Plant stem cell extraction method

Abstract

The invention relates to the technical field of stem cell extraction, in particular to a plant stem cell extraction method. Comprising the following steps: SS01, grinding, taking freeze-dried plant tissues, carrying out fine grinding and wall breaking to obtain tissue fine powder, S02, extracting, putting the tissue fine powder obtained in the step SS01 into a subcritical extractor, extracting to obtain an extraction mixed solution, SS03, purifying, separating and purifying the extraction mixed solution obtained in the step SS02 to obtain a tissue stock solution, SS04, preparing a stock solution, sS03, carrying out centrifugal separation on the tissue stock solution obtained in the step SS03, taking a lower turbid solution to obtain a stem cell stock solution, SS05, extracting, and carrying out sampling extraction on the stem cell stock solution obtained in the step SS04 by using plant stem cell extraction equipment in different periods, in different regions and in different times. The stem cell extraction equipment disclosed by the invention has the beneficial effects that during sampling, through the arrangement of structures such as the back-cleaning pipe, the back-cleaning flow channel and the back-cleaning hole, the stem cell extraction equipment can realize self-cleaning of the extraction equipment when the stem cell extraction equipment is used for sampling in different regions and times or in different times.

Description

Technical Field

[0001] The present invention relates to the technical field of stem cell extraction, in particular to a method for extracting plant stem cells. Background Art

[0002] Plant stem cells are the origin of all plant cells, and plants are formed by the division of plant stem cells.

[0003] In the prior art, there have been various forms of plant stem cell extraction methods and extraction equipment. For example, the patent document with publication number CN111869564A discloses a method for extracting plant stem cells by ultra-low temperature freeze-drying and grinding. The above method uses freeze-dried plant tissue for cell extraction and then culture the plant stem cells, so that the cells can fully absorb the nutrients in the culture medium, thereby improving the culture efficiency. The freeze-dried plant tissue is used for culture, and the demand for native plant tissue is low, which is conducive to maintaining ecological balance. For example, the patent document with publication number CN218435736U discloses a plant stem cell extraction device. The above device scheme adopts a method of extracting the plant stem cells by using ... A connecting channel is provided inside to connect the accommodating chamber with the sampling bottle. The sampling end of the sampling tube is used for sampling, and the sample outlet end is connected with the sampling bottle. The piston movement causes negative pressure to be generated in the accommodating chamber. At the same time, the sampling bottle and the sampling tube generate negative pressure to suck the sample into the sampling bottle. The sampling bottle is used to hold plant stem cell samples. However, when the above-mentioned stem cell extraction device performs zoned, sequential or time-divided sampling, the sampling device does not have a self-cleaning function, which leads to problems of cell mixing and cell contamination during stem cell sampling. In addition, the existing sampling device cannot achieve an automatic homogenization effect on the sampled stem cells during sampling. Based on this, the present invention provides a plant stem cell extraction method to solve the problems raised in the above-mentioned background technology. Summary of the invention

[0004] The present invention aims at the technical problems existing in the prior art and provides a plant stem cell extraction method to solve the problem that when the existing stem cell extraction equipment performs zoned, phased or timed sampling, the sampling equipment does not have a self-cleaning function, which leads to easy cell mixing and cell contamination during stem cell sampling, and the existing sampling equipment cannot achieve an automatic homogenization effect on the sampled stem cells during sampling.

[0005] The technical solution of the present invention to solve the above technical problem is as follows: a plant stem cell extraction method, comprising the following steps: SS01, grinding, taking freeze-dried plant tissue and grinding it to break the wall to obtain fine tissue powder; S02, extraction, placing the tissue fine powder obtained in step SS01 into a subcritical extractor for extraction to obtain an extraction mixture; SS03, purification, separating and purifying the extraction mixture in step SS02 to obtain a tissue stock solution; SS04, stock solution preparation, centrifuging the tissue stock solution obtained in step SS03, taking out the turbid liquid layer, and obtaining the stem cell stock solution; SS05, extraction, using plant stem cell extraction equipment to extract the stem cell stock solution in step SS04 in different time periods, zones and times, and then extract multiple tubes of stem cell stock solution; The plant stem cell extraction device comprises a frame, a reverse cleaning pipe is fixedly installed inside the frame, a rotating pipe driven by a motor a is rotatably connected to the peripheral side of the reverse cleaning pipe, an extraction pipe is fixedly connected to the peripheral side of the rotating pipe, a reverse cleaning flow channel is fixedly provided at the axial position of the reverse cleaning pipe, a liquid storage mechanism connected to the reverse cleaning flow channel is installed on the top surface of the frame, a horizontally arranged reverse cleaning hole and an extraction flow channel are respectively provided inside the reverse cleaning pipe, both ends of the extraction flow channel are open, a liquid guide ring cavity is fixedly provided inside the rotating pipe, a positioning ring is rotatably connected to the top surface of the positioning ring, a liquid guide pipe is fixedly connected to the top surface of the positioning ring, the peripheral side of the liquid guide pipe is fixedly connected to the reverse cleaning pipe, and the inner wall of the liquid guide pipe A sample outlet tube is slidably connected, and an elastic driving component for driving the sample outlet tube to rise and fall is installed on the surface of the frame; one end of the sample outlet tube is fixedly connected to a fixed sealing cover, and a negative pressure pump is installed on the top surface of the frame, and the port of the negative pressure pump is fixedly connected to the fixed sealing cover through a corrugated connecting pipe, and the peripheral side surface of the fixed sealing cover is rotatably connected to a rotary sealing cover, and the peripheral side surface of the rotating tube is rotatably connected to a driven rotary ring, and a vibration guide component is installed on the surface of the frame, and the port of the vibration guide component is transmission-connected to a vibration frame, and the inner wall of the vibration frame is rotationally connected to a rotary ring driven by a motor b, and a group of sample storage modules distributed in a circular array are installed inside the vibration frame, and the port of the vibration guide component is transmission-connected to a guide module, and the sample storage module is driven by the guide module.

[0006] The beneficial effects of the present invention are: 1) The present invention provides a reverse cleaning tube, a reverse cleaning channel, a reverse cleaning hole and other structures during sampling, so that the stem cell extraction device can achieve self-cleaning when performing zoned, phased or timed sampling. The realization of the self-cleaning function effectively solves the problem of cell mixing and cell contamination that is easy to occur during stem cell sampling due to the lack of reverse cleaning function in traditional stem cell extraction devices. In addition, the provision of a rotating component enables the extraction device to achieve automatic homogenization of the extracted stem cells after the stem cells are extracted.

[0007] 2) When the present invention is working, through the arrangement of structures such as the semi-guide gear, the return spring, and the guide tooth plate, after the motor c outputs the rotation speed, the vibration frame and the rotating ring can reciprocate within the set stroke. By realizing the reciprocating effect of the vibration frame and the rotating ring within the set stroke, the stem cells extracted into the stem cell storage tube are effectively homogenized. Through the homogenization, the concentration uniformity of the stem cells in the stem cell storage tube is effectively improved. Moreover, through the vibration storage arrangement in the stem cell storage tube after the stem cells are extracted, the precipitation of the extracted stem cells can be effectively avoided.

[0008] 3) When the present invention performs the extraction operation of stem cell stock solution, the extraction tube and the extraction flow channel are immersed in the stem cell stock solution. During the extraction, the extraction tube is connected to the extraction flow channel. During the extraction, under the action of the negative pressure pump, the stem cell stock solution enters the designated stem cell storage test tube through the extraction flow channel, the extraction tube and the liquid guide tube for storage. When the stem cell stock solution is extracted, the tube is rotated and the extraction tube is connected to the back-clearing hole. After the extraction tube is connected to the back-clearing hole, the waste liquid storage tube is rotated to the bottom of the sample outlet tube. Subsequently, the pump body sends physiological cleaning saline into the back-clearing flow channel. After the physiological cleaning saline is injected into the back-clearing flow channel, it finally enters the extraction tube and removes the residual saline in the extraction tube. The stem cell stock solution is cleaned and brushed. When the extraction tube is cleaned with physiological cleaning saline for a specified time, the pump body is reversed and the physiological cleaning saline remaining in the extraction tube and the back-cleaning flow channel is refluxed. After the reflux of the physiological cleaning saline is completed, the liquid inlet port of the extraction tube is staggered with the extraction flow channel and the back-cleaning hole. When the next stem cell stock solution is required, the liquid inlet port of the extraction tube is reconnected with the extraction flow channel, thereby facilitating the next stem cell stock solution extraction operation. Through the above-mentioned back-cleaning process setting, on the one hand, the extraction tube can be reused, and on the other hand, the interference rate of the residual stem cell liquid on the extraction liquid during each extraction can be effectively reduced.

[0009] Based on the above technical solution, the present invention can also be improved as follows.

[0010] Furthermore, the liquid storage mechanism includes a liquid storage tank fixedly connected to the frame, physiological cleaning saline is stored in the liquid storage tank, a pump body is fixedly installed on the top surface of the liquid storage tank, the liquid outlet port of the pump body is fixedly connected to the anti-clearing flow channel through a pipeline, an electric control box is fixedly installed on the side of the liquid storage tank, a battery is built in the electric control box, a single-chip microcomputer is fixedly installed on the end surface of the electric control box, and the port of the battery is electrically connected to the single-chip microcomputer.

[0011] The beneficial effect of adopting the above further scheme is that, when working, a sufficient amount of physiological cleaning saline is stored inside the liquid storage tank, and the physiological cleaning saline is stored to clean the inner wall of the extraction tube. The battery is used to supply power to the power using mechanism in the device, and the single-chip microcomputer is used to control the working status of the corresponding power mechanism in the device in real time.

[0012] Furthermore, a sealing ring is fixedly provided at the rotating connection between the anti-cleaning pipe and the rotating pipe, the bottom axis of the extraction pipe is parallel to the axis of the anti-cleaning hole, and the extraction flow channel is an inverted L-shaped structure with rounded corners.

[0013] The beneficial effect of adopting the above further solution is that, when in use, the sealing ring is provided to effectively ensure the sealing of the rotating connection between the anti-cleaning pipe and the rotating pipe.

[0014] Furthermore, the elastic driving component includes an active pressure frame and an elastic pressure frame respectively, a group of pressure push rods are installed between the relative surfaces of the active pressure frame and the frame body, a group of guide rods slidably connected to the active pressure frame are fixedly installed on the top surface of the elastic pressure frame, and a pressure spring is sleeved on the peripheral side surface of each guide rod and at the position corresponding to the active pressure frame and the elastic pressure frame, and the inner wall of the elastic pressure frame is fixedly connected to the fixed sealing cover and the sample outlet tube respectively.

[0015] The beneficial effect of adopting the above further scheme is that when it is necessary to replace the stem cell storage tube, the pressure push rod drives the fixed cover to be fully lifted, thereby providing sufficient replacement space for the stem cell storage tube. When an unused stem cell storage tube moves to the bottom of the fixed cover, the bottom surface of the screw cover is fully fitted with the top of the stem cell storage tube under the driving action of the pressure push rod. When the stem cell storage tube reciprocates within the set stroke under the driving action of the vibration frame, the pressure spring is set so that the screw cover can move up and down synchronously with the stem cell storage tube. By setting the rotary connection of the screw cap at the fixed cap, when the stem cell storage tube is driven by the outer gear ring, the screw cap can rotate synchronously with the stem cell storage tube.

[0016] Furthermore, the vibration guide components respectively include a motor c fixed on the surface of the frame and a guide gear plate vertically arranged and fixed on the top surface of the vibration frame, the inner wall of the guide gear plate is slidably connected to the frame, the inner wall of the frame is rotatably connected with a flat shaft, the circumferential side surfaces of the flat shaft are respectively fixedly installed with intermediate bevel gears and semi-guide gears, the output shaft end of the motor c is fixedly installed with an active bevel gear transmission-connected to the intermediate bevel gear, and the circumferential side surfaces of the semi-guide gears are transmission-connected to the guide gear plate.

[0017] Furthermore, the circumferential side surface of the semi-guide gear is symmetrically provided with a toothed portion and a toothless portion, the full arc of the toothed portion and the toothless portion is 180°, the circumferential side surface of the toothed portion is transmission-connected to the guide tooth plate, a vertically arranged guide groove is fixedly provided inside the guide tooth plate, and a guide block slidably connected to the guide groove is fixedly installed on the surface of the frame.

[0018] Furthermore, the guide module includes a shaft ring fixedly connected to the frame, the inner wall of the shaft ring is rotatably connected to an outer gear ring, and the peripheral side surface of the outer gear ring is fixedly mounted with a driven bevel gear ring transmission-connected to the intermediate bevel gear.

[0019] Furthermore, the sample storage module includes a bracket fixedly connected to a rotating ring, a vertically arranged guide column is fixedly installed on the bottom surface of the bracket, the peripheral side surface of the guide column is slidably connected to the driven rotating ring, and a return spring is sleeved on the peripheral side surface of the guide column and the position corresponding to the bracket and the driven rotating ring; the inner wall of the bracket in one of the sample storage modules is rotatably connected to a waste liquid storage tube with an open top, and the inner walls of the brackets in the other sample storage modules are rotatably connected to a support tube with an open top, the peripheral sides of the support tube and the waste liquid storage tube are fixedly installed with an internal gear sleeve transmission-connected to an external gear ring, and a stem cell storage tube is clamped inside the support tube.

[0020] The beneficial effect of adopting the above further scheme is that, during operation, by setting structures such as semi-guide gears, return springs, and guide tooth plates, after the motor C outputs a rotation speed, the vibration frame and the rotating ring can reciprocate within a set stroke. By realizing the reciprocating effect of the vibration frame and the rotating ring within the set stroke, the stem cells extracted into the stem cell storage tube are effectively homogenized. Through the homogenization, the concentration uniformity of the stem cells in the stem cell storage tube is effectively improved. Moreover, by setting the vibration storage in the stem cell storage tube after the stem cells are extracted, the precipitation of the extracted stem cells can be effectively avoided.

[0021] Furthermore, the tooth height of the inner gear sleeve is 5 to 10 times the tooth height of the outer gear ring.

[0022] The beneficial effect of adopting the above further scheme is that by setting the height difference of the teeth of the inner gear sleeve and the outer gear ring, the outer gear ring can continuously and effectively transmit the inner gear sleeve when the bracket and the inner gear ring move back and forth up and down, and by setting the rotational state of the stem cell storage tube during stem cell storage, the adhesion or adhesion rate of stem cells to the inner wall of the stem cell storage tube can be effectively reduced, thereby facilitating the complete discharge of stem cells from the inside of the stem cell storage tube after storage.

[0023] Furthermore, driven gear rings are fixedly installed on the circumferential side surfaces of the rotating tube and the rotating ring, and transmission gears are fixedly installed on the output shaft ends of the motor a and the motor b. The circumferential side surfaces of the two transmission gears are respectively connected to the driven gear rings at the corresponding positions, and the tooth height of the transmission gear at the motor b is 6 to 10 times the tooth height of the driven gear ring at the rotating ring. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a schematic diagram of the overall structure of the plant stem cell extraction device used in the plant stem cell extraction method of the present invention; Figure 2 For the present invention Figure 1 A schematic diagram of the local enlarged structure at point A in the middle; Figure 3 For the present invention Figure 1 A schematic diagram of the local enlarged structure at B in the middle; Figure 4 For the present invention Figure 1 A schematic diagram of the cross-sectional structure of; Figure 5 For the present invention Figure 4 A schematic diagram of the local enlarged structure at C in the middle; Figure 6 For the present invention Figure 4 A schematic diagram of the local enlarged structure at D in the middle; Figure 7 It is a schematic diagram of the cross-sectional structure of the reverse cleaning pipe and the pipe rotation of the present invention; Figure 8 It is a schematic diagram of the structure of the rotating tube and the inner gear sleeve of the present invention; Fig. 9 It is a schematic structural diagram of the negative pressure pump and the guide rod of the present invention; Fig.10 It is a schematic diagram of the cross-sectional structure of the sample tube of the present invention; Fig.11 It is a schematic diagram of the process structure of the plant stem cell extraction method of the present invention.

[0025] In the accompanying drawings, the components represented by the reference numerals are listed as follows: 1. Plant stem cell extraction equipment; 2. Frame; 3. Anti-cleaning tube; 4. Motor a; 5. Rotating tube; 6. Extraction tube; 7. Anti-cleaning hole; 8. Extraction flow channel; 9. Liquid guide ring cavity; 10. Positioning ring; 11. Liquid guide tube; 12. Sample outlet tube; 13. Fixed sealing cover; 14. Negative pressure pump; 15. Rotary sealing cover; 16. Driven rotary ring; 17. Vibration frame; 18. Motor b; 19. Rotary ring; 20. Liquid storage tank; 21. Pump body; 22. Electric control box ; 23. Single chip microcomputer; 24. Active pressure frame; 25. Elastic pressure frame; 26. Pressure push rod; 27. Guide rod; 28. Pressure spring; 29. ​​Motor c; 30. Guide gear plate; 31. Flat shaft; 32. Intermediate bevel gear; 33. Semi-guide gear; 34. Shaft ring; 35. Outer gear ring; 36. Driven bevel gear ring; 37. Bracket; 38. Guide column; 39. Return spring; 40. Waste liquid storage pipe; 41. Bracket; 42. Inner gear sleeve. DETAILED DESCRIPTION

[0026] The principles and features of the present invention are described below in conjunction with the accompanying drawings. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0027] The present invention provides the following preferred embodiments like Figure 1-11As shown, the plant stem cell extraction method comprises the following steps: SS01, grinding, taking freeze-dried plant tissue and grinding it to break the wall to obtain fine tissue powder; S02, extraction, placing the tissue fine powder obtained in step SS01 into a subcritical extractor for extraction to obtain an extraction mixture; SS03, purification, separating and purifying the extraction mixture in step SS02 to obtain a tissue stock solution; SS04, stock solution preparation, centrifuging the tissue stock solution obtained in step SS03, taking out the turbid liquid layer, and obtaining the stem cell stock solution; SS05, extraction, using the plant stem cell extraction device 1 to extract the stem cell stock solution in step SS04 in different time periods, zones and times, and then extract multiple tubes of stem cell stock solution; The plant stem cell extraction device 1 comprises a frame 2, a reverse cleaning pipe 3 is fixedly installed inside the frame 2, and a rotating pipe 5 driven by a motor a4 is rotatably connected to the peripheral side of the reverse cleaning pipe 3; A sealing ring is fixedly provided at the rotation connection between the back-cleaning pipe 3 and the rotating pipe 5 , and the axis of the bottom end of the extraction pipe 6 is parallel to the axis of the back-cleaning hole 7 .

[0028] When in use, the sealing ring is provided to effectively ensure the sealing of the rotating connection between the anti-cleaning pipe 3 and the rotating pipe 5; The peripheral side of the rotating tube 5 is fixedly connected to an extraction tube 6, the axis position of the back-cleaning tube 3 is fixedly provided with a back-cleaning flow channel, and the top surface of the frame 2 is equipped with a liquid storage mechanism connected to the back-cleaning flow channel; The liquid storage mechanism includes a liquid storage tank 20 fixedly connected to the frame 2, and physiological cleaning saline is stored in the liquid storage tank 20. A pump body 21 is fixedly installed on the top surface of the liquid storage tank 20, and the liquid outlet port of the pump body 21 is fixedly connected to the anti-clearing flow channel through a pipeline. An electric control box 22 is fixedly installed on the side of the liquid storage tank 20, and a battery is built in the electric control box 22. A single-chip microcomputer 23 is fixedly installed on the end surface of the electric control box 22, and the port of the battery is electrically connected to the single-chip microcomputer 23.

[0029] During operation, the liquid storage tank 20 stores a sufficient amount of physiological cleaning saline solution, which is used to clean the inner wall of the extraction tube 6. The storage battery is used to supply power to the power using mechanism in the device, and the single chip microcomputer 23 is used to control the working state of the corresponding power mechanism in the device in real time. The interior of the anti-cleaning pipe 3 is provided with a horizontal anti-cleaning hole 7 and an extraction channel 8, respectively. The extraction channel 8 is an inverted L-shaped structure with rounded corners, and both ends of the extraction channel 8 are open; When the stem cell stock solution is extracted, the extraction tube 6 and the extraction channel 8 are immersed in the stem cell stock solution. During the extraction, the extraction tube 6 is connected with the extraction channel 8. During the extraction, under the action of the negative pressure pump 14, the stem cell stock solution enters the designated stem cell storage tube through the extraction channel 8, the extraction tube 6 and the liquid guide tube 11 for storage; When the stem cell stock solution is extracted, the rotating tube 5 rotates and connects the extraction tube 6 with the back-cleaning hole 7. When the extraction tube 6 is connected with the back-cleaning hole 7, the waste liquid storage tube 40 rotates to the bottom of the sample outlet tube 12. Then, the pump body 21 sends physiological cleaning saline into the back-cleaning channel. After the physiological cleaning saline is injected into the back-cleaning channel, it finally enters the extraction tube 6 and cleans the residual stem cell stock solution in the extraction tube 6. When the extraction tube 6 is cleaned with the physiological cleaning saline for a specified time, the pump body 21 reverses and refluxes the residual physiological cleaning saline in the extraction tube 6 and the back-cleaning channel. After the reflux of the physiological cleaning saline is completed, the liquid inlet port of the extraction tube 6 is staggered with the extraction channel 8 and the back-cleaning hole 7. When the next extraction of the stem cell stock solution is required, the liquid inlet port of the extraction tube 6 is reconnected with the extraction channel 8, so as to facilitate the next extraction of the stem cell stock solution. Through the above-mentioned backwashing process setting, on the one hand, the extraction tube 6 can be reused, and on the other hand, the interference rate of the residual stem cell liquid on the extraction liquid during each extraction can be effectively reduced; A liquid guide ring cavity 9 is fixedly provided inside the rotating tube 5, and a positioning ring 10 is rotatably connected to the top surface of the liquid guide ring cavity 9. A liquid guide tube 11 is fixedly connected to the top surface of the positioning ring 10, and the peripheral side surface of the liquid guide tube 11 is fixedly connected to the anti-cleaning tube 3; The inner wall of the liquid guiding tube 11 is slidably connected with a sample outlet tube 12, and the surface of the frame 2 is equipped with an elastic driving component for driving the sample outlet tube 12 to rise and fall; The elastic driving components include an active pressure frame 24 and an elastic pressure frame 25. A group of pressure push rods 26 are installed between the active pressure frame 24 and the relative surface of the frame body 2. A group of guide rods 27 slidably connected to the active pressure frame 24 are fixedly installed on the top surface of the elastic pressure frame 25. A pressure spring 28 is sleeved on the circumferential side surface of each guide rod 27 and corresponding to the position between the active pressure frame 24 and the elastic pressure frame 25. The inner wall of the elastic pressure frame 25 is fixedly connected to the fixed sealing cover 13 and the sample outlet tube 12 respectively.

[0030] When the stem cell storage tube needs to be replaced, the pressure push rod 26 drives the fixed cover 13 to be fully lifted, thereby providing sufficient replacement space for the stem cell storage tube. When an unused stem cell storage tube moves to the bottom of the fixed cover 13, the bottom surface of the screw cover 15 is fully fitted with the top of the stem cell storage tube under the driving action of the pressure push rod 26. When the stem cell storage tube reciprocates within the set stroke under the driving action of the vibration frame 17, the pressure spring 28 is set to enable the screw cover 15 to move up and down synchronously with the stem cell storage tube. The rotary sealing cover 15 is connected to the fixed sealing cover 13 so that when the stem cell storage tube is driven by the outer gear ring 35, the rotary sealing cover 15 can rotate synchronously with the stem cell storage tube. One end of the sample outlet tube 12 is fixedly connected to a fixed sealing cover 13, a negative pressure pump 14 is installed on the top surface of the frame 2, and the port of the negative pressure pump 14 is fixedly connected to the fixed sealing cover 13 through a corrugated connecting pipe, the peripheral side surface of the fixed sealing cover 13 is rotatably connected to a rotary sealing cover 15, and the peripheral side surface of the rotating tube 5 is rotatably connected to a driven rotary ring 16, a vibration guide component is installed on the surface of the frame 2, and the port of the vibration guide component is transmission-connected to a vibration frame 17, and the inner wall of the vibration frame 17 is rotationally connected to a rotary ring 19 driven by a motor b18, and a group of sample storage modules distributed in a circular array are installed inside the vibration frame 17, and the port of the vibration guide component is transmission-connected to a guide module, and the sample storage module is driven by the guide module.

[0031] The vibration guide components include a motor c29 fixed on the surface of the frame 2 and a guide gear plate 30 vertically arranged and fixed on the top surface of the vibration frame 17. The inner wall of the guide gear plate 30 is slidably connected to the frame 2. The inner wall of the frame 2 is rotatably connected to a flat shaft 31. The circumferential side surfaces of the flat shaft 31 are respectively fixedly installed with intermediate bevel teeth 32 and semi-guide gears 33. The output shaft end of the motor c29 is fixedly installed with active bevel teeth that are transmission-connected to the intermediate bevel teeth 32. The circumferential side surfaces of the semi-guide gears 33 are transmission-connected to the guide gear plate 30.

[0032] The circumferential side surface of the semi-guide gear 33 is symmetrically provided with a toothed portion and a toothless portion, and the full arc of the toothed portion and the toothless portion is 180°. The circumferential side surface of the toothed portion is transmission-connected to the guide gear plate 30, and a vertically-disposed guide groove is fixedly provided inside the guide gear plate 30, and a guide block slidably connected to the guide groove is fixedly installed on the surface of the frame 2.

[0033] The guide module includes a shaft ring 34 fixedly connected to the frame 2 , an outer gear ring 35 is rotatably connected to the inner wall of the shaft ring 34 , and a driven bevel gear ring 36 drivingly connected to the intermediate bevel gear 32 is fixedly mounted on the peripheral side of the outer gear ring 35 .

[0034] The sample storage module includes a bracket 37 fixedly connected to the rotating ring 19, a vertically arranged guide column 38 is fixedly installed on the bottom surface of the bracket 37, the peripheral side surface of the guide column 38 is slidably connected to the driven rotating ring 16, and a return spring 39 is sleeved on the peripheral side surface of the guide column 38 and the position corresponding to the bracket 37 and the driven rotating ring 16. The inner wall of the bracket 37 in one sample storage module is rotatably connected to a waste liquid storage tube 40 with an open top, and the inner walls of the bracket 37 in other sample storage modules are rotatably connected to a support tube 41 with an open top, and the peripheral sides of the support tube 41 and the waste liquid storage tube 40 are fixedly installed with an inner gear sleeve 42 that is transmission-connected to the outer gear ring 35, the tooth height of the inner gear sleeve 42 is 8 times the tooth height of the outer gear ring 35, and a stem cell storage test tube is clamped inside the support tube 41.

[0035] During operation, through the arrangement of structures such as the semi-guide gear 33, the return spring 39, and the guide gear plate 30, after the motor c29 outputs the rotation speed, the vibration frame 17 and the rotating ring 19 can reciprocate within the set stroke. By realizing the reciprocating effect of the vibration frame 17 and the rotating ring 19 within the set stroke, the stem cells extracted into the stem cell storage tube are effectively homogenized. Through the homogenization, the concentration uniformity of the stem cells in the stem cell storage tube is effectively improved. Moreover, through the vibration storage arrangement in the stem cell storage tube after the stem cell extraction, the precipitation of the extracted stem cells can be effectively avoided.

[0036] By setting the height difference between the teeth of the inner gear sleeve 42 and the outer gear ring 35, the outer gear ring 35 can continuously and effectively transmit the inner gear sleeve 42 when the support tube 41 and the inner gear ring move back and forth up and down. By setting the rotation state of the stem cell storage tube during stem cell storage, the adhesion or adhesion rate of stem cells to the inner wall of the stem cell storage tube can be effectively reduced, thereby facilitating the complete discharge of stem cells from the inside of the stem cell storage tube after storage.

[0037] The circumferential side surfaces of the rotating tube 5 and the rotating ring 19 are fixedly installed with driven gear rings, and the output shaft ends of the motors a4 and b18 are fixedly installed with transmission gears. The circumferential side surfaces of the two transmission gears are respectively connected to the driven gear rings at the corresponding positions. The tooth height of the transmission gear at the motor b18 is 8 times the tooth height of the driven gear ring at the rotating ring 19.

[0038] In summary: The beneficial effects of the present invention are specifically embodied in The present invention arranges structures such as a reverse cleaning tube, a reverse cleaning flow channel, and a reverse cleaning hole during sampling, so that the stem cell extraction device can achieve self-cleaning when performing zoned, phased, or time-divided sampling. The realization of the self-cleaning function effectively solves the problem that traditional stem cell extraction devices are prone to cell mixing and cell contamination during stem cell sampling due to the lack of a reverse cleaning function. In addition, the arrangement of a rotating component enables the extraction device to achieve automatic homogenization of the extracted stem cells after the stem cells are extracted.

Claims

1. A method for extracting plant stem cells, characterized in that: The following steps are involved: SS01, grinding, taking freeze-dried plant tissue and grinding it to break the wall to obtain fine tissue powder; S02, extraction, placing the tissue fine powder obtained in step SS01 into a subcritical extractor for extraction to obtain an extraction mixture; SS03, purification, separating and purifying the extraction mixture in step SS02 to obtain a tissue stock solution; SS04, stock solution preparation, centrifuging the tissue stock solution obtained in step SS03, taking out the turbid liquid layer, and obtaining the stem cell stock solution; SS05, extraction, using the plant stem cell extraction equipment (1) to extract the stem cell stock solution in step SS04 in different time periods, zones and times, and then extract multiple tubes of stem cell stock solution; The plant stem cell extraction device (1) comprises a frame (2), a vibration guide component is installed on the surface of the frame (2), the port of the vibration guide component is connected to a vibration frame (17) in a transmission manner, the inner wall of the vibration frame (17) is rotatably connected to a rotating ring (19) driven by a motor b (18), a group of sample storage modules distributed in a circular array are installed inside the vibration frame (17), the port of the vibration guide component is connected to a guide module in a transmission manner, and the sample storage module is driven by the guide module.

2. The plant stem cell extraction method according to claim 1, characterized in that: The interior of the anti-cleaning pipe (3) is provided with a horizontally arranged anti-cleaning hole (7) and an extraction flow channel (8), both ends of the extraction flow channel (8) are open, and the interior of the rotating tube (5) is fixedly provided with a liquid guide ring cavity (9), and the top surface of the liquid guide ring cavity (9) is rotatably connected to a positioning ring (10).

3. The plant stem cell extraction method according to claim 2, characterized in that: The top surface of the positioning ring (10) is fixedly connected to a liquid guide tube (11), the peripheral side surface of the liquid guide tube (11) is fixedly connected to the anti-cleaning tube (3), the inner wall of the liquid guide tube (11) is slidably connected to a sample outlet tube (12), and the surface of the frame (2) is installed with an elastic driving component for driving the sample outlet tube (12) to rise and fall; One end of the sample outlet tube (12) is fixedly connected to a fixed sealing cover (13); a negative pressure pump (14) is installed on the top surface of the frame (2); a port of the negative pressure pump (14) is fixedly connected to the fixed sealing cover (13) via a corrugated connecting pipe; a peripheral side surface of the fixed sealing cover (13) is rotatably connected to a rotary sealing cover (15); and a peripheral side surface of the rotary tube (5) is rotatably connected to a driven rotary ring (16); The liquid storage mechanism comprises a liquid storage box (20) fixedly connected to the frame (2), wherein physiological cleaning saline is stored in the liquid storage box (20), a pump body (21) is fixedly mounted on the top surface of the liquid storage box (20), a liquid outlet port of the pump body (21) is fixedly connected to a reverse cleaning channel via a pipeline, an electric control box (22) is fixedly mounted on the side surface of the liquid storage box (20), a storage battery is built in the electric control box (22), a single-chip microcomputer (23) is fixedly mounted on the end surface of the electric control box (22), and a port of the storage battery is electrically connected to the single-chip microcomputer (23); A reverse cleaning pipe (3) is fixedly installed inside the frame (2); a rotating pipe (5) driven by a motor a (4) is rotatably connected to the peripheral side of the reverse cleaning pipe (3); an extraction pipe (6) is fixedly connected to the peripheral side of the rotating pipe (5); a reverse cleaning flow channel is fixedly opened at the axial position of the reverse cleaning pipe (3); and a liquid storage mechanism connected to the reverse cleaning flow channel is installed on the top surface of the frame (2); A sealing ring is fixedly provided at the rotational connection between the anti-cleaning pipe (3) and the rotating pipe (5); the bottom axis of the extraction pipe (6) is parallel to the axis of the anti-cleaning hole (7); and the extraction flow channel (8) is an inverted L-shaped structure with rounded corners; The elastic driving component comprises an active pressure frame (24) and an elastic pressure frame (25), a group of pressure push rods (26) are installed between the active pressure frame (24) and the relative surface of the frame body (2), and a group of guide rods (27) slidably connected to the active pressure frame (24) are fixedly installed on the top surface of the elastic pressure frame (25); The vibration guide components respectively include a motor c (29) fixed to the surface of the frame (2) and a guide tooth plate (30) vertically arranged and fixed to the top surface of the vibration frame (17); the inner wall of the guide tooth plate (30) is slidably connected to the frame (2); the inner wall of the frame (2) is rotatably connected to a flat shaft (31); and the peripheral side surfaces of the flat shaft (31) are respectively fixedly mounted with intermediate bevel teeth (32) and a semi-guide gear (33); The circumferential side surface of the semi-guide gear (33) is symmetrically provided with a toothed portion and a toothless portion, the full arc of the toothed portion and the toothless portion is 180°, the circumferential side surface of the toothed portion is transmission-connected to the guide tooth plate (30), a vertically-disposed guide groove is fixedly provided inside the guide tooth plate (30), and a guide block slidably connected to the guide groove is fixedly mounted on the surface of the frame (2); The guide module comprises a shaft ring (34) fixedly connected to the frame (2); an outer gear ring (35) is rotatably connected to the inner wall of the shaft ring (34); and a driven bevel gear ring (36) drivingly connected to the intermediate bevel gear (32) is fixedly mounted on the peripheral side surface of the outer gear ring (35).

4. The method for extracting plant stem cells according to claim 3, characterized in that: A pressure spring (28) is sleeved on the peripheral side surface of each guide rod (27) and at a position corresponding to the active pressure frame (24) and the elastic pressure frame (25), and the inner wall of the elastic pressure frame (25) is fixedly connected to the fixed sealing cover (13) and the sample outlet tube (12) respectively.

5. The method for extracting plant stem cells according to claim 3, characterized in that: The output shaft end of the motor c (29) is fixedly mounted with a driving bevel gear that is transmission-connected to the intermediate bevel gear (32), and the peripheral side surface of the semi-guide gear (33) is transmission-connected to the guide gear plate (30).

6. The method for extracting plant stem cells according to claim 5, characterized in that: The sample storage module comprises a bracket (37) fixedly connected to the rotating ring (19), a vertically arranged guide column (38) being fixedly mounted on the bottom surface of the bracket (37), and a peripheral side surface of the guide column (38) being slidably connected to the driven rotating ring (16).

7. The method for extracting plant stem cells according to claim 6, characterized in that: A return spring (39) is sleeved on the peripheral side surface of the guide column (38) and at a position corresponding to the bracket (37) and the driven rotating ring (16); the inner wall of the bracket (37) in one of the sample storage modules is rotatably connected to a waste liquid storage tube (40) with an opening at the top, and the inner walls of the brackets (37) in the other sample storage modules are all rotatably connected to a support tube (41) with an opening at the top.

8. The method for extracting plant stem cells according to claim 7, characterized in that: An inner gear sleeve (42) drivingly connected to the outer gear ring (35) is fixedly mounted on the peripheral side surfaces of the support cylinder (41) and the waste liquid storage tube (40), and a stem cell storage test tube is clamped inside the support cylinder (41).

9. The method for extracting plant stem cells according to claim 8, characterized in that: The tooth height of the inner gear sleeve (42) is 5 to 10 times the tooth height of the outer gear ring (35).

10. The method for extracting plant stem cells according to claim 3, characterized in that: The circumferential side surfaces of the rotating tube (5) and the rotating ring (19) are both fixedly mounted with driven gear rings, and the output shaft ends of the motor a (4) and the motor b (18) are both fixedly mounted with transmission gears, and the circumferential side surfaces of the two transmission gears are respectively connected to the driven gear rings at corresponding positions, and the tooth height of the transmission gear at the motor b (18) is 6 to 10 times the tooth height of the driven gear ring at the rotating ring (19).

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