Multi-stage dehydration device for stem cell vitrification cryopreservation

Abstract

The utility model discloses a multi-stage dehydration device for stem cell vitrification cryopreservation, which comprises a base, the top of the base is provided with a containing groove, the containing groove is internally provided with a storage pipe for storing stem cell suspension and a [-shaped frame, the top of the [-shaped frame is provided with a lifting assembly, and the lifting assembly is provided with a water inlet and a water outlet. The movable end of the lifting assembly is fixedly provided with an injection pipe, the outer wall of the injection pipe is fixedly communicated with a peristaltic pump through a pipeline, through linkage control of the lifting assembly and the peristaltic pump, automatic conveying of a dehydrating agent and accurate lifting of the injection pipe are achieved, manual pipetting or container replacement is not needed, the operation efficiency is remarkably improved, and the labor intensity of workers is reduced. The quick plugging design of the quick-plugging male connector and the quick-plugging female connector supports switching of multiple groups of dehydrating agent storage tanks as required, ensures continuous gradient progression of dehydrating agents with different concentrations, realizes gradual dehydration of stem cell suspension in a gradient concentration environment, realizes smooth transition of the concentration of the dehydrating agent, and is beneficial to further improvement of the survival rate of stem cells.

Description

Technical Field

[0001] This utility model relates to the field of biomedical cryopreservation technology, specifically a multi-stage dehydration device for vitrification cryopreservation of stem cells. Background Technology

[0002] As one of the core technologies in modern cryobiology and regenerative medicine, stem cell vitrification cryopreservation technology provides key support for the long-term preservation and clinical application of cell resources. Vitrification cryopreservation involves mixing cell suspension with a high concentration of cryoprotectant to form an amorphous vitrified structure at ultra-low temperatures. The key is to avoid ice crystal formation through dehydration, thereby reducing mechanical damage.

[0003] In existing technologies, single-stage dehydration is widely used due to its ease of operation. However, single-stage dehydration cannot achieve a smooth transition in concentration, resulting in low cell survival rates. Therefore, we need to propose a multi-stage dehydration device for vitrification cryopreservation of stem cells. Utility Model Content

[0004] The purpose of this invention is to provide a multi-stage dehydration device for vitrification and cryopreservation of stem cells. By setting up a lifting component to drive the injection tube to automatically insert into the storage tube, and combining the peristaltic pump for precise flow control, the dehydrating agent can be slowly and quantitatively injected. By setting up several sets of dehydrating agent storage tanks and matching quick-connect connectors, dehydrating agent solutions of different concentrations can be flexibly connected to achieve gradual dehydration of stem cell suspensions in a gradient concentration environment, thereby solving the problems mentioned in the background art.

[0005] To achieve the above objectives, this utility model provides the following technical solution:

[0006] A multi-stage dehydration device for vitrification cryopreservation of stem cells includes: a base, the top of which has a receiving groove, and the inside of the receiving groove is provided with a storage tube for storing stem cell suspension;

[0007] A U-shaped frame, the top of which is provided with a lifting assembly, an injection tube is fixedly installed on the movable end of the lifting assembly, a peristaltic pump is fixedly connected to the outer wall of the injection tube through a pipe, a flexible tube is fixedly connected to one end of the peristaltic pump, and a quick-connect male connector is fixedly connected to one end of the flexible tube.

[0008] Several sets of dehydrating agent storage tanks are fixedly connected to one side wall of the base. A connecting pipe is fixedly connected to the top of the dehydrating agent storage tank. A quick-connect female connector is fixedly connected to the top of the connecting pipe. The quick-connect male connector is inserted into the top of the quick-connect female connector.

[0009] Preferably, the lifting assembly includes an electric push rod, which is fixedly installed on the top of the C-shaped frame. The bottom end of the telescopic end of the electric push rod passes through the C-shaped frame and is fixedly connected to a lifting plate. The injection tube is fixedly installed at the bottom of the lifting plate. Two sets of limiting rods are slidably inserted into the top of the C-shaped frame. The bottom ends of both sets of limiting rods pass through the C-shaped frame and are fixedly connected to the top of the lifting plate.

[0010] Preferably, the interior of the receiving tank is provided with a low-speed centrifugation assembly for centrifuging the stem cell suspension inside the storage tube. The low-speed centrifugation assembly includes a drive motor, a turntable, and a clamping mechanism for positioning the storage tube. The bottom of the turntable is connected to the drive motor via a rotating shaft.

[0011] Preferably, the clamping mechanism includes a fixed plate, a threaded sleeve, a screw, a knob, and a clamping block;

[0012] The fixing plate is fixedly installed on the top of the turntable, the threaded sleeve is fixedly embedded inside the fixing plate, the screw is threadedly connected to the inside of the threaded sleeve, one end of the screw passes through the threaded sleeve and is rotatably connected to one side of the clamping block, and the other end of the screw is fixedly connected to the knob.

[0013] Preferably, it also includes a guide rod, which is slidably inserted into the interior of the fixing plate, and one end of the guide rod passes through the fixing plate and is fixedly connected to one side of the clamping block.

[0014] Preferably, the drive motor is fixedly installed at the bottom of the base, and one end of the output shaft of the drive motor is fixedly connected to the bottom end of the rotating shaft.

[0015] Preferably, the top of the storage tube is provided with a sealing cap, the sealing cap having a through hole for the injection tube to pass through, and an elastic sealing ring being embedded in the inner wall of the through hole.

[0016] Compared with the prior art, the beneficial effects of this utility model are:

[0017] This invention achieves automatic delivery of dehydrating agent and precise lifting and lowering of injection tube through the linkage control of lifting components and peristaltic pump, eliminating the need for manual liquid transfer or container replacement, significantly improving operational efficiency. The quick-connect male and female connectors allow for rapid insertion and removal, supporting on-demand switching of multiple dehydrating agent storage tanks, ensuring a continuous gradient of different concentrations of dehydrating agent. This enables the gradual dehydration of stem cell suspensions in a gradient concentration environment, achieving a smooth transition of dehydrating agent concentration, which is beneficial for further improving stem cell survival rate. Attached Figure Description

[0018] Figure 1 This is a schematic diagram of the structure of this utility model;

[0019] Figure 2This is a schematic diagram of the axial side structure of this utility model;

[0020] Figure 3 This is a schematic diagram of the structure of the low-speed centrifugal assembly of this utility model;

[0021] Figure 4 This is a schematic diagram of the lifting assembly and injection tube of this utility model.

[0022] In the diagram: 1. Base; 2. Receiving tank; 3. Storage tube; 4. Low-speed centrifugal assembly; 401. Drive motor; 402. Turntable; 403. Clamping mechanism; 4031. Fixing plate; 4032. Threaded sleeve; 4033. Screw; 4034. Knob; 4035. Clamping block; 4036. Guide rod; 5. C-shaped frame; 6. Lifting assembly; 601. Electric push rod; 602. Lifting plate; 603. Limiting rod; 7. Injection tube; 8. Peristaltic pump; 9. Flexible tube; 10. Quick-connect male connector; 11. Dehydrating agent storage tank; 12. Through pipe; 13. Quick-connect female connector; 14. Sealing cap. Detailed Implementation

[0023] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0024] Please see Figure 1-4 This utility model provides a technical solution:

[0025] A multi-stage dehydration device for vitrification cryopreservation of stem cells includes: a base 1, a receiving groove 2 on the top of the base 1, and a storage tube 3 for storing stem cell suspension inside the receiving groove 2. The storage tube 3 is made of transparent polypropylene with a wall thickness of 2-3 mm, is resistant to low temperatures down to -196℃, and its inner wall is hydrophilic to reduce cell adhesion.

[0026] The frame 5 has a lifting assembly 6 on its top. An injection tube 7 is fixedly installed on the movable end of the lifting assembly 6. A peristaltic pump 8 is fixedly connected to the outer wall of the injection tube 7 through a pipe. A flexible tube 9 is fixedly connected to one end of the peristaltic pump 8. A quick-connect male connector 10 is fixedly connected to one end of the flexible tube 9. The peristaltic pump 8 adopts a multi-channel roller structure with a speed range of 0.1-100 rpm and a flow accuracy of ±1%. The dehydrating agent is delivered without pulsation through the flexible tube 9. The flexible tube 9 is made of platinum-iridium alloy spring-reinforced silicone tubing, which is resistant to chemical corrosion and kinking.

[0027] Several sets of dehydrating agent storage tanks 11 are fixedly connected to one side wall of the base 1. The top of the dehydrating agent storage tank 11 is fixedly connected to a pipe 12, and the top of the pipe 12 is fixedly connected to a quick-connect female connector 13. The quick-connect male connector 10 is inserted into the top of the quick-connect female connector 13. The quick-connect male connector 10 and the quick-connect female connector 13 are of Luer interface standard. The built-in one-way valve prevents liquid backflow and ensures convenient operation. There are a total of 4 sets of dehydrating agent storage tanks 11, which store DMSO-glucose composite protective agent solutions with concentrations of 10%, 20%, 30%, and 40% respectively. The tank volume is 500mL and is made of borosilicate glass. The inner diameter of the top pipe 12 is 6mm. The outer wall is wrapped with a heat tracing cable to maintain a constant solution temperature (4℃). Each tank is connected to the quick-connect female connector 13 through an independent valve, which supports manual or programmed switching to realize flexible control of the gradient dehydration process.

[0028] The lifting assembly 6 includes an electric push rod 601, which is driven by a stepper motor with a stroke accuracy of ±0.1mm to ensure controllable insertion depth of the injection tube 7. The electric push rod 601 is fixedly installed on the top of the C-shaped frame 5. The bottom end of the telescopic end of the electric push rod 601 passes through the C-shaped frame 5 and is fixedly connected to a lifting plate 602. The lifting plate 602 is made of aluminum alloy with an anodized surface to reduce weight and improve corrosion resistance. The injection tube 7 is fixedly installed on the bottom of the lifting plate 602. Two sets of limit rods 603 are slidably inserted into the top of the C-shaped frame 5. The bottom ends of both sets of limit rods 603 pass through the C-shaped frame 5 and are fixedly connected to the top of the lifting plate 602. The limit rods 603 are stainless steel optical shafts to eliminate sway error during the lifting process of the injection tube 7.

[0029] The interior of the receiving tank 2 is equipped with a low-speed centrifugation assembly 4 for centrifuging the stem cell suspension inside the storage tube 3. The low-speed centrifugation assembly 4 includes a drive motor 401, a turntable 402, and a clamping mechanism 403 for positioning the storage tube 3. The bottom of the turntable 402 is connected to the drive motor 401 via a rotating shaft.

[0030] The clamping mechanism 403 includes a fixed plate 4031, a threaded sleeve 4032, a screw 4033, a knob 4034, and a clamping block 4035. The fixed plate 4031 is fixedly installed on the top of the turntable 402. The threaded sleeve 4032 is fixedly embedded inside the fixed plate 4031. The screw 4033 is threadedly connected to the inside of the threaded sleeve 4032. One end of the screw 4033 passes through the threaded sleeve 4032 and is rotatably connected to one side of the clamping block 4035. The other end of the screw 4033 is fixedly connected to the knob 4034.

[0031] It also includes a guide rod 4036, which is slidably inserted into the interior of the fixing plate 4031. One end of the guide rod 4036 passes through the fixing plate 4031 and is fixedly connected to one side of the clamping block 4035. By setting the guide rod 4036, the clamping block 4035 is limited, making its movement process more stable and providing a stable clamping force for the storage tube 3.

[0032] The drive motor 401 is fixedly installed at the bottom of the base 1. One end of the output shaft of the drive motor 401 is fixedly connected to the bottom end of the rotating shaft. The drive motor 401 is a brushless DC motor with a speed range of 100-3000rpm. Smooth speed regulation is achieved through a frequency converter. The turntable 402 is made of aluminum alloy and has an anti-corrosion coating sprayed on its surface.

[0033] The top of the storage tube 3 is provided with a sealing cap 14. The sealing cap 14 has a through hole for the injection tube 7 to pass through, and the inner wall of the through hole is embedded with an elastic sealing ring. The elastic sealing ring in the through hole of the sealing cap 14 adopts a double-layer structure. The outer layer is a rigid polytetrafluoroethylene support ring, and the inner layer is a fluororubber sealing lip, which is resistant to organic solvents such as DMSO. The gap between the turntable 402 and the receiving tank 2 can be equipped with a negative pressure adsorption device to maintain the internal pressure ≤-50Pa and prevent the intrusion of external microorganisms.

[0034] Inject the stem cell suspension into the storage tube 3, tighten the sealing cap 14 and insert it into the receiving groove 2. Rotate the screw 4033 by the knob 4034 of the clamping mechanism 403 to push the clamping block 4035 to slide along the guide rod 4036, fix the storage tube 3 in the center of the turntable 402, start the drive motor 401, drive the turntable 402 to rotate at 300 rpm for 5 minutes, so that the stem cells settle to the bottom of the storage tube 3 and reduce the volume of the supernatant.

[0035] First stage of dehydration: Insert the quick-connect male connector 10 into the quick-connect female connector 13 of the 10% concentration dehydrating agent storage tank 11, start the peristaltic pump 8 to inject the dehydrating agent at a flow rate of 2ml / min, and simultaneously start the electric push rod 601 to slowly press down the injection tube 7 so that the liquid level rises at a speed ≤0.5mm / s to avoid impact damage to cells.

[0036] After completing the current concentration dehydration, turn off the peristaltic pump 8, replace the quick-connect male connector 10 with the next concentration tank, and repeat the above operation until the final concentration of 40% is reached. The dehydration time for each stage is dynamically adjusted according to the cell type. After dehydration is completed, lift the injection tube 7 using the lifting assembly 6, remove the storage tube 3, and quickly transfer it to the liquid nitrogen tank to complete vitrification.

[0037] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A multi-stage dehydration device for vitrification cryopreservation of stem cells, characterized in that: include: The base (1) has a receiving groove (2) on its top, and a storage tube (3) for storing stem cell suspension is provided inside the receiving groove (2). A U-shaped frame (5) is provided with a lifting assembly (6) on the top of the U-shaped frame (5). An injection tube (7) is fixedly installed on the movable end of the lifting assembly (6). A peristaltic pump (8) is fixedly connected to the outer wall of the injection tube (7) through a pipe. A flexible tube (9) is fixedly connected to one end of the peristaltic pump (8). A quick-connect male connector (10) is fixedly connected to one end of the flexible tube (9). Several sets of dehydrating agent storage tanks (11) are fixedly connected to one side wall of the base (1). A connecting pipe (12) is fixedly connected to the top of the dehydrating agent storage tank (11). A quick-connect female connector (13) is fixedly connected to the top of the connecting pipe (12). A quick-connect male connector (10) is inserted into the top of the quick-connect female connector (13).

2. The multi-stage dehydration device for vitrification cryopreservation of stem cells according to claim 1, characterized in that: The lifting assembly (6) includes an electric push rod (601), which is fixedly installed on the top of the shaped frame (5). The bottom end of the telescopic end of the electric push rod (601) passes through the shaped frame (5) and is fixedly connected to the lifting plate (602). The injection tube (7) is fixedly installed on the bottom of the lifting plate (602). Two sets of limiting rods (603) are slidably inserted into the top of the shaped frame (5). The bottom ends of the two sets of limiting rods (603) pass through the shaped frame (5) and are fixedly connected to the top of the lifting plate (602).

3. The multi-stage dehydration device for vitrification cryopreservation of stem cells according to claim 1, characterized in that: The container (2) is equipped with a low-speed centrifugation assembly (4) for centrifuging the stem cell suspension inside the storage tube (3). The low-speed centrifugation assembly (4) includes a drive motor (401), a turntable (402), and a clamping mechanism (403) for positioning the storage tube (3). The bottom of the turntable (402) is connected to the drive motor (401) via a rotating shaft.

4. The multi-stage dehydration device for vitrification cryopreservation of stem cells according to claim 3, characterized in that: The clamping mechanism (403) includes a fixed plate (4031), a threaded sleeve (4032), a screw (4033), a knob (4034), and a clamping block (4035); The fixing plate (4031) is fixedly installed on the top of the turntable (402), the threaded sleeve (4032) is fixedly embedded inside the fixing plate (4031), the screw (4033) is threadedly connected to the inside of the threaded sleeve (4032), one end of the screw (4033) passes through the threaded sleeve (4032) and is rotatably connected to one side of the clamping block (4035), and the other end of the screw (4033) is fixedly connected to the knob (4034).

5. A multi-stage dehydration device for vitrification cryopreservation of stem cells according to claim 4, characterized in that: It also includes a guide rod (4036), which is slidably inserted into the interior of the fixing plate (4031), and one end of the guide rod (4036) passes through the fixing plate (4031) and is fixedly connected to one side of the clamping block (4035).

6. The multi-stage dehydration device for vitrification cryopreservation of stem cells according to claim 5, characterized in that: The drive motor (401) is fixedly installed at the bottom of the base (1), and one end of the output shaft of the drive motor (401) is fixedly connected to the bottom end of the rotating shaft.

7. The multi-stage dehydration device for vitrification cryopreservation of stem cells according to claim 1, characterized in that: The top of the storage tube (3) is provided with a sealing cap (14), and the sealing cap (14) has a through hole for the injection tube (7) to pass through, and the inner wall of the through hole is embedded with an elastic sealing ring.

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