by continuously adjusting and recycling culture medium components, reducing waste accumulation and maintaining optimal conditions, thereby extending the culture period and improving product quality.
Smart Images
Figure 2026081586000001_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to a cell culture system and a cell culture method.
Background Art
[0002] In recent years, in addition to pharmaceuticals mainly composed of conventional synthetic chemical substances, the use of biopharmaceuticals, which are pharmaceuticals originating from biological materials produced using biotechnology techniques, has been spreading. Among biopharmaceuticals, particularly remarkable ones are cell products such as antibodies. Although these biopharmaceuticals have very high effects, they are expensive, which is an issue. Also, since they are products from organisms, there is a possibility that variations in quality during the manufacturing process may be larger compared to conventional pharmaceuticals. Therefore, there is a demand for the development of a system that can culture cells in a larger amount at once and manufacture products at a low cost, and a culture system that can stably maintain the culture environment and improve the quality of products by maintaining the quality of cells (see, for example, Patent Document 1).
Prior Art Documents
Patent Documents
[0003]
Patent Document 1
Summary of the Invention
Problems to be Solved by the Invention
[0004] An object of the present invention is to provide a cell culture system and a cell culture method capable of culturing cells for a long period.
Means for Solving the Problems
[0005] [1] A cell culture system comprising: a culture tank configured for culturing cells in a culture medium; a first storage tank configured for storing a component adjustment solution for adjusting the components of the culture medium; a component adjustment means configured for adjusting the components of the culture medium using the component adjustment solution, returning the adjusted culture medium to the culture tank, and returning the used component adjustment solution to the first storage tank; a component adjustment solution discharge channel configured for discharging the component adjustment solution from the first storage tank; a second storage tank configured for storing the component adjustment solution; and a component adjustment solution supply channel configured for supplying the component adjustment solution from the second storage tank to the first storage tank.
[0006] [2] The cell culture system according to [1], further comprising: a component adjustment solution detector provided in a first storage tank and configured to detect the amount of component adjustment solution; and a fluid machine configured to supply component adjustment solution from a second storage tank to the first storage tank when the amount of component adjustment solution in the first storage tank decreases.
[0007] [3] The cell culture system according to [2], wherein the component adjustment solution detector is configured to detect the liquid level of the component adjustment solution.
[0008] [4] The cell culture system according to [2] or [3] wherein the amount of component-adjusted solution in the first storage tank is reduced by discharging the component-adjusted solution from the first storage tank into the component-adjusted solution discharge channel.
[0009] [5] A cell culture system according to any one of [1] to [4], wherein the component preparation means comprises a separation membrane.
[0010] [6] The cell culture system according to [5], wherein the separation membrane is a hollow fiber membrane.
[0011] [7] The cell culture system according to [5] or [6], further comprising: a culture medium supply channel configured to supply culture medium from a culture tank to a first surface of a separation membrane; a culture medium return channel configured to return the culture medium in contact with the first surface of the separation membrane to the culture tank; a component adjustment solution supply channel configured to supply component adjustment solution from a first storage tank to a second surface of the separation membrane; and a component adjustment solution return channel configured to return the component adjustment solution in contact with the second surface of the separation membrane to the first storage tank.
[0012] [8] A cell culture method comprising culturing cells in a culture medium in a culture tank, adjusting the components of the culture medium using a component adjusting solution to adjust the components of the culture medium stored in a first storage tank, returning the culture medium with adjusted components to the culture tank, returning the used component adjusting solution to the first storage tank, discharging the component adjusting solution from the first storage tank, and supplying the component adjusting solution from the second storage tank to the first storage tank.
[0013] [9] The cell culture method according to [8], wherein the amount of component adjustment solution in the first storage tank is detected, and if the amount of component adjustment solution in the first storage tank decreases, the component adjustment solution is transferred from the second storage tank to the first storage tank.
[0014]
[10] The cell culture method according to [9], wherein the amount of component-adjusting solution in the first storage tank is detected by detecting the liquid level of the component-adjusting solution in the first storage tank.
[0015]
[11] A cell culture method according to any one of [8] to
[10] , wherein the components of the culture medium are prepared via a separation membrane.
[0016]
[12] The cell culture method according to
[11] , wherein the separation membrane is a hollow fiber membrane.
[0017]
[13] The cell culture method according to
[11] or
[12] , wherein, in adjusting the components of the culture medium, the culture medium is sent from the culture tank to the first surface of the separation membrane, the culture medium in contact with the first surface of the separation membrane is returned to the culture tank, the component adjustment solution is sent from the first storage tank to the second surface of the separation membrane, and the component adjustment solution in contact with the second surface of the separation membrane is returned to the first storage tank.
Advantages of the Invention
[0018] According to the present invention, it is possible to provide a cell culture system and a cell culture method capable of culturing cells for a long period of time.
Brief Description of the Drawings
[0019] [Figure 1] It is a schematic diagram of a cell culture system according to an embodiment. [Figure 2] It is a flowchart of a cell culture method according to an embodiment. [Figure 3] It is a flowchart of a cell culture method according to an embodiment.
Modes for Carrying Out the Invention
[0020] Hereinafter, the present invention will be described in detail in accordance with specific embodiments. However, the present invention is not limited to the following embodiments, and can be implemented in any form without departing from the spirit of the present invention.
[0021] As shown in FIG. 1, the cell culture system according to the embodiment includes a culture tank 1 configured to culture cells in a culture solution 9, a first storage tank 2 configured to store a component adjustment solution 10 for adjusting the components of the culture solution 9, a component adjustment means 30 configured to adjust the components of the culture solution 9 using the component adjustment solution 10, return the culture solution 9 with adjusted components to the culture tank 1, and return the used component adjustment solution 10 to the first storage tank 2, a component adjustment solution discharge channel 21 configured to discharge the component adjustment solution 10 from the first storage tank 2, a second storage tank 40 configured to store the component adjustment solution 10, and a component adjustment solution supply channel 41 configured to supply the component adjustment solution from the second storage tank 40 to the first storage tank 2.
[0022] The component adjustment means 30 includes, for example, a separation membrane 3. The separation membrane 3 has a first surface and a second surface facing the first surface. At least a part of the components of the liquid in contact with the first surface of the separation membrane 3 permeate from the first surface to the second surface and are discharged from the second surface. At least a part of the components of the liquid in contact with the second surface of the separation membrane 3 permeate from the second surface to the first surface and are discharged from the first surface. The separation membrane 3 is, for example, a semipermeable membrane. The separation membrane 3 is, for example, a hollow fiber membrane. The separation membrane 3 may be contained in a housing and constitute a separation membrane module.
[0023] The component adjustment means 30 includes, for example, a culture solution feed channel 4 configured to send the culture solution 9 from the culture tank 1 to the first surface of the separation membrane 3, a culture solution return channel 5 configured to return the culture solution 9 in contact with the first surface of the separation membrane 3 to the culture tank 1, a component adjustment solution feed channel 6 configured to send the component adjustment solution 10 from the first storage tank 2 to the second surface of the separation membrane 3, and a component adjustment solution return channel 7 configured to return the component adjustment solution 10 in contact with the second surface of the separation membrane 3 to the first storage tank 2.
[0024] The shape and material of the culture tank 1 are not particularly limited as long as cells can be cultured inside the culture tank 1. The culture tank 1 can be a dish, a beaker, or a bottle. The material of the culture tank 1 can be a resin such as polystyrene and glass such as silica. The cells cultured in the culture tank 1 are not particularly limited, but are, for example, mammalian cells. The cells may be undifferentiated cells or differentiated cells. The cells may be cultured for the purpose of recovering cell products. Examples of cell products include proteins, nucleic acids, and viruses. Examples of proteins include antibodies. The cells may be transfected with factors to produce a desired cell product.
[0025] The cells may be cells harvested from biological tissue by enzymatic treatment, blood-derived cells, mesenchymal stem cells, ES cells, and iPS cells. The cells cultured in culture tank 1 may be of one type or multiple types. The cells cultured in culture tank 1 may be cultured in adherent culture or in suspension culture. When performing adherent culture, feeder cells may be cultured on the bottom surface of culture tank 1, and the desired cells may be cultured on the feeder cells.
[0026] The culture medium 9 is not particularly limited as long as it is suitable for the cells cultured in the culture vessel 1. For example, if the cells cultured in the culture vessel 1 are stem cells, a stem cell medium is used, and if the cells are differentiated cells, a differentiated cell medium is used. For example, if the cells are blood-derived cells, a blood-derived cell medium is used, and if the cells are mesenchymal stem cells, a mesenchymal stem cell medium is used. The culture medium 9 contains culture components to maintain cell survival. Examples of culture components include amino acids, vitamins, and inorganic salts.
[0027] Examples of amino acids include essential amino acids, conditionally essential amino acids, and non-essential amino acids. Examples of essential amino acids include histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. Examples of conditionally essential amino acids include arginine, cysteine, and tyrosine. Examples of non-essential amino acids include glycine, glutamine, and serine. Examples of vitamins include the B vitamins. Examples of inorganic salts include calcium salts, magnesium salts, potassium salts, and sodium salts.
[0028] Cells cultured in culture tank 1 excrete waste products into the culture medium. Examples of cellular waste products include carbon dioxide and uric acid. When cells are cultured in culture tank 1, the culture components contained in the culture medium 9 are consumed by the cells, and the concentration of culture components in the culture medium 9 decreases. Also, when cells are cultured in culture tank 1, the cells excrete waste products into the culture medium 9, and the concentration of waste products in the culture medium 9 increases.
[0029] The component adjustment solution 10 may contain at least one of the culture components contained in the culture medium 9. The component adjustment solution 10 may contain the same culture components as the culture medium 9. Unused component adjustment solution 10 may contain culture components at the same concentration as unused culture medium 9, for example. When the culture medium 9 is sent from the culture tank 1 to the first surface of the separation membrane 3 via the culture medium supply channel 4, and the component adjustment solution 10 is sent from the first storage tank 2 to the second surface of the separation membrane 3 via the component adjustment solution supply channel 6, components are exchanged between the culture medium 9 and the component adjustment solution 10 due to the concentration gradient resulting from the difference in concentration between the components contained in the culture medium 9 and the components contained in the component adjustment solution 10.
[0030] Specifically, in the culture medium 9, as cell culture progresses, the concentration of culture components decreases, so the culture components contained in the component adjustment solution 10 permeate through the separation membrane 3 and move into the culture medium 9. Also, as cell culture progresses in the culture medium 9, the concentration of waste products increases, so the waste products contained in the culture medium 9 permeate through the separation membrane 3 and move into the component adjustment solution 10. The culture medium 9, supplied with culture components by the separation membrane 3, is returned to the culture tank 1 via the culture medium return channel 5. The component adjustment solution 10, supplied with waste products by the separation membrane 3, is returned to the first storage tank 2 via the component adjustment solution return channel 7.
[0031] The culture medium supply channel 4 is equipped with a fluid machine 14 for supplying, for example, the culture medium 9 in the culture tank 1 to the first surface of the separation membrane 3. The component adjustment liquid supply channel 6 is equipped with a fluid machine 16 for supplying, for example, the component adjustment liquid 10 in the first storage tank 2 to the second surface of the separation membrane 3. The component adjustment liquid return channel 7 is equipped with a fluid machine 17 for supplying, for example, the component adjustment liquid 10 in contact with the second surface of the separation membrane 3 to the first storage tank 2. The fluid machine is, for example, a pump. Examples of pumps include peristaltic pumps and diaphragm pumps.
[0032] As cell culture progresses and component exchange between the culture medium 9 and the component adjustment solution 10 in the separation membrane 3 increases, the concentration of waste products in the component adjustment solution 10 rises. When the concentration of waste products in the component adjustment solution 10 rises, the concentration gradient due to the difference between the concentration of waste products in the culture medium 9 and the concentration of waste products in the component adjustment solution 10 decreases. As a result, the amount of waste products from the culture medium 9 that moves to the component adjustment solution 10 in the separation membrane 3 decreases, and waste products may accumulate in the culture medium 9.
[0033] In contrast, the cell culture system according to the embodiment discharges the used component adjustment solution 10 from the first storage tank 2 via the component adjustment solution discharge channel 21, thereby reducing the amount of used component adjustment solution 10 in the first storage tank 2. Furthermore, the cell culture system according to the embodiment supplies fresh component adjustment solution 10 from the second storage tank 40 to the first storage tank 2 via the component adjustment solution supply channel 41. As a result, the cell culture system according to the embodiment can reduce the concentration of waste products in the component adjustment solution 10 in the first storage tank 2. Therefore, since the cell culture system according to the embodiment can reduce the concentration of waste products in the culture medium 9 for a long period of time, it is possible to culture cells in the culture tank 1 for a long period of time.
[0034] The component adjustment solution discharge channel 21 is provided with, for example, a fluid machine 22 for discharging the component adjustment solution 10 used from the first storage tank 2. The component adjustment solution supply channel 41 is provided with, for example, a fluid machine 42 for sending the component adjustment solution 10 from the second storage tank 40 to the first storage tank 2. The first storage tank 2 may be provided with a component adjustment solution detector 50 configured to detect the amount of component adjustment solution 10 in the first storage tank 2. The component adjustment solution detector 50 may be configured to detect the liquid level of the component adjustment solution 10 in the first storage tank 2. The fluid machine 42 may be configured to send component adjustment solution 10 from the second storage tank 40 to the first storage tank 2 when the amount of component adjustment solution 10 in the first storage tank 2 decreases. The cell culture system according to the embodiment may further include a control device 100 configured to control the fluid machines 14, 16, 17, 22, 42 and the component adjustment solution detector 50.
[0035] Next, the cell culture method according to the embodiment will be described with reference to the flowcharts shown in Figures 2 and 3.
[0036] In step S101, the culture medium 9 is placed in the culture tank 1 and cell culture is started. In step S102, the control device 100 controls the fluid machine 14 to send the culture medium 9 in the culture tank 1 to the first surface of the separation membrane 3 via the culture medium supply channel 4, and returns the culture medium 9 that is in contact with the first surface of the separation membrane 3 to the culture tank 1 via the culture medium return channel 5, thereby perfusion of the culture medium 9 within the culture tank 1, the module including the culture medium supply channel 4 and the separation membrane 3, and the culture medium return channel 5. In step S103, the control device 100 controls the fluid machines 16 and 17 to send the component adjustment liquid 10 in the first storage tank 2 to the second surface of the separation membrane 3 via the component adjustment liquid supply channel 6, and returns the component adjustment liquid 10 that has come into contact with the second surface of the separation membrane 3 to the first storage tank 2 via the component adjustment liquid return channel 7, thereby recirculating the component adjustment liquid 10 within the module including the first storage tank 2, the component adjustment liquid supply channel 6, the separation membrane 3, and the component adjustment liquid return channel 7.
[0037] In step S104, the control device 100 controls the component adjustment liquid detector 50 to detect the amount of component adjustment liquid 10 in the first storage tank 2. For example, the control device 100 controls the component adjustment liquid detector 50 to detect the height of the liquid level of the component adjustment liquid 10 in the first storage tank 2. In step S105, the control device 100 controls the fluid machines 16 and 17 to adjust the flow rate of the component adjustment liquid 10 in the component adjustment liquid supply channel 6 and the flow rate of the component adjustment liquid 10 in the component adjustment liquid return channel 7 so that the amount of component adjustment liquid 10 in the first storage tank 2 remains constant. For example, the control device 100 controls the fluid machines 16 and 17 to adjust the flow rate of the component adjustment liquid 10 in the component adjustment liquid supply channel 6 and the flow rate of the component adjustment liquid 10 in the component adjustment liquid return channel 7 so that the height of the liquid level of the component adjustment liquid 10 in the first storage tank 2 remains constant.
[0038] In step S106, the control device 100 determines whether or not to start discharging the component adjustment solution 10 from the first storage tank 2 after a predetermined time has elapsed since the start of cell culture in the culture tank 1. The predetermined time is, for example, the time from the start of cell culture in the culture tank 1 until the concentration of waste products in the culture medium 9 exceeds a predetermined concentration. Alternatively, the predetermined time is, for example, the time from the start of cell culture in the culture tank 1 until the concentration of waste products in the component adjustment solution 10 exceeds a predetermined concentration. The predetermined time can be obtained in advance. If it is determined that the discharge of the component adjustment solution 10 from the first storage tank 2 should be started, the process proceeds to step S107. If it is determined that the discharge of the component adjustment solution 10 from the first storage tank 2 should not be started, the process returns to step S104.
[0039] In step S107, the control device 100 controls the fluid machine 22 to discharge the used component adjustment liquid 10 from the first storage tank 2 via the component adjustment liquid discharge channel 21. In step S108, the control device 100 determines whether to stop the discharge of a predetermined amount of the component adjustment liquid 10 used from the first storage tank 2 by the fluid machine 22. The predetermined amount is determined in advance. The amount of component adjustment liquid 10 discharged may be measured by the fluid machine 22 or by the component adjustment liquid detector 50. If it is determined to stop the discharge of the component adjustment liquid 10 from the first storage tank 2, the process proceeds to step S109. If it is determined not to stop the discharge of the component adjustment liquid 10 from the first storage tank 2, the discharge of the component adjustment liquid 10 from the first storage tank 2 continues. In step S109, the control device 100 controls the fluid machine 22 to stop the discharge of the component adjustment liquid 10 from the first storage tank 2.
[0040] In step S110, the control device 100 controls the fluid machine 42 to send the component adjustment liquid 10 from the second storage tank 40 to the first storage tank 2 via the component adjustment liquid supply channel 41. In step S111, the control device 100 controls the component adjustment liquid detector 50 to detect the amount of component adjustment liquid 10 in the first storage tank 2. For example, the control device 100 controls the component adjustment liquid detector 50 to detect the height of the liquid level of the component adjustment liquid 10 in the first storage tank 2. In step S112, the control device 100 determines whether the amount of component adjustment liquid 10 in the first storage tank 2 has reached a predetermined amount and whether to stop supplying the component adjustment liquid 10 to the first storage tank 2. The predetermined amount is, for example, the same amount as the amount of component adjustment liquid 10 in the first storage tank 2 before the component adjustment liquid 10 used in step S107 is discharged from the first storage tank 2. For example, the control device 100 determines whether to stop supplying the component adjustment liquid 10 to the first storage tank 2 when the liquid level of the component adjustment liquid 10 in the first storage tank 2 reaches a predetermined height. The predetermined height is, for example, the same height as the liquid level of the component adjustment liquid 10 in the first storage tank 2 before the component adjustment liquid 10 used in step S107 is discharged from the first storage tank 2. If it is determined to stop supplying the component adjustment liquid 10 to the first storage tank 2, the process proceeds to step S113. If it is determined not to stop supplying the component adjustment liquid 10 to the first storage tank 2, the process returns to step S111.
[0041] In step S113, the control device 100 controls the fluid machine 42 to stop the supply of the component adjustment solution 10 from the second storage tank 40 to the first storage tank 2. In step S114, the control device 100 determines whether to stop the culture because the amount of culture medium 9 perfusing the culture tank 1, the culture medium supply channel 4, the module including the separation membrane 3, and the culture medium return channel 5 has reached a predetermined amount. The amount of perfused culture medium 9 is measured, for example, by the fluid machine 14. If it is determined to stop the culture, the cell culture in the culture tank 1 is terminated. If it is determined not to stop the culture, the process returns to step S104.
[0042] Although the present invention has been described above by embodiments, the descriptions and drawings that constitute part of this disclosure should not be understood as limiting the invention. Various alternative embodiments, examples, and operational techniques should become apparent to those skilled in the art from this disclosure. It should be understood that the present invention encompasses various embodiments and the like that are not described herein. [Explanation of Symbols]
[0043] 1...Culture tank, 2...First storage tank, 3...Separation membrane, 4...Culture medium supply channel, 5...Culture medium return channel, 6...Component adjustment solution supply channel, 7...Component adjustment solution return channel, 9...Culture medium, 10...Component adjustment solution, 14, 16, 17, 22, 42...Fluid machinery, 21...Component adjustment solution discharge channel, 30...Component adjustment means, 40...Second storage tank, 41...Component adjustment solution supply channel, 50...Component adjustment solution detector, 100...Control device
Claims
1. A culture vessel configured for culturing cells in a culture medium, A first storage tank configured to store a component adjustment solution for adjusting the components of the culture medium, A component adjustment means configured to adjust the components of the culture medium using the component adjustment solution, return the culture medium with the adjusted components to the culture tank, and return the used component adjustment solution to the first storage tank, A component adjustment liquid discharge channel configured to discharge the component adjustment liquid from the first storage tank, A second storage tank configured to store the aforementioned component adjustment liquid, A component adjustment liquid supply channel configured to supply the component adjustment liquid from the second storage tank to the first storage tank, A cell culture system equipped with the following features.
2. A component adjustment liquid detector is provided in the first storage tank and configured to detect the amount of the component adjustment liquid, A fluid machine configured to transfer the component adjustment liquid from the second storage tank to the first storage tank when the component adjustment liquid in the first storage tank decreases, The cell culture system according to claim 1, further comprising:
3. The cell culture system according to claim 2, wherein the component adjustment solution detector is configured to detect the liquid level of the component adjustment solution.
4. The cell culture system according to claim 2, wherein the amount of the component adjusting solution in the first storage tank is reduced by discharging the component adjusting solution from the first storage tank to the component adjusting solution discharge channel.
5. The cell culture system according to claim 1, wherein the component adjustment means comprises a separation membrane.
6. The cell culture system according to claim 5, wherein the separation membrane is a hollow fiber membrane.
7. The aforementioned component adjustment means A culture medium supply channel configured to deliver the culture medium from the culture tank to the first surface of the separation membrane, A culture medium return channel is configured to return the culture medium in contact with the first surface of the separation membrane back to the culture tank, A component adjustment liquid supply channel configured to supply the component adjustment liquid from the first storage tank to the second surface of the separation membrane, A component adjustment liquid return channel is configured to return the component adjustment liquid in contact with the second surface of the separation membrane back to the first storage tank, The cell culture system according to claim 5, further comprising:
8. Culturing cells in the culture medium within a culture vessel, The components of the culture medium stored in the first storage tank are adjusted using a component adjusting solution, the culture medium with adjusted components is returned to the culture tank, and the component adjusting solution used is returned to the first storage tank. Discharging the component adjustment liquid from the first storage tank, The component adjustment liquid is supplied from the second storage tank to the first storage tank, A cell culture method, including the following.