A cell culture device and method

Abstract

The present invention discloses a cell culture device and method, including a glove box, a test tube rotating seat assembly, a liquid extraction assembly and a transmission assembly, wherein a culture chamber is provided in the glove box, an observation window and a discharge port are respectively provided on one side surface of the glove box, a column is integrally formed on the other side surface of the glove box, the test tube rotating seat assembly is arranged on the bottom wall of the culture chamber, the liquid extraction assembly is arranged on the surface of the column, and the transmission assembly is arranged inside the column. Thus, the device can synchronously drive multiple groups of test tubes to rotate synchronously to implement synchronous low-speed centrifugation operation, and at the same time, it can also synchronously drive multiple groups of liquid extraction needles to extract culture fluid from multiple groups of test tubes. This innovative design not only realizes the function of extracting culture fluid during low-speed centrifugation, but also effectively reduces the operation frequency, greatly improves work efficiency, and significantly reduces the labor intensity of the operator. In addition, this synchronous operation also ensures the maintenance of the cell aggregation effect, and the use effect is good.

Description

Technical Field

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

[0002] Somatic cell culture is to remove relevant tissues from the animal body, disperse them into single cells and then place them in a suitable culture medium to allow these cells to grow and proliferate. It needs to be cultured in a sterile environment to avoid bacterial contamination. It is divided into adherent cells and suspended cells. During the culture process of suspended cells, the suspension needs to be kept mixed to avoid cell sedimentation.

[0003] In the existing suspension cell culture process, the culture medium needs to be replaced after a period of time. When replacing the culture medium, the culture tube is taken out of the incubator and then put back into the incubator after the replacement is completed. This can easily cause the suspension to be contaminated by bacteria, which is not conducive to cell culture.

[0004] To solve the above problems, a patent application document with publication number CN113214970B discloses an adaptive regulating somatic cell culture device and a culture method thereof. Although the device effectively solves the above problems, it still shows some limitations in practical applications.

[0005] Specifically, since multiple groups of test tubes are fixed on the chassis, the centrifuge motor drives the chassis to rotate to achieve low-speed centrifugation, thereby causing the cells to aggregate at the bottom of the test tubes. However, this design hinders the liquid extraction operation during the rotation of the chassis, and the culture medium can only be extracted through the liquid extraction tube after the chassis stops rotating. Furthermore, each time the culture medium in a group of test tubes is extracted, in order to maintain the cell aggregation effect, the centrifuge motor needs to be started again to drive the chassis to rotate to achieve low-speed centrifugation. This frequent low-speed centrifugation and liquid extraction operation not only reduces work efficiency, but also increases the labor intensity of the operator. Summary of the invention

[0006] The present invention aims to solve one of the technical problems in the related art at least to a certain extent.

[0007] To this end, the purpose of the present invention is to propose a cell culture device and method, which can synchronously drive multiple groups of test tubes to rotate synchronously to implement synchronous low-speed centrifugation operations. At the same time, it can also synchronously drive multiple groups of extraction needles to extract culture fluid from multiple groups of test tubes. This innovative design not only realizes the function of extracting culture fluid during low-speed centrifugation, but also effectively reduces the operation frequency, greatly improves work efficiency, and significantly reduces the labor intensity of the operator. In addition, this synchronous operation also ensures the maintenance of the cell aggregation effect, and the use effect is good.

[0008] To achieve the above-mentioned purpose, the present invention proposes a cell culture device, comprising a glove box, a test tube rotating seat assembly, a liquid pumping assembly and a transmission assembly, wherein a culture chamber is provided in the glove box, an observation window and a discharge port are respectively provided on one side surface of the glove box, and are arranged up and down, and a column is integrally formed on the other side surface of the glove box and is connected to the interior of the glove box; the test tube rotating seat assembly is arranged on the bottom wall of the culture chamber, and one end of the test tube rotating seat assembly penetrates into the interior of the glove box; the liquid pumping assembly is arranged on the surface of the column, and one end of the liquid pumping assembly penetrates into the interior of the culture chamber and corresponds to the position of the test tube rotating seat assembly; the transmission assembly is arranged in the interior of the column, one end of the transmission assembly is connected to one end of the liquid pumping assembly, and the other end of the transmission assembly is connected to one end of the test tube rotating seat assembly that penetrates into the interior of the glove box.

[0009] A cell culture device of the present invention can synchronously drive multiple groups of test tubes to rotate synchronously to implement synchronous low-speed centrifugation operations. At the same time, it can also synchronously drive multiple groups of extraction needles to extract culture fluid from the multiple groups of test tubes. This innovative design not only realizes the function of extracting culture fluid during low-speed centrifugation, but also effectively reduces the operation frequency, greatly improves work efficiency, and significantly reduces the labor intensity of the operator. In addition, this synchronous operation also ensures the maintenance of the cell aggregation effect, and the use effect is good.

[0010] In addition, the cell culture device proposed in the application may also have the following additional technical features:

[0011] Specifically, the test tube rotating seat assembly includes a fixed seat, a rotating seat, a test tube, a middle gear, a side gear, a large transmission gear and a toothed synchronous belt, wherein the fixed seat is fixedly connected to the bottom wall of the culture chamber, the rotating seat is evenly rotationally connected to the top of the fixed seat, the test tubes are respectively clamped and fixed on the top of the rotating seat, the middle gear is rotationally connected to the inner wall of the glove box and corresponds to the position of the fixed seat, the side gear is evenly rotationally connected to the inner wall of the glove box and is located on the outside of the middle gear, multiple groups of side gears respectively correspond to the positions of multiple groups of rotating seats and are connected, multiple groups of side gears are respectively meshed with the middle gear, the large transmission gear is fixedly connected to the outer surface of one end of the central axis of the middle gear and is located on one side of the bottom of the middle gear, and the large transmission gear is connected to one end of the transmission assembly through the toothed synchronous belt.

[0012] Specifically, the liquid extraction component includes a needle tube mechanism, an extraction mechanism and a drainage tube, wherein the needle tube mechanism is arranged on the surface of the column and connected to one end of the transmission component, the extraction mechanism is arranged on the top of the column and connected to one end of the needle tube mechanism through the drainage tube, and one end of the needle tube mechanism penetrates into the interior of the culture chamber and corresponds to the position of the test tube.

[0013] Specifically, the needle tube mechanism includes a lifting slide, a connecting seat, a liquid separation seat, a main liquid tube and a liquid extraction needle, wherein the lifting slide is slidably connected to the surface of the column, one end of the lifting slide passes through the interior of the column and is connected to one end of the transmission component, the other end of the lifting slide is fixedly connected to the connecting seat, the liquid separation seat is fixedly connected to the inner wall of the connecting seat, one end of the main liquid tube is arranged on the top of the liquid separation seat and is connected to the interior of the liquid separation seat, the other end of the main liquid tube passes through the top of the lifting slide and is connected to one end of the extraction mechanism through the drain pipe, the liquid extraction needles are evenly arranged at the bottom of the liquid separation seat and are connected to the interior of the liquid separation seat, and one end of multiple groups of liquid extraction needles respectively pass through the interior of the culture chamber and correspond to the positions of the test tubes.

[0014] Specifically, the extraction mechanism includes a mounting plate frame, a transmission side gear, a one-way transmission, a threaded screw, a horizontal slide, a liquid extraction pipe, a piston, a liquid inlet end, a liquid discharge end, a valve, a rod frame and a toothed portion, wherein the mounting plate frame is fixedly connected to the top of the column, the transmission side gear, the one-way transmission and the threaded screw are rotatably connected to the surface of the mounting plate frame in sequence and are connected to each other, the horizontal slide is threadedly connected to the outer surface of the threaded screw and is slidably connected to the inner wall of the mounting plate frame, the liquid extraction pipe is clamped and fixed to the surface of the mounting plate frame and is located on one side of the horizontal slide, the piston is slidably connected to the inner wall of the liquid extraction pipe, and the center of the piston One end of the shaft passes through the outside of the liquid pumping tube and is fixedly connected to the surface of one end of the horizontal slide. The liquid inlet end is arranged at the top of the end of the liquid pumping tube close to the horizontal slide and is communicated with the interior of the liquid pumping tube. The liquid inlet end is connected to one end of the main liquid pipe through the liquid discharge pipe. The liquid discharge end is arranged at the bottom of the end of the liquid pumping tube close to the horizontal slide and is communicated with the interior of the liquid pumping tube. The valve is arranged on the surface of the liquid discharge end. The rod frame is fixedly connected to the top of the lifting slide and is located on one side of the transmission side gear. The tooth portion is arranged on the surface of the rod frame, and the teeth on the tooth portion are meshed with the transmission side gear.

[0015] Specifically, the extraction mechanism also includes a disconnecting mechanism, which includes a connecting slide and a fixing bolt, wherein two ends of the connecting slide are respectively slidably connected to one end surface of the one-way transmission and one end surface of the threaded screw, and the two ends of the connecting slide are respectively fixedly connected to one end surface of the one-way transmission and one end surface of the threaded screw through the fixing bolt; a key groove and a key are respectively provided at the inner wall of the connecting slide corresponding to the positions of one end surface of the one-way transmission and one end surface of the threaded screw, the key is slidably connected to the inner wall of the key groove, and the surface of the key and the inner wall of the key groove are respectively provided with fixing holes that are compatible with the outer dimensions of the fixing bolt.

[0016] Specifically, the disconnect mechanism further includes a driving disk arranged on the surface of one end of the threaded screw.

[0017] Specifically, the transmission assembly includes a dual-axis motor, an upper reciprocating screw rod, a lower connecting shaft rod and a transmission gear, wherein the dual-axis motor is fixedly connected to the inner wall of the column, the upper reciprocating screw rod and the lower connecting shaft rod are respectively rotatably connected to the inner wall of the column and are located on the outside of the dual-axis motor, one end of the upper reciprocating screw rod and one end of the lower connecting shaft rod are respectively fixedly connected to the output end of the dual-axis motor, and the transmission gear is fixedly connected to the surface of one end of the lower connecting shaft rod away from the dual-axis motor, and is connected to the transmission gearwheel through the toothed synchronous belt.

[0018] Specifically, the transmission assembly also includes a PLC controller, which is arranged on the surface of the glove box and located on one side of the top of the observation window. The PLC controller is electrically connected to an external power supply and is electrically connected to the dual-axis motor.

[0019] The method for using the above-mentioned cell culture device comprises the following steps:

[0020] S1. Preparation of suspension: adding a certain amount of antibiotics to the culture medium, taking animal embryos or organs and tissues, cutting the materials into pieces, and treating them with trypsin or collagenase to form dispersed single cells, transferring the treated cells into the culture medium to prepare a cell suspension of a certain concentration, and transferring the cell suspension to the test tube;

[0021] S2 culture: when the internal environment of the glove box is stable, the test tube containing the cell suspension in S1 is placed on the top of the rotating seat, and then the dual-axis motor is started. The forward and reverse rotation of the dual-axis motor synchronously links the forward and reverse rotation of the test tube. The forward and reverse rotation of the test tube makes the internal cell suspension fully mixed to avoid stratification;

[0022] S3, replacing the culture medium: after a period of cultivation, when the nutrients in the cell suspension are insufficient, the dual-axis motor is started, the dual-axis motor is operated to link the rotation seat to rotate, and the rotation of the rotation seat synchronously drives the test tube to rotate, thereby centrifuging the cell suspension at a low speed, so that the cells gather at the bottom of the test tube. At the same time, the operation of the dual-axis motor also links the liquid extraction needle to descend, and after the liquid extraction needle descends into the test tube, the liquid in the test tube is automatically extracted to realize the drainage operation. After the drainage is completed, new culture medium is added to the main liquid tube to realize the liquid addition operation;

[0023] S4. Complete the culture: Check and replace the culture medium regularly to allow the cells to complete multiple proliferations until the cell culture goal is achieved.

[0024] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the following description of the embodiments in conjunction with the accompanying drawings, in which:

[0026] Figure 1 This is a schematic diagram of the structure of a cell culture device and method of the present invention;

[0027] Figure 2 A schematic diagram of the structure of a culture chamber in a cell culture device and method of the present invention;

[0028] Figure 3 It is a schematic structural diagram of a test tube rotating seat assembly in a cell culture device and method of the present invention;

[0029] Figure 4 It is a schematic diagram of the structure of a fixing seat in a cell culture device and method of the present invention;

[0030] Figure 5 It is a schematic diagram of the structure of a liquid extraction component in a cell culture device and method of the present invention;

[0031] Figure 6 It is a schematic diagram of the structure of an extraction mechanism in a cell culture device and method of the present invention;

[0032] Figure 7 It is a schematic diagram of the structure of a transmission component in a cell culture device and method of the present invention;

[0033] Figure 8 It is a schematic diagram of the structure of a disconnection mechanism in a cell culture device and method of the present invention;

[0034] Fig. 9The figure is a schematic diagram of the structure of a one-way transmission device in a cell culture device and method of the present invention.

[0035] As shown in the figure:

[0036] 1. Glove box; 2. Test tube rotating seat assembly; 4. Liquid extraction assembly; 5. Transmission assembly;

[0037] 11. Cultivation chamber; 12. Observation window; 13. Discharge port; 14. Column;

[0038] 21. Fixed seat; 22. Rotating seat; 23. Test tube; 24. Middle gear; 25. Side gear; 26. Transmission gear; 27. Toothed synchronous belt;

[0039] 41. needle tube mechanism; 42. extraction mechanism; 43. drainage tube;

[0040] 411, lifting slide seat; 412, connecting seat; 413, liquid separation seat; 414, main liquid pipe; 415, liquid extraction needle;

[0041] 421, mounting plate frame; 422, transmission side gear; 423, one-way transmission device; 424, threaded screw; 425, horizontal slide seat; 426, liquid extraction pipe; 427, piston; 428, liquid inlet end; 429, liquid discharge end; 4210, valve; 4211, rod frame; 4212, tooth portion;

[0042] 6. Disconnection mechanism; 61. Connecting slide; 62. Fixing bolt;

[0043] 4241, driver disk;

[0044] 51. Double-axis motor; 52. Upper reciprocating screw rod; 53. Lower connecting shaft rod; 54. Transmission gear;

[0045] 7.PLC controller. DETAILED DESCRIPTION

[0046] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements with the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and are not to be construed as limitations of the present invention. On the contrary, embodiments of the present invention include all changes, modifications and equivalents that fall within the spirit and connotation of the appended claims.

[0047] A cell culture device and method according to an embodiment of the present invention will be described below with reference to the accompanying drawings.

[0048] like Figure 1-Figure 9As shown, a cell culture device according to an embodiment of the present invention comprises a glove box 1, a test tube rotating seat assembly 2, a liquid extraction assembly 4 and a transmission assembly 5, wherein a culture chamber 11 is provided in the glove box 1, and an observation window 12 and a discharge port 13 are respectively provided on one side surface of the glove box 1, and are arranged up and down, and a column 14 is integrally formed on the other side surface of the glove box 1 and is connected to the interior of the glove box 1; the test tube rotating seat assembly 2 is arranged on the bottom wall of the culture chamber 11, and one end of the test tube rotating seat assembly 2 penetrates into the interior of the glove box 1; the liquid extraction assembly 4 is arranged on the surface of the column 14, and one end of the liquid extraction assembly 4 penetrates into the interior of the culture chamber 11 and corresponds to the position of the test tube rotating seat assembly 2; the transmission assembly 5 is arranged in the interior of the column 14, one end of the transmission assembly 5 is connected to one end of the liquid extraction assembly 4, and the other end of the transmission assembly 5 is connected to one end of the test tube rotating seat assembly 2 penetrating into the interior of the glove box 1.

[0049] It should be noted that the glove box 1 described in this embodiment has a built-in sterilization lamp (not shown in the figure).

[0050] It should also be noted that a sealing door (not shown in the figure) is hingedly fixed at the discharge port 13.

[0051] Specifically, when in use, the sealing door is opened, a plurality of test tubes 23 are placed on the test tube rotating seat assembly 2, and then the sealing door is closed.

[0052] Then, the transmission assembly 5 is operated, and the operation of the transmission assembly 5 drives the test tube rotating seat assembly 2 to operate, and the operation of the test tube rotating seat assembly 2 drives the test tube 23 to rotate forward and reversely. The forward and reverse rotation of the test tube 23 allows the internal cell suspension to be fully mixed to avoid stratification.

[0053] After a period of cultivation, when the nutrients in the cell suspension are insufficient, the transmission component 5 is operated, and the transmission component 5 drives the test tube rotating seat component 2 to operate, and the test tube rotating seat component 2 drives the test tube 23 to rotate unidirectionally, thereby centrifuging the cell suspension at a low speed, so that the cells gather at the bottom of the test tube 23. At the same time, the operation of the transmission component 5 also links the operation of the liquid extraction component 4, and the liquid extraction component 4 synchronously extracts the culture fluid inside the test tube 23 when the test tube 23 rotates. After the extraction is completed, the transmission component 5 is stopped, and then new culture fluid is injected into the test tube 23 through the liquid extraction component 4.

[0054] In one embodiment of the present invention, Figure 4As shown, the test tube rotating seat assembly 2 includes a fixed seat 21, a rotating seat 22, a test tube 23, a middle gear 24, a side gear 25, a transmission gear 26 and a toothed synchronous belt 27, wherein the fixed seat 21 is fixedly connected to the inner bottom wall of the culture chamber 11, the rotating seat 22 is evenly rotatably connected to the top of the fixed seat 21, the test tubes 23 are respectively clamped and fixed on the top of the rotating seat 22, the middle gear 24 is rotatably connected to the inner wall of the glove box 1 and corresponds to the position of the fixed seat 21, the side gear 25 is evenly rotatably connected to the inner wall of the glove box 1 and is located on the outside of the middle gear 24, multiple groups of side gears 25 respectively correspond to the positions of multiple groups of rotating seats 22 and are connected, and multiple groups of side gears 25 are respectively meshed with the middle gear 24, the transmission gear 26 is fixedly connected to the outer surface of one end of the central axis of the middle gear 24 and is located on one side of the bottom of the middle gear 24, and the transmission gear 26 is connected to one end of the transmission assembly 5 through a toothed synchronous belt 27.

[0055] It should be noted that the test tube rotating seat assembly 2 described in this embodiment can drive multiple groups of test tubes 23 to rotate synchronously, realize synchronous low-speed centrifugation operation, and does not affect the extraction of culture fluid, effectively improves the operating efficiency, and has a good use effect.

[0056] Specifically, the transmission assembly 5 drives the large transmission gear 26 to rotate through the toothed synchronous belt 27, the rotation of the large transmission gear 26 synchronously drives the middle gear 24 to rotate, the rotation of the middle gear 24 synchronously drives the multiple sets of side gears 25 to rotate, the rotation of the multiple sets of side gears 25 synchronously drives the rotation of the multiple sets of rotating seats 22, the rotation of the multiple sets of rotating seats 22 synchronously drives the multiple sets of test tubes 23 to rotate, thereby realizing synchronous low-speed centrifugation operation.

[0057] In one embodiment of the present invention, Figure 3 As shown, the liquid extraction component 4 includes a needle tube mechanism 41, an extraction mechanism 42 and a drainage tube 43, wherein the needle tube mechanism 41 is arranged on the surface of the column 14 and is connected to one end of the transmission component 5, the extraction mechanism 42 is arranged on the top of the column 14, and is connected to one end of the needle tube mechanism 41 through the drainage tube 43, and one end of the needle tube mechanism 41 penetrates into the interior of the culture chamber 11 and corresponds to the position of the test tube 23.

[0058] It should be noted that the extraction mechanism 42 described in this embodiment is provided with a guide frame (not shown in the figure) for guiding the drainage pipe 43. The guide frame is provided to prevent the drainage pipe 43 from being entangled and bent, thereby affecting the conveying effect.

[0059] Specifically, the structure and connection relationship of the liquid extraction component 4 are further explained. When the needle tube mechanism 41 descends, the extraction mechanism 42 cooperates with the extraction needle tube mechanism 41 to extract the culture fluid inside the test tube 23 through the drainage tube 43.

[0060] In one embodiment of the present invention, Figure 1 , Figure 3 and Figure 5 As shown, the needle tube mechanism 41 includes a lifting slide 411, a connecting seat 412, a liquid separation seat 413, a main liquid tube 414 and a liquid extraction needle 415, wherein the lifting slide 411 is slidably connected to the surface of the column 14, one end of the lifting slide 411 penetrates into the interior of the column 14 and is connected to one end of the transmission component 5, the other end of the lifting slide 411 is fixedly connected to the connecting seat 412, the liquid separation seat 413 is fixedly connected to the inner wall of the connecting seat 412, one end of the main liquid tube 414 is arranged on the top of the liquid separation seat 413 and is connected to the interior of the liquid separation seat 413, the other end of the main liquid tube 414 penetrates out of the top of the lifting slide 411, and is connected to one end of the extraction mechanism 42 through the drain pipe 43, the liquid extraction needles 415 are evenly arranged at the bottom of the liquid separation seat 413, and are connected to the interior of the liquid separation seat 413, and one end of multiple groups of liquid extraction needles 415 respectively penetrate into the interior of the culture chamber 11 and correspond to the positions of the test tubes 23.

[0061] It should be noted that the surface of the column 14 described in this embodiment is provided with a limiting sliding groove for facilitating the vertical lifting of the lifting slide 411.

[0062] It should also be noted that a guide seat is provided at the top of the glove box 1 corresponding to the position of the liquid extraction needle 415, the liquid extraction needle 415 is slidably connected to the inner wall of the guide seat, and a sealing ring is provided at the connection with the inner wall of the guide seat.

[0063] Specifically, the lifting slide 411 descends along the surface of the column 14 driven by the transmission component 5. The descent of the lifting slide 411 synchronously drives the connecting seat 412, the liquid separation seat 413, the main liquid tube 414 and the liquid extraction needle 415 to descend, until the liquid extraction needle 415 enters the set position inside the test tube 23, and the lifting slide 411 automatically rises and resets under the drive of the transmission component 5.

[0064] When the culture fluid is extracted, the drainage tube 43 is separated from the main liquid tube 414 , and then new culture fluid is injected into the main liquid tube 414 . The culture fluid finally drips from the extraction needle 415 into the test tube 23 .

[0065] In one embodiment of the present invention, Figure 1 , Figure 3 , Figure 5-Figure 6As shown, the extraction mechanism 42 includes a mounting plate frame 421, a transmission side gear 422, a one-way transmission device 423, a threaded screw 424, a horizontal slide 425, a liquid extraction pipe 426, a piston 427, a liquid inlet end 428, a liquid discharge end 429, a valve 4210, a rod frame 4211 and a tooth portion 4212, wherein the mounting plate frame 421 is fixedly connected to the top of the column 14, the transmission side gear 422, the one-way transmission device 423 and the threaded screw 424 are rotatably connected to the surface of the mounting plate frame 421 in sequence and are connected to each other, the horizontal slide 425 is threadedly connected to the outer surface of the threaded screw 424 and is slidably connected to the inner wall of the mounting plate frame 421, the liquid extraction pipe 426 is clamped and fixed to the surface of the mounting plate frame 421 and is located on one side of the horizontal slide 425, and the piston 427 is slidably connected to the liquid extraction pipe 4 26 inner wall, one end of the central axis of the piston 427 passes through the outside of the liquid extraction tube 426 and is fixedly connected to the surface of one end of the horizontal slide 425, the liquid inlet terminal 428 is arranged at the top of one end of the liquid extraction tube 426 close to the horizontal slide 425, and is communicated with the inside of the liquid extraction tube 426, the liquid inlet terminal 428 is connected to one end of the main liquid pipe 414 through the liquid discharge pipe 43, the liquid discharge terminal 429 is arranged at the bottom of one end of the liquid extraction tube 426 close to the horizontal slide 425, and is communicated with the inside of the liquid extraction tube 426, the valve 4210 is arranged on the surface of the liquid discharge terminal 429, the rod frame 4211 is fixedly connected to the top of the lifting slide 411, and is located on one side of the transmission side gear 422, the tooth portion 4212 is arranged on the surface of the rod frame 4211, and the teeth on the tooth portion 4212 are meshed with the transmission side gear 422.

[0066] It should be noted that the one-way transmission 423 described in this embodiment is a ratchet one-way transmission, which is a prior art and will not be described in detail here.

[0067] It should also be noted that a limiting groove is provided on the mounting plate frame 421 to facilitate the horizontal sliding of the horizontal slide seat 425 .

[0068] Specifically, when in use, the lifting slide 411 descends, synchronously driving the rod frame 4211 and the tooth portion 4212 to descend, the teeth on the tooth portion 4212 descend and synchronously driving the transmission side gear 422 to rotate in the opposite direction, the transmission side gear 422 rotates and synchronously drives the threaded screw 424 to rotate through the one-way transmission device 423, the threaded screw 424 rotates and synchronously drives the horizontal slide 425 to move horizontally toward the side of the liquid extraction tube 426, the movement of the horizontal slide 425 synchronously drives the piston 427 to move toward the end away from the liquid inlet end 428, and generates a suction force, and the culture fluid inside the test tube 23 is pumped into the liquid extraction tube 426 under the cooperation of the liquid inlet end 428, the discharge tube 43, the main liquid tube 414, the liquid separation seat 413 and the liquid extraction needle 415.

[0069] The lifting slide 411 is reset and synchronously drives the rod frame 4211 and the tooth part 4212 to rise. Since a one-way transmission device 423 is provided, the positive rotation of the transmission side gear 422 will not drive the threaded screw 424 to rotate, thereby ensuring the stable position of the piston 427. When discharging the culture fluid inside the suction tube 426, just open the valve 4210 on the surface of the discharge end 429.

[0070] In one embodiment of the present invention, Figure 6 and Figure 8 As shown, the extraction mechanism 42 also includes a disconnecting mechanism 6, which includes a connecting slide 61 and a fixing bolt 62, wherein two ends of the connecting slide 61 are respectively slidably connected to one end surface of the one-way transmission 423 and one end surface of the threaded screw 424, and the two ends of the connecting slide 61 are respectively fixedly connected to one end surface of the one-way transmission 423 and one end surface of the threaded screw 424 through the fixing bolt 62; a key groove and a key are respectively provided at the positions of the inner wall of the connecting slide 61 corresponding to the positions of one end surface of the one-way transmission 423 and one end surface of the threaded screw 424, the key is slidably connected to the inner wall of the key groove, and the surface of the key and the inner wall of the key groove are respectively provided with fixing holes that are compatible with the outer dimensions of the fixing bolt 62.

[0071] It should be noted that the disconnection mechanism 6 described in this embodiment is used to cut off the connection between the one-way transmission device 423 and the threaded screw 424, which is convenient for disassembly and assembly and drives the piston 427 to reset, and has a good use effect.

[0072] Specifically, when the connection is cut off, the fixing bolt 62 connecting the connecting slide 61 and one end of the threaded screw 424 is removed, and then the connecting slide 61 is slid toward one end of the one-way transmission 423 until the connecting slide 61 is separated from one end of the threaded screw 424.

[0073] In one embodiment of the present invention, Figure 8 As shown, the disconnect mechanism 6 also includes a drive disk 4241 arranged on one end surface of the threaded screw 424 .

[0074] Specifically, the drive disk 4241 is provided to facilitate the rotation of the threaded screw 424, thereby improving the operating efficiency and achieving good use effect.

[0075] In one embodiment of the present invention, Figure 3-Figure 5 and Figure 7As shown, the transmission assembly 5 includes a dual-axis motor 51, an upper reciprocating screw rod 52, a lower connecting shaft rod 53 and a transmission gear 54, wherein the dual-axis motor 51 is fixedly connected to the inner wall of the column 14, the upper reciprocating screw rod 52 and the lower connecting shaft rod 53 are respectively rotatably connected to the inner wall of the column 14, and are located on the outside of the dual-axis motor 51, one end of the upper reciprocating screw rod 52 and one end of the lower connecting shaft rod 53 are respectively fixedly connected to the output end of the dual-axis motor 51, and the transmission gear 54 is fixedly connected to the surface of one end of the lower connecting shaft rod 53 away from the dual-axis motor 51, and is connected to the transmission gear 26 through a toothed synchronous belt 27.

[0076] It should be noted that the length of the reciprocating guide groove on the surface of the upper reciprocating screw rod 52 described in this embodiment is compatible with the stroke of the liquid extraction needle 415. When the lifting slide 411 moves to the top of the reciprocating guide groove, the liquid extraction needle 415 moves to the set position above the top of the test tube 23. When the lifting slide 411 moves to the bottom of the reciprocating guide groove, the liquid extraction needle 415 moves to the set position inside the test tube 23.

[0077] Specifically, the structure and connection relationship of the transmission assembly 5 are further explained. The dual-axis motor 51 is powered on and runs. The dual-axis motor 51 rotates and synchronously drives the upper reciprocating screw 52 and the lower connecting shaft 53 to rotate. The upper reciprocating screw 52 rotates and synchronously drives the lifting slide 411 to reciprocate up and down along the surface of the column 14. The lower connecting shaft 53 rotates and synchronously drives the transmission gear 54 to rotate. The transmission gear 54 rotates and synchronously drives the transmission large gear 26 to rotate through the toothed synchronous belt 27.

[0078] In one embodiment of the present invention, Figure 1 As shown, the transmission assembly 5 also includes a PLC controller 7, which is arranged on the surface of the glove box 1 and located on the top side of the observation window 12. The PLC controller 7 is electrically connected to an external power supply, and the PLC controller 7 is electrically connected to the dual-axis motor 51.

[0079] It should be noted that the structure and control relationship of the PLC controller 7 described in this embodiment are all prior art, so they will not be described in detail here.

[0080] It should also be noted that the germicidal lamp is arranged in parallel on the circuit between the PLC controller 7 and the dual-axis motor 51 .

[0081] Specifically, the PLC controller 7 is provided to facilitate the control of the dual-axis motor 51 to run according to the set program, and the use effect is good.

[0082] The method for using the above-mentioned cell culture device comprises the following steps:

[0083] S1. Preparation of suspension: adding a certain amount of antibiotics to the culture medium, taking animal embryos or organs and tissues, cutting the materials into pieces, and treating them with trypsin or collagenase to form dispersed single cells, transferring the treated cells into the culture medium to prepare a cell suspension of a certain concentration, and transferring the cell suspension into the test tube 23;

[0084] S2 culture: After the internal environment of the glove box 1 is stable, the test tube 23 containing the cell suspension in S1 is placed on the top of the rotating seat 22, and then the double-axis motor 51 is started. The forward and reverse rotation of the double-axis motor 51 synchronously links the forward and reverse rotation of the test tube 23. The forward and reverse rotation of the test tube 23 allows the internal cell suspension to be fully mixed to avoid stratification;

[0085] S3, replacing the culture medium: after a period of cultivation, when the nutrients in the cell suspension are insufficient, the dual-axis motor 51 is started, and the dual-axis motor 51 is operated to link the rotating seat 22 to rotate, and the rotating seat 22 rotates synchronously to drive the test tube 23 to rotate, so as to centrifuge the cell suspension at a low speed, so that the cells gather at the bottom of the test tube 23. At the same time, the operation of the dual-axis motor 51 also links the liquid extraction needle 415 to descend. After the liquid extraction needle 415 descends to the inside of the test tube 23, the liquid inside the test tube 23 is automatically extracted to realize the drainage operation. After the drainage is completed, new culture medium is added to the main liquid tube 414 to realize the liquid addition operation;

[0086] S4. Complete the culture: Check and replace the culture medium regularly to allow the cells to complete multiple proliferations until the cell culture goal is achieved.

[0087] In summary, a cell culture device and method according to an embodiment of the present invention can synchronously drive multiple groups of test tubes 23 to rotate synchronously to implement synchronous low-speed centrifugation operations. At the same time, it can also synchronously drive multiple groups of extraction needles 415 to extract the culture fluid inside the multiple groups of test tubes 23. This innovative design not only realizes the function of extracting culture fluid during low-speed centrifugation, but also effectively reduces the operation frequency, greatly improves work efficiency, and significantly reduces the labor intensity of the operator. In addition, this synchronous operation also ensures the maintenance of the cell aggregation effect, and the use effect is good.

[0088] In the description of this specification, the terms "first" and "second" are used for descriptive purposes only and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present invention, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0089] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0090] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and deform the above embodiments within the scope of the present invention.

Claims

1. A cell culture device, characterized in that: It comprises a glove box (1), a test tube rotating seat assembly (2), a liquid extraction assembly (4) and a transmission assembly (5), wherein: The glove box (1) is provided with a culture chamber (11), one side surface of the glove box (1) is provided with an observation window (12) and a discharge port (13), which are arranged up and down, and the other side surface of the glove box (1) is provided with a column (14) which is integrally formed and communicates with the interior of the glove box (1); The test tube rotating seat assembly (2) is arranged on the inner bottom wall of the culture chamber (11), and one end of the test tube rotating seat assembly (2) penetrates into the interior of the glove box (1); The liquid extraction component (4) is arranged on the surface of the column (14), one end of the liquid extraction component (4) penetrates into the interior of the culture chamber (11) and corresponds to the position of the test tube rotating seat component (2); The transmission assembly (5) is arranged inside the column (14), one end of the transmission assembly (5) is connected to one end of the liquid extraction assembly (4), and the other end of the transmission assembly (5) is connected to one end of the test tube rotating seat assembly (2) that penetrates into the glove box (1); The test tube rotating seat assembly (2) comprises a fixed seat (21), a rotating seat (22), a test tube (23), a middle gear (24), a side gear (25), a transmission gear (26) and a toothed synchronous belt (27), wherein: The fixed seat (21) is fixedly connected to the inner bottom wall of the culture chamber (11); the rotating seat (22) is evenly rotatably connected to the top of the fixed seat (21); the test tubes (23) are respectively clamped and fixed to the top of the rotating seat (22); the middle gear (24) is rotatably connected to the inner wall of the glove box (1) and corresponds to the position of the fixed seat (21); the side gear (25) is evenly rotatably connected to the inner wall of the glove box (1) and is located outside the middle gear (24); multiple groups of side gears (25) correspond to the positions of multiple groups of rotating seats (22) and are connected; multiple groups of side gears (25) are respectively meshed with the middle gear (24); the transmission gear (26) is fixedly connected to the outer surface of one end of the central axis of the middle gear (24) and is located on one side of the bottom of the middle gear (24); the transmission gear (26) is connected to one end of the transmission assembly (5) through the toothed synchronous belt (27); The liquid extraction assembly (4) comprises a needle tube mechanism (41), an extraction mechanism (42) and a liquid discharge tube (43), wherein: The needle tube mechanism (41) is arranged on the surface of the column (14) and is connected to one end of the transmission assembly (5); the extraction mechanism (42) is arranged on the top of the column (14) and is connected to one end of the needle tube mechanism (41) through the drainage tube (43); one end of the needle tube mechanism (41) penetrates into the culture chamber (11) and corresponds to the position of the test tube (23); The needle tube mechanism (41) comprises a lifting slide seat (411), a connecting seat (412), a liquid separation seat (413), a main liquid tube (414) and a liquid extraction needle (415), wherein: The lifting slide (411) is slidably connected to the surface of the column (14); one end of the lifting slide (411) penetrates into the interior of the column (14) and is connected to one end of the transmission component (5); the other end of the lifting slide (411) is fixedly connected to the connecting seat (412); the liquid separation seat (413) is fixedly connected to the inner wall of the connecting seat (412); one end of the main liquid pipe (414) is arranged on the top of the liquid separation seat (413) and is connected to the liquid separation seat (413) is connected to the inside, the other end of the main liquid pipe (414) passes through the top of the lifting slide (411) and is connected to one end of the extraction mechanism (42) through the drainage pipe (43), the liquid extraction needles (415) are evenly arranged at the bottom of the liquid separation seat (413) and are connected to the inside of the liquid separation seat (413), and one end of multiple groups of liquid extraction needles (415) respectively passes through the inside of the culture chamber (11) and corresponds to the position of the test tube (23); The transmission assembly (5) comprises a double-shaft motor (51), an upper reciprocating screw rod (52), a lower connecting shaft rod (53) and a transmission gear (54), wherein: The dual-axis motor (51) is fixedly connected to the inner wall of the column (14); the upper reciprocating screw rod (52) and the lower connecting shaft rod (53) are respectively rotatably connected to the inner wall of the column (14) and are located outside the dual-axis motor (51); one end of the upper reciprocating screw rod (52) and one end of the lower connecting shaft rod (53) are respectively fixedly connected to the output end of the dual-axis motor (51); the transmission gear (54) is fixedly connected to the surface of one end of the lower connecting shaft rod (53) away from the dual-axis motor (51) and is connected to the transmission gear (26) via the toothed synchronous belt (27).

2. The cell culture device according to claim 1, characterized in that: The extraction mechanism (42) comprises a mounting plate frame (421), a transmission side gear (422), a one-way transmission device (423), a threaded screw (424), a horizontal slide seat (425), a liquid extraction pipe (426), a piston (427), a liquid inlet end (428), a liquid discharge end (429), a valve (4210), a rod frame (4211) and a toothed portion (4212), wherein: The mounting plate frame (421) is fixedly connected to the top of the column (14); the transmission side gear (422), the one-way transmission device (423) and the threaded screw (424) are rotatably connected to the surface of the mounting plate frame (421) in sequence and are connected to each other; the horizontal slide (425) is threadedly connected to the outer surface of the threaded screw (424) and is slidably connected to the inner wall of the mounting plate frame (421); the liquid extraction tube (426) is clamped and fixed to the surface of the mounting plate frame (421) and is located on one side of the horizontal slide (425); the piston (427) is slidably connected to the inner wall of the liquid extraction tube (426); one end of the central axis of the piston (427) passes through the outside of the liquid extraction tube (426) and is fixedly connected to the surface of one end of the horizontal slide (425); the liquid inlet end (428) is arranged on The liquid extraction pipe (426) is located at the top of one end of the horizontal slide (425) and is in communication with the interior of the liquid extraction pipe (426); the liquid inlet end (428) is connected to one end of the main liquid pipe (414) through the liquid discharge pipe (43); the liquid discharge end (429) is located at the bottom of one end of the liquid extraction pipe (426) located at the horizontal slide (425) and is in communication with the interior of the liquid extraction pipe (426); the valve (4210) is located on the surface of the liquid discharge end (429); the rod frame (4211) is fixedly connected to the top of the lifting slide (411) and is located on one side of the transmission side gear (422); the tooth portion (4212) is located on the surface of the rod frame (4211); the teeth on the tooth portion (4212) are meshed with the transmission side gear (422).

3. The cell culture device according to claim 2, characterized in that: The extraction mechanism (42) further comprises a disconnection mechanism (6), wherein the disconnection mechanism (6) comprises a connecting slide cylinder (61) and a fixing bolt (62), wherein: The two ends of the connecting slide cylinder (61) are respectively slidably connected to one end surface of the one-way transmission device (423) and one end surface of the threaded screw rod (424), and the two ends of the connecting slide cylinder (61) are respectively fixedly connected to one end surface of the one-way transmission device (423) and one end surface of the threaded screw rod (424) via the fixing bolt (62); A keyway and a key are respectively provided at positions corresponding to the inner wall of the connecting slide (61) and the one end surface of the one-way transmission device (423) and the one end surface of the threaded screw (424). The key is slidably connected to the inner wall of the keyway. The surface of the key and the inner wall of the keyway are respectively provided with fixing holes that match the outer dimensions of the fixing bolt (62).

4. The cell culture device according to claim 3, characterized in that: The disconnection mechanism (6) further comprises a driving disk (4241) arranged on the surface of one end of the threaded screw rod (424).

5. The cell culture device according to claim 2, characterized in that: The transmission assembly (5) further comprises a PLC controller (7), wherein the PLC controller (7) is arranged on the surface of the glove box (1) and is located on one side of the top of the observation window (12), wherein the PLC controller (7) is electrically connected to an external power source, and wherein the PLC controller (7) is electrically connected to the dual-axis motor (51).

6. A method for using a cell culture device according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1. preparing a suspension: adding a certain amount of antibiotics to the culture medium, taking animal embryos or organs or tissues, cutting the materials into pieces, and treating them with trypsin or collagenase to form dispersed single cells, transferring the treated cells into the culture medium to prepare a cell suspension of a certain concentration, and transferring the cell suspension to the test tube (23); S2 culture: when the internal environment of the glove box (1) is stable, the test tube (23) containing the cell suspension in S1 is placed on the top of the rotating seat (22), and then the dual-axis motor (51) is started. The forward and reverse rotation of the dual-axis motor (51) synchronously links the forward and reverse rotation of the test tube (23). The forward and reverse rotation of the test tube (23) allows the internal cell suspension to be fully mixed to avoid stratification; S3, replacing the culture medium: after a period of culture, when the nutrients in the cell suspension are insufficient, the dual-axis motor (51) is started, and the dual-axis motor (51) is operated to link the rotating seat (22) to rotate, and the rotating seat (22) rotates synchronously to drive the test tube (23) to rotate, thereby centrifuging the cell suspension at a low speed, so that the cells gather at the bottom of the test tube (23). At the same time, the operation of the dual-axis motor (51) also links the liquid extraction needle (415) to descend. After the liquid extraction needle (415) descends into the test tube (23), the liquid in the test tube (23) is automatically extracted to realize the liquid drainage operation. After the liquid drainage is completed, new culture medium is added to the main liquid tube (414) to realize the liquid addition operation; S4. Complete the culture: Check and replace the culture medium regularly to allow the cells to complete multiple proliferations until the cell culture goal is achieved.

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