Extraction operation method of snail stem cells

Abstract

The invention relates to the technical field of biological extraction, and particularly discloses a snail stem cell extraction operation method which comprises the steps of snail pretreatment, stem cell extraction, tissue enzymolysis treatment, cell culture and stem cell storage. By integrating enzymolysis, stirring, magnetic force, electric wave and other operations acting on a single sample, the extraction efficiency of snail stem cells is greatly improved, the consistency and representativeness of the extraction process are optimized, magnetic stirring and electric wave emission are combined, the enzymolysis process is accelerated by utilizing the force, stem cell release and automatic control of equipment are promoted, and the extraction efficiency of snail stem cells is improved. By means of precise driving of the electric cylinder and the motor, the operation process is simplified, possible errors caused by manual intervention are reduced, and the operation precision and the intelligent level of equipment are remarkably improved.

Description

Technical Field

[0001] The invention relates to the technical field of biological extraction, and in particular to an extraction operation method of snail stem cells. Background Art

[0002] At present, most snail stem cell extraction technologies remain at the traditional centrifugation stage, lacking efficient, automated, and convenient operation methods. Some studies have tried to use micro-magnetic stirrers to accelerate enzymatic hydrolysis reactions, but there are still limitations such as difficulty in full automation and inability to achieve batch operations.

[0003] Existing snail stem cell extraction technology has some major drawbacks:

[0004] 1. Low extraction efficiency: Traditional methods rely on manual operation and cumbersome centrifugation, which makes it difficult to ensure enzymatic efficiency and stem cell separation rate. It is time-consuming and labor-intensive, and difficult to achieve large-scale production.

[0005] 2. The operation is complicated and time-consuming: The extraction process is cumbersome, the operator's technical requirements are high, and environmental factors such as temperature and humidity need to be strictly controlled. The extraction time is also long, which leads to a decrease in the activity of stem cells.

[0006] 3. Low degree of automation: The extraction equipment mainly relies on manual control and lacks automated devices, resulting in low efficiency and difficulty in meeting large-scale cell resource needs.

[0007] Therefore, the present invention aims to provide an efficient, automated and convenient snail stem cell extraction method and equipment, which can effectively improve the efficiency of stem cell extraction, reduce the difficulty of operation, and realize batch operation, so as to meet the demand for snail stem cell resources in the scientific research and medical fields. Summary of the invention

[0008] In view of the above situation, in order to overcome the defects of the prior art, the present invention provides a method for extracting snail stem cells to solve the above-mentioned technical defects.

[0009] To achieve the above objectives, the present invention is implemented by the following technical scheme: a method for extracting snail stem cells, comprising the following steps:

[0010] Step 1: Snail pretreatment: Select healthy and active snails as the extraction object, let the snail rest and remove the debris in the body;

[0011] Step 2: Stem cell extraction: Using sterile surgical instruments, extract tissue samples from specific parts of the snail;

[0012] Step 3: Tissue enzymatic hydrolysis: The extracted tissue sample is sent to an extraction device for enzymatic hydrolysis to separate the stem cells, and the stem cells are further purified through centrifugation and filtration steps;

[0013] Step 4: Cell culture: Use a culture medium suitable for stem cell growth to inoculate the extracted stem cells into a culture dish and maintain a constant temperature, humidity and CO2 concentration;

[0014] Step 5: Stem cell storage: cryopreserve the cultured stem cells using a cryoprotectant containing a protective agent and gradually cool them down according to the freezing procedure;

[0015] Specifically, the extraction equipment includes an extraction frame, a top frame is fixedly arranged on the top of the extraction frame, and a rotating frame is rotatably arranged inside the extraction frame, a driving frame is fixedly arranged on the bottom of the extraction frame, and a rotating motor for driving the rotating frame to rotate is arranged inside the driving frame, and the output shaft of the driving motor is fixedly connected to the middle part of the rotating frame, a plurality of separation tanks are fixedly arranged on the top of the rotating frame, and culture tanks are rotatably arranged inside the plurality of separation tanks, and stirrers are placed inside the plurality of culture tanks; after the enzyme solution is sent into the culture tank, an enzymatic hydrolysis reaction occurs between the snail extracted tissue sample and the enzyme solution inside the culture tank, at this time, the rotating frame is controlled to rotate 60 degrees, the culture tank is rotated to just above the first magnetic stirrer, the movable frame at the bottom of the first magnetic stirrer is controlled to move upward, the magnetic stirrer is used to control the stirrer inside the culture tank to rotate, and the enzyme solution and the snail extracted tissue sample inside the culture tank are magnetically stirred to make the enzyme fully contact with the tissue sample.

[0016] Furthermore, servo electric cylinders are fixedly provided on both sides of the bottom of the top frame, a lifting frame is movably provided below the top frame, and driving ends of two servo electric cylinders are fixedly connected to the top of the lifting frame, a material guide pipe is fixedly provided on one side of the top of the lifting frame, and a material extraction pipe is also fixedly provided on the other side of the top of the lifting frame, and the top ends of the material guide pipe and the material extraction pipe extend to the top of the top frame.

[0017] Furthermore, two movable frames are provided on one side of the inner wall of the extraction frame for sliding up and down through an electric slide, and magnetic stirrers are fixedly provided on the top of the two movable frames, a plurality of through grooves matching the separation tank are provided at the bottom of the rotating frame, a micro electric cylinder 1 is fixedly provided on one side of the top of the lifting frame, a driving end of the micro electric cylinder 1 extends to the bottom of the lifting frame, and a radio frequency wave generator is fixedly provided on the driving end of the micro electric cylinder 1.

[0018] Furthermore, a fixed extraction tube is fixedly provided on one side of the top of the lifting frame, and the bottom end of the fixed extraction tube extends to the bottom of the lifting frame, a movable extraction tube is slidably provided on the bottom end of the fixed extraction tube, and a servo electric cylinder 2 is fixedly provided on one side of the top of the lifting frame, and the driving end of the servo electric cylinder 2 is fixedly connected to one side of the movable extraction tube.

[0019] Furthermore, two micro electric cylinders 2 are fixedly arranged on one side of the top of the lifting frame, and a connecting frame is fixedly arranged on the top of the driving ends of the two micro electric cylinders 2, a centrifugal motor is fixedly arranged on the bottom of the connecting frame, the bottom end of the output shaft of the centrifugal motor extends to the bottom of the lifting frame, and a driving gear is fixedly arranged on the bottom end of the output shaft of the centrifugal motor, an external tooth groove matching the driving gear is arranged on the top of the outer peripheral surface of the culture tank, centrifugal holes are arranged on both sides of the inner wall of the culture tank, micro electric cylinders 3 are fixedly arranged on both sides of the inner wall of the separation tank, and closing plates are fixedly arranged on the driving ends of the micro electric cylinders 3 on both sides, and the closing plates on both sides are respectively fitted with the centrifugal holes on both sides of the inner wall of the culture tank.

[0020] Furthermore, a discharge port is provided at the bottom of the rotating frame, and the interiors of the plurality of separation tanks are communicated with the interior of the discharge port.

[0021] Furthermore, the method for using the extraction device in step 3 is as follows:

[0022] After the enzyme solution is sent into the culture tank, an enzymatic hydrolysis reaction occurs between the snail extracted tissue sample and the enzyme solution in the culture tank. At this time, the rotating frame is controlled to rotate 60 degrees, and the culture tank is rotated to the top of the first magnetic stirrer. The movable frame at the bottom of the first magnetic stirrer is controlled to move upward, and the magnetic stirrer is used to control the stirring bar in the culture tank to rotate, and the enzyme solution and the snail extracted tissue sample in the culture tank are magnetically stirred, so that the enzyme and the tissue sample are fully in contact;

[0023] When the enzymatic hydrolysis reaction proceeds to a certain extent, the rotating frame is then controlled to rotate 60 degrees, so that the enzyme solution and snail extracted tissue sample stirred by the magnetic force are moved to the top of the second magnetic stirrer. At this time, the stirring bar inside the culture tank is continued to be controlled by the second magnetic stirrer to rotate at a low speed. At the same time, the driving end of the micro-electric cylinder 1 controls the radio frequency wave generator to move downward, and the radio frequency wave generator is turned on to generate electric waves for the enzyme solution and the extracted tissue sample inside the culture tank, so as to generate tiny vibrations inside the tissue, increase the contact area between the enzyme and the tissue, and thus accelerate the enzymatic hydrolysis reaction.

[0024] After completing the separation of stem cells inside the snail tissue sample, the rotating frame is controlled to rotate 60 degrees so that the separation tank is rotated to the bottom of the driving gear, and the driving end of the second micro-electric cylinder is used to control the connecting frame to move downward so that the surface of the driving gear and the outer peripheral surface of the culture tank are meshed for transmission, and the output shaft of the centrifugal motor is used in conjunction with the driving gear to control the culture tank to rotate inside the separation tank, and the stem cells inside the separation tank are centrifugally filtered. The rotating frame is then controlled to rotate 60 degrees so that the culture tank is rotated to the bottom of the extraction tube and the fixed extraction tube, and the driving end of the second servo electric cylinder is used to control the movable extraction tube to move downward, and the bottom end of the movable extraction tube is used to extract the stem cells inside the culture tank, and then the cleaning agent is introduced into the interior of the culture tank through the extraction tube to rinse the interior of the culture tank, and the culture tank is continued to be used to automatically extract stem cells from the snail tissue sample.

[0025] The beneficial effects achieved by the present invention using the above structure are as follows:

[0026] 1. The present invention integrates multiple operations such as enzymatic hydrolysis, stirring, magnetic force, and radio waves on a single sample, which not only greatly improves the extraction efficiency of snail stem cells, but also optimizes the consistency and representativeness of the extraction process. The magnetic stirring and radio wave emission are combined to accelerate the enzymatic hydrolysis process and promote the release of stem cells. The automated control of the equipment, such as the precise drive of the electric cylinder and the motor, simplifies the operation process, reduces the possible errors caused by manual intervention, and significantly improves the accuracy of the operation and the intelligence level of the equipment.

[0027] 2. In the present invention, when extracting stem cells from tissue samples, a magnetic stirrer is used to control the stirring bar inside the culture tank to rotate, and the enzyme solution and the snail extracted tissue sample inside the culture tank are magnetically stirred, so that the enzyme is fully in contact with the tissue sample, and the enzymatic hydrolysis efficiency is improved. At the same time, stirring can also accelerate the diffusion of the enzyme in the solution, so that the enzyme is more evenly distributed in the tissue sample, thereby accelerating the degradation of the extracellular matrix and the release of stem cells. In addition, stirring can avoid the situation where the enzyme concentration is too high or too low locally in the centrifuge tube, and ensure that the enzymatic hydrolysis reaction is carried out evenly throughout the sample, which helps to improve the consistency of enzymatic hydrolysis and reduce the problem of insufficient enzymatic hydrolysis or excessive enzymatic hydrolysis of some tissues due to concentration differences. In addition, magnetic stirring can make the solution temperature in the culture tank more uniform, avoid local overheating or overcooling, thereby ensuring the activity and stability of the enzyme.

[0028] 3. The present invention combines magnetic stirring with radio wave assistance, and utilizes a radio frequency generator to generate radio waves for the enzyme solution and the extracted tissue sample inside the culture tank, so as to generate tiny vibrations inside the tissue, increase the contact area between the enzyme and the tissue, and thus accelerate the enzymatic hydrolysis reaction. At the same time, the radio waves can also stimulate the cells to a certain extent, causing the water in the cells to heat up rapidly and generate steam pressure, thereby destroying the cell structure, promoting the separation of stem cells, and helping to release stem cells from the tissue, thereby greatly improving the extraction efficiency of stem cells inside the snail tissue sample. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0030] Figure 1 This is a flow chart of a snail stem cell extraction method according to an embodiment of the present invention;

[0031] Figure 2 A schematic diagram of the structure of an extraction device according to an embodiment of the present invention;

[0032] Figure 3 Extract an exploded view of the device structure for an embodiment of the present invention;

[0033] Figure 4 A bottom view of the lifting frame structure according to an embodiment of the present invention;

[0034] Figure 5 A top view of the lifting frame structure according to an embodiment of the present invention;

[0035] Figure 6 A schematic diagram of the structure of the frame and the rotating frame is extracted for an embodiment of the present invention;

[0036] Figure 7 A schematic diagram of the internal structure of a rack is extracted for an embodiment of the present invention;

[0037] Figure 8 It is a schematic diagram of the separation tank and culture tank structure according to an embodiment of the present invention.

[0038] In the figure, 1. extraction rack; 2. rotating rack; 3. separation tank; 4. culture tank; 5. lifting rack; 6. servo electric cylinder 1; 7. material guide tube; 8. material extraction tube; 9. radio frequency wave generator; 10. movable rack; 11. magnetic stirrer; 12. stirring bar; 13. top rack; 14. driving rack; 15. fixed extraction tube; 16. movable extraction tube; 17. servo electric cylinder 2; 18. connecting rack; 19. micro electric cylinder 1; 20. micro electric cylinder 2; 21. centrifugal motor; 22. driving gear; 23. micro electric cylinder 3; 24. closing plate; 25. discharge port. DETAILED DESCRIPTION

[0039] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0040] In the description of the present invention, it is necessary to understand that the terms "upper", "lower", "front", "back", "left", "right", "top", "bottom", "inside", "outside", etc., indicating the orientation or position relationship are based on the orientation or position relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention.

[0041] Embodiment 1:

[0042] See also Figures 1 to 8 As shown, a method for extracting snail stem cells comprises the following steps:

[0043] Step 1: Snail pretreatment: Select healthy and active snails as the extraction object, let the snail rest, remove the debris in the body, and ensure that the cells are in the best condition during extraction;

[0044] Step 2: Stem cell extraction: Using sterile surgical instruments, extract tissue samples from specific parts of the snail (such as the foot);

[0045] Step 3: Tissue enzymatic hydrolysis: The extracted tissue sample is sent to the extraction equipment for enzymatic hydrolysis to separate the stem cells, and the stem cells are further purified through centrifugation, filtration and other steps;

[0046] Step 4: Cell culture: Use a culture medium suitable for stem cell growth, inoculate the extracted stem cells into a culture dish, maintain a constant temperature, humidity and CO2 concentration to promote cell proliferation, regularly observe cell growth, and replace the culture medium in a timely manner;

[0047] Step 5. Stem cell storage: Cryopreserve the cultured stem cells using a freezing solution containing a protective agent and gradually cool them down according to the freezing procedure; design an intelligent collection and storage system to optimize storage environment parameters through detection and analysis modules, environmental control modules, etc., to maintain the survival rate and functionality of stem cells.

[0048] Embodiment 2:

[0049] Specifically, the extraction device includes an extraction frame 1, a top frame 13 is fixedly arranged on the top of the extraction frame 1, and a rotating frame 2 is rotatably arranged inside the extraction frame 1, a driving frame 14 is fixedly arranged on the bottom of the extraction frame 1, and a rotating motor for driving the rotating frame 2 to rotate is arranged inside the driving frame 14, and the output shaft of the driving motor is fixedly connected to the middle part of the rotating frame 2, a plurality of separation tanks 3 are fixedly arranged on the top of the rotating frame 2, and a culture tank 4 is rotatably arranged inside the plurality of separation tanks 3, and a stirring bar 12 is placed inside the plurality of culture tanks 4;

[0050] Furthermore, servo electric cylinders 6 are fixedly provided on both sides of the bottom of the top frame 13, a lifting frame 5 is movably provided below the top frame 13, and the driving ends of the two servo electric cylinders 6 are fixedly connected to the top of the lifting frame 5, a material guide pipe 7 is fixedly provided on one side of the top of the lifting frame 5, and a material extraction pipe 8 is also fixedly provided on the other side of the top of the lifting frame 5, and the top ends of the material guide pipe 7 and the material extraction pipe 8 are extended to the top of the top frame 13, and enzyme liquid is fed into the interior of the culture tank 4 through the material guide pipe 7, so that the snail's extracted tissue sample is subjected to enzymatic hydrolysis treatment in the culture tank 4, and the amount of enzyme liquid introduced is dynamically set and adjusted according to the size of the snail's extracted tissue sample; after completing the stem cell extraction operation on the snail's extracted tissue sample, the interior of the culture tank 4 is automatically flushed through the material extraction pipe 8 to ensure the cleanliness of the interior of the culture tank 4 without manual auxiliary operation;

[0051] Furthermore, two movable frames 10 are provided on one side of the inner wall of the extraction frame 1 for sliding up and down through an electric slide, and magnetic stirrers 11 are fixedly provided on the top of the two movable frames 10, a plurality of through grooves matching the separation tank 3 are provided at the bottom of the rotating frame 2, a micro electric cylinder 19 is fixedly provided on one side of the top of the lifting frame 5, the driving end of the micro electric cylinder 19 extends to the bottom of the lifting frame 5, and a radio frequency wave generator 9 is fixedly provided on the driving end of the micro electric cylinder 19.

[0052] It should be noted that after the enzyme solution is sent into the culture tank 4, an enzymatic reaction occurs between the snail extracted tissue sample and the enzyme solution in the culture tank 4. At this time, the rotating frame 2 is controlled to rotate 60 degrees, and the culture tank 4 is rotated to the top of the first magnetic stirrer 11. The movable frame 10 at the bottom of the first magnetic stirrer 11 is controlled to move upward, and the magnetic stirrer 11 is used to control the stirrer 12 in the culture tank 4 to rotate, and the enzyme solution and the snail extracted tissue sample in the culture tank 4 are magnetically stirred, so that the enzyme and the tissue sample are fully in contact, and the enzymatic hydrolysis efficiency is improved. At the same time, stirring can also accelerate the diffusion of the enzyme in the solution, so that the enzyme is more evenly distributed in the tissue sample, thereby accelerating the degradation of the extracellular matrix and the release of stem cells. In addition, stirring can avoid the situation where the enzyme concentration is too high or too low locally in the centrifuge tube, and ensure that the enzymatic hydrolysis reaction is carried out evenly throughout the sample, which helps to improve the consistency of the enzymatic hydrolysis, reduce the problem of insufficient enzymatic hydrolysis or excessive enzymatic hydrolysis of some tissues due to concentration differences, and through magnetic stirring, the culture tank can be 4, the solution temperature is more uniform, avoiding local overheating or overcooling, thereby ensuring the activity and stability of the enzyme; when the enzymatic reaction reaches a certain extent, the rotating frame 2 is then controlled to rotate 60 degrees, so that the enzyme solution and the snail extracted tissue sample that have been magnetically stirred are moved to the top of the second magnetic stirrer 11, and at this time, the second magnetic stirrer 11 continues to control the stirring bar 12 inside the culture tank 4 to rotate at a low speed, and at the same time, the driving end of the micro-electric cylinder 19 controls the radio frequency wave generator 9 to move downward, and the radio frequency wave generator 9 is turned on to generate radio waves for the enzyme solution and the extracted tissue sample inside the culture tank 4, so that tiny vibrations are generated inside the tissue, increasing the contact area between the enzyme and the tissue, thereby accelerating the enzymatic reaction, and at the same time, the radio waves can also have a certain stimulating effect on the cells, so that the water in the cells is quickly heated up, generating steam pressure, thereby destroying the cell structure, promoting the separation of stem cells, and helping to release stem cells from the tissue, thereby greatly improving the extraction efficiency of stem cells inside the snail extracted tissue sample.

[0053] Furthermore, a fixed extraction tube 15 is fixedly provided on one side of the top of the lifting frame 5, and the bottom end of the fixed extraction tube 15 extends to the bottom of the lifting frame 5, and a movable extraction tube 16 is slidably provided on the bottom end of the fixed extraction tube 15, and a servo electric cylinder 2 17 is fixedly provided on one side of the top of the lifting frame 5, and the driving end of the servo electric cylinder 2 17 is fixedly connected to one side of the movable extraction tube 16;

[0054] Furthermore, two micro electric cylinders 20 are fixedly provided on one side of the top of the lifting frame 5, and a connecting frame 18 is fixedly provided on the top of the driving ends of the two micro electric cylinders 20, and a centrifugal motor 21 is fixedly provided on the bottom of the connecting frame 18. The bottom end of the output shaft of the centrifugal motor 21 extends to the bottom of the lifting frame 5, and a driving gear 22 is fixedly provided on the bottom end of the output shaft of the centrifugal motor 21. The top of the outer peripheral surface of the culture tank 4 is provided with an external tooth groove matching the driving gear 22, and centrifugal holes are provided on both sides of the inner wall of the culture tank 4. Micro electric cylinders 3 23 are fixedly provided on both sides of the inner wall of the separation tank 3, and closing plates 24 are fixedly provided on the driving ends of the micro electric cylinders 3 23 on both sides, and the closing plates 24 on both sides are respectively fitted with the centrifugal holes on both sides of the inner wall of the culture tank 4.

[0055] Furthermore, a discharge port 25 is provided at the bottom of the rotating frame 2, and the interiors of the plurality of separation tanks 3 are connected to the interior of the discharge port 25;

[0056] It should be noted that after completing the separation of stem cells inside the snail tissue sample, the rotating frame 2 is controlled to rotate 60 degrees, so that the separation tank 3 rotates to the position directly below the driving gear 22, and the driving end of the micro-electric cylinder 20 is used to control the connecting frame 18 to move downward, so that the surface of the driving gear 22 and the outer peripheral surface of the culture tank 4 are meshed and transmitted, and the output shaft of the centrifugal motor 21 is used to cooperate with the driving gear 22 to control the culture tank 4 to rotate inside the separation tank 3, and the stem cells inside the separation tank 3 are centrifugally filtered, and then the rotating frame 2 is controlled to rotate 60 degrees, so that the culture tank 4 rotates to the position directly below the extraction tube 8 and the fixed extraction tube 15, and the driving end of the servo electric cylinder 2 17 is used to control the movable extraction tube 16 to move downward, and the stem cells inside the culture tank 4 are extracted using the bottom end of the movable extraction tube 16, and then the extraction tube 8 is used to pass the cleaning agent into the interior of the culture tank 4 to rinse the interior of the culture tank 4, and the culture tank 4 is continued to be used to automatically extract stem cells from the snail tissue sample, which greatly improves the efficiency of stem cell extraction from the snail tissue sample.

[0057] Embodiment 3:

[0058] Specifically, the method for using the extraction device is as follows:

[0059] After the enzyme solution is sent into the culture tank 4, an enzymatic hydrolysis reaction occurs between the snail extracted tissue sample and the enzyme solution in the culture tank 4. At this time, the rotating frame 2 is controlled to rotate 60 degrees, and the culture tank 4 is rotated to the top of the first magnetic stirrer 11. The movable frame 10 at the bottom of the first magnetic stirrer 11 is controlled to move upward, and the magnetic stirrer 11 is used to control the stirring bar 12 in the culture tank 4 to rotate, and the enzyme solution and the snail extracted tissue sample in the culture tank 4 are magnetically stirred, so that the enzyme and the tissue sample are fully in contact;

[0060] When the enzymatic hydrolysis reaction proceeds to a certain extent, the rotating frame 2 is then controlled to rotate 60 degrees, so that the enzyme solution and snail extracted tissue sample that have been stirred by the magnetic force are moved to the top of the second magnetic stirrer 11. At this time, the stirring bar 12 inside the culture tank 4 is continued to be controlled by the second magnetic stirrer 11 to rotate at a low speed. At the same time, the driving end of the micro-electric cylinder 19 controls the radio frequency wave generator 9 to move downward, and the radio frequency wave generator 9 is turned on to generate radio waves for the enzyme solution and the extracted tissue sample inside the culture tank 4, so that tiny vibrations are generated inside the tissue, increasing the contact area between the enzyme and the tissue, thereby accelerating the enzymatic hydrolysis reaction.

[0061] After completing the separation of stem cells inside the snail tissue sample, the rotating frame 2 is controlled to rotate 60 degrees, so that the separation tank 3 rotates to the position directly below the driving gear 22, and the driving end of the micro-electric cylinder 20 is used to control the connecting frame 18 to move downward, so that the surface of the driving gear 22 and the outer peripheral surface of the culture tank 4 are meshed and transmitted, and the output shaft of the centrifugal motor 21 is used to cooperate with the driving gear 22 to control the culture tank 4 to rotate inside the separation tank 3, and the stem cells inside the separation tank 3 are centrifugally filtered, and then the rotating frame 2 is controlled to rotate 60 degrees, so that the culture tank 4 rotates to the position directly below the extraction tube 8 and the fixed extraction tube 15, and the driving end of the servo electric cylinder 2 17 is used to control the movable extraction tube 16 to move downward, and the stem cells inside the culture tank 4 are extracted using the bottom end of the movable extraction tube 16, and then the extraction tube 8 is used to pass the cleaning agent into the interior of the culture tank 4 to rinse the interior of the culture tank 4, and the culture tank 4 is continued to be used to automatically extract stem cells from the snail tissue sample.

[0062] Meanwhile, the contents not described in detail in this specification belong to the prior art known to those skilled in the art.

[0063] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0064] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the attached claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims are included in the present invention. Any figure mark in the claims should not be regarded as limiting the claims involved.

[0065] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A method for extracting snail stem cells, characterized in that: The following steps are involved: Step 1: Snail pretreatment: Select healthy and active snails as the extraction object, let the snail rest and remove the debris in the body; Step 2: Stem cell extraction: Using sterile surgical instruments, extract tissue samples from specific parts of the snail; Step 3: Tissue enzymatic hydrolysis: The extracted tissue sample is sent to an extraction device for enzymatic hydrolysis to separate the stem cells, and the stem cells are further purified through centrifugation and filtration steps; Step 4: Cell culture: Use a culture medium suitable for stem cell growth to inoculate the extracted stem cells into a culture dish and maintain a constant temperature, humidity and CO2 concentration; Step 5: Stem cell storage: cryopreserve the cultured stem cells using a cryoprotectant containing a protective agent and gradually cool them down according to the freezing procedure; Specifically, the extraction device comprises an extraction frame (1), a top frame (13) is fixedly arranged on the top of the extraction frame (1), and a rotating frame (2) is rotatably arranged inside the extraction frame (1), a driving frame (14) is fixedly arranged on the bottom of the extraction frame (1), and a rotating motor for driving the rotating frame (2) to rotate is arranged inside the driving frame (14), and the output shaft of the driving motor is fixedly connected to the middle part of the rotating frame (2), a plurality of separation tanks (3) are fixedly arranged on the top of the rotating frame (2), and a culture tank (4) is rotatably arranged inside the plurality of separation tanks (3), and the plurality of culture tanks (4) are all provided with a stirrer (12); after the enzyme solution is sent into the culture tank (4), an enzymatic hydrolysis reaction occurs between the snail tissue sample extracted from the culture tank (4) and the enzyme solution, at which time the rotating frame (2) is controlled to rotate 60 degrees, the culture tank (4) is rotated to the top of the first magnetic stirrer (11), the movable frame (10) at the bottom of the first magnetic stirrer (11) is controlled to move upward, and the magnetic stirrer (11) is used to control the stirrer (12) inside the culture tank (4) to rotate, and the enzyme solution and the snail tissue sample inside the culture tank (4) are magnetically stirred, so that the enzyme and the tissue sample are fully in contact.

2. The method for extracting snail stem cells according to claim 1, characterized in that: A servo electric cylinder (6) is fixedly arranged on both sides of the bottom of the top frame (13); a lifting frame (5) is movably arranged below the top frame (13); and the driving ends of the two servo electric cylinders (6) are fixedly connected to the top of the lifting frame (5); a material guide pipe (7) is fixedly arranged on one side of the top of the lifting frame (5); and a material extraction pipe (8) is also fixedly arranged on the other side of the top of the lifting frame (5); and the top ends of the material guide pipe (7) and the material extraction pipe (8) extend to the top of the top frame (13).

3. The method for extracting snail stem cells according to claim 2, characterized in that: One side of the inner wall of the extraction frame (1) is also provided with two movable frames (10) which slide up and down via an electric slide, and the tops of the two movable frames (10) are fixedly provided with magnetic stirrers (11), the bottom of the rotating frame (2) is provided with a plurality of through grooves matching the separation tank (3), and one side of the top of the lifting frame (5) is fixedly provided with a micro electric cylinder (19), the driving end of the micro electric cylinder (19) extends to the bottom of the lifting frame (5), and the driving end of the micro electric cylinder (19) is fixedly provided with a radio frequency wave generator (9).

4. The method for extracting snail stem cells according to claim 2, characterized in that: A fixed extraction tube (15) is also fixedly provided on one side of the top of the lifting frame (5), and the bottom end of the fixed extraction tube (15) extends to the bottom of the lifting frame (5), and a movable extraction tube (16) is slidably provided on the bottom end of the fixed extraction tube (15) up and down, and a servo electric cylinder 2 (17) is fixedly provided on one side of the top of the lifting frame (5), and the driving end of the servo electric cylinder 2 (17) is fixedly connected to one side of the movable extraction tube (16).

5. The method for extracting snail stem cells according to claim 2, characterized in that: Two micro electric cylinders (20) are fixedly arranged on one side of the top of the lifting frame (5), and the top ends of the driving ends of the two micro electric cylinders (20) are fixedly arranged with a connecting frame (18), and a centrifugal motor (21) is fixedly arranged at the bottom of the connecting frame (18), and the bottom end of the output shaft of the centrifugal motor (21) extends to the bottom of the lifting frame (5), and a driving gear (22) is fixedly arranged at the bottom end of the output shaft of the centrifugal motor (21), and the top of the outer peripheral surface of the culture tank (4) is provided with an external tooth groove matching the driving gear (22), and centrifugal holes are arranged on both sides of the inner wall of the culture tank (4), and micro electric cylinders (23) are fixedly arranged on both sides of the inner wall of the separation tank (3), and the driving ends of the micro electric cylinders (23) on both sides are fixedly arranged with closing plates (24), and the closing plates (24) on both sides are respectively fitted with the centrifugal holes on both sides of the inner wall of the culture tank (4).

6. The method for extracting snail stem cells according to claim 1, characterized in that: A discharge port (25) is provided at the bottom of the rotating frame (2), and the interiors of the plurality of separation tanks (3) are connected to the interior of the discharge port (25).

7. A method for extracting snail stem cells according to any one of claims 1 to 6, characterized in that: The method for using the device extracted in step 3 is as follows: After the enzyme solution is introduced into the culture tank (4), an enzymatic hydrolysis reaction occurs between the snail extracted tissue sample and the enzyme solution in the culture tank (4). At this time, the rotating frame (2) is controlled to rotate 60 degrees, and the culture tank (4) is rotated to the top of the first magnetic stirrer (11). The movable frame (10) at the bottom of the first magnetic stirrer (11) is controlled to move upward, and the magnetic stirrer (11) is used to control the stirring bar (12) in the culture tank (4) to rotate, so as to magnetically stir the enzyme solution and the snail extracted tissue sample in the culture tank (4), so that the enzyme and the tissue sample are fully in contact; When the enzymatic hydrolysis reaction proceeds to a certain extent, the rotating frame (2) is then controlled to rotate 60 degrees, so that the enzyme solution and the snail extracted tissue sample that have been stirred by the magnetic force are moved to the top of the second magnetic stirrer (11). At this time, the stirring bar (12) inside the culture tank (4) is continuously controlled by the second magnetic stirrer (11) to rotate at a low speed. At the same time, the driving end of the micro-electric cylinder (19) controls the radio frequency wave generator (9) to move downward, and the radio frequency wave generator (9) is turned on to generate radio waves for the enzyme solution and the extracted tissue sample inside the culture tank (4), so as to generate tiny vibrations inside the tissue, increase the contact area between the enzyme and the tissue, and thus accelerate the enzymatic hydrolysis reaction. After the separation of stem cells in the snail tissue sample is completed, the rotating frame (2) is controlled to rotate 60 degrees, so that the separation tank (3) is rotated to the position directly below the driving gear (22), and the driving end of the second micro-electric cylinder (20) is used to control the connecting frame (18) to move downward, so that the surface of the driving gear (22) and the outer peripheral surface of the culture tank (4) are meshed and transmitted, and the output shaft of the centrifugal motor (21) is used to cooperate with the driving gear (22) to control the culture tank (4) to rotate inside the separation tank (3), so as to centrifugally filter the stem cells inside the separation tank (3). The rotating frame (2) is controlled to rotate 60 degrees, so that the culture tank (4) is rotated to the position directly below the extraction tube (8) and the fixed extraction tube (15), and the driving end of the servo electric cylinder (17) is used to control the movable extraction tube (16) to move downward, and the bottom end of the movable extraction tube (16) is used to extract the stem cells inside the culture tank (4), and then the extraction tube (8) is used to pass a cleaning agent into the interior of the culture tank (4) to rinse the interior of the culture tank (4), and the culture tank (4) is continued to be used to extract the snail tissue sample to perform the automatic extraction operation of stem cells.

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