Efficient NK cell culture device and culture method technology thereof

Abstract

The invention relates to the technical field of cell culture, and particularly discloses an efficient NK cell culture device and a culture method thereof, the efficient NK cell culture device comprises a culture box body, a preheating box body and a servo motor are fixedly mounted on the adjacent outer side walls of the culture box body respectively, a culture frame assembly is arranged in the culture box body, and a sealing assembly is arranged in the preheating box body; the placing platform is installed in the connecting seat through the connecting mechanism, the connecting seat is intermittently driven by the servo motor to drive the placing platform to adjust the position in the incubator body and keep the placing platform in a horizontal state, a sealing plate on the preheating box body is rotated, and the sealing plate drives the movable block and the butt joint block to move towards the placing platform through the transmission mechanism. And then, the placing platform is moved into the preheating box body from the connecting base. In the moving process, the placing platform is attached to the culture box body, so that the dissipation of heat and carbon dioxide in the culture box body can be reduced, the stability of the environment in the culture box body is ensured, and the adverse effect on cell culture proliferation is reduced.

Description

Technical Field

[0001] The present invention relates to the field of cell culture technology, and in particular to a NK cell efficient culture device and a culture method thereof. Background Art

[0002] NK cells, or natural killer cells, are important immune cells in the body and belong to the lymphocyte category. They are mainly derived from bone marrow hematopoietic stem cells. After maturing in the bone marrow, they enter the peripheral blood and lymphatic tissues. By culturing NK cells, we can simulate the in vivo environment in vitro and deeply explore how NK cells recognize and kill target cells, as well as the regulatory effects of various cytokines and signal molecules on NK cell functions.

[0003] There are three main ways to culture NK cells at present. No matter which method is used, it is inseparable from the cooperation of the incubator. The incubator can provide the required temperature, carbon dioxide and nutrients for the cell culture and proliferation process, ensuring that the NK cells in the culture bottle are well cultured and proliferated. When using the incubator, preheating the box and adjusting the carbon dioxide concentration in advance can reduce the impact of environmental changes. However, when placing the culture bottle in the existing incubator, opening the door will still cause a large amount of heat and carbon dioxide to escape, causing a large change in the internal environment. In this way, the cells will still produce a stress response due to the sudden change in the environment, and be affected by the changes in the activity of the intracellular enzyme, thereby interfering with the normal physiological process of the cells and bringing adverse consequences to the metabolism and growth of the cells.

[0004] Therefore, it is necessary to provide an efficient NK cell culture device and a culture method technology thereof to solve the above problems. Summary of the invention

[0005] The main purpose of the present invention is to provide a NK cell efficient culture device and culture method technology, which can effectively solve the problems in the background technology.

[0006] To achieve the above object, the technical solution adopted by the present invention is:

[0007] A highly efficient NK cell culture device comprises a culture box, wherein adjacent outer side walls of the culture box are respectively fixedly mounted with a preheating box and a servo motor, a culture rack assembly is arranged inside the culture box, and a sealing assembly is arranged inside the preheating box;

[0008] The culture rack assembly includes a driving shaft, a mounting seat, a connecting seat, a connecting mechanism and a placement platform, wherein the driving shaft is rotatably connected to the inside of the culture box, the mounting seat is fixedly connected to the outside of one end of the driving shaft located in the culture box, the number of the connecting seat, the connecting mechanism and the placement platform are all set to multiple, the connecting seat and the placement platform are detachably installed through the connecting mechanism, the driving shaft is fixedly connected to the output shaft of the servo motor through a coupling, and a counterweight is provided at the bottom of the placement platform;

[0009] The sealing assembly includes a sealing plate, a fixed plate, a transmission mechanism, a movable block, a docking block and a docking mechanism. The sealing plate is hinged to the preheating box, the sealing plate is fixedly connected to the fixed plate, the movable block and the docking block are both slidably connected to the inside of the preheating box, the fixed plate and the movable block are synchronously moved by the transmission mechanism, the docking block is symmetrically fixedly connected to a push rod on one side of the movable block, and the docking mechanism is used to realize the taking of the placement platform.

[0010] As a further improvement of the above solution, handles are symmetrically fixedly connected to the outer wall of the culture box body, and universal wheels are evenly installed on the bottom of the culture box body and the preheating box body.

[0011] As a further improvement of the above scheme, the connecting mechanism includes a symmetrically arranged movable seat and an installation shaft, the installation shaft is rotatably connected to the interior of the connecting seat, the movable seats are fixedly connected to the installation shaft, the outer wall of the placement platform is symmetrically fixedly connected with an installation bar, and the installation bar is slidably connected to the interior of the movable seat.

[0012] As a further improvement of the above scheme, the interior of the movable seat is slidably connected with a push block, a spring 1 is fixedly connected between the push block and the inner wall of the movable seat, the interior of the movable seat is symmetrically slidably connected with a stopper, the interior of the movable seat is symmetrically fixedly connected with a fixed block, the stopper is slidably connected to the outer side of the fixed block, and a spring 2 is fixedly connected between the inner wall of the stopper and the fixed block.

[0013] As a further improvement of the above solution, the transmission mechanism includes a slider symmetrically slidably connected to the preheating box body, a connecting rod 2 is rotatably connected between the slider and the movable block, and a connecting rod 1 is rotatably connected between the slider and the fixed plate.

[0014] As a further improvement of the above scheme, the docking mechanism includes a limiting rod one symmetrically fixedly connected to the outer wall of the docking block, the movable block is slidably connected to the outside of the two limiting rods one, and a spring three is symmetrically fixedly connected between the movable block and the docking block, and the spring three is sleeved on the outside of the limiting rod one.

[0015] As a further improvement of the above scheme, the docking block is symmetrically and slidably connected inside with a clamping block and two limit rods, the two limit rods are fixedly connected to the clamping block, a spring four is fixedly connected between the clamping block and the inner wall of the docking block, the movable block is symmetrically provided with wedge-shaped grooves, and the clamping block is slidably connected to the inside of the wedge-shaped grooves.

[0016] As a further improvement of the above solution, magnets are fixedly connected to the opposite ends of the placement platform and the docking block, and card slots adapted to the card blocks are symmetrically provided on the outer side wall of the placement platform.

[0017] The NK cell culture method and technology comprises the following steps:

[0018] Step 1: Obtain peripheral blood samples from healthy donors by venous blood sampling, prepare lymphocyte separation fluid, and carefully layer the peripheral blood samples on the lymphocyte separation fluid;

[0019] Step 2: Perform centrifugation at a specific speed and time to separate the different components in the blood into layers according to density. After the PBMC is located at the interface between the plasma and the separation fluid, use a pipette to carefully aspirate the PBMC layer and transfer it to a new sterile centrifuge tube.

[0020] Step 3: Inoculate the isolated PBMC into a suitable culture bottle;

[0021] Step 4: Select serum-free medium or medium containing specific ingredients to meet the growth requirements of NK cells;

[0022] Step 5: Place the culture container inoculated with PBMC at ℃ and %CO 2 Cultured in a cell culture incubator;

[0023] Step 6: Add trophoblast cells to promote the growth and activation of NK cells;

[0024] Step 7: Regularly observe the morphology, density and activity of cells under a microscope;

[0025] Step 8: When the NK cells reach a certain number or activity, separate and collect the NK cells using centrifugation.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] 1. The placement platform is installed in the connection seat through a connection mechanism. The connection seat is intermittently driven by the servo motor to drive the placement platform to adjust its position in the incubator and keep it in a horizontal state. When placing the cell culture bottle, the sealing plate on the preheating box is rotated, and the movable block and the docking block are driven to move toward the placement platform through the transmission mechanism to grab the placement platform. Then the placement platform is moved from the connection seat into the preheating box. During the movement, the placement platform fits with the incubator, which can reduce the heat and carbon dioxide dissipation in the incubator, ensure the stability of the incubator environment, and reduce the adverse effects on cell culture and proliferation.

[0028] 2. The movable block moves to push the docking block toward the placement platform, and the push rod pushes the push block to release the clamping of the mounting strip in the movable seat. Then the movable block continues to push the docking block to make the clamping block enter the clamping slot, and the docking block is magnetically connected to the placement platform with the help of a magnet. Then the movable block drives the docking block to move in the opposite direction, and the movable block moves first to make the clamping block clamped in the magnet, so that the placement platform can be pulled out of the connection seat more stably, thereby ensuring the smooth removal of the placement platform, avoiding shaking or instability during the removal process, and reducing the impact on the cell culture bottle placed on the platform. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0030] Figure 1 It is a schematic diagram of the overall structure of the present invention;

[0031] Figure 2 It is a structural schematic diagram of the culture box of the present invention;

[0032] Figure 3 It is a schematic diagram of the internal structure of the culture box and the preheating box of the present invention;

[0033] Figure 4 It is a structural schematic diagram of the culture rack assembly of the present invention;

[0034] Figure 5 This is a schematic diagram of the internal structure of the connecting socket of the present invention;

[0035] Figure 6 It is a schematic cross-sectional structure diagram of a part of the movable seat of the present invention;

[0036] Figure 7 It is a schematic diagram of the cross-sectional structure of the placement platform of the present invention;

[0037] Figure 8It is a structural schematic diagram of the sealing assembly of the present invention;

[0038] Fig. 9 It is a schematic diagram of the cross-sectional structure of the movable block of the present invention;

[0039] Fig.10 For the present invention Fig. 9 The structural diagram at A in the middle;

[0040] Fig.11 It is a structural schematic diagram of the transmission mechanism of the present invention.

[0041] In the figure: 1, incubator; 2, preheating box; 3, universal wheel; 4, handle; 5, servo motor; 6, incubator assembly; 61, drive shaft; 62, mounting seat; 63, connecting seat; 64, connecting mechanism; 641, movable seat; 642, mounting shaft; 643, mounting bar; 644, push block; 645, spring 1; 646, stop block; 647, spring 2; 648, fixed block; 65, placement platform; 66. Magnet; 67. Slot; 7. Sealing assembly; 71. Sealing plate; 72. Fixed plate; 73. Transmission mechanism; 731. Sliding block; 732. Connecting rod one; 733. Connecting rod two; 74. Movable block; 75. Docking block; 76. Push rod; 77. Docking mechanism; 771. Limit rod one; 772. Block; 773. Spring three; 774. Limit rod two; 775. Spring four; 776. Wedge groove. DETAILED DESCRIPTION

[0042] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0043] See also Figures 1 to 11 As shown, the present invention provides an embodiment: an efficient NK cell culture device, comprising a culture box 1, a preheating box 2 and a servo motor 5 are respectively fixedly installed on the adjacent outer side walls of the culture box 1, a culture rack assembly 6 is arranged inside the culture box 1, and a sealing assembly 7 is arranged inside the preheating box 2;

[0044] The culture rack assembly 6 includes a driving shaft 61, a mounting seat 62, a connecting seat 63, a connecting mechanism 64 and a placement platform 65. The driving shaft 61 is rotatably connected to the inside of the culture box 1, and the mounting seat 62 is fixedly connected to the outer side of one end of the driving shaft 61 located in the culture box 1. The number of the connecting seat 63, the connecting mechanism 64 and the placement platform 65 is set to multiple. The connecting seat 63 and the placement platform 65 are detachably installed through the connecting mechanism 64. The driving shaft 61 and the output shaft of the servo motor 5 are fixedly connected through a coupling, and a counterweight block is arranged at the bottom of the placement platform 65.

[0045] The sealing assembly 7 includes a sealing plate 71, a fixed plate 72, a transmission mechanism 73, a movable block 74, a docking block 75 and a docking mechanism 77. The sealing plate 71 is hinged to the preheating box 2, the sealing plate 71 is fixedly connected to the fixed plate 72, the movable block 74 and the docking block 75 are both slidably connected to the inside of the preheating box 2, the fixed plate 72 and the movable block 74 are synchronously moved by the transmission mechanism 73, the docking block 75 is symmetrically fixedly connected to a push rod 76 on one side of the movable block 74, and the docking mechanism 77 is used to realize the taking of the placement platform 65.

[0046] In practical application, the embodiment of the present invention is firstly Figure 3 and Figure 4 As shown, the culture box 1 provides the main space environment for cell culture. In the culture rack assembly 6 inside the culture box 1, the driving shaft 61 rotates in the culture box 1, and the mounting seat 62 is fixed to the outer side of one end of the driving shaft 61 located in the culture box 1. A plurality of connecting seats 63, connecting mechanisms 64 and a placement platform 65 cooperate with each other. The connecting seat 63 and the placement platform 65 are detachably installed through the connecting mechanism 64, and the driving shaft 61 is fixedly connected to the output shaft of the servo motor 5 through a coupling. The counterweight block at the bottom of the placement platform 65 ensures its stability.

[0047] like Figure 3 and Figure 8As shown in the figure, in the sealing assembly 7 inside the preheating box 2, the sealing plate 71 is hinged to the preheating box 2 and fixedly connected to the fixed plate 72, the movable block 74 and the docking block 75 slide inside the preheating box 2, the fixed plate 72 and the movable block 74 are synchronously moved by the transmission mechanism 73, and the side of the docking block 75 away from the movable block 74 is symmetrically fixedly connected with a push rod 76, and the docking mechanism 77 is used to realize the taking of the placement platform 65. In actual operation, when it is necessary to place a cell culture bottle, the sealing plate 71 on the preheating box 2 can be rotated, and the sealing plate 71 is driven by the transmission mechanism 73 to move. The movable block 74 and the docking block 75 move toward the placement platform 65 to capture the placement platform 65, and then move the placement platform 65 from the connecting seat 63 into the preheating box 2. During the movement, the placement platform 65 fits with the culture box 1, which can reduce the heat and carbon dioxide dissipation in the culture box 1, ensure the stability of the environment in the culture box 1, and reduce the adverse effects on cell culture and proliferation. The servo motor 5 intermittently drives the drive shaft 61 to rotate, thereby driving the placement platform 65 to adjust its position in the culture box 1 and maintain a horizontal state, providing more suitable conditions for cell culture.

[0048] like Figure 1 As shown, handles 4 are symmetrically fixedly connected to the outer wall of the culture box body 1, and universal wheels 3 are evenly installed on the bottom of the culture box body 1 and the preheating box body 2.

[0049] In actual application of the embodiment of the present invention, the handles 4 symmetrically fixedly connected on the outer wall of the culture box 1 are convenient for the operator to grasp and apply force when the culture device needs to be moved; and the universal wheels 3 evenly installed on the bottom of the culture box 1 and the preheating box 2 enable the entire device to be flexibly moved between different positions, which is convenient for adjusting the position of the device according to actual needs, and providing a more convenient operating environment for the culture of NK cells.

[0050] like Figure 5 and Figure 6 As shown, the connecting mechanism 64 includes a symmetrically arranged movable seat 641 and a mounting shaft 642, the mounting shaft 642 is rotatably connected to the inside of the connecting seat 63, the movable seats 641 are fixedly connected to the mounting shaft 642, the outer wall of the placement platform 65 is symmetrically fixedly connected with a mounting bar 643, the mounting bar 643 is slidably connected to the inside of the movable seat 641, the inside of the movable seat 641 is slidably connected with a push block 644, a spring 1 645 is fixedly connected between the push block 644 and the inner wall of the movable seat 641, the inside of the movable seat 641 is symmetrically slidably connected with a stopper 646, the inside of the movable seat 641 is symmetrically fixedly connected with a fixed block 648, the stopper 646 is slidably connected to the outer side of the fixed block 648, and a spring 2 647 is fixedly connected between the inner wall of the stopper 646 and the fixed block 648.

[0051] When the embodiment of the present invention is actually applied, the mounting shaft 642 is rotatably connected to the inside of the connecting seat 63, the symmetrically arranged movable seat 641 is fixedly connected to the mounting shaft 642, and the mounting strip 643 symmetrically fixed on the outer wall of the placement platform 65 can be slidably connected to the inside of the movable seat 641 to achieve a preliminary connection between the placement platform 65 and the connecting seat 63. When the mounting strip 643 is inserted into the movable seat 641, the push block 644, under the action of the spring 1 645, plays a certain blocking and fixing role on the stopper 646 to prevent the stopper 646 from easily falling out, ensuring that the stopper 646 symmetrically slidably connected inside the movable seat 641, under the action of the spring 2 647, enhances the fixing effect of the mounting strip 643, and the fixed block 648 provides a stable support for the sliding of the stopper 646, ensuring the stability of the connection between the placement platform 65 and the connecting seat 63, providing a stable placement platform 65 for cell culture, and even when the device moves or is subjected to a certain external force, it can ensure that the placement platform 65 will not be easily separated from the connecting seat 63, thereby ensuring the smooth progress of the cell culture process.

[0052] like Figure 8 and Fig.11 As shown, the transmission mechanism 73 includes a slider 731 symmetrically slidably connected to the preheating box 2, a second connecting rod 733 is rotatably connected between the slider 731 and the movable block 74, and a first connecting rod 732 is rotatably connected between the slider 731 and the fixed plate 72.

[0053] In actual application of the embodiment of the present invention, when the rotating sealing plate 71 drives the fixed plate 72 to move, the fixed plate 72 pushes the slider 731 to slide in the preheating box 2 through the connecting rod 1 732, and the movement of the slider 731 drives the movable block 74 to move through the connecting rod 2 733, so that the rotation of the sealing plate 71 can be accurately converted into the linear motion of the movable block 74, thereby realizing the pushing of the docking block 75, and then completing the grabbing operation of the placement platform 65, ensuring the accuracy and stability of the operation, making the actions of the various components coordinated and consistent, and providing a reliable guarantee for the efficient operation of the cell culture device.

[0054] like Figures 8 to 10As shown, the docking mechanism 77 includes a limiting rod 1 771 symmetrically fixedly connected to the outer side wall of the docking block 75, the movable block 74 is slidably connected to the outer side of the two limiting rods 1 771, a spring 3 773 is symmetrically fixedly connected between the movable block 74 and the docking block 75, and the spring 3 773 is sleeved on the outer side of the limiting rod 1 771, and a clamping block 772 and a limiting rod 2 774 are symmetrically slidably connected inside the docking block 75, and the limiting rod 2 774 is fixedly connected to the clamping block 772, and a spring 4 775 is fixedly connected between the clamping block 772 and the inner wall of the docking block 75, a wedge-shaped groove 776 is symmetrically opened inside the movable block 74, and the clamping block 772 is slidably connected to the inside of the wedge-shaped groove 776, and the opposite end of the placement platform 65 and the docking block 75 is fixedly connected with a magnet 66, and a clamping groove 67 adapted to the clamping block 772 is symmetrically opened on the outer side wall of the placement platform 65.

[0055] When the embodiment of the present invention is actually used, the limiting rod 1 771 symmetrically fixedly connected to the outer wall of the docking block 75 provides a guide for the sliding of the movable block 74. The movable block 74 slides on the outside of the two limiting rods 1 771 and is connected to the docking block 75 through the spring 3 773. The spring 3 773 is sleeved on the outside of the limiting rod 1 771 to ensure that the movable block 74 has a certain elastic buffer during the movement. The block 772 and the limiting rod 2 774 symmetrically slidably connected inside the docking block 75 cooperate with each other. The limiting rod 2 774 ensures the stable sliding of the block 772. The block 772 can move flexibly inside the docking block 75 under the action of the spring 4 775. When the movable block 74 pushes the docking block 75 to be placed flat, When the platform 65 approaches, the block 772 is squeezed and contracted under the action of the wedge-shaped groove 776. When the block 772 is aligned with the groove 67 on the placement platform 65, the block 772 pops out into the groove 67 under the action of the spring four 775, and the magnetic connection between the docking block 75 and the placement platform 65 is achieved with the help of the magnet 66. Then the movable block 74 drives the docking block 75 to move in the opposite direction. The movable block 74 moves first to make the block 772 more firmly connected in the magnet 66, so that the placement platform 65 can be pulled out of the connecting seat 63 more stably, ensuring the smooth removal of the placement platform 65, avoiding shaking or instability during the removal process, and reducing the impact on the cell culture bottle placed on the platform.

[0056] The present invention provides another embodiment:

[0057] A NK cell culture method technology is now proposed, comprising the following steps:

[0058] Step 1: Obtain peripheral blood samples from healthy donors by venous blood sampling, prepare lymphocyte separation fluid, and carefully layer the peripheral blood samples on the lymphocyte separation fluid;

[0059] Step 2: Perform centrifugation at a specific speed and time to separate the different components in the blood into layers according to density. After the PBMC is located at the interface between the plasma and the separation fluid, use a pipette to carefully aspirate the PBMC layer and transfer it to a new sterile centrifuge tube.

[0060] Step 3: Inoculate the isolated PBMC into a suitable culture bottle;

[0061] Step 4: Select serum-free medium or medium containing specific ingredients to meet the growth requirements of NK cells;

[0062] Step 5: Place the culture container inoculated with PBMC at 37°C and 5% CO 2 Cultured in a cell culture incubator;

[0063] Step 6: Add trophoblast cells to promote the growth and activation of NK cells;

[0064] Step 7: Regularly observe the morphology, density and activity of cells under a microscope;

[0065] Step 8: When the NK cells reach a certain number or activity, separate and collect the NK cells using centrifugation.

[0066] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "including one..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0067] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A highly efficient NK cell culture device, comprising a culture box (1), characterized in that: A preheating box (2) and a servo motor (5) are fixedly mounted on adjacent outer side walls of the culture box (1), a culture rack assembly (6) is arranged inside the culture box (1), and a sealing assembly (7) is arranged inside the preheating box (2); The culture rack assembly (6) comprises a driving shaft (61), a mounting seat (62), a connecting seat (63), a connecting mechanism (64) and a placement platform (65); the driving shaft (61) is rotatably connected to the interior of the culture box (1); the mounting seat (62) is fixedly connected to the outer side of one end of the driving shaft (61) located in the culture box (1); the number of the connecting seat (63), the connecting mechanism (64) and the placement platform (65) is set to be multiple; the connecting seat (63) and the placement platform (65) are detachably mounted via the connecting mechanism (64); the driving shaft (61) and the output shaft of the servo motor (5) are fixedly connected via a coupling; and a counterweight block is provided at the bottom of the placement platform (65); The sealing assembly (7) comprises a sealing plate (71), a fixed plate (72), a transmission mechanism (73), a movable block (74), a docking block (75) and a docking mechanism (77); the sealing plate (71) is hinged to the preheating box (2); the sealing plate (71) is fixedly connected to the fixed plate (72); the movable block (74) and the docking block (75) are both slidably connected to the inside of the preheating box (2); the fixed plate (72) and the movable block (74) are synchronously moved by the transmission mechanism (73); the docking block (75) is symmetrically fixedly connected to a push rod (76) on one side of the movable block (74); and the docking mechanism (77) is used to realize the taking of the placement platform (65).

2. The NK cell efficient culture device according to claim 1, characterized in that: A handle (4) is symmetrically fixedly connected to the outer wall of the culture box (1), and universal wheels (3) are evenly installed on the bottom of the culture box (1) and the preheating box (2).

3. The NK cell efficient culture device according to claim 2, characterized in that: The connecting mechanism (64) comprises a symmetrically arranged movable seat (641) and a mounting shaft (642), wherein the mounting shaft (642) is rotatably connected to the interior of the connecting seat (63), and the movable seats (641) are fixedly connected to the mounting shaft (642). The outer side wall of the placement platform (65) is symmetrically fixedly connected with a mounting bar (643), and the mounting bar (643) is slidably connected to the interior of the movable seat (641).

4. The NK cell efficient culture device according to claim 3, characterized in that: The interior of the movable seat (641) is slidably connected with a push block (644), and a spring 1 (645) is fixedly connected between the push block (644) and the inner wall of the movable seat (641). The interior of the movable seat (641) is symmetrically slidably connected with a stopper (646), and the interior of the movable seat (641) is symmetrically fixedly connected with a fixed block (648). The stopper (646) is slidably connected to the outer side of the fixed block (648), and a spring 2 (647) is fixedly connected between the inner wall of the stopper (646) and the fixed block (648).

5. The NK cell efficient culture device according to claim 2, characterized in that: The transmission mechanism (73) comprises a slider (731) symmetrically slidably connected to the preheating box (2); a second connecting rod (733) is rotatably connected between the slider (731) and the movable block (74); and a first connecting rod (732) is rotatably connected between the slider (731) and the fixed plate (72).

6. The NK cell efficient culture device according to claim 2, characterized in that: The docking mechanism (77) comprises a limiting rod (771) symmetrically fixedly connected to the outer wall of the docking block (75); the movable block (74) is slidably connected to the outer sides of the two limiting rods (771); a spring (773) is symmetrically fixedly connected between the movable block (74) and the docking block (75); and the spring (773) is sleeved on the outer side of the limiting rod (771).

7. The NK cell efficient culture device according to claim 3, characterized in that: The docking block (75) is symmetrically and slidably connected with a clamping block (772) and a second limiting rod (774) inside. The second limiting rod (774) is fixedly connected to the clamping block (772). A fourth spring (775) is fixedly connected between the clamping block (772) and the inner wall of the docking block (75). The movable block (74) is symmetrically provided with a wedge-shaped groove (776) inside. The clamping block (772) is slidably connected to the inside of the wedge-shaped groove (776).

8. The NK cell efficient culture device according to claim 3, characterized in that: The placement platform (65) and the opposite end of the docking block (75) are both fixedly connected with a magnet (66), and the outer side wall of the placement platform (65) is symmetrically provided with a clamping groove (67) adapted to the clamping block (772).

9. A method and technology for culturing NK cells, wherein the method and technology is based on any one of the NK cell efficient culturing devices in claims 1-8, characterized in that: The following steps are involved: Step 1: Obtain peripheral blood samples from healthy donors by venous blood sampling, prepare lymphocyte separation fluid, and carefully layer the peripheral blood samples on the lymphocyte separation fluid; Step 2: Perform centrifugation at a specific speed and time to separate the different components in the blood into layers according to density. After the PBMC is located at the interface between the plasma and the separation fluid, use a pipette to carefully aspirate the PBMC layer and transfer it to a new sterile centrifuge tube. Step 3: Inoculate the isolated PBMC into a suitable culture bottle; Step 4: Select serum-free medium or medium containing specific ingredients to meet the growth requirements of NK cells; Step 5: Place the culture container inoculated with PBMC in a cell culture incubator at 37°C and 5% CO2; Step 6: Add trophoblasts to promote the growth and activation of NK cells; Step 7: Regularly observe the morphology, density and activity of cells under a microscope; Step 8: When the NK cells reach a certain number or activity, separate and collect the NK cells using centrifugation.

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