Closed liquid supplementing device for suspension cell culture
By using a closed-loop fluid replenishment device with a transfer unit and a peristaltic pump system, the problems of easy fluid detachment, large errors, and high risk of contamination in existing technologies are solved, achieving closed, precise, and efficient fluid replenishment for cell culture, which is suitable for large-scale culture.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- QINGDAO SINO-CELL BIOMEDICINE CO LTD
- Filing Date
- 2025-03-28
- Publication Date
- 2026-05-08
AI Technical Summary
Current cell culture procedures involve fluid replenishment, which is prone to detachment, has large errors, is complicated, and carries a high risk of contamination, making them unsuitable for large-scale culture.
A closed-loop fluid replenishment device is used, including a transfer device and a peristaltic pump. The culture medium bottle and culture bag are connected through an interface device consisting of a medical threaded conversion cap, a one-way valve and a filter. The peristaltic pump is used to achieve unidirectional and uniform pumping, and the device is combined with a scale and controller to achieve precise control.
It enables closed, precise, and efficient fluid replenishment operations, reduces the risk of contamination, and improves the efficiency and success rate of cell culture, making it suitable for large-scale culture.
Smart Images
Figure CN224212667U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of cell culture technology, and more specifically to a closed-loop fluid replenishment device for suspension cell culture. Background Technology
[0002] This section is intended to provide background or context for the embodiments of this application as set forth in the claims. The description herein is not intended to imply that it is prior art that has been disclosed, simply because it is included in this section.
[0003] Cell culture technology is a technique that simulates the in vivo environment in vitro, enabling cells to survive, grow, multiply, and maintain their structure and function. It plays a vital role in basic research, drug screening and toxicity testing, biopharmaceutical production, regenerative medicine, and tissue engineering, and has broad application prospects.
[0004] In cell culture, fluid replenishment is a crucial step, directly impacting the efficiency and quality of the culture. Currently, the most common method for fluid replenishment in cell culture is to use a syringe syringe. The syringe is placed in a syringe holder, connected to the inlet of the cell culture bag, and the culture medium is poured into the syringe.
[0005] The above-mentioned manual refill method is prone to detachment at the syringe and culture bag connection, and the use of pipettes or pouring according to the culture bottle scale can easily cause large errors. It requires multiple pours and a lot of personnel interaction, making the operation complicated and unsuitable for large-scale culture. It is also easy to spill when pouring culture medium, and the risk of cell and culture medium contamination is high due to repeated open operation. Summary of the Invention
[0006] The purpose of this invention is to provide a closed-loop fluid replenishment device for suspension cell culture, enabling closed, precise, and efficient fluid replenishment during the cell culture process.
[0007] This application discloses a closed-loop fluid replenishment device for suspension cell culture, including a transfer device and a peristaltic pump:
[0008] The transfer device includes a refill tube, a first interface device, and a second interface device. One end of the refill tube is connected to a culture medium bottle through the first interface device, which consists of a medical threaded conversion cap, a one-way valve, and a filter. The other end of the refill tube is connected to a culture bag through the second interface device. A closed system is formed between the culture medium bottle, the transfer device, and the culture bag.
[0009] The peristaltic pump is used to pump the liquid in the culture medium bottle into the culture bag in a unidirectional and uniform manner. When in use, the replenishment tube is placed in the peristaltic pump.
[0010] In a preferred embodiment, in the first interface device, the medical threaded conversion cap is sized to match the outlet of the culture medium bottle and is fitted onto the outlet of the culture medium bottle; the cap of the medical threaded conversion cap has two round holes, through which one end of the infusion tube and one end of the one-way valve are respectively connected on the outside of the cap, and the other end of the one-way valve is connected to the filter;
[0011] The one-way valve is used to control the air flow to flow only from the outside to the inside of the culture medium bottle; the filter is used to filter impurities in the air.
[0012] In a preferred embodiment, the one-way valve is composed of multiple sections of pipes of different diameters connected together, and one of the multiple sections of pipes of different diameters has a circular one-way valve disc inside.
[0013] In a preferred embodiment, the one-way valve is composed of three pipe sections of different diameters connected together, and the middle one of the three pipe sections of different diameters has a circular one-way valve disc inside.
[0014] The filter is composed of a connecting pipe, a disc-shaped structure and an inlet round pipe connected in sequence, and the disc-shaped structure contains a filter membrane.
[0015] The one-way valve is connected to the connecting pipe of the filter.
[0016] In a preferred embodiment, a scale is also included;
[0017] The scale is used to weigh the culture bag in real time during the replenishment process and monitor the mass change of the liquid inside the culture bag.
[0018] In a preferred embodiment, a controller is also included, electrically connected to the scale and the peristaltic pump, for receiving a mass signal from the scale and sending a stop signal to stop the peristaltic pump when the mass of the culture bag reaches the expected mass.
[0019] In a preferred embodiment, the peristaltic pump receives a mass signal from the scale and stops operating when the mass of the culture bag reaches the expected mass; the peristaltic pump has a flow control function and adjusts the flow control according to the mass signal.
[0020] In a preferred embodiment, the second interface device includes a Luer connector, a PVC heat-sealed pipe, or a sterile pipe fitting machine.
[0021] In a preferred embodiment, the peristaltic pump has the function of controlling the flow rate.
[0022] In a preferred embodiment, the peristaltic pump has the function of controlling the rotational speed and pumping time.
[0023] In a preferred embodiment, the peristaltic pump has the function of controlling the pumping volume.
[0024] In a preferred embodiment, the peristaltic pump has the function of controlling the rotational speed, pumping time, and pumping volume.
[0025] In a preferred embodiment, the fluid replenishment tube comprises a silicone tube, a sterile polyvinyl chloride tube, or a thermoplastic elastomer tube.
[0026] In a preferred embodiment, the transfer device further includes a first water-stop clamp, and the culture bag further includes a second water-stop clamp;
[0027] The first water-stop clamp is used to control the flow and stop of liquid in the replenishment tube, and is clamped on the replenishment tube during use;
[0028] The second water-stop clamp is used to control the flow and stop of liquid in the inlet connection pipe of the culture bag, and is clamped on the inlet connection pipe of the culture bag during use.
[0029] In a preferred embodiment, a first support frame is further included for placing the culture medium bottle and the peristaltic pump, wherein the culture medium bottle is positioned upside down on the first support frame during use.
[0030] In a preferred embodiment, the system further includes a first support frame and a second support frame, wherein the first support frame is used to place the culture medium bottle and the peristaltic pump, and the culture medium bottle is hung upside down on the first support frame during use;
[0031] The second support frame is used to support the replenishment tube, thereby ensuring that the outlet of the replenishment tube and the inlet of the culture bag are in a relative position.
[0032] In a preferred embodiment, the first support frame is a floor-standing metal support frame;
[0033] In a preferred embodiment, the second support frame is a small acrylic support frame.
[0034] In a preferred embodiment, the culture bag is placed in a Class A environment, and the peristaltic pump, culture medium bottle, and first support frame are placed in a Class C environment.
[0035] In a preferred embodiment, the culture bag, peristaltic pump, culture medium bottle, and first support frame are placed in a Class A environment.
[0036] In a preferred embodiment, the culture bag and the second support frame are placed in a Class A environment, while the peristaltic pump, culture medium bottle, and the first support frame are placed in a Class C environment.
[0037] In a preferred embodiment, the culture bag, the second support frame, the peristaltic pump, the culture medium bottle, and the first support frame are placed in a Class A environment.
[0038] The main differences and effects of this utility model embodiment compared with the prior art are as follows:
[0039] This application has good sealing properties and a low risk of contamination;
[0040] Furthermore, this application provides precise and easily controllable fluid replacement;
[0041] Furthermore, this application is easy to operate and significantly improves the efficiency and success rate of cell culture;
[0042] Furthermore, this application has broad applicability, applicable to large-scale cell culture and to all culture bags available on the market.
[0043] It should be understood that, within the scope of this utility model, the above-described technical features of this utility model and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description
[0044] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0045] Figure 1 This is a schematic diagram of a cell culture fluid replenishment device according to one embodiment of this application.
[0046] Figure 2 This is a schematic diagram of the structure of a liquid transfer device according to one embodiment of this application.
[0047] Figure 3 This is a schematic diagram of the peristaltic pump according to one embodiment of this application.
[0048] Figure 4 This is a schematic diagram of a culture bag according to one embodiment of this application.
[0049] Figure 5 This is a schematic diagram of a Luer connector according to one embodiment of this application.
[0050] The labels in each of the attached figures are as follows:
[0051] 1-Transfer Unit
[0052] 101-Replenishment tubing;
[0053] 102-Medical threaded adapter cap;
[0054] 103 - Check valve;
[0055] 104-Filter;
[0056] 105-Ruhr joint;
[0057] 106 - First waterstop clamp;
[0058] 2-Peristaltic pump;
[0059] 3-Cultural culture bottle;
[0060] 4-Cultivation bag;
[0061] 401 - Inlet connecting pipe;
[0062] 5-Scale;
[0063] 6-Controller;
[0064] 7-First support frame. Detailed Implementation
[0065] In the following description, many technical details are presented to help the reader better understand this application. However, those skilled in the art will understand that the technical solutions claimed in this application can be implemented even without these technical details and various variations and modifications based on the following embodiments.
[0066] To make the objectives, technical solutions, and advantages of this application clearer, the embodiments of this application will be described in further detail below with reference to the accompanying drawings.
[0067] This application discloses a closed-loop fluid replenishment device for suspension cell culture, the structure of which is as follows: Figure 1 As shown, it includes a transfer device 1 and a peristaltic pump 2:
[0068] Transfer device 1, such as Figure 2 As shown, it includes a refill tube 101, a first interface device and a second interface device. One end of the refill tube 101 is connected to the culture medium bottle 3 through the first interface device, which is composed of a medical threaded conversion cap 102, a one-way valve 103 and a filter 104. The other end of the refill tube 101 is connected to the culture bag 4 through the second interface device. A closed system is formed between the culture medium bottle 3, the transfer device 1 and the culture bag 4.
[0069] In one embodiment, the transfer device 1 is reusable.
[0070] Peristaltic pump 2, such as Figure 3 As shown, the liquid in the culture medium bottle 3 is pumped into the culture bag 4 in a unidirectional and uniform manner. During use, the replenishment tube 101 is placed in the peristaltic pump 2.
[0071] This application is simple to operate, reduces personnel interaction, and is suitable for large-scale culture. The culture medium flows in the replenishment tube 101 throughout the process, without contact with the outside world to avoid introducing contaminants. Furthermore, the peristaltic pump 2 is used to control the one-way replenishment, avoiding cross-contamination between cell suspension and culture medium, which significantly improves the efficiency and success rate of cell culture.
[0072] In one embodiment, in the first interface device, the medical threaded conversion cap 102 is sized to match the outlet of the culture medium bottle 3 and is fitted onto the outlet of the culture medium bottle 3; the cap of the medical threaded conversion cap 102 has two round holes, through which one end of the infusion tube 101 and one end of the one-way valve 103 are respectively connected to the outside of the cap, and the other end of the one-way valve 103 is connected to the filter 104; the one-way valve 103 is used to control the air to flow only from the outside to the inside of the culture medium bottle 3; the filter 104 is used to filter impurities in the air.
[0073] In one embodiment, the one-way valve 103 is composed of multiple sections of pipes with different diameters connected together, and one of the multiple sections of pipes with different diameters has a circular one-way valve disc inside.
[0074] In one embodiment, the one-way valve 103 is composed of three pipes of different diameters connected together, and the middle one of the three pipes of different diameters has a circular one-way valve flap inside; the filter 104 is composed of a connecting pipe, a disc-shaped structure and an inlet circular pipe connected in sequence, and the disc-shaped structure has a filter membrane inside.
[0075] The one-way valve 103 is connected to the filter 104 via a connecting pipe.
[0076] In one embodiment, filter 104 uses a filter head with a finer aperture of 0.22µm or less, such as a filter head with an even smaller aperture of 0.2µm.
[0077] In one embodiment, a scale 5 is also included;
[0078] Weigher 5 is used to weigh the culture bag 4 in real time during the replenishment process and monitor the mass change of the liquid inside the culture bag 4.
[0079] In one embodiment, the scale 5 is an electronic scale; the electronic scale can accurately control the volume of liquid replenishment, ensuring the accuracy of liquid replenishment.
[0080] In one embodiment, such as Figure 1 As shown, it also includes a controller 6, which is electrically connected to the scale 5 and the peristaltic pump 2, for receiving mass signals from the scale 5 and sending a stop signal to stop the peristaltic pump 2 when the mass of the culture bag 4 reaches the expected mass.
[0081] In one embodiment, the peristaltic pump 2 receives a mass signal from the scale 5 and stops operating when the mass of the culture bag 4 reaches the expected mass; the peristaltic pump 2 has a flow control function and adjusts the flow control according to the mass signal.
[0082] In one embodiment, such as Figure 4 As shown, the culture bag 4 is rectangular, with a capacity of 500-1500ml. A short tube interface is located in the middle of one of the shorter sides of the culture bag 4. Next to the short tube interface is a connecting tube 401 at the inlet of the culture bag 4. During use, the connecting tube 401 is connected to the replenishment tube 101. This device is also applicable to all types of culture bags; the size of the first interface device can be adjusted according to the type of culture bag.
[0083] In one embodiment, the second interface device includes a Luer connector 105, a PVC heat-sealing tube, or a sterile connector. The Luer connector 105 is used to ensure a tight connection between the infusion tube 101 and the culture bag 4, ensuring that the seamless connection will not come loose. The sterile connector is also used to connect the infusion tube 101 and the culture bag 4, which can better ensure the tightness and airtightness of the connection.
[0084] In one embodiment, such as Figure 5 As shown, Luer connector 105 is a double male Luer connector made of hard plastic.
[0085] In one embodiment, the peristaltic pump 2 has the function of controlling the flow rate.
[0086] In one embodiment, the peristaltic pump 2 has the function of controlling the rotational speed and pumping time.
[0087] In one embodiment, the peristaltic pump 2 has the function of controlling the pumping volume.
[0088] In one embodiment, the peristaltic pump 2 has the function of controlling the rotational speed, pumping time, and pumping volume.
[0089] In one embodiment, the peristaltic pump 2 has the function of precisely controlling the rotation speed and pumping time, and will not produce backflow, thereby realizing the unidirectional and uniform pumping of the culture medium, ensuring the stability and accuracy of the replenishment process, while avoiding cross-contamination between cell suspension and culture medium.
[0090] In one embodiment, the parameters of the peristaltic pump 2 include a rotational speed of 80-100 rpm.
[0091] In one embodiment, the fluid replenishment tube 101 includes a silicone tube, a sterile polyvinyl chloride tube, or a thermoplastic elastomer tube, and can be selected from various diameters and lengths, with the length range including 50-150cm.
[0092] In one embodiment, different models of culture medium bottles 3 and different diameter infusion tubes 101 can be adapted by replacing the medical thread conversion caps 102 of different specifications.
[0093] In one embodiment, the culture medium bottle 3 is a medical bottle with rounded corners and a cuboid shape.
[0094] In one embodiment, the transfer device 1 further includes a first water-stop clamp 106, and the culture bag 4 further includes a second water-stop clamp (not shown in the figure). The first water-stop clamp 106 is used to control the flow and stop of liquid in the replenishment tube 101 and is clamped on the replenishment tube 101 during use.
[0095] The second water-stop clamp is used to control the flow and stop of liquid in the inlet connection pipe 401 of the culture bag 4. It is clamped on the inlet connection pipe 401 of the culture bag 4 during use.
[0096] In one embodiment, it further includes a first support frame 7 and a second support frame (not shown in the figure). The first support frame 7 is used to place the culture medium bottle 3 and the peristaltic pump 2. When in use, the culture medium bottle 3 is hung upside down on the first support frame 7.
[0097] The second support frame is used to support the replenishment tube 101, thereby ensuring that the output port of the replenishment tube 101 and the inlet of the culture bag 4 are in a relative position.
[0098] In one embodiment, the first support frame 7 is a floor-mounted metal support frame;
[0099] In one embodiment, the second support frame is a small acrylic support frame.
[0100] In one embodiment, the culture bag 4 is placed in a Class A environment, and the peristaltic pump 2, culture medium bottle 3, and first support frame 7 are placed in a Class C environment.
[0101] In one embodiment, the culture bag 4, the peristaltic pump 2, the culture medium bottle 3, and the first support frame 7 are placed in a Class A environment.
[0102] In one embodiment, the culture bag 4 and the second support frame are placed in a Class A environment, while the peristaltic pump 2, the culture medium bottle 3, and the first support frame 7 are placed in a Class C environment.
[0103] In one embodiment, the culture bag 4, the second support frame, the peristaltic pump 2, the culture medium bottle 3, and the first support frame 7 are placed in a Class A environment.
[0104] Although this application pertains to a closed-loop fluid replenishment device for suspension cell culture, it may also be applicable to other cell types, such as non-suspension immune cells, etc., which are also within the scope of this application.
[0105] Example 1
[0106] In one embodiment, the device of this application can be used to perform cell rehydration as follows: replace the cap of the culture medium bottle 3 with the medical threaded conversion cap 102 of the transfer device 1, and place the culture medium bottle 3 on the first support frame 7. Before rehydration, the culture bag 4 contains a small amount of cell suspension, such as 0 to 200 ml, and the required rehydration volume is 200 to 500 ml. Clamp the first stop clamp 106 on the rehydration tube 101 and the second stop clamp on the inlet connecting tube 401 of the culture bag 4, then connect the tubing of the transfer device 1, turn on the peristaltic pump 2, and open or remove the first stop clamp 106 and the second stop clamp.
[0107] The culture process takes approximately 10-20 days. During the culture, the device does not need to be continuously connected, as it is easy to connect, store, and disassemble, and can be used flexibly as needed for replenishment. The progress of replenishment can be observed using the scale 5. After replenishment is complete, stop the peristaltic pump 2, clamp the first stop clamp 106 onto the replenishment tube 101 and the second stop clamp onto the inlet connection tube 401 of the culture bag 4, open the second interface device, and remove the culture bag 4. Place the culture medium bottle 3 and the transfer device 1 back into the refrigerator, and simultaneously return the culture bag 4 to the incubator. Generally, one culture medium bottle 3 is used to replenish one culture bag 4, because different cells cannot use different culture media to avoid cross-contamination. Each bottle can be replenished two or three times.
[0108] It should be noted that in the claims and specification of this patent, relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one" does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.
[0109] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.
Claims
1. A closed-loop fluid replenishment device for suspension cell culture, characterized in that, Includes a transfer device (1) and a peristaltic pump (2): The transfer device (1) includes a replenishment tube (101), a first interface device, and a second interface device. One end of the replenishment tube (101) is connected to the culture medium bottle (3) through the first interface device, which consists of a medical threaded conversion cap (102), a one-way valve (103), and a filter (104). The other end of the replenishment tube (101) is connected to the culture bag (4) through the second interface device. A closed system is formed between the culture medium bottle (3), the transfer device (1), and the culture bag (4). The peristaltic pump (2) is used to pump the liquid in the culture medium bottle (3) into the culture bag (4) in a unidirectional and uniform manner. When in use, the replenishment tube (101) is placed in the peristaltic pump (2).
2. The apparatus according to claim 1, characterized in that, In the first interface device, the medical threaded conversion cap (102) is sized to match the outlet of the culture medium bottle (3) and is fitted onto the outlet of the culture medium bottle (3); the cap of the medical threaded conversion cap (102) has two round holes, through which one end of the infusion tube (101) and one end of the one-way valve (103) are respectively connected on the outside of the cap, and the other end of the one-way valve (103) is connected to the filter (104); The one-way valve (103) is used to control the air to flow only from the outside to the inside of the culture medium bottle (3); the filter (104) is used to filter impurities in the air.
3. The apparatus according to claim 1, characterized in that, The one-way valve (103) is composed of multiple sections of pipes with different diameters connected together, and one of the multiple sections of pipes with different diameters has a circular one-way valve disc inside. The filter (104) is composed of a connecting pipe, a disc-shaped structure and an inlet round pipe connected in sequence, and the disc-shaped structure contains a filter membrane. The one-way valve (103) is connected to the connecting pipe of the filter (104).
4. The apparatus according to claim 1, characterized in that, It also includes scales (5); The scale (5) is used to weigh the mass of the culture bag (4) in real time during the replenishment process and monitor the mass change of the liquid in the culture bag (4).
5. The apparatus according to claim 4, characterized in that, It also includes a controller (6) electrically connected to the scale (5) and the peristaltic pump (2), for receiving a mass signal from the scale (5) and sending a stop signal to stop the peristaltic pump (2) when the mass of the culture bag (4) reaches the expected mass.
6. The apparatus according to claim 1, characterized in that, The second interface device includes a Luer connector (105), a PVC heat-sealed pipe, or a sterile pipe fitting machine.
7. The peristaltic pump (2) according to claim 1 has the function of controlling the flow rate.
8. The apparatus according to claim 1, characterized in that, The fluid replenishment tube (101) includes a silicone tube, a sterile polyvinyl chloride tube, or a thermoplastic elastomer tube.
9. The apparatus according to claim 1, characterized in that, The transfer device (1) further includes a first water-stop clamp (106), and the culture bag (4) further includes a second water-stop clamp; The first water-stopping clamp (106) is used to control the flow and stop of liquid in the replenishment tube (101), and is clamped on the replenishment tube (101) during use; The second water-stopping clamp is used to control the flow and stop of liquid in the inlet connecting pipe (401) of the culture bag (4), and is clamped on the inlet connecting pipe (401) of the culture bag (4) during use.
10. The apparatus according to claim 1, characterized in that, It also includes a first support frame (7) and a second support frame. The first support frame (7) is used to place the culture medium bottle (3) and the peristaltic pump (2). When in use, the culture medium bottle (3) is hung upside down on the first support frame (7). The second support frame is used to support the replenishment tube (101) to ensure that the outlet of the replenishment tube (101) and the inlet of the culture bag (4) are in a relative position.