A multi-layer culture device suitable for cell suspension culture
By designing a multilayer culture vessel suitable for suspension culture, and using a float to control the position of the extraction tube and a sealed system, the problem of leakage of suspension cells when changing the culture medium was solved, which improved production efficiency and reduced the risk of contamination, and realized the large-scale culture and sealed operation of suspension cells.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- SHANGHAI ANGECON BIOTECH
- Filing Date
- 2025-08-05
- Publication Date
- 2026-07-07
AI Technical Summary
Most existing multilayer culture vessels are suitable for the large-scale production of adherent cells, but suspension cells are prone to overflow when the culture medium is changed, which cannot meet the needs of suspension culture. In addition, traditional culture containers are small in scale, cumbersome to operate, and increase the risk of contamination.
A multilayer culture device suitable for suspension culture was designed, comprising an inlet tube assembly, an outlet tube assembly, and multiple stacked culture container units. The position of the suction tube is controlled by a float to ensure that cells do not flow out when changing the culture medium, and the liquid addition and replacement operations are carried out in a closed system.
It improves the production efficiency of suspended cells, enables the harvesting of a large number of cells at once, reduces the risk of contamination, and ensures the airtightness of the culture process and the ease of operation.
Smart Images
Figure CN224467815U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to a cell culture device, and more particularly to a multilayer culture device suitable for cell suspension culture, belonging to the field of cell culture technology. Background Technology
[0002] Cell culture vessels are devices or containers used for in vitro cell culture. They are typically made of glass, plastic, or stainless steel and are broadly classified into adherent culture and suspension culture based on cell type and experimental requirements. Common culture vessels include culture dishes, multi-well plates, T-flasks, shake flasks, and bioreactors, which play a crucial role in cell growth, proliferation, and metabolic research. Adherent culture vessels usually have specially treated surfaces to promote cell adhesion, while suspension culture vessels are designed for cells that do not require adhesion, such as certain tumor cells, hybridoma cells, or CHO cells used in biopharmaceuticals.
[0003] Many cell types, such as neural stem cells, are suitable for suspension or static culture, but not for adherent culture or stirred culture in bioreactors. Most commercially available multilayer culture devices are suitable for the large-scale production of adherent cells; when the culture medium is changed, the cells adhere to the container walls and do not flow out with the medium. However, for cells growing in suspension, because they cannot adhere, traditional multilayer culture devices cause cells to flow out with the medium during culture changes, failing to meet the required culture conditions.
[0004] Currently, most culture containers used for suspension culture of cells are single-layer culture vessels, culture flasks, and multi-well plates with low adsorption treatment. These have small culture scales, are cumbersome to operate, and increase the risk of contamination. Traditional multi-layer culture vessels are only suitable for adherent cell culture and cannot meet the needs of medium change operations for suspension culture of cells. Summary of the Invention
[0005] The purpose of this invention is to provide a multilayer culture device suitable for cell suspension culture, which greatly improves the production efficiency of suspension cells and allows for the harvesting of a large number of cells at once. At the same time, it enables the addition and replacement of culture medium during cell culture to be carried out in a closed system, avoiding contamination during the culture process.
[0006] The present invention adopts the following technical solution:
[0007] A multilayer culture device suitable for cell suspension culture includes an inlet tube assembly, an outlet tube assembly, and multiple stacked culture container units. Each culture container unit has a channel 6 with a connecting hole 10 at its upper part. The sidewall of each culture container unit has an inlet 5 and an outlet 4. An outlet tube 11 extends from the outlet 4 into the culture container unit, and a float 12 is provided on the outlet tube 11. Each culture container unit has at least one channel 6 with a connecting hole 10 at its upper part, communicating with the interior of the culture container unit. When multiple culture container units are stacked, the channels 6 of each culture container unit are stacked and connected sequentially. A vent 3 is provided above the channel 6 of the topmost culture container unit. The inlet tube assembly includes a main inlet tube 7 and multiple branch inlet tubes respectively connected to the inlet 5. The outlet tube assembly includes a main outlet tube 1 and multiple branch outlet tubes respectively connected to the outlet 4.
[0008] In this technical solution, when changing the culture medium, side A is first flipped upwards. The float 12 controls the position of the suction port of the suction tube 11 to always be below the liquid surface. Since the cells are denser than the culture medium, they will sink to the bottom. This allows only the upper layer of culture medium to be suctioned, making it more convenient to change the culture medium in cell suspension culture. At the same time, multiple culture container units are stacked together to form a whole, which greatly improves the production efficiency of suspension cells compared to existing cell suspension culture, allowing for the harvesting of a large number of cells at once. Moreover, the addition and replacement of culture medium during cell culture are all carried out in a closed system, avoiding contamination during the culture process.
[0009] Preferably, the culture container unit has a flat rectangular structure.
[0010] Furthermore, the inlet 5 and outlet 4 are located on the same side A of the culture container unit. When the culture medium solution is drawn out through the outlet pipe, side A is flipped upward.
[0011] Furthermore, the breathing port 3 has a pair located near the side A, and when the side A is flipped upward, the breathing port 3 is higher than the liquid level inside the culture container unit.
[0012] Preferably, the main inlet pipe 7 is provided with a pipe clamp 9 and a filter 8;
[0013] Furthermore, the inlet and outlet branches are equipped with pipe clamps.
[0014] Preferably, the main inlet pipe 7 is connected to multiple inlet branch pipes via a ball, and the main outlet pipe 1 is connected to multiple outlet branch pipes via a ball.
[0015] Furthermore, the multilayer culture vessel is provided with several bases 2 for support on its side A. When the culture medium solution is injected into the multilayer culture vessel in the initial stage, A is flipped downwards so that the liquid level of the culture medium solution in each culture container unit quickly reaches the same level through the channel 6 and the connecting hole 10. At this time, the base 2 can protect the liquid inlet 5 and the liquid outlet 4.
[0016] Furthermore, after the initial stage, the cell inoculation solution is injected into the multilayer culture vessel through the channel 6, the lid of the breathing port 3 is closed.
[0017] Furthermore, the cell inoculation solution is a mixture of cells and culture medium.
[0018] The beneficial effects of this utility model are as follows:
[0019] 1) This utility model is a multilayer cell culture device, the surface of the culture area is a low adsorption treatment surface, which is suitable for large-scale culture of suspended cells.
[0020] 2) When changing the culture medium for cells in suspension culture, the present invention can ensure that the cells will not flow out with the culture medium by setting up a float.
[0021] 3) In this invention, the culture system is kept in a sealed state during the entire process of cell collection, from cell inoculation to cell collection at the end of culture, including the addition and replacement of culture medium, which greatly reduces the risk of contamination. Attached Figure Description
[0022] Figure 1 This is a schematic diagram of the structure of the multilayer culture device of this utility model, which is suitable for cell suspension culture.
[0023] Figure 2 This is a schematic diagram of the structure of a single culture container unit.
[0024] In the diagram, 1. liquid outlet pipe, 2. base, 3. breather, 4. liquid outlet, 5. liquid inlet, 6. channel, 7. liquid inlet pipe, 8. filter, 9. pipe clamp, 10. connecting hole, 11. liquid extraction pipe, 12. float. Detailed Implementation
[0025] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.
[0026] See Figure 1 A multilayer culture device suitable for cell suspension culture includes an inlet tube assembly, an outlet tube assembly, and multiple stacked culture container units; wherein Figure 2A culture vessel unit is shown, with each layer having a basically identical structure, except for the first layer, which has a vent 3 that allows air to pass through but isolates the liquid. The inner surface of each culture vessel layer is treated with a low-adsorption treatment to ensure that cells cultured in suspension do not adhere to the vessel walls.
[0027] See Figure 2 The culture container unit is provided with a channel 6, and the upper part of the channel 6 is provided with a connecting hole 10; through the channel 6 and the connecting hole, it can be ensured that the interior of each culture container unit can be connected with the outside air when it is lying horizontally.
[0028] See Figure 2 The culture container unit has an inlet 5 and an outlet 4 on its side wall; the outlet 4 extends into the culture container unit with a suction pipe 11, and the suction pipe 11 is equipped with a float 12; the culture container unit has at least one channel 6, and the upper part of the channel 6 is equipped with a connecting hole 10 that communicates with the inside of the culture container unit; after multiple culture container units are stacked, the channels 6 of each culture container unit are stacked and connected to each other in sequence, and the topmost culture container unit has a breathing port 3 above its channel 6.
[0029] See Figure 1 The inlet pipe assembly includes a main inlet pipe 7 and multiple inlet branch pipes respectively connected to the inlet port 5; the outlet pipe assembly includes a main outlet pipe 1 and multiple outlet branch pipes respectively connected to the outlet port 4.
[0030] When changing the culture medium, first flip side A upwards. The float 12 controls the position of the suction port of the suction tube 11 to always be below the set position on the liquid surface. Since the cells are denser than the culture medium, they will sink to the bottom. This allows only the upper layer of culture medium to be suctioned, making medium replacement in cell suspension culture much more convenient. Simultaneously, multiple culture container units are stacked together as a single unit, significantly improving the production efficiency of suspension cells compared to existing cell suspension cultures, allowing for the harvesting of a large number of cells at once. Furthermore, the addition and replacement of culture medium during cell culture are all performed in a closed system, avoiding contamination during the culture process.
[0031] In this embodiment, see Figure 2 The culture container unit has a flat rectangular structure.
[0032] In this embodiment, see Figure 1-2 The inlet 5 and outlet 4 are located on the same side A of the culture container unit. When the culture medium solution is drawn out through the outlet pipe, side A is flipped upward.
[0033] In this embodiment, referring to 1-2, the breathing port 3 is a pair located near the side A. When the side A is flipped upward, the breathing port 3 is higher than the liquid level inside the culture container unit.
[0034] In this embodiment, see Figure 1 The main inlet pipe 7 is equipped with a pipe clamp 9 and a filter 8; furthermore, the inlet branch pipe and the outlet branch pipe are equipped with pipe clamps. The filter 8 can ensure the sterility of the culture medium during the culture medium addition process.
[0035] See also Figure 1 The main inlet pipe 7 is connected to multiple inlet branch pipes via a ball, and the main outlet pipe 1 is connected to multiple outlet branch pipes via a ball.
[0036] See also Figure 1 The multilayer culture vessel has several bases 2 on its side A for support. When the culture medium solution is injected into the multilayer culture vessel in the initial stage, A is flipped downward so that the liquid level of the culture medium solution in each culture container unit can quickly reach the same level through the channel 6 and the connecting hole 10. At this time, the base 2 can protect the liquid inlet 5 and the liquid outlet 4.
[0037] In this embodiment, after the initial stage of injecting cell seeding solution (cells and culture medium) into the multilayer culture vessel through the channel 6, the cap of the breathing port 3 is closed.
[0038] How to use:
[0039] S1. Assembly and Connection: Assembly should be performed in a Class A environment, and all components must be sterile. Connect the inlet and outlet tubes to the inlet and outlet ports respectively, and clamp all tubing with tube clamps.
[0040] S2. Inoculation: Open the vent and add the predetermined amount of cell suspension into the multilayer culture vessel through the channel below the vent. Tighten the vent cap and place the front end of the multilayer culture vessel downwards. After the liquid levels in each layer are consistent, place the multilayer culture vessel flat on the work surface. The liquid will then be evenly distributed to each layer of the multilayer culture vessel. Gently shake to mix.
[0041] S3. Culture Medium Addition: Connect the container containing the culture medium to the inlet pipe, open all clamps on the inlet pipe, and use a peristaltic pump to pump the culture medium into the multilayer culture apparatus. After the culture medium addition is complete, close all clamps on the inlet pipe. The amount of culture medium added to each layer can be controlled by opening and closing the clamps.
[0042] S4. Culture Medium Replacement: Place the multilayer culture vessel rear-side down on the work surface and allow the cells to settle to the bottom. The float in the internal aspiration tube will float on the surface of the liquid. Connect the external outlet tube to the vacuum pump or peristaltic pump, open all clamps on all outlet tubes, and start the pump. The upper layer of culture medium will be aspirated; since the cells are at the bottom, they will not be aspirated. After the culture medium has been aspirated, close all clamps on the outlet tubes. The amount of culture medium aspirated from each layer can be controlled by opening and closing the clamps.
[0043] S5. Add culture medium again: See the procedure for adding culture medium in S3.
[0044] S6. Cell Harvesting: After cell culture is complete, place the culture container face down on the work surface. Connect the outlet tube to the vacuum pump and extract the cells from the multilayer culture vessel through the outlet tube.
[0045] This invention relates to a multilayer cell culture device. The surface of the culture area is a low-adsorption treatment surface, which is suitable for large-scale culture of suspension cells. When changing the culture medium for suspension cultured cells, the float design ensures that the cells will not flow out with the culture medium. From cell inoculation until cell collection at the end of the culture, the culture system is in a sealed state during the entire process of adding and changing the culture medium, which greatly reduces the risk of contamination.
[0046] The above are preferred embodiments of the present utility model. Those skilled in the art can make various changes or improvements based on this. Without departing from the overall concept of the present utility model, these changes or improvements should all fall within the scope of protection claimed by the present utility model.
Claims
1. A multilayer culture device suitable for cell suspension culture, characterized in that: It includes an inlet pipe assembly, an outlet pipe assembly, and multiple stacked culture container units; The culture container unit is provided with a channel (6), and the upper part of the channel (6) is provided with a connecting hole (10); The culture container unit has an inlet (5) and an outlet (4) on its side wall; the outlet (4) extends into the culture container unit into a suction pipe (11), and the suction pipe (11) is equipped with a float (12); The culture container unit is provided with at least one channel (6), and the upper part of the channel (6) is provided with a connecting hole (10) that communicates with the inside of the culture container unit; after multiple culture container units are stacked, the channels (6) of each culture container unit are stacked and connected to each other in sequence, and the channel (6) of the topmost culture container unit is provided with a breathing port (3) above it. The inlet pipe assembly includes a main inlet pipe (7) and multiple inlet branch pipes that are respectively connected to the inlet port (5); The liquid outlet assembly includes a main liquid outlet pipe (1) and multiple branch liquid outlet pipes that are respectively connected to the liquid outlet (4).
2. The multilayer culture vessel for cell suspension culture as described in claim 1, characterized in that: The culture container unit has a flat rectangular structure.
3. The multilayer culture vessel for cell suspension culture as described in claim 2, characterized in that: The inlet (5) and outlet (4) are located on the same side A of the culture container unit. When the culture medium solution is drawn out through the outlet pipe, the side A is flipped upward.
4. The multilayer culture vessel for cell suspension culture as described in claim 3, characterized in that: The breathing port (3) is a pair located near the side A. When the side A is flipped upward, the breathing port (3) is higher than the liquid level inside the culture container unit.
5. The multilayer culture vessel for cell suspension culture as described in claim 1, characterized in that: The main inlet pipe (7) is equipped with a pipe clamp (9) and a filter (8).
6. The multilayer culture vessel for cell suspension culture as described in claim 5, characterized in that: The inlet and outlet branches are equipped with pipe clamps.
7. The multilayer culture vessel for cell suspension culture as described in claim 1, characterized in that: The main inlet pipe (7) is connected to multiple inlet branch pipes via a sphere, and the main outlet pipe (1) is connected to multiple outlet branch pipes via a sphere.
8. The multilayer culture vessel for cell suspension culture as described in claim 3, characterized in that: The multilayer culture vessel is provided with several bases (2) for support on its side A. When the culture medium solution is injected into the multilayer culture vessel in the initial stage, the A is flipped downward so that the liquid surface of the culture medium solution in each culture container unit quickly reaches the same level through the channel (6) and the connecting hole (10). At this time, the base (2) can protect the liquid inlet (5) and the liquid outlet (4).
9. The multilayer culture vessel for cell suspension culture as described in claim 8, characterized in that: After the initial stage, the cell seeding solution is injected into the multilayer culture vessel through the channel (6), the lid of the breathing port (3) is closed.
10. The multilayer culture vessel for cell suspension culture as described in claim 8, characterized in that: The cell inoculation solution is a mixture of cells and culture medium.