An integrated automatic exhaust sample freezing device

Abstract

The present invention discloses an integrated automatic exhaust sample freezing device, which belongs to the technical field of umbilical cord blood and cell freezing and recovery. It aims to solve the problems that existing freezing technology cannot realize multiple freezing and recovery of the same umbilical cord blood or cells, and there is a risk of contamination caused by manual operation and transfer during operation, as well as manual repetitive exhaust and exhaust difficulty. The device comprises a mixing bag for preventing local DMSO concentration from being too high to kill cells, an input component for inputting cells and freezing solution into an input pipeline, and an output component for outputting a mixture of cells and freezing solution. The input component and the output component are both connected to the mixing bag. The device can realize multiple uses of the same umbilical cord blood or cells, is convenient for sample quality inspection, improves the efficiency of cell use, greatly reduces the risk of contamination, facilitates the injection of cell mixture, solves the problems of manual repetitive exhaust and exhaust difficulty, and is very convenient for exhausting the gas in the cell freezing tube and introducing the cell mixture in the mixing bag.

Description

Technical Field

[0001] The present invention relates to the technical field of umbilical cord blood and cell freezing and resuscitation, and in particular to an integrated automatic exhaust type sample freezing device. Background Art

[0002] With the advancement of biological technology, cell therapy is rapidly developing and gaining increasing acceptance worldwide. Stem cell research and clinical application are the core areas of cell therapy. In recent years, stem cell transplantation has been gradually applied to treat malignant hematological diseases, aplastic anemia, advanced solid tumors, heart disease, nervous system damage, tissue and organ repair, diabetes, and vascular disease. Stem cells also have powerful health and disease prevention functions, enhancing immunity, repairing diseased or aging tissues, and prolonging life. Stem cells come from a wide range of sources, primarily umbilical cord blood, bone marrow, and peripheral blood. In 2001, the Ministry of Health approved the establishment of an umbilical cord blood hematopoietic stem cell bank. Cryopreservation, thawing, and preventing contamination during manipulation are key to successful cell therapy. In vitro cell culture has also been widely used in various biological research fields. In in vitro cell culture, how to avoid repeated freezing and thawing and ensure multiple uses of the same cord blood or cells has become a key research topic.

[0003] Currently, most national institutions for cell therapy use single-chamber cryopreservation bags for cryopreservation. If cryopreservation is done in cryopreservation bags, a portion of umbilical cord blood or cells can only be used once. Repeated freezing and thawing will reduce cell activity and increase the risk of contamination, which in turn greatly reduces the benefits that umbilical cord blood or cells can bring. If split cell input and output are used, manual operation is required, which directly increases the probability of contamination during manual connection and transfer, and also increases the time it takes for cells to enter the cryogenic equipment for cryopreservation, ultimately affecting cell activity. During the entire transfer process, the gas in the cryopreservation bag cannot be discharged, and the cryopreservation bag needs to be manually squeezed repeatedly to exhaust. The exhaust process and time are relatively long, which seriously affects cell activity. Summary of the Invention

[0004] The present invention aims to solve the problems that existing cryopreservation technology cannot realize multiple cryopreservation and resuscitation of the same umbilical cord blood or cells, and there is a risk of contamination due to manual operation and transfer during the operation, as well as manual repetitive exhaust and difficulty in exhausting.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions: 1. An integrated automatic exhaust sample freezing device includes a mixing bag for pre-treatment of cell freezing to prevent direct mixing in a single-chamber freezing tube, which may cause excessive local DMSO concentration to kill cells, an input component for inputting cells and freezing solution, and an output component for outputting a mixture of cells and freezing solution. The input component and the output component are both connected to the mixing bag.

[0006] Preferably, the input component includes a first input pipe, a second input pipe, and a freezing liquid injection head. One end of the first input pipe is connected to the mixing bag, and the other end of the first input pipe is connected to the freezing liquid injection head and the second input pipe through a Y-shaped three-way interface. A first switch card is provided on the first input pipe, and the end of the second input pipe not connected to the Y-shaped three-way interface is bifurcated into multiple branch pipes through a first multi-way brancher. Each branch pipe has a Luer interface at one end of the pipe mouth, and the Luer interface can be of various types. Each branch pipe is provided with a flow regulator.

[0007] Preferably, the output component includes a first output pipeline, a cell freezing tube, and an exhaust filter. One end of the first output pipeline is connected to the mixing bag, and the other end is bifurcated into multiple branch pipelines through a second multi-way brancher. A second switch card is provided on the first output pipeline. Each branch pipeline is connected to a cell freezing tube. One end of each cell freezing tube is provided with a puncture port and the other end is provided with an exhaust pipeline. The ends of the exhaust pipelines not connected to the cell freezing tubes are respectively connected to one of the exhaust filters.

[0008] Preferably, the connecting end of each branch pipe and the cell freezing tube is at the same end as the exhaust pipe.

[0009] Preferably, the connecting end of each branch pipe and the cell freezing tube is at the same end as the puncture port.

[0010] Preferably, the output component includes a first output pipe, a cell cryopreservation tube formed by connecting multiple cell cryopreservation tubes through heat sealing, and an exhaust filter. One end of the first output pipe is connected to the mixing bag, and the other end is connected to the cell cryopreservation tube. A second switch card is provided on the first output pipe, and each cell cryopreservation tube that constitutes the cell cryopreservation tube is provided with a puncture port. The exhaust filter is connected to the end of the cell cryopreservation tube that is not connected to the first output pipe through the exhaust pipe, and a third switch card is provided on the exhaust pipe.

[0011] Preferably, a fourth switch card is provided on the first output pipe, the exhaust pipe is connected to the first output pipe through a Y-shaped three-way interface, the other end of the exhaust pipe is connected to the exhaust filter, the exhaust pipe is provided with a third switch card, the first output pipe is provided with a second switch card on the pipe connected to the mixing bag and the Y-shaped three-way interface, and the fourth switch card is provided on the pipe connected to the first output pipe and the cell freezing tube and the Y-shaped three-way interface.

[0012] Preferably, the exhaust filter is an automatic exhaust filter or a manual syringe.

[0013] Preferably, an automatic exhaust filter cap is provided on the automatic exhaust filter.

[0014] Preferably, a connection to an exhaust filter can be omitted.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] 1. By setting up multiple separate cell cryopreservation tubes or heat-sealing the cell cryopreservation tubes to connect with the mixing bag, and each cell cryopreservation tube has a separate puncture port, the same cord blood or cells can be used multiple times, and the sample quality inspection is convenient, thereby improving the efficiency of cell use.

[0017] 2. The mixing bag is directly connected to the cell cryopreservation tube, which avoids the manual transfer process between the mixing bag and the cell cryopreservation tube, greatly reduces the risk of contamination and shortens the transfer time.

[0018] 3. If the freezing solution and cells are directly mixed in the cell cryopreservation tube, the local DMSO concentration will be too high, which will kill the cells. The mixing bag is used as a pretreatment for cell freezing to prevent direct mixing in the single-chamber cryopreservation tube, which will cause the local DMSO concentration to be too high and kill the cells.

[0019] 4. The automatic exhaust filter can exhaust the gas in the cell cryopreservation tube to maintain the normal atmospheric pressure inside, which is convenient for injecting the cell mixture and solves the problems of manual repetitive exhaust and exhaust difficulty. It can easily exhaust the gas in the cell cryopreservation tube and smoothly introduce the cell mixture in the mixing bag. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a structural diagram of an integrated automatic exhaust sample freezing device according to a first embodiment of the present invention;

[0021] Figure 2 This is a structural diagram of an integrated automatic exhaust sample freezing device according to a second embodiment of the present invention;

[0022] Figure 3 This is a structural diagram of an integrated automatic exhaust sample freezing device according to a third embodiment of the present invention;

[0023] Figure 4 Schematic diagram of the structure of an integrated automatic exhaust sample freezing device according to a fourth embodiment of the present invention.

[0024] Description of reference numerals:

[0025] In the figure: mixing bag 1; input component 2; first input pipe 21; Y-type three-way interface 22; second input pipe 23; first multi-way branch 24; first flow regulator 25; first Luer interface 26; freezing liquid injection head 27; second flow regulator 28; first switch card 29; output component 3; first output pipe 31; second switch card 32; second multi-way branch 33; puncture port 34; cell freezing tube 35; third switch card 36; automatic exhaust filter 37; automatic exhaust filter cap 38; exhaust pipe 39; fourth switch card 310; manual syringe 311. DETAILED DESCRIPTION

[0026] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0027] It should be noted that, in the description of the present invention, terms such as "center", "upper", "lower", "horizontal", and "inner" indicating directions or positional relationships are based on the directions or positional relationships shown in the accompanying drawings. This is only for the convenience of description and does not indicate or imply that the device or element must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it cannot be understood as a limitation on the present invention.

[0028] Furthermore, it should be noted that, in the description of the present invention, unless otherwise expressly specified or limited, the terms "disposed," "installed," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integral connections; they may refer to mechanical connections; they may refer to direct connections, indirect connections through an intermediate medium, or internal connections between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0029] Example:

[0030] like Figure 1 As shown, the first embodiment of the present invention is an integrated automatic exhaust sample freezing device, comprising a mixing bag 1 for pre-treatment of cell freezing to prevent direct mixing in a single-chamber freezing tube, which may cause excessive local DMSO concentration to kill cells, an input component 2 for inputting cells and freezing solution into an input pipeline, and an output component 3 for outputting a mixture of cells and freezing solution. Both the input component 2 and the output component 3 are connected to the mixing bag 1.

[0031] The input component 2 includes a first input pipe 21, a second input pipe 23, and a freezing liquid injection head 27. One end of the first input pipe 21 is connected to the mixing bag 1, and the other end of the first input pipe 21 is connected to the freezing liquid injection head 27 and the second input pipe 23 through a "Y"-shaped three-way interface. The first input pipe 21 is provided with a first switch card 29 for controlling the safety of the pipeline. The end of the second input pipe 23 not connected to the three-way interface is forked into a first branch pipe and a second branch pipe through a first multi-way brancher 24. The first branch pipe and the second branch pipe are both provided with a Luer interface at one end of the pipe mouth. The Luer interface can be of various types. The first branch pipe is provided with a first flow regulator 25, and the second branch pipe is provided with a second flow regulator 28.

[0032] The output assembly 3 includes a first output pipeline 31, a cell freezing tube 35, and an automatic exhaust filter 37. One end of the first output pipeline 31 is connected to the mixing bag 1, and the other end is bifurcated into multiple branch pipelines through a second multi-way brancher 33. A second switch card 32 is provided on the first output pipeline 31. Each branch pipeline is connected to a cell freezing tube 35 through a heat seal. One end of each cell freezing tube 35 is provided with a puncture port 34 and the other end is provided with an exhaust pipe 39. The exhaust pipe 39 is not connected to one end of the cell freezing tube 35. They are respectively connected to an automatic exhaust filter 37, and each automatic exhaust filter 37 is provided with an automatic exhaust filter cap 38. The automatic exhaust filter 37 is used to automatically exhaust the gas in the cell freezing tube 35, so that the internal pressure remains at normal atmospheric pressure, which is convenient for injecting cell mixture and reducing manual repetitive exhaust. The gas in the cell freezing tube 35 can be easily exhausted and the cell mixture in the mixing bag 1 can be smoothly introduced. The connecting end of each branch pipe and the cell freezing tube 35 is at the same end as the exhaust pipe 39.

[0033] Figure 2 The difference between the integrated automatic exhaust sample freezing device of the second embodiment shown and the integrated automatic exhaust sample freezing device of the first embodiment lies in that the connection position of each branch pipe and the cell freezing tube 35 is different. The connecting end of each branch pipe of the second embodiment and the cell freezing tube 35 is at the same end as the puncture port 34.

[0034] Figure 3The integrated automatic exhaust sample freezing device of the third embodiment shown is different from the integrated automatic exhaust sample freezing device of the first embodiment in that the output components are different. The output component 3 of the third embodiment includes a first output pipe 31, a cell freezing tube 35 formed by heat-sealing multiple cell freezing tubes, and an automatic exhaust filter 37. One end of the first output pipe 31 is connected to the mixing bag 1, and the other end is connected to the cell freezing tube 35. A second switch card 32 is provided on the first output pipe 31, and each cell freezing tube that constitutes the cell freezing tube 35 is provided with a puncture port 34. The automatic exhaust filter 37 is connected to the end of the cell freezing tube 35 that is not connected to the first output pipe 31 through the exhaust pipe 39. An automatic exhaust filter cap 38 is provided at the left end of the automatic exhaust filter 37, and a third switch card 36 is provided on the exhaust pipe 39.

[0035] Figure 4 The difference between the integrated automatic exhaust sample freezing device of the fourth embodiment shown and the third embodiment is that a fourth switch card 310 is provided on the first output pipe 31, the exhaust pipe 39 is connected to the first output pipe 31 through a Y-shaped three-way interface, and the other end of the exhaust pipe 39 is connected to the automatic exhaust filter 37. A third switch card 36 is provided on the exhaust pipe 39, and a second switch card 32 is provided on the pipe connecting the first output pipe 31 with the mixing bag 1 and the Y-shaped three-way interface, and a fourth switch card 310 is provided on the pipe connecting the first output pipe 31 with the cell freezing tube 35 and the Y-shaped three-way interface.

[0036] How to use the first and second embodiments:

[0037] First, close the second switch card 32, open the first switch card, close the second flow regulator 28, and open the first flow regulator 25 (open the flow regulator of the pipeline with the Luer interface on it, and close the flow regulators of other pipelines). Transfer the pretreated umbilical cord blood hematopoietic stem cells into the mixing bag 1 through the Luer interface, place the mixing bag 1 on a small shaker at 4°C for precooling to 4°C, inject the freezing solution into the mixing bag 1 through the freezing solution injection head 27, close the first switch card on the first input pipeline 21, and evenly mix the liquid in the mixing bag 1.

[0038] Open the second switch card 32 and the third switch card 36, and open the automatic exhaust filter cap 38 on the automatic exhaust filter 37, so that the mixed liquid in the mixing bag 1 flows through the first output pipe 31, passes through the second multi-way branch 33, and then flows into each independent cell freezing tube 35 through multiple branch pipes. Through the automatic exhaust adjustment of the automatic exhaust filter 37, the cell freezing tube 35 is filled with liquid without air and bubbles, and after the exhaust pipe 39 of the cell freezing tube 35 is filled with liquid, the third checkpoint 36 is closed.

[0039] Use scissors to cut the branch pipe from the middle of the heat-sealed seal line between the exhaust pipe 39. Place the cell freezing tube 35 filled with liquid in an adapted freezing aluminum box, and place it in a low-temperature device for freezing after programmed cooling.

[0040] The usage of the third embodiment:

[0041] First, close the second switch card 32, open the first switch card, close the second flow regulator 28, and open the first flow regulator 25 (open the flow regulator of the pipeline with the Luer interface on it, and close the flow regulators of other pipelines). Transfer the pretreated umbilical cord blood hematopoietic stem cells into the mixing bag 1 through the Luer interface, place the mixing bag 1 on a small shaker at 4°C for precooling to 4°C, inject the freezing solution into the mixing bag 1 through the freezing solution injection head 27, close the first switch card on the first input pipeline 21, and evenly mix the liquid in the mixing bag 1.

[0042] Open the second switch card 32 and the third switch card 36, open the automatic exhaust filter cap 38 on the automatic exhaust filter 37, and allow the mixed liquid in the mixing bag 1 to flow into the cell freezing tube 35 through the first output pipe 31. Through the automatic exhaust adjustment of the automatic exhaust filter 37, the cell freezing tube 35 is filled with liquid without air and bubbles, and after the exhaust pipe 39 of the cell freezing tube 35 is filled with liquid, the third checkpoint 36 is closed.

[0043] The usage of the fourth embodiment:

[0044] First, close the second switch card 32, open the first switch card, close the second flow regulator 28, and open the first flow regulator 25 (open the flow regulator of the pipeline with the Luer interface on it, and close the flow regulators of other pipelines). Transfer the pretreated umbilical cord blood hematopoietic stem cells into the mixing bag 1 through the Luer interface, place the mixing bag 1 on a small shaker at 4°C for precooling to 4°C, inject the freezing solution into the mixing bag 1 through the freezing solution injection head 27, close the first switch card on the first input pipeline 21, and evenly mix the liquid in the mixing bag 1.

[0045] Close the second switch card 32, open the fourth switch card 310 and the third switch card 36, and adjust the cell freezing tube 35 through the manual syringe 311 to be in a vacuum state. Close the fourth switch card 310, open the second switch card 32, and allow the mixed liquid in the mixing bag 1 to flow into the cell freezing tube 35 through the first output pipe 31. Through the exhaust adjustment of the manual syringe 311, the cell freezing tube 35 is filled with liquid without air and bubbles, and after the exhaust pipe 39 of the cell freezing tube 35 is filled with liquid, close the third switch 36.

[0046] In summary, the present invention can realize multiple freezing and resuscitation of the same umbilical cord blood or cells, and is convenient for sample quality inspection, thereby improving the efficiency of cell use, avoiding the manual transfer process between the mixing bag and the cell cryopreservation tube, greatly reducing the risk of contamination, and shortening the transfer time. It can prevent direct mixing in the single-chamber cryopreservation tube, which may cause excessive local DMSO concentration to poison the cells, and can maintain normal atmospheric pressure inside the cell cryopreservation tube, making it convenient to inject the cell mixture and reduce manual repetitive exhaust. The gas in the cell cryopreservation tube can be easily discharged and the cell mixture can be smoothly introduced into the mixing bag. The above can effectively solve the problems that the existing freezing technology cannot realize multiple uses of the same umbilical cord blood or cells, and that manual connection and transfer operation pollution and manual repetitive exhaust are difficult during the operation.

[0047] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. An integrated automatic exhaust sample freezing device, characterized by: The invention comprises a mixing bag (1) for pre-treating cells for freezing to prevent direct mixing in a single-chamber cryopreservation tube from causing excessive local DMSO concentration to kill cells, an input component (2) for inputting cells and a freezing solution into an input pipe, and an output component (3) for outputting a mixture of cells and the freezing solution, wherein the input component (2) and the output component (3) are both connected to the mixing bag (1); The input assembly (2) comprises a first input pipe (21), a second input pipe (23), and a cryopreservation liquid injection head (27); one end of the first input pipe (21) is in communication with the mixing bag (1); the other end of the first input pipe (21) is connected to the cryopreservation liquid injection head (27) and the second input pipe (23) via a Y-shaped three-way interface (22); a first switch card (29) is provided on the first input pipe (21); the end of the second input pipe (23) not connected to the Y-shaped three-way interface (22) is bifurcated into a plurality of branch pipes via a first multi-way brancher (24); a Luer interface is provided at one end of the pipe opening of each branch pipe, and the Luer interface can be of various types; and a flow regulator is provided on each branch pipe; The output assembly (3) includes a first output pipeline (31), a cell freezing tube (35), and an exhaust filter. One end of the first output pipeline (31) is connected to the mixing bag (1), and the other end is bifurcated into a plurality of branch pipelines through a second multi-way brancher (33). A second switch card (32) is provided on the first output pipeline (31). Each branch pipeline is connected to a cell freezing tube (35). One end of each cell freezing tube (35) is provided with a puncture port (34) and the other end is provided with an exhaust pipeline (39). The ends of the exhaust pipelines (39) not connected to the cell freezing tube (35) are respectively connected to one of the exhaust filters. The exhaust filter is an automatic exhaust filter (37) or a manual syringe (311); an automatic exhaust filter cap (38) is provided on the automatic exhaust filter (37); and the connection with the exhaust filter can be cancelled.

2. The integrated automatic exhaust sample freezing device according to claim 1, characterized in that: The communicating end of each branch pipe and the cell freezing tube (35) is at the same end as the exhaust pipe (39).

3. The integrated automatic exhaust sample freezing device according to claim 1, characterized in that: The communicating end of each branch pipe and the cell freezing tube (35) is located at the same end as the puncture port (34).

4. The integrated automatic exhaust sample freezing device according to claim 1, characterized in that: The output assembly (3) includes a first output pipe (31), a cell cryopreservation tube (35) formed by connecting a plurality of cell cryopreservation tubes through heat sealing, and an exhaust filter. One end of the first output pipe (31) is connected to the mixing bag (1), and the other end is connected to the cell cryopreservation tube (35). A second switch card (32) is provided on the first output pipe (31), and each cell cryopreservation tube constituting the cell cryopreservation tube (35) is provided with a puncture port (34). The exhaust filter is connected to the end of the cell cryopreservation tube (35) not connected to the first output pipe (31) through an exhaust pipe (39), and a third switch card (36) is provided on the exhaust pipe (39).

5. The integrated automatic exhaust sample freezing device according to claim 4, characterized in that: A fourth switch card (310) is further provided on the first output pipe (31), the exhaust pipe (39) is connected to the first output pipe (31) through a Y-shaped three-way interface, the other end of the exhaust pipe (39) is connected to an exhaust filter, a third switch card (36) is provided on the exhaust pipe (39), a second switch card (32) is provided on the pipe connecting the first output pipe (31) to the mixing bag (1) and the Y-shaped three-way interface, and a fourth switch card (310) is provided on the pipe connecting the first output pipe (31) to the cell freezing tube (35) and the Y-shaped three-way interface.

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