A disposable cell separation device and a cell separation apparatus

Abstract

The application discloses a disposable cell separation device, which comprises a transfer container, a rotary joint and a separation container connected with the rotary joint, the transfer container is communicated with the separation container through a pipeline, the transfer container is provided with a ventilation connecting part for generating positive pressure or negative pressure in the inside of the transfer container, the pipeline is provided with a first docking part for connecting an initial container, a second docking part for connecting a buffer solution container and a third docking part for connecting a collection container. The application also discloses a disposable cell separation device. In the feeding and discharging process, the material does not directly pass through the positive and negative pressure device and is not repeatedly extruded by the positive and negative pressure device, so that cell damage in the feeding and discharging conveying process is avoided and the cell yield and activity are greatly improved. The disposable cell separation device and the cell separation device have the advantages of avoiding cell damage caused by the suction device and greatly improving the cell yield and activity.

Description

Technical Field

[0001] This invention relates to the field of cell drug preparation technology, and in particular to a disposable cell separation device and cell separation apparatus. Background Technology

[0002] Cell isolation from suspensions (such as blood or bone marrow) is becoming increasingly common in medical treatments. For this purpose, cells are removed from the patient, isolated to provide the desired target cells, and then stimulated / treated / expanded before being introduced into the same or different patients. For example, PBMC isolation, or peripheral blood mononuclear cell isolation, has long been a practice in this field. Human peripheral blood contains lymphocytes, monocytes, dendritic cells, and a small number of other cells. The isolation, screening, and separation of PBMCs using blood cell separation tubing systems can yield large quantities of immune cells for cell therapy. Early laboratories typically used manual methods, injecting the suspension to be separated into a separation container, which was then mounted on a centrifuge. After centrifugation, the separated cells were harvested using syringes or other instruments. Now, to avoid cell contamination, fully automated cell centrifuges and accompanying disposable consumables have gradually emerged. These disposable consumables are containers, tubing, and other equipment that come into direct contact with the cells; their interiors are sealed and do not come into contact with the external environment, and they are packaged in sealed sterile bags before use.

[0003] When using disposable consumables for cell separation, the suspension to be separated and the necessary solution must be injected into the disposable consumable in a sterile environment, or a container containing the suspension can be attached to the disposable product. The entire disposable consumable is then installed on a centrifuge for fully automated cell centrifugation. This effectively avoids the contamination problems associated with manual operation. However, existing tubing systems for blood cell separation directly pump the cell fluid using a peristaltic pump. For example, patent application CN202121194589.5 describes a tubing system for blood separation that directly delivers the cell fluid using a peristaltic pump. However, peristaltic pumps deliver cell fluid by squeezing a flexible tube, which can easily cause cell damage, reducing cell yield and viability. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to overcome the shortcomings of the prior art and provide a disposable cell separation device and cell separation apparatus that avoids cell damage caused by aspiration devices and greatly improves cell yield and viability.

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0006] A disposable cell separation device includes a transfer container, a rotary joint, and a separation container connected to the rotary joint. The transfer container and the separation container are connected by a pipeline. The transfer container is provided with a venting connection part to generate positive or negative pressure by allowing air to pass through it. The pipeline is provided with a first docking part for connecting to an initial container, a second docking part for connecting to a buffer solution container, and a third docking part for connecting to a collection container.

[0007] As a further improvement to the above technical solution:

[0008] The first docking part is docked to an initial container, and / or the second docking part is docked to a buffer container, and / or the third docking part is docked to a collection container.

[0009] The collection container is equipped with a sampling interface and / or a first gas filter.

[0010] The pipeline is equipped with a fourth docking section for connecting waste liquid containers.

[0011] The fourth docking part is connected to a waste liquid container.

[0012] The pipeline is provided with a fifth docking section for connecting a transition container and / or a sixth docking section for connecting a separation liquid container.

[0013] The fifth docking part is connected to a transition container.

[0014] The sixth docking section is connected to a separation liquid container.

[0015] A demagnetizing component is provided between the first docking part and the transfer container.

[0016] The pipeline is connected to the bottom of the transfer container.

[0017] The pipeline is also connected to the top of the transfer container.

[0018] The ventilation connection is located on the side wall or top wall of the transfer container.

[0019] The ventilation connection is connected to a positive and negative pressure device.

[0020] The positive and negative pressure device includes a second gas filter and a gas pipe, wherein the second gas filter is connected to the transfer container through the gas pipe.

[0021] An intermediate container is provided between the trachea and the transfer container.

[0022] The transfer container is a rigid container.

[0023] The separation container is connected to the pipeline via a rotary joint.

[0024] The rotary joint includes an inner column and a rotary sleeve. The rotary sleeve is rotatably fitted outside the inner column. The rotary sleeve and the inner column are provided with a connection channel that maintains communication during relative rotation. The pipeline is connected to the connection channel of the rotary joint, and the separation container is connected to the connection channel of the rotary sleeve.

[0025] The inner column is provided with a first connecting part communicating with its connecting channel, the pipeline is connected to the first connecting part, the rotating sleeve is provided with a second connecting part communicating with its connecting channel, and the separation container is connected to the second connecting part.

[0026] A cell separation device includes a mounting panel and the aforementioned disposable cell separation apparatus. The mounting panel is provided with multiple mounting parts, multiple clamping valves, and multiple fixed pipe fittings. The transfer container is detachably mounted on the corresponding mounting part, the pipeline is fixed on the fixed pipe fitting, and the clamping valve is clamped on the corresponding pipe position.

[0027] Compared with the prior art, the advantages of the present invention are as follows:

[0028] The disposable cell separation device of the present invention prevents materials from directly passing through the positive and negative pressure devices during the feeding and discharging process, thus avoiding cell damage during the feeding and discharging process and significantly improving cell yield and viability.

[0029] The cell separation device of this invention features a disposable cell separation apparatus that is detachably mounted on the installation panel. After use, the disposable cell separation apparatus is disposed of. Separation containers, as well as initial containers, buffer containers, and other containers, are housed within the separation device or mounted on hooks. This design allows the clamp valve to be reused multiple times, rather than being used only with the disposable cell separation apparatus, thus saving costs. Attached Figure Description

[0030] Figure 1 This is a schematic diagram of the structure of the disposable cell separation device of the present invention.

[0031] Figure 2 This is a schematic diagram of the collection container of the disposable cell separation device of the present invention.

[0032] Figure 3 This is a schematic diagram of the rotary joint of the disposable cell separation device of the present invention.

[0033] Figure 4 This is a diagram showing the combination of the disposable cell separation device of the present invention and the peristaltic pump.

[0034] Figure 5 This is the book Figure 4 Enlarged view of point A in the middle.

[0035] Figure 6 This is a packaging diagram of the disposable cell separation device of the present invention.

[0036] Figure 7 This is a schematic diagram of the packaging bag for the disposable cell separation device of the present invention.

[0037] Figure 8 This is a front view structural diagram of the cell separation device of the present invention.

[0038] The labels in the diagram represent:

[0039] 1. Transfer container; 10. Piping; 101. Sixth docking part; 102. Demagnetizing component; 11. Ventilation connection part; 12. Rotary joint; 121. Inner column; 122. Rotating sleeve; 123. First connection part; 124. Second connection part; 13. Peristaltic pump; 14. Packaging bag; 2. Separation container; 3. Initial container; 31. First docking part; 4. Separation liquid container; 41. Second docking part; 5. Collection container; 51. Third docking part; 52. Sampling interface; 53. First gas filter; 6. Waste liquid container; 61. Fourth docking part; 7. Positive and negative pressure device; 70. Gas pipe; 71. Second gas filter; 72. Intermediate container; 8. Transition container; 81. Fifth docking part; 9. Mounting panel; 91. Mounting part; 92. Pinch valve; 93. Fixed pipe fitting. Detailed Implementation

[0040] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0041] Example 1:

[0042] Figures 1 to 7 An embodiment of the disposable cell separation device of the present invention is shown. This disposable cell separation device includes a transfer container 1, a rotary joint 12, and a separation container 2 connected to the rotary joint 12. The transfer container 1 and the separation container 2 are connected by a pipe 10. The transfer container 1 is provided with a venting connection part 11 to generate positive or negative pressure by allowing air to pass through it. The pipe 10 is provided with a first docking part 101 for connecting to an initial container, a second docking part 31 for connecting to a buffer solution container 3, and a third docking part 51 for connecting to a collection container 5. The second docking part 31 is shown for the initial container.

[0043] When using, install this disposable cell separation device on such as Figure 4The mounting panel 9 shown has a clamp valve 92 that clamps onto the corresponding pipe positions, controlling the opening and closing of the corresponding pipe section 10. During cell separation, in a sterile environment, the initial container containing blood is connected to the first docking part 101, the buffer solution container 3 containing buffer solution is connected to the second docking part 31, the collection container 5 is connected to the third docking part 51, and the positive and negative pressure device 7 is connected to the venting connection part 11. The pipes 10 between the initial container and the transfer container 1, the buffer solution container 3 and the transfer container 1, the separation container 2 and the transfer container 1, and the collection container 5 and the transfer container 1 are clamped and disconnected by the clamp valve 92. Then, the positive and negative pressure device 7 is used to evacuate the transfer container 1 to generate negative pressure. Then, the... The clamp valve 92 opens the tubing 10 between the initial container and the transfer container 1 and / or the buffer solution container 3 and the transfer container 1. Under negative pressure, the blood in the initial container and / or the buffer solution in the buffer solution container 3 enter the transfer container 1. Then, the clamp valve 92 clamps and disconnects the tubing 10 between the initial container and the transfer container 1, and between the buffer solution container 3 and the transfer container 1. The tubing 10 between the transfer container 1 and the separation container 2 is then opened. The positive and negative pressure device 7 inflates the transfer container 1 to generate positive pressure, allowing the material in the transfer container 1 to enter the separation container 2.

[0044] After blood separation in separation container 2 is completed, the tubing 10 between the initial container and transfer container 1, buffer container 3 and transfer container 1, collection container 5 and transfer container 1, and separation container 2 and transfer container 1 is clamped and closed by clamp valve 92. Then, the transfer container 1 is evacuated by positive and negative pressure device 7 to generate negative pressure. Next, the tubing 10 between separation container 2 and transfer container 1 is released and opened by clamp valve 92. Under the action of negative pressure, the target cells in separation container 2 enter transfer container 1. Then, the tubing 10 between separation container 2 and transfer container 1 is clamped and disconnected by clamp valve 92, and the tubing 10 between transfer container 1 and collection container 5 is released and opened. Then, the transfer container 1 is inflated by positive and negative pressure device 7 to generate positive pressure, so that the target cells in transfer container 1 enter collection container 5. In this way, the materials (blood, cells) do not pass directly through the positive and negative pressure device 7 during the feeding and discharging process, and are not subjected to the repeated squeezing action of the positive and negative pressure device 7, thus avoiding cell damage during the feeding and discharging process and greatly improving cell yield and viability.

[0045] In this embodiment, the first docking part 31 is docked with an initial container, and / or the second docking part 31 is docked with a buffer solution container 3, and / or the third docking part 51 is docked with a collection container 5. Thus, in subsequent use, it is not necessary to prepare an initial container that can dock with the first docking part 31, a buffer solution container 3 that can dock with the second docking part 31, or a collection container 5 that can dock with the third docking part 51.

[0046] In this embodiment, the collection container 5 is provided with a sampling interface 52 and / or a first gas filter 53. The sampling interface 52 facilitates sampling and detection. The first gas filter 53 is used to balance the gas pressure inside the collection container 5.

[0047] In this embodiment, the pipeline 10 is provided with a fourth docking part 61 for connecting the waste liquid container 6. In this way, it is not necessary to prepare a waste liquid container 6 that can dock with the fourth docking part 61 in subsequent use.

[0048] In this embodiment, the fourth docking part 61 is docked with the waste liquid container 6. During the separation process, the separation container 2 docks the waste liquid container 6 with the fourth docking part 61, which can be used to collect the waste liquid discharged from the separation container 2.

[0049] In this embodiment, the pipeline 10 is provided with a fifth docking part 81 for connecting the transition container 8. Since the initial container holds several times more blood than the separation container 2, and separation can generally be completed in one step, a fifth docking part 81 is provided for connecting the transition container 8. During use, the transition container 8 is connected to the fifth docking part 81. In this way, the blood in the initial container can be divided into multiple portions and output sequentially. One portion of blood is output into the separation container 2 at a time, and the separation container 2 separates one portion of blood at a time. After each separation, the target cells from the separation container 2 are first output to the transition container 8 for unified collection. After all portions of blood have been separated, the target cell fluid collected in the transition container 8 is returned to the separation container 2 for further separation.

[0050] In this embodiment, the fifth docking part 81 is docked with the transition container 8. Therefore, in subsequent use, it is not necessary to prepare a transition container 8 that can dock with the fifth docking part 81.

[0051] In this embodiment, the pipeline 10 is provided with a sixth docking part 41 for connecting the separation liquid container 4. The sixth docking part 41 is used to connect with the separation liquid container 4, so that in subsequent use, it is not necessary to prepare a separation liquid container 4 that can dock with the sixth docking part 41.

[0052] In this embodiment, the sixth docking part 41 is docked with the separation liquid container 4. During the separation process, the separation liquid container 2 docks with the sixth docking part 41 to provide separation liquid into the separation container 2.

[0053] In this embodiment, a demagnetizing component 102 is provided between the first docking part 101 and the pipeline 10. Since the cell fluid contained in the cell fluid container (initial container) generally contains magnetic beads, it is necessary to remove the magnetic beads during discharge. Therefore, a demagnetizing component 102 is provided between the sixth docking part 41 and the pipeline 10 to remove the magnetic beads in the cells.

[0054] In this embodiment, pipe 10 is connected to the bottom of transfer container 1. Pipe 10 is also connected to the top of transfer container 1.

[0055] Pipeline 10 connects the top and bottom of transfer container 1. Since transfer container 2 is required for both inlet (material flows from the initial container into separation container 2) and outlet (material flows from separation container 2 into collection container 5), during inlet, liquid enters transfer container 1 from the initial container via pipeline 10 and the top of transfer container 1, then flows through the bottom of transfer container 1 and pipeline 10 into separation container 2. During outlet, liquid enters transfer container 1 from separation container 2 via pipeline 10 and the top of transfer container 1, then flows through the bottom of transfer container 1 and pipeline 10 into collection container 5. Therefore, both inlet and outlet flow through the top and bottom of transfer container 1, preventing liquid from entering from the bottom. If the outlet of separation container 2 enters from the bottom of transfer container 1, a large number of air bubbles will be drawn into transfer container 1, and the bursting of these bubbles will adversely affect the cells.

[0056] In this embodiment, the venting connection 11 is provided on the side wall or top wall of the transfer container 1. In this way, during the positive pressure process, gas enters through the side wall or top wall of the transfer container 1. If the venting connection 11 is provided at the bottom of the transfer container 1, when liquid enters the transfer container 1, the liquid may enter the positive and negative pressure device 7 through the venting connection 11 or even leak.

[0057] In this embodiment, the ventilation connection 11 is connected to a positive and negative pressure device 7. The positive and negative pressure device 7 includes a second gas filter 71 and an air pipe 70. The second gas filter 71 is connected to the transfer container 1 through the air pipe 70. In use, the air pipe 70 is placed in the peristaltic pump 13 (e.g., Figure 4 and Figure 5 As shown, when the peristaltic pump 13 is working, it can supply or evacuate air to the transfer container 1 through the air tube 70, thereby achieving positive or negative pressure inside the transfer container 1. The second gas filter 71 is used to filter impurities in the gas to prevent internal contamination of the culture vessel. In addition, when the peristaltic pump 13 clamps the air tube 70, squeezing the air tube 70 may generate particles. Therefore, the second gas filter 71 can be placed between the peristaltic pump 13 and the air connection part 11.

[0058] In this embodiment, an intermediate container 72 is provided between the trachea 70 and the transfer container 1. The intermediate container 72 is used to add a separating fluid to the separation container 2 during cell concentration. During the centrifugation and layering process in the separation container 2, the separating fluid is located at the bottom of the separation container 2, cushioning the cell fluid between the cell solution and the inner wall of the separation container 2. This prevents the target cells from being directly squeezed against the inner wall of the separation container 2 during centrifugation, thereby significantly reducing the physical damage to the target cells during centrifugation and resulting in a high yield and large concentrated volume of target cells. The separating fluid can be human albumin or other liquids with a density greater than that of the target cells that do not damage the target cells.

[0059] In this embodiment, the transfer container 1 is a rigid container. This ensures that the transfer container 1 will not deform under positive or negative pressure conditions.

[0060] In this embodiment, the separation container 2 is connected to the pipeline 10 via a rotary joint 12. This prevents the pipeline 10 from becoming entangled during the rotation of the separation container 2.

[0061] In this embodiment, as Figure 3 As shown, the rotary joint 12 includes an inner column 121 and a rotating sleeve 122. The rotating sleeve 122 is rotatably fitted over the inner column 121. A connection channel is provided between the rotating sleeve 122 and the inner column 121 to maintain communication during relative rotation. The pipeline 10 communicates with the connection channel of the rotary joint 12, and the separation container 2 communicates with the connection channel of the rotating sleeve 122. Thus, when the separation container 2 rotates, the pipeline 10 can remain stationary with the inner column 121 and will not become entangled.

[0062] In this embodiment, the inner column 121 is provided with a first connecting part 123 communicating with its connecting channel, the pipeline 10 is connected to the first connecting part 123, the rotating sleeve 122 is provided with a second connecting part 124 communicating with its connecting channel, and the separation container 2 is connected to the second connecting part 124.

[0063] In this embodiment, as Figure 6 and Figure 7 As shown, the disposable cell culture equipment also includes a packaging bag 14, in which each container and tubing 10 is packaged.

[0064] In this embodiment, the pipeline 10 and the container can be connected by a Luer taper, a two-way connector, etc., that is, the connection part is a Luer taper, a two-way connector, etc., or a connection device can be used to connect the pipeline 10 and the container, such as a hot melt welding machine.

[0065] Example 2:

[0066] Figure 8An embodiment of the cell separation device of the present invention is shown. This embodiment includes a mounting panel 9 and a disposable cell separation device as described in Embodiment 1. The mounting panel 9 is provided with multiple mounting parts 91, multiple clamp valves 92, and multiple fixing fittings 93. A transfer container 1 is detachably mounted on a corresponding mounting part 91, a pipeline 10 is fixed to a fixing fitting 93, and a clamp valve 92 is clamped onto a corresponding pipe position. The disposable cell separation device is detachably mounted on the mounting panel 9 and is disposed of after use. The separation container 2, as well as the initial container, buffer container 3, and other containers, are disposed inside the separation device or on hooks. In this way, the clamp valve 92 is not used once with the disposable cell separation device and can be used multiple times, saving costs.

[0067] While the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the invention. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the scope of the present invention. Therefore, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the scope of the present invention, should fall within the protection scope of the present invention.

Claims

1. A disposable cell separation device, characterized in that: The container includes a transfer container (1), a rotary joint (12), and a separation container (2) connected to the rotary joint (12). The transfer container (1) and the separation container (2) are connected by a pipe (10). The transfer container (1) is provided with a venting connection (11) to generate positive or negative pressure by allowing air to pass through it. The pipe (10) is provided with a first docking part (101) for connecting the initial container, a second docking part (31) for connecting the buffer solution container (3), and a third docking part (51) for connecting the collection container (5). The pipe (10) is connected to the bottom of the transfer container (1) and also to the top of the transfer container (1). The venting connection (11) is located on the side wall or top wall of the transfer container (1). The transfer container (1) is... A rigid container; the separation container (2) is connected to the pipeline (10) via a rotary joint (12). The rotary joint (12) includes an inner column (121) and a rotating sleeve (122). The rotating sleeve (122) and the inner column (121) are provided with a connection channel that maintains communication when rotating relative to each other. The pipeline (10) is connected to the connection channel of the rotary joint (12). The separation container (2) is connected to the connection channel of the rotating sleeve (122). The inner column (121) is provided with a first connection part (123) that communicates with its connection channel. The pipeline (10) is connected to the first connection part (123). The rotating sleeve (122) is provided with a second connection part (124) that communicates with its connection channel. The separation container (2) is connected to the second connection part (124).

2. The disposable cell separation device according to claim 1, characterized in that: The first docking part (101) is docked with an initial container, and / or the second docking part (31) is docked with a buffer container (3), and / or the third docking part (51) is docked with a collection container (5).

3. The disposable cell separation device according to claim 2, characterized in that: The collection container (5) is provided with a sampling interface (52) and / or a first gas filter (53).

4. The disposable cell separation device according to claim 1, characterized in that: The pipeline (10) is provided with a fourth docking part (61) for connecting the waste liquid container (6).

5. The disposable cell separation device according to claim 4, characterized in that: The fourth docking part (61) is docked with a waste liquid container (6).

6. The disposable cell separation device according to claim 1, characterized in that: The pipeline (10) is provided with a fifth docking part (81) for connecting the transition container (8) and / or a sixth docking part (41) for connecting the separation liquid container (4).

7. The disposable cell separation device according to claim 6, characterized in that: The fifth docking part (81) is docked with a transition container (8).

8. The disposable cell separation device according to claim 6, characterized in that: The sixth docking part (41) is docked with a separation liquid container (4).

9. The disposable cell separation device according to claim 1, characterized in that: A demagnetizing component (102) is provided between the first docking part (101) and the transfer container (1).

10. The disposable cell separation device according to any one of claims 1 to 9, characterized in that: The ventilation connection (11) is connected to a positive and negative pressure device (7).

11. The disposable cell separation device according to claim 10, characterized in that: The positive and negative pressure device (7) includes a second gas filter (71) and a gas pipe (70), wherein the second gas filter (71) is connected to the transfer container (1) through the gas pipe (70).

12. The disposable cell separation device according to claim 11, characterized in that: An intermediate container (72) is provided between the trachea (70) and the transfer container (1).

13. The disposable cell separation device according to any one of claims 1 to 9, characterized in that: The rotating sleeve (122) is rotatably mounted outside the inner column (121).

14. A cell separation device, characterized in that: The device includes a mounting panel (9) and a disposable cell separation apparatus according to any one of claims 1 to 13. The mounting panel (9) is provided with a plurality of mounting parts (91), a plurality of clamp valves (92) and a plurality of fixed pipe fittings (93). The transfer container (1) is detachably mounted on the corresponding mounting part (91), the pipeline (10) is fixed on the fixed pipe fitting (93), and the clamp valve (92) is clamped on the corresponding pipe position.

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