Mononuclear cell separation bag

Abstract

The utility model discloses a mononuclear cell separation bag, the open end of a whole blood sample bag is provided with a liquid one-way flow conduit and is connected with a blood sample transfer conduit, a plasma transfer conduit and a blood cell transfer conduit, the whole blood sample bag is connected with a plasma bag through the plasma transfer conduit, the open end of the plasma bag is provided with the liquid one-way flow conduit, and the blood cell transfer conduit is connected with the blood cell transfer conduit. The blood sample transfer catheter is connected with the plasma transfer catheter and the blood cell transfer catheter, the other end of the blood cell transfer catheter is connected with the blood cell transfer catheter, the other end of the blood cell transfer catheter is connected with the blood cell transfer bag, the open end of the blood cell transfer bag is provided with a sample extraction port, and the blood sample transfer catheter, the plasma transfer catheter and the blood cell transfer catheter are provided with disposable pipeline plugging devices and pipeline temporary sealing devices. The disposable closed container is used, so that the centrifugal tube is prevented from being frequently replaced, and meanwhile, the experiment failure caused by sample pollution due to the fact that hands of operators or articles pass through the upper part of the open centrifugal tube is also avoided.

Description

Technical Field

[0001] This utility model relates to the field of human peripheral blood mononuclear cell extraction, specifically a mononuclear cell separation bag. Background Technology

[0002] Peripheral blood mononuclear cells (PBMCs) are cells with a single nucleus found in peripheral blood, primarily including lymphocytes and monocytes. PBMCs are an important component of the immune system and directly participate in tumor immunity. The volume, morphology, and specific gravity of PBMCs differ from other cells in peripheral blood. The main method for isolating PBMCs is density gradient centrifugation.

[0003] Currently, centrifuge tubes are commonly used in laboratories as separation containers for peripheral blood mononuclear cell (PBMC) extraction. The following risks (or deficiencies) exist during the operation: (1) During PBMC extraction, the centrifuge tube opening is open. Due to negligence or a lack of aseptic technique, personnel may allow hands or other objects to pass over the open centrifuge tube, increasing the risk of contamination and potentially leading to experimental failure. (2) When adding peripheral blood samples to centrifuge tubes containing Ficoll solution, the blood must be added slowly to avoid breaking the boundary between Ficoll and peripheral blood. In this process, the sedimentation of the samples often varies depending on the order in which they are added. (3) When slowly adding peripheral blood samples to centrifuge tubes containing Ficoll, the operator must carefully control the blood flow rate. Poor flow rate control can break the Ficoll-blood separation, leading to laboratory failure. Mastering the skill of adding peripheral blood to centrifuge tubes containing Ficoll requires extensive experimental practice. (4) The peripheral blood plasma obtained by centrifugation needs to be inactivated by centrifugation in a 56°C water bath to obtain autologous peripheral blood serum. This operation process carries the risk of autologous serum contamination due to improper sealing of the centrifuge tube caused by negligence.

[0004] This invention builds upon previous density gradient centrifugation experiments on peripheral blood mononuclear cells using centrifuge tubes. By employing disposable, sealed containers, it avoids frequent tube replacements and prevents sample contamination caused by operator hands or objects passing over open centrifuge tubes, thus preventing experimental failure. The invention features a rationally designed mononuclear cell separation bag, controlling the liquid flow rate by regulating the tube diameter to prevent the Ficoll and blood sample from separating due to excessively rapid sample addition, thereby reducing the difficulty of experimental operation. During the experiment, the obtained autologous peripheral blood plasma is transferred through tubing to a separate bag, which is double-sealed using heat sealing and a specially designed sealing system, thus avoiding the risk of autologous serum contamination due to operational negligence. Utility Model Content

[0005] The technical problems to be solved by this utility model are: (1) During the mononuclear cell extraction experiment, the centrifuge tube opening is open. Due to negligence or a lack of awareness of aseptic operation, the experimenter may cause hands or other items to pass over the open centrifuge tube during the experiment, increasing the risk of contamination and leading to experimental failure; (2) When adding peripheral blood samples to centrifuge tubes containing Ficoll solution, the peripheral blood needs to be added slowly to avoid breaking the boundary between Ficoll and peripheral blood. In this operation, the sedimentation of the samples often varies depending on the order in which the peripheral blood samples are added; (3) When adding peripheral blood samples slowly to centrifuge tubes containing Ficoll, the operator must carefully control the blood flow rate. There is a risk that poor flow rate control may break the stratification between Ficoll and blood, causing laboratory failure. The experimental skill of adding peripheral blood to centrifuge tubes containing Ficoll requires a lot of experimental operation to master. (4) The peripheral blood plasma obtained by centrifugation needs to be inactivated by centrifugation in a 56°C water bath to obtain autologous peripheral blood serum. This operation process carries the risk of autologous serum contamination due to improper sealing of the centrifuge tube caused by negligence.

[0006] To solve the above-mentioned technical problems, the present invention adopts the following technical means:

[0007] A mononuclear cell separation bag includes a whole blood sample bag, a plasma bag, and a mononuclear cell bag. The whole blood sample bag has a liquid one-way flow conduit at its open end. There are three liquid one-way flow conduits, each connected to a blood sample transfer conduit, a plasma transfer conduit, and a blood cell transfer conduit, respectively. The whole blood sample bag and the plasma bag are connected via a plasma transfer conduit. The plasma bag has a liquid one-way flow conduit at its open end. There are two liquid one-way flow conduits, each connected to a plasma transfer conduit and an inactivated serum transfer conduit, respectively. One end of the inactivated serum transfer conduit is connected to the plasma bag, and the other end is connected to an inactivated serum container bag. The inactivated serum container bag has a sample extraction port at its open end. The whole blood sample bag and the mononuclear cell bag are connected via a blood cell transfer conduit. The blood sample transfer conduit, plasma transfer conduit, and blood cell transfer conduit are equipped with a disposable pipe sealing device and a temporary pipe closure device.

[0008] To address the shortcomings of the aforementioned peripheral blood mononuclear cell extraction experiments, the purpose of this invention is to provide a sealed peripheral blood mononuclear cell density gradient centrifugation extraction multi-segment bag. Its function is to avoid the risk of sample contamination caused by objects passing over the open centrifuge tube. This invention features a specially designed peripheral blood density gradient centrifugation bag that controls the flow rate by adjusting the tube diameter, preventing the mononuclear cell extraction experiment from failing due to the disruption of the layer interface caused by excessively fast flow, thus reducing the experimental difficulty. Furthermore, in addition to using heat-sealing to seal the tubing, this invention employs a specially designed tubing sealing device to further ensure double protection of the tubing seal.

[0009] As a preferred embodiment, a further technical solution of this utility model is:

[0010] The blood sample transfer catheter is located in the middle of the opening of the whole blood sample bag, the plasma transfer catheter is located on the right side of the blood sample transfer catheter, and the blood cell transfer catheter is located on the left side of the blood sample transfer catheter. One end of the blood sample transfer catheter is provided with a liquid filling port, and the liquid filling port is wrapped with a sterile plastic seal bag.

[0011] The opening end of the mononuclear cell bag is provided with a one-way liquid flow conduit, which is connected to a blood cell transfer conduit, a liquid infusion conduit, a peripheral blood mononuclear cell bag, and a liquid bag. There are three one-way liquid flow conduits: the middle one-way liquid flow conduit is connected to the liquid infusion conduit, the right one-way liquid flow conduit is connected to the blood cell transfer conduit, and the left one-way liquid flow conduit is connected to the peripheral blood mononuclear cell bag and the liquid bag. The opening end of the mononuclear cell bag is provided with suspension holes on both the left and right sides.

[0012] The disposable pipe sealing device includes a liquid sample pipe, a liquid pipe reinforcement, a pipe sealing slider, a pipe sealing slider base, a sealing slider limiting slot, and a sealing slider limiting device. The disposable pipe sealing device is wrapped in a transparent sterile packaging sleeve.

[0013] The sample extraction port is wrapped in a sterile plastic seal bag. The sample extraction port consists of a bag body, an inner base on both sides of the bag body, an outer base, a conduit connecting the bag body, a sealing plug that isolates the conduit from the outside world, and a dust cap for the sample extraction port. The dust cap is equipped with a dust cap handle.

[0014] The inner sidewall of the liquid unidirectional flow conduit is provided with biomimetic valves. There are multiple biomimetic valves, which are evenly arranged inside the sidewall of the conduit. The biomimetic valves stand up or tilt depending on the direction of liquid flow.

[0015] The temporary pipe closure device includes sealing teeth, fixing teeth, limiting teeth, a base, and a pipe. The base can be deformed to allow the limiting teeth and fixing teeth to engage and disengage. The sealing teeth on both sides of the pipe achieve temporary closure by squeezing the pipe walls on both sides.

[0016] This invention relates to a mononuclear cell separation bag. The bag employs a multi-unit design, comprising a whole blood separation module, a plasma inactivation module, and a mononuclear cell extraction module. After the blood sample for mononuclear cell extraction is transferred into the multi-unit bag system, the extraction and separation of mononuclear cells are performed within the system. This eliminates the need for frequent centrifuge tube changes, reducing the risk of contamination due to inadequate aseptic technique. Furthermore, it avoids inconsistent cell separation and sedimentation in different centrifuge tubes caused by inconsistent experimental procedures during the addition of Ficoll separation solution to dilute the blood. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of a specific embodiment of the present invention.

[0018] Figure 2 This is a schematic diagram of the structure of a whole blood sample bag according to a specific embodiment of the present invention.

[0019] Figure 3 This is a schematic diagram of the structure of a plasma bag according to a specific embodiment of the present invention.

[0020] Figure 4 This is a schematic diagram of a single nucleated cell bag structure according to a specific embodiment of the present invention.

[0021] Figure 5 This is a schematic diagram of a disposable pipe sealing device according to a specific embodiment of the present invention.

[0022] Figure 6 This is a schematic diagram of a disposable pipe sealing device according to a specific embodiment of the present invention.

[0023] Figure 7 This is a schematic diagram of the sample extraction port structure of a specific embodiment of the present invention.

[0024] Figure 8 This is a side view of a dust cap according to a specific embodiment of the present invention.

[0025] Figure 9 This is a schematic diagram of the structure of a dust cap according to a specific embodiment of the present invention.

[0026] Figure 10 This is a diagram showing the working state of a liquid unidirectional flow conduit according to a specific embodiment of the present invention.

[0027] Figure 11 This is a diagram showing another working state of the liquid unidirectional flow conduit according to a specific embodiment of the present invention.

[0028] Figure 12 This is a schematic diagram of a temporary pipe closure device according to a specific embodiment of the present invention.

[0029] Figure 13 This is a cross-sectional view of a temporary pipe closure device according to a specific embodiment of the present invention.

[0030] Explanation of reference numerals in the attached diagram: 1. Whole blood sample bag; 2. Plasma bag; 3. Mononuclear cell bag; 4. Disposable tubing sealing device; 5. Sample extraction port; 6. One-way flow tubing; 7. Liquid filling port; 8. Sterile plastic seal bag; 9. Temporary tubing closure device;

[0031] 11. Blood sample transfer catheter; 12. Plasma transfer catheter; 13. Blood cell transfer catheter;

[0032] 21. Inactivated serum container bag; 22. Inactivated serum transfer catheter;

[0033] 31. Peripheral blood mononuclear cell bag; 32. Liquid bag; 33. Liquid infusion tubing; 34. Suspension port for mononuclear cell bag;

[0034] 41: Liquid sample pipeline; 42: Liquid pipeline reinforcement; 43: Pipeline plugging slider; 44: Pipeline plugging slider base; 45: Plug-in slider limiting slot; 46: Plug-in slider limiting device; 47: Transparent sterile packaging sleeve;

[0035] 52. Bag body; 53. Inner base; 54. Outer base; 55. Tubing; 56. Rubber stopper; 57. Dust cap; 58. Dust cap handle;

[0036] 91. Sealing tooth; 92. Fixing tooth; 93. Limiting tooth; 94. Base; 95. Pipeline. Detailed Implementation

[0037] The present invention will be further described below with reference to the embodiments. Specific Implementation

[0038] See Figure 5 , Figure 6The working instructions for the disposable pipe sealing device 4 are as follows: By pressing the pipe sealing slider 43 on the disposable pipe sealing device 4, the sealing slider limiting device 46 on the pipe sealing slider 43 pops up and locks into the sealing slider limiting slot 45 on the pipe sealing slider 44. At this time, the sealing slider 43 completely seals the pipeline through compression on both sides, and the liquid on both sides of the liquid sample pipeline 41 no longer flows. In the pipe sealing device 4, because the liquid sample pipeline 41 is a flexible tube, in order to enhance the pipe sealing effect, the liquid sample pipeline is appropriately thickened inside the pipe sealing device 4 for liquid pipeline reinforcement 42, to ensure the pipe sealing effect and enhance the strength and toughness of the pipeline to avoid breakage. At the same time, the entire pipe sealing device 4 is wrapped by a transparent sterile packaging sleeve 47 to ensure the integrity of the device and isolate it from the outside world.

[0039] See Figure 7 , Figure 8 , Figure 9 The working instructions for sample extraction port 5 are as follows: Sample extraction port 5 consists of a sterile plastic-sealed bag 8 wrapped around the sample extraction port, a bag body 51, an inner base 52 and an outer base 53 located on both sides of the bag body, a conduit 54 connecting the bag body, a sealing plug 55 isolating the conduit from the outside environment, a dust cap 56 for the sample extraction port, and a dust cap handle 57 connected to the dust cap. When it is necessary to use a syringe to extract the sample from the bag inside the sample extraction port 5, first remove the sterile plastic-sealed bag 8 inside the biosafety cabinet. Then, hold the sample extraction port 5 on the sample bag body with one hand, and pull out the dust cap handle 57 with the other hand to pull the dust cap 56 off the extraction port 5 and place it on the biosafety cabinet surface to expose the sample extraction port 5. Sterilize the rubber stopper on the sample extraction port 5 with the outer flame of an alcohol lamp. Then, holding the syringe, insert the syringe needle through the rubber stopper 55 into the sample bag to extract the sample to be extracted. After the sampling operation is completed, the outer surface of the rubber stopper is disinfected again with the outer flame of the alcohol lamp. Then, the dust cap 56 is put back on the sample sampling port 5 by using the handle 57 to hold the dust cap.

[0040] See Figure 10 , Figure 11 The working principle of the unidirectional liquid flow conduit 6 is as follows: When a liquid sample flows through the liquid flow control valve 6, if the liquid flow direction is consistent with the set direction of the control valve, the bionic valve 62 attached to the side wall 61 of the conduit is tilted, allowing the liquid to pass smoothly through the flow control valve. If the liquid flow direction is opposite to the set direction of the control valve, multiple, multi-layered bionic valves 62 stand up, mimicking the working state of venous valves and atrioventricular valves, to prevent the liquid from flowing through.

[0041] See Figure 12 , Figure 13The working principle of the temporary pipe closure device 9 is as follows: The temporary pipe closure device 9 is a conventional plastic part. The base 94 has a certain deformation capacity, and the insertion and release of the limiting tooth 93 and the fixing tooth 92 are achieved by the deformation of the base 94. The temporary pipe closure device 9 is designed with multiple different tooth positions to accommodate conduits of different diameters. When it is necessary to close the pipe, the pipe closure limiting tooth 93 is pressed into the pipe closure limiting fixing tooth 92 and fixed in the required tooth position. At this time, the sealing teeth 91 on both sides of the pipe achieve temporary closure of the pipe by squeezing the pipe walls on both sides of the pipe (lumen) 95. When it is necessary to reopen the passage, the limiting tooth 93 is disengaged from the tooth position by withdrawing the fixing tooth 92 outward. The sealing teeth 91 on both sides of the pipe wall no longer squeeze the pipe (lumen) 95, thus realizing the re-opening of the pipe.

[0042] See Figure 1 , Figure 2 , Figure 3 , Figure 4 The process involves uniformly spraying disinfectant onto the outer packaging of the human peripheral blood mononuclear cell density gradient centrifugation extraction multi-unit bag, removing the outer packaging near the biosafety cabinet, and placing the bag inside. The collection container containing the peripheral blood sample is then placed inside the biosafety cabinet after surface disinfection. The sterile plastic seal 8 connecting the liquid filling port to the whole blood sample bag 1 is removed, exposing the liquid filling port 7. The dust cap on the liquid filling port 7 is unscrewed, and the port is connected to a syringe. The peripheral blood sample is transferred into the whole blood sample bag using the syringe. After the blood sample is transferred into the whole blood sample bag 1, the liquid filling port 7 is separated from the syringe, disinfected with the outer flame of an alcohol lamp, and the dust cap is screwed back on. The temporary sealing device 9 connecting the liquid filling port to the whole blood sample bag 1 is closed, temporarily sealing the liquid filling pipeline. The human peripheral blood mononuclear cell density gradient centrifugation extraction multi-unit bag is placed in a centrifuge to separate blood cells and plasma.

[0043] After centrifugation, remove the multi-bag system, check the tubing, and confirm that the temporary sealing device 9 connecting the mononuclear cell bag 3 to the whole blood sample bag 1 is closed, while the temporary sealing device 9 connecting the plasma bag 2 to the whole blood sample bag 1 is open. Use a plasma clamp to press the bag tightly at the boundary between the plasma and blood cells in the whole blood sample bag 1. At this time, the blood cell layer and the plasma layer will separate due to the pressure of the bag. Transfer the plasma from the upper layer of the whole blood sample bag 1 to the plasma transfer catheter 12 connected to the whole blood sample bag 1 through the plasma transfer catheter 12. Then, use the disposable tubing sealing device 4 on the plasma transfer catheter 12 to seal the tubing. At this time, the whole blood sample bag 1 and the plasma bag 2 are no longer connected. Remove the whole blood sample bag 1 from the plasma clamp, and use a heat sealer to heat seal multiple sections of the plasma transfer catheter 12 between the two disposable tubing sealing devices 4. Separate the plasma bag from the multi-bag system.

[0044] The temporary sealing device 9 on the inactivated serum transfer catheter 22 is used to place the plasma bag 2 containing plasma in a 56°C water bath for plasma inactivation. The autologous inactivated serum container is not placed in the water bath. After inactivation, the plasma is centrifuged to precipitate the denatured substances at the bottom of the bag. The serum is then separated from the precipitate using a separator. The temporary sealing device 9 is then opened, and the inactivated serum transfer catheter 22 is reopened to transfer the inactivated serum into the inactivated serum bag 21. Next, the disposable catheter sealing devices 4 at both ends of the inactivated serum transfer catheter 22 are used to seal the catheter. Multiple sections of the inactivated serum transfer catheter 22 are then heat-sealed between the disposable catheter sealing devices 4. The inactivated serum bag is then detached from the plasma bag system. The inactivated serum bag can be labeled and placed directly in a refrigerator. When needed, it can be drawn from the sample extraction port 5 using a syringe, or it can be drawn once using a syringe and transferred to a new centrifuge tube for later use.

[0045] Place the multi-bag system (without plasma bag 2) back into the biosafety cabinet. Unscrew the dust cap on the liquid inlet 7 and connect it to the syringe. Reopen the temporary closure device 9 connecting the tubing to the liquid inlet on the whole blood sample bag 1. Add physiological saline through the liquid inlet 7, then close the disposable tubing closure device 4 on the tubing connected to the liquid inlet 7. Place the whole blood sample bag 1 on a shaker to resuspend the blood cells. Dilute and resuspend the concentrated blood cells. After adding physiological saline, close the disposable tubing closure device 4 on the tubing connected to the liquid inlet 7.

[0046] The resuspended blood cells are transferred to the mononuclear cell bag 3 via the blood cell transfer catheter 13. Then, the disposable tubing sealing devices 4 at both ends of the cell transfer catheter 13 are closed, and the tubing sections are heat-sealed. The whole blood sample bag 1 is then removed. To prevent rapid injection of Ficoll into the bag from disrupting the separation between Ficoll and blood cells, the diameter of the liquid infusion catheter 33 is controlled between 0.2 mm and 1 mm, preferably 0.5 mm. The mononuclear cell bag 3 has a conical bottom design. The mononuclear cell bag 3 is suspended upright on the biosafety counter through the suspension hole 34. After a period of settling, the syringe is connected to the liquid infusion port on the mononuclear cell bag 3, and Ficoll flows in through the syringe via the unidirectional liquid flow catheter 6. Due to the limitation of the liquid infusion catheter 33's diameter, the Ficoll is slowly injected into the bottom of the cell bag 3. During this process, the boundary between Ficoll and blood cells is clear, and the separation is not easily disrupted.

[0047] Upon completion of Ficoll injection, close the disposable tubing closure device 4 on the fluid infusion tubing 33, remove the mononuclear cell bag 3 from its hanging position, and slowly, gently, and vertically transfer it into a centrifuge for density gradient centrifugation. After density gradient centrifugation, carefully transfer the mononuclear cell bag 3 to the plasma separator, separating the liquid layer of diluted plasma from the underlying mononuclear cell layer using the separator. Ensure that the temporary tubing closure device 9 connecting the mononuclear cell bag 31 is closed and the temporary tubing closure device 9 on the liquid bag 32 is open, transfer the liquid from the upper layer of the density gradient centrifugation into the liquid bag 32, and close the temporary tubing closure device 9. Move the mononuclear cell bag 3 to separate the mononuclear cell layer from the Ficoll layer and blood cell layer below the bag. Gently knead the bag to resuspend the mononuclear cells attached to the inner surface of the bag in the liquid environment. Open the temporary tubing closure device 9 connecting the mononuclear cell bag 31 and transfer the suspension rich in mononuclear cells into the mononuclear cell bag 31. Close the temporary sealing device 9 of the pipe connected to the mononuclear cell container 31 and heat seal the pipe to separate the mononuclear cell container 31 from the mononuclear cell bag.

[0048] The cell suspension in the mononuclear cell container 31 is extracted through the sample extraction port using a syringe and then subjected to subsequent experiments such as washing, centrifugation, counting, cell cryopreservation, or cell culture.

[0049] This technology features a simple structure, providing an independent, high-density cell separation and extraction system. This reduces the need for constant centrifuge tube replacement during single-nuclear cell separation. It also avoids the waste of valuable biological samples due to microbial contamination caused by improper aseptic technique, such as operating above open centrifuge tubes. Furthermore, this invention eliminates the differences in cell sedimentation rates caused by sequential operation when using multiple centrifuge tubes for density gradient centrifugation. It also reduces the experimental difficulty of breaking Ficoll stratification during the addition of blood samples in Ficoll density gradient centrifugation.

[0050] By using this single-cell separation and extraction multi-sack, the experimental difficulty is reduced, the training intensity of experimental operation is reduced, and the experimental efficiency is improved. In the process of using this utility model, the risk of sample contamination caused by experimental operation is also reduced, and the number of repeated experiments caused by sample contamination is reduced. In addition, this utility model also reduces the use of centrifuge tubes and pipettes, saving time and money.

[0051] Since the above description is only a specific embodiment of the present utility model, the protection of the present utility model is not limited thereto. Any equivalent changes or substitutions of the technical features of the present technical solution that can be conceived by those skilled in the art are covered within the protection scope of the present utility model.

Claims

1. A mononuclear cell separation bag, comprising a whole blood sample bag, a plasma bag, and a mononuclear cell bag; characterized in that: The whole blood sample bag has a liquid one-way flow conduit at its opening end. There are three liquid one-way flow conduits, which are respectively connected to a blood sample transfer conduit, a plasma transfer conduit, and a blood cell transfer conduit. The whole blood sample bag and the plasma bag are connected via a plasma transfer conduit. The plasma bag has a liquid one-way flow conduit at its opening end. There are two liquid one-way flow conduits, which are respectively connected to a plasma transfer conduit and an inactivated serum transfer conduit. One end of the inactivated serum transfer conduit is connected to the plasma bag, and the other end is connected to an inactivated serum container bag. The inactivated serum container bag has a sample extraction port at its opening end. The whole blood sample bag and the mononuclear cell bag are connected via a blood cell transfer conduit. The blood sample transfer conduit, plasma transfer conduit, and blood cell transfer conduit are equipped with disposable pipe sealing devices and temporary pipe sealing devices.

2. The mononuclear cell separation bag according to claim 1, characterized in that: The blood sample transfer catheter is located in the middle of the opening of the whole blood sample bag, the plasma transfer catheter is located on the right side of the blood sample transfer catheter, and the blood cell transfer catheter is located on the left side of the blood sample transfer catheter. One end of the blood sample transfer catheter is provided with a liquid filling port, and the liquid filling port is wrapped with a sterile plastic seal bag.

3. The mononuclear cell separation bag according to claim 1, characterized in that: The opening end of the mononuclear cell bag is provided with a one-way liquid flow conduit, which is connected to a blood cell transfer conduit, a liquid infusion conduit, a peripheral blood mononuclear cell bag, and a liquid bag. There are three one-way liquid flow conduits: the middle one-way liquid flow conduit is connected to the liquid infusion conduit, the right one-way liquid flow conduit is connected to the blood cell transfer conduit, and the left one-way liquid flow conduit is connected to the peripheral blood mononuclear cell bag and the liquid bag. The opening end of the mononuclear cell bag is provided with suspension holes on both the left and right sides.

4. The mononuclear cell separation bag according to claim 1, characterized in that: The disposable pipe sealing device includes a liquid sample pipe, a liquid pipe reinforcement, a pipe sealing slider, a pipe sealing slider base, a sealing slider limiting slot, and a sealing slider limiting device. The disposable pipe sealing device is wrapped in a transparent sterile packaging sleeve.

5. A mononuclear cell separation bag according to claim 1, characterized in that: The sample extraction port is wrapped in a sterile plastic seal bag. The sample extraction port consists of a bag body, an inner base on both sides of the bag body, an outer base, a conduit connecting the bag body, a sealing plug that isolates the conduit from the outside world, and a dust cap for the sample extraction port. The dust cap is equipped with a dust cap handle.

6. A mononuclear cell separation bag according to claim 1, characterized in that: The inner sidewall of the liquid unidirectional flow conduit is provided with biomimetic valves. There are multiple biomimetic valves, which are evenly arranged inside the sidewall of the conduit. The biomimetic valves stand up or tilt depending on the direction of liquid flow.

7. A mononuclear cell separation bag according to claim 1, characterized in that: The temporary pipe closure device includes sealing teeth, fixing teeth, limiting teeth, a base, and a pipe. The base can be deformed to allow the limiting teeth and fixing teeth to engage and disengage. The sealing teeth on both sides of the pipe achieve temporary closure by squeezing the pipe walls on both sides.

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