A method for extracting stem cells

Abstract

The invention discloses a stem cell extraction method in the technical field of stem cell extraction, comprising a shaking incubator, wherein a placing tray is installed at the front opening of the shaking incubator, two installation boxes are arranged on the front side of the shaking incubator, the two installation boxes are connected by a first connecting rod, a clamping mechanism is arranged at the bottom end of the installation box, the clamping mechanism is used to clamp the top opening of triangular flasks of different sizes, a plurality of clamping rollers are arranged at the top of the placing tray through a supporting mechanism, the supporting mechanism is used to drive two adjacent clamping rollers to move away from each other when the clamping mechanism moves downward, a driving mechanism is arranged at one end of the two installation boxes that are away from each other, the driving mechanism is used to drive the clamping mechanism to transport the triangular flask through the installation box, and the problem that the existing equipment for stem cell extraction does not have the automatic picking and placing function of the triangular flask and the self-adjusting function of the triangular flask rack is solved.

Description

Technical Field

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

[0002] Stem cells are a type of cell with unlimited or immortal self-renewal capabilities and can produce at least one type of highly differentiated daughter cells; stem cell extraction is the process of separating stem cells from certain tissues or organs of the human body, such as extracting them from tissues such as the placenta, umbilical cord, fat, etc.; the extraction steps of adipose stem cells include: sampling, digestion, centrifugation and counting, inoculation and culture, and observation.

[0003] In the existing technology, the inoculated experimental cells are placed in a triangular flask, and then the staff manually places the triangular flasks one by one in a shaking incubator. When a large number of cultures are carried out, the staff needs to regularly take and place the triangular flasks one by one, which is a large workload and consumes a lot of labor, increasing the labor cost; secondly, when different batches of stem cells are extracted, the sizes of the triangular flasks used are different, and the triangular flask racks inside the shaking incubator need to be replaced one by one before placing the triangular flasks, further increasing the workload of the staff, and the replacement steps are very cumbersome, which reduces the efficiency of stem cell extraction and brings inconvenience to the stem cell extraction work. The existing equipment for stem cell extraction does not have the automatic taking and placing function of the triangular flasks and the self-adjustment function of the triangular flask rack.

[0004] Based on this, the present invention designs a stem cell extraction method to solve the above problems. Summary of the invention

[0005] The object of the present invention is to provide a stem cell extraction device and method with a function of automatically taking and placing a triangular flask and a self-adjusting function of a triangular flask stand, so as to solve the problems raised in the above background technology.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] A method for extracting stem cells, the specific steps of the method are:

[0008] Step 1: Take human adipose tissue removed by clinical sterilization and wash it thoroughly with PBS solution containing 100U / ml penicillin and 100U / ml streptomycin on a clean bench;

[0009] Step 2: Use ophthalmic scissors to cut off the blood vessels and fascia tissue visible to the naked eye, transfer to a centrifuge tube, cut the tissue into 1mm³ paste, wash with sterile PBS, add mixed collagenase, and digest at 37℃ for 30-40min until the fat tissue becomes chyme, and add an equal volume of complete culture medium (DMEM+10%FBS+1% double antibody) to terminate the digestion;

[0010] Step 3: Pour the mixture into a 200-mesh cell strainer, pipette through the sieve, collect the filtrate in a sterile culture dish, and centrifuge at 800 rpm / min for 5 min;

[0011] Step 4: Discard the supernatant, add 3 ml of complete culture medium to resuspend the cells for inoculation, and then place them in a conical flask. Use a culture auxiliary device to culture the conical flasks in batches at a constant temperature of 5% CO2 and 37°C. Change the medium for the first time after 48 hours.

[0012] The culture auxiliary device comprises a shaking incubator, a placing tray is installed at the front opening of the shaking incubator, two installation boxes are arranged on the front side of the shaking incubator, the two installation boxes are connected by a first connecting rod, a clamping mechanism is arranged at the bottom end of the installation box, the clamping mechanism is used to clamp the top openings of triangular flasks of different sizes, a plurality of clamping rollers are arranged at the top of the placing tray through a supporting mechanism, the supporting mechanism is used to drive two adjacent clamping rollers to move away from each other when the clamping mechanism moves downward, a driving mechanism is arranged at one end of the two installation boxes that are away from each other, the driving mechanism is used to drive the clamping mechanism to transport the triangular flask through the installation box, a discharging mechanism is arranged at the lower side of the installation box on the right side, the discharging mechanism is used to arrange the triangular flasks neatly and corresponds to the clamping mechanism;

[0013] As a further solution of the present invention, the clamping mechanism includes mounting brackets arranged at the lower sides of the mounting boxes, the top of the mounting brackets is fixedly mounted with a second connecting rod, the top of the second connecting rod passes through the mounting box and is slidably connected thereto, a clamping box is arranged at the bottom of the front and rear ends of the mounting bracket, mounting rods are fixedly mounted at the tops of the left and right ends of the clamping box, the tops of the mounting rods pass through the mounting bracket and are slidably connected thereto, a C-shaped frame is slidably connected to the tops of the front and rear ends of the mounting box, the tops of the mounting rods pass through the ends of the C-shaped frame and are slidably connected thereto, an electric clamp is installed at the middle part of the top of the mounting box, the side walls of the electric clamp are connected to the adjacent C-shaped frames by extension springs, and a telescopic plate is arranged at the bottom of the clamping box by a telescopic mechanism, and the telescopic mechanism is used to drive the telescopic plate to gradually retract into the clamping box when the adjacent clamping boxes on both sides are close to each other;

[0014] As a further solution of the present invention, the telescopic mechanism includes connecting rings fixedly installed at both ends of the left and right ends of the two clamping boxes away from each other, and the bottoms of the four ends of the mounting frame are fixedly installed with pull ropes, and the other ends of the pull ropes are connected to the bottom ends of the telescopic plates, and the top ends of the telescopic plates are serrated, and the side walls of the clamping boxes are adapted to the shapes of the telescopic plates and are slidably connected thereto;

[0015] As a further solution of the present invention, two limit plates are fixedly installed at both ends of the mounting rod, and the limit plates are respectively fitted with the upper and lower ends of the mounting frame, and there is a certain gap between the bottom end of the mounting frame and the top end of the clamping box;

[0016] As a further solution of the present invention, the discharging mechanism includes a discharging platform arranged on the right side of the front end of the storage tray, a first conveyor is installed in the middle of the discharging platform, and two first partitions are arranged on the top of the discharging platform, the left ends of the first partitions are fixedly installed with first racks, the top of the left end of the discharging platform is rotatably connected with a first gear, the two first racks are respectively located on the left and right sides of the first gear and meshed with it, a plurality of limit blocks are fixedly installed on the top of the discharging platform, the middle parts of the limit blocks are slidably connected with limit rods, the ends of the limit rods are fixedly connected to the side walls of the first partition, and the first partition is connected to the plurality of limit blocks through compression springs;

[0017] As a further solution of the present invention, a material receiving platform is provided on the left side of the front end of the storage tray, a second conveyor is installed in the middle of the material receiving platform, and two second partitions are fixedly installed on the top of the material receiving platform;

[0018] As a further solution of the present invention, the supporting mechanism comprises a plurality of U-shaped frames fixedly installed on the top of the left and right ends of the storage tray, the inner sides of the U-shaped frames are open and closed with two sliding grooves, the left and right ends of the clamping roller are fixedly installed with sliders, the sliders are located on the inner sides of the sliding grooves and are connected to the inside of the sliding grooves through compression springs. A second rack is fixedly installed on the side of the slider away from the clamping roller, and the side of the U-shaped frame away from the clamping roller is rotatably connected to the second gear, and two adjacent second racks are respectively located on the upper and lower sides of the second gear and meshed with the second gear. A vertical rod is installed on the side of the rear end of the U-shaped frame away from the clamping roller, and a pressing block is slidably connected to the outer side of the top end of the vertical rod, and the front end of the pressing block is hingedly connected to a hinge rod, and the other end of the hinge rod is hingedly connected to the rear end of the second rack on the upper side, and a pressing mechanism is provided at the bottom end of the first connecting rod, and the pressing mechanism is used to continuously push the hinge rod to a horizontal state when the installation box moves downward;

[0019] As a further solution of the present invention, the pressing mechanism comprises two support plates fixedly mounted at the rear end of the first connecting rod, and the rear end of the support plates is slidably connected to the pressing rod via a tension spring;

[0020] As a further solution of the present invention, the driving mechanism includes a connecting frame on which two mounting boxes are fixedly installed at one end away from each other, electric slide rails are arranged on the left and right sides of the front end of the storage tray, and driving columns are installed on the top ends of the electric slide rails. The top ends of the driving columns pass through the front end of the connecting frame and are slidably connected thereto, and electric push rods are fixedly installed on the front ends of the driving columns, and the output ends of the electric push rods are connected to the front end bottom of the connecting frame.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The present invention provides a clamping mechanism, and when the mounting frame contacts the top of the triangular flask, the mounting frame stops moving, and does not hinder the movement of the mounting box, thereby realizing the clamping operation of triangular flasks of different sizes and heights, greatly increasing the applicability of the device; the clamping mechanism simultaneously clamps a plurality of triangular flasks during operation, so that the triangular flasks can be carried and placed in a row, greatly shortening the operation steps and improving the placement efficiency of the triangular flasks; when the clamping mechanism at the bottom end of the right mounting box clamps another batch of triangular flasks, the clamping mechanism at the bottom end of the left mounting box clamps the triangular flasks at the top of the storage tray, thereby realizing loading during the unloading process, further improving the work efficiency, and solving the problem that the existing equipment for stem cell extraction does not have the function of automatically taking and placing triangular flasks.

[0023] The present invention sets a telescopic mechanism. Taking a large-sized triangular bottle as an example, the diameter of the bottle mouth of the triangular bottle is relatively large, and the distance that the two clamps move towards each other is relatively short. At the same time, a smaller part of the telescopic plate is received in the clamping box. At this time, the telescopic plate plays a role of extending the clamping box, increasing the contact surface between the clamping mechanism and the bottle mouth of the triangular bottle, making the clamping more stable. When the triangular bottle is small in size, the diameter of the bottle mouth of the triangular bottle is relatively small and the bottle mouth is relatively short. Similar to the above, a larger part of the telescopic plate is received in the clamping box, thereby preventing the clamping box and the telescopic plate from clamping the inclined surface of the triangular bottle body and causing it to slip when clamping the triangular bottle, thereby increasing the practicality of the present device.

[0024] The present invention provides a supporting mechanism. When the triangular flasks are placed in a row, the supporting mechanism drives two adjacent clamping rollers to move away from each other. When the triangular flasks are placed, the supporting mechanism drives the clamping rollers to move again and clamp the triangular flasks. The device can fix triangular flasks of different sizes, so that the staff does not need to replace the fixing device on the top of the storage tray before work, which greatly increases the applicability of the device, saves the cost of production and processing, and solves the problem that the existing equipment for stem cell extraction does not have the self-adjusting function of the triangular flask rack. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is a flow chart of the method of the present invention;

[0026] Figure 2 It is a schematic diagram of the overall structure of the present invention;

[0027] Figure 3 This is a schematic diagram of the structure of the present invention from a front side perspective;

[0028] Figure 4 It is a schematic diagram of the connection structure between the shaking incubator and the storage tray in the present invention;

[0029] Figure 5 It is a schematic diagram of a local structure from a top-down perspective in the present invention;

[0030] Figure 6 For the present invention Figure 5 The enlarged structural diagram at A in the middle;

[0031] Figure 7 It is a schematic diagram of the partial structure of the present invention from a side perspective;

[0032] Figure 8 For the present invention Figure 7 The enlarged structural diagram at B in the middle;

[0033] Fig. 9 It is a schematic diagram of the connection structure between the installation box and the clamping box in the present invention;

[0034] Fig.10 It is a schematic diagram of the connection structure between the mounting frame and the mounting rod in the present invention;

[0035] Fig.11 For the present invention Fig.10 The enlarged structural diagram at C in the middle;

[0036] Fig.12 This is a schematic diagram of the connection structure between the electric clamp and the C-shaped frame in the present invention;

[0037] Fig.13 It is a schematic diagram of the connection structure between the clamping box and the telescopic plate in the present invention.

[0038] In the accompanying drawings, the components represented by the reference numerals are listed as follows:

[0039] 1. Shaking incubator; 2. Storage tray; 3. Mounting box; 4. First connecting rod; 5. Clamping roller; 6. Mounting frame; 7. Second connecting rod; 8. Clamping box; 9. Mounting rod; 10. C-shaped frame; 11. Electric clamp; 12. Telescopic plate; 13. Connecting ring; 14. Pull rope; 15. Limiting plate; 16. Discharge table; 17. First conveyor; 18. First partition; 19. First rack; 20. First Gear; 21. Limit block; 22. Limit rod; 23. Material receiving platform; 24. Second conveyor; 25. Second partition; 26. U-shaped frame; 27. Slide; 28. Slider; 29. ​​Second rack; 30. Second gear; 31. Vertical pole; 32. Pressing block; 33. Articulated rod; 34. Support plate; 35. Pressing rod; 36. Connecting frame; 37. Electric slide rail; 38. Driving column; 39. Electric push rod. DETAILED DESCRIPTION

[0040] See also Figure 1-Figure 13 , the present invention provides a technical solution:

[0041] A method for extracting stem cells, characterized in that the specific steps of the method are:

[0042] Step 1: Take human adipose tissue removed by clinical sterilization and wash it thoroughly with PBS solution containing 100U / ml penicillin and 100U / ml streptomycin on a clean bench;

[0043] Step 2: Use ophthalmic scissors to cut off the blood vessels and fascia tissue visible to the naked eye, transfer to a centrifuge tube, cut the tissue into 1mm³ paste, wash with sterile PBS, add mixed collagenase, and digest at 37℃ for 30-40min until the fat tissue becomes chyme, and add an equal volume of complete culture medium (DMEM+10%FBS+1% double antibody) to terminate the digestion;

[0044] Step 3: Pour the mixture into a 200-mesh cell strainer, pipette through the sieve, collect the filtrate in a sterile culture dish, and centrifuge at 800 rpm / min for 5 min;

[0045] Step 4: Discard the supernatant, add 3 ml of complete culture medium to resuspend the cells for inoculation, and then place them in a conical flask. Use a culture auxiliary device to culture the conical flasks in batches at a constant temperature of 5% CO2 and 37°C. Change the medium for the first time after 48 hours.

[0046] A culture auxiliary device comprises a shaking incubator 1, a placing tray 2 is installed at the front opening of the shaking incubator 1, two installation boxes 3 are arranged on the front side of the shaking incubator 1, the two installation boxes 3 are connected by a first connecting rod 4, a clamping mechanism is arranged at the bottom end of the installation box 3, the clamping mechanism is used to clamp the top opening of triangular flasks of different sizes, a plurality of clamping rollers 5 are arranged at the top of the placing tray 2 through a supporting mechanism, the supporting mechanism is used to drive two adjacent clamping rollers 5 to move away from each other when the clamping mechanism moves downward, a driving mechanism is arranged at one end of the two installation boxes 3 that are away from each other, the driving mechanism is used to drive the clamping mechanism to transport the triangular flask through the installation box 3, a discharging mechanism is arranged at the lower side of the right installation box 3, the discharging mechanism is used to arrange the triangular flasks neatly and corresponds to the clamping mechanism;

[0047] The clamping mechanism includes a mounting frame 6 provided at the lower side of the mounting box 3, a second connecting rod 7 is fixedly installed on the top of the mounting frame 6, and the top of the second connecting rod 7 passes through the mounting box 3 and is slidably connected thereto, a clamping box 8 is provided at the bottom of the front and rear ends of the mounting frame 6, a mounting rod 9 is fixedly installed on the top of the left and right ends of the clamping box 8, and the top of the mounting rod 9 passes through the mounting frame 6 and is slidably connected thereto, a C-shaped frame 10 is slidably connected to the top of the front and rear ends of the mounting box 3, and the top of the mounting rod 9 passes through the end of the C-shaped frame 10 and is slidably connected thereto, an electric clamp 11 is installed in the middle of the top of the mounting box 3, and the side walls of the electric clamp 11 are connected to the adjacent C-shaped frame 10 through a tension spring, and a telescopic plate 12 is provided at the bottom of the clamping box 8 through a telescopic mechanism, and the telescopic mechanism is used to drive the telescopic plate 12 to gradually retract into the clamping box 8 when the adjacent clamping boxes 8 on both sides are close to each other;

[0048] During operation, the driving mechanism starts to drive the two installation boxes 3 to move downward synchronously. The installation box 3 moves downward and drives the installation frame 6 and the clamping box 8 to move downward through the second connecting rod 7. The right installation frame 6 is blocked by the top of the triangular bottle during the downward movement. At this time, the right installation frame 6 and the clamping box 8 can no longer move downward, and the installation box 3 continues to move downward. The second connecting rod 7 slides on the inner side of the installation box 3. When the installation box 3 no longer moves downward, the electric clamp 11 is started. The electric clamp 11 starts to pull the two C-shaped frames 10 closer to each other through the tension spring. The two C-shaped frames 10 approach each other and drive the two clamping boxes 8 closer to each other through the installation rod 9. The two clamping boxes 8 are blocked by the mouth of the triangular bottle during the process of approaching each other, so that The two clamping boxes 8 cannot move any more, and at the same time, the electric clamping claw 11 continues to move to pull the tension spring connected thereto to stretch, until the two clamping claws of the electric clamping claw 11 cannot get close to each other, at this time, the two clamping boxes 8 clamp and fix the bottle mouth of the triangular bottle, in preparation for the subsequent transportation of the triangular bottle. The clamping mechanism clamps multiple triangular bottles at the same time during operation, so that the triangular bottles can be transported and placed in a row, which greatly shortens the operation steps and improves the placement efficiency of the triangular bottles; by setting the clamping mechanism, when the mounting frame 6 contacts the top of the triangular bottle, it stops moving, and will not hinder the movement of the mounting box 3, thereby realizing the clamping operation of triangular bottles of different sizes and heights, greatly increasing the applicability of the device;

[0049] After the clamping mechanism clamps the triangular flask, the driving mechanism starts to drive the installation box 3 to return to the initial state, and then drives the installation box 3 to move to the left until the right installation box 3 moves to the top center position of the placement tray 2, and then the installation box 3 moves downward, and the installation box 3 moves downward to drive a row of triangular flasks to move downward synchronously through the clamping mechanism. In the process of the triangular flask moving downward, the supporting mechanism drives the two adjacent clamping rollers 5 to move away from each other, so that the bottom end of the triangular flask can pass between the clamping rollers 5 until the triangular flask is placed on the top of the placement tray 2. At this time, the clamping mechanism releases the triangular flask and moves it upward to restore the initial position. At the same time, the supporting mechanism starts to drive the two adjacent clamping rollers 5 to move closer to each other and fix the triangular flask on the top of the placement tray 2, thereby realizing the placement and fixing operation of the triangular flask. No manual operation is required during the operation, which greatly saves labor. The simultaneous operation of a row of triangular flasks greatly improves the efficiency of stem cell lifting.

[0050] When a row of flasks is placed, the shaking incubator 1 drives the placement tray 2 to move forward a certain distance, so that the latter group of clamping rollers 5 moves forward to the bottom of the installation box 3, thereby preparing for the placement operation of the next row of flasks. When the placement tray 2 is full, the placement tray 2 is received into the shaking incubator 1 for constant temperature culture. When the culture time is over, the placement tray 2 is extended from the opening of the shaking incubator 1 again. At this time, the driving mechanism starts to drive the installation box 3 to move again. When the clamping mechanism at the bottom end of the right installation box 3 clamps another batch of flasks, the clamping mechanism at the bottom end of the left installation box 3 clamps the flasks at the top of the placement tray 2, thereby realizing loading during the unloading process, further improving work efficiency, and solving the problem that the existing equipment for stem cell extraction does not have the automatic flask picking and placing function and the flask rack self-adjustment function.

[0051] As a further solution of the present invention, the telescopic mechanism includes a connecting ring 13 fixedly installed at both ends of the left and right sides of the two clamping boxes 8 away from each other, and a pull rope 14 is fixedly installed at the bottom of the four ends of the mounting frame 6, and the other end of the pull rope 14 is connected to the bottom end of the telescopic plate 12, and the top of the telescopic plate 12 is serrated, and the side wall of the clamping box 8 is adapted to the shape of the telescopic plate 12 and is slidably connected thereto;

[0052] During operation, when two adjacent clamping boxes 8 approach each other, the clamping box 8 drives the telescopic plate 12 to move synchronously. At the same time, the pull rope 14 pulls the telescopic plate 12 to move toward the inside of the clamping box 8 under the guidance of the connecting ring 13. Taking the large-sized triangular bottle as an example, the bottle mouth diameter of the triangular bottle is larger, and the distance that the two clamps move closer to each other is shorter. At the same time, a smaller part of the telescopic plate 12 is received into the clamping box 8. At this time, the telescopic plate 12 plays a role in extending the clamping box 8, increasing the contact surface between the clamping mechanism and the bottle mouth of the triangular bottle, making the clamping more stable. When the triangular bottle is small in size, the bottle mouth diameter of the triangular bottle is smaller and the bottle mouth is shorter. Similarly to the above, a larger part of the telescopic plate 12 is received into the clamping box 8, thereby avoiding the clamping box 8 and the telescopic plate 12 from clamping the inclined surface of the triangular bottle body and causing it to slide when clamping the triangular bottle, thereby increasing the practicality of the device.

[0053] As a further solution of the present invention, two limit plates 15 are fixedly installed at both ends of the mounting rod 9, and the limit plates 15 are respectively fitted with the upper and lower ends of the mounting frame 6, and there is a certain gap between the bottom end of the mounting frame 6 and the top end of the clamping box 8;

[0054] When working, the edge of the bottle mouth of the triangular flask is convex. By setting a gap between the mounting frame 6 and the clamping box 8, it is avoided that the clamping box 8 only clamps the convex part of the bottle mouth edge when clamping the triangular flask, and the stability of the triangular flask when moving is further increased.

[0055] As a further solution of the present invention, the discharging mechanism includes a discharging platform 16 arranged on the right side of the front end of the storage tray 2, a first conveyor 17 is installed in the middle of the discharging platform 16, and two first partitions 18 are arranged on the top of the discharging platform 16, and the left ends of the first partitions 18 are fixedly installed with first racks 19, and the top of the left end of the discharging platform 16 is rotatably connected with a first gear 20, and the two first racks 19 are respectively located on the left and right sides of the first gear 20 and meshed with it, and a plurality of limit blocks 21 are fixedly installed on the top of the discharging platform 16, and the middle parts of the limit blocks 21 are slidably connected with limit rods 22, and the ends of the limit rods 22 are fixedly connected to the side walls of the first partition 18, and the first partition 18 is connected to the plurality of limit blocks 21 through compression springs;

[0056] During operation, the first conveyor 17 drives a number of triangular bottles to be transported from the right side to the left end of the discharge table 16. When the triangular bottles move to the left, they are guided by the two first partitions 18, so that the triangular bottles are neatly arranged at the top center of the discharge table 16, preparing for the subsequent clamping of the clamping mechanism; when the first triangular bottle passes through between the two first partitions 18, the triangular bottle squeezes the two first partitions 18 away from each other, and the movement of the two partitions drives the first rack 19 to move. The two first racks 19 are restricted by the first gear 20 and move synchronously, so that the two first partitions 18 move the same distance when they are separated, further ensuring that the triangular bottles can be accurately clamped subsequently.

[0057] As a further solution of the present invention, a receiving platform 23 is provided on the left side of the front end of the storage tray 2, a second conveyor 24 is installed in the middle of the receiving platform 23, and two second partitions 25 are fixedly installed on the top of the receiving platform 23;

[0058] During operation, the clamping mechanism at the bottom end of the left installation box 3 clamps the triangular flasks that have been cultured on the top of the placement tray 2 and transports them to the left until the triangular flasks are transported to the top of the receiving table 23. Then the installation box 3 moves downward and places a row of triangular flasks that have been cultured on the top of the receiving table 23 through the clamping mechanism. At this time, the second conveyor 24 drives the triangular flasks to move slowly to the left and transport them, and proceeds to the next step.

[0059] As a further solution of the present invention, the support mechanism includes a plurality of U-shaped frames 26 fixedly installed on the top of the left and right ends of the storage tray 2, and two slide grooves 27 are opened and closed on the inner side of the U-shaped frame 26. Slide blocks 28 are fixedly installed on the left and right ends of the clamping roller 5. The slide blocks 28 are located on the inner side of the slide groove 27 and are connected to the inside thereof through a compression spring. A second rack 29 is fixedly installed on the side of the slide block 28 away from the clamping roller 5. The side of the U-shaped frame 26 away from the clamping roller 5 is rotatably connected to a second gear 30, and two adjacent second gears are connected to each other. The racks 29 are respectively located at the upper and lower sides of the second gear 30 and meshed therewith. A vertical rod 31 is installed on the side of the rear end of the U-shaped frame 26 away from the clamping roller 5. A pressing block 32 is slidably connected to the outer side of the top of the vertical rod 31. The front end of the pressing block 32 is hingedly connected to a hinge rod 33. The other end of the hinge rod 33 is hingedly connected to the rear end of the second rack 29 on the upper side. A pressing mechanism is provided at the bottom end of the first connecting rod 4. The pressing mechanism is used to continuously push the hinge rod 33 to a horizontal state when the installation box 3 moves downward;

[0060] During operation, the clamping mechanism clamps a row of triangular bottles and moves them to the top of the storage tray 2. At this time, the installation box 3 moves downward, driving the triangular bottles to move downward synchronously. During the downward movement of the installation box 3, the pressing mechanism pushes the pressing block 32 to move downward, and the pressing block 32 soon cannot move downward any more. During the downward movement of the pressing block 32, the rear end of the hinged rod 33 is pushed downward. At this time, the front end of the hinged rod 33 pushes the upper second rack 29 to move forward, and the upper second rack 29 moves forward to push the front slider 28 to slide forward. The front slider 28 moves forward to drive the front clamping roller 5 to move forward. At the same time, the upper second rack 29 moves forward and drives the lower second rack 29 to move backward through the second gear 30. The lower second rack 29 moves backward to push the rear slider 28 to move backward. The rear slider 28 moves backward to push the rear clamping roller 5 to move backward, thereby realizing that the two adjacent clamping rollers 5 move away from each other.

[0061] When the triangular bottle is placed and the clamping mechanism is reset upward, the pressing mechanism no longer squeezes the pressing block 32. At this time, the slider 28 drives the two adjacent clamping rollers 5 to approach each other under the push of the compression spring until the two clamping rollers 5 are blocked by the triangular bottle body at the same time, thereby fixing the triangular bottle on the top of the storage tray 2. No manual operation is required during the fixing process, which saves labor. By arranging the clamping roller 5 and the slider 28, the device can fix triangular bottles of different sizes, so that the staff does not have to replace the fixing equipment on the top of the storage tray 2 before work, which greatly increases the applicability of the device and saves the cost of production and processing.

[0062] As a further solution of the present invention, the pressing mechanism includes two support plates 34 fixedly mounted at the rear end of the first connecting rod 4, and the rear end of the support plate 34 is slidably connected to a pressing rod 35 via a tension spring;

[0063] During operation, the installation box 3 and the first connecting rod 4 move downward, driving the support plate 34 to move downward. The support plate 34 moves downward, driving the pressing rod 35 to move downward. During the downward movement of the pressing rod 35, the pressing block 32 is squeezed to move downward. When the pressing block 32 can no longer move downward, the pressing rod 35 can no longer move downward. At this time, the support plate 34 slides downward on the outside of the pressing rod 35.

[0064] As a further solution of the present invention, the driving mechanism includes a connecting frame 36 on which two ends of the two mounting boxes 3 are fixedly mounted away from each other, electric slide rails 37 are arranged on the left and right sides of the front end of the storage tray 2, and driving columns 38 are installed on the top ends of the electric slide rails 37. The top ends of the driving columns 38 pass through the front end of the connecting frame 36 and are slidably connected thereto, and electric push rods 39 are fixedly installed on the front ends of the driving columns 38, and the output ends of the electric push rods 39 are connected to the bottom of the front end of the connecting frame 36.

[0065] During operation, the electric push rod 39 starts to drive the connecting frame 36 to move downward, and the downward movement of the connecting frame 36 drives the two installation boxes 3 and the first connecting rod 4 to move downward synchronously, thereby providing power for the downward placement of the clamping box 8 of the clamping mechanism; the electric slide rail 37 starts to drive the driving column 38 to move left synchronously, and the driving column 38 moves leftward and drives the clamping mechanism to move left through the connecting frame 36 and the installation box 3, thereby providing power for the transportation and movement of the triangular bottle.

Claims

1. A method for extracting stem cells, characterized in that: The specific steps of this method are: Step 1: Take human adipose tissue removed by clinical sterilization and wash it thoroughly with PBS solution containing 100U / ml penicillin and 100U / ml streptomycin on a clean bench; Step 2: Use ophthalmic scissors to cut off the blood vessels and fascia tissue visible to the naked eye, transfer to a centrifuge tube, cut the tissue into 1mm³ paste, wash with sterile PBS, add mixed collagenase, and digest at 37℃ for 30-40min until the fat tissue becomes chyme, and add an equal volume of complete culture medium to terminate the digestion; Step 3: Pour the mixture into a 200-mesh cell strainer, pipette through the sieve, collect the filtrate in a sterile culture dish, and centrifuge at 800 rpm / min for 5 min; Step 4: Discard the supernatant, add 3 ml of complete culture medium to resuspend the cells for inoculation, and then place them in a conical flask. Use a culture auxiliary device to culture the conical flasks in batches at a constant temperature of 5% CO2 and 37°C. Change the medium for the first time after 48 hours. The culture auxiliary device comprises a shaking incubator (1), a storage tray (2) is installed at the front opening of the shaking incubator (1), two installation boxes (3) are arranged on the front side of the shaking incubator (1), the two installation boxes (3) are connected by a first connecting rod (4), a clamping mechanism is arranged at the bottom end of the installation box (3), the clamping mechanism is used to clamp the top opening of triangular flasks of different sizes, a plurality of clamping rollers (5) are arranged at the top of the storage tray (2) through a supporting mechanism, the supporting mechanism is used to drive two adjacent clamping rollers (5) to move away from each other when the clamping mechanism moves downward, a driving mechanism is arranged at one end of the two installation boxes (3) that are away from each other, the driving mechanism is used to drive the clamping mechanism to transport the triangular flasks through the installation box (3), and a discharge mechanism is arranged at the lower side of the right installation box (3), the discharge mechanism is used to arrange the triangular flasks in an orderly manner and corresponds to the clamping mechanism; The clamping mechanism comprises a mounting frame (6) provided at the lower side of the mounting box (3); a second connecting rod (7) is fixedly installed at the top of the mounting frame (6); the top of the second connecting rod (7) passes through the mounting box (3) and is slidably connected thereto; a clamping box (8) is provided at the bottom of the front and rear ends of the mounting frame (6); a mounting rod (9) is fixedly installed at the top of the left and right ends of the clamping box (8); the top of the mounting rod (9) passes through the mounting frame (6) and is slidably connected thereto; the top of the front and rear ends of the mounting box (3) are A C-shaped frame (10) is slidably connected, the top end of the mounting rod (9) passes through the end of the C-shaped frame (10) and is slidably connected thereto, an electric clamp (11) is installed in the middle of the top end of the mounting box (3), the side walls of the electric clamp (11) are connected to the adjacent C-shaped frame (10) through a tension spring, and a telescopic plate (12) is provided at the bottom end of the clamping box (8) through a telescopic mechanism, and the telescopic mechanism is used to drive the telescopic plate (12) to gradually retract into the clamping box (8) when the adjacent clamping boxes (8) on both sides are close to each other; The telescopic mechanism comprises connecting rings (13) fixedly mounted on both left and right ends of a side of the two clamping boxes (8) that are away from each other, and pull ropes (14) fixedly mounted on the bottoms of the four ends of the mounting frame (6), the other ends of the pull ropes (14) are connected to the bottom ends of the telescopic plates (12), the top ends of the telescopic plates (12) are sawtooth-shaped, and the side walls of the clamping boxes (8) are adapted to the shapes of the telescopic plates (12) and are slidably connected thereto; The support mechanism comprises a plurality of U-shaped frames (26) fixedly mounted on the top of both left and right ends of the storage tray (2), the inner side of each of the U-shaped frames (26) is provided with two slide grooves (27) for opening and closing, and sliders (28) are fixedly mounted on both left and right ends of the clamping roller (5), the sliders (28) are located on the inner side of the slide grooves (27) and are connected to the inside of the slide grooves (27) via compression springs, a second rack (29) is fixedly mounted on the side of the slider (28) away from the clamping roller (5), and a second gear (30) is rotatably connected to the side of the U-shaped frame (26) away from the clamping roller (5), and two adjacent second racks (29) are connected to each other. The U-shaped frame (26) is respectively located on the upper and lower sides of the second gear (30) and meshed therewith. A vertical rod (31) is installed on the side of the rear end of the U-shaped frame (26) away from the clamping roller (5). A pressing block (32) is slidably connected to the outer side of the top end of the vertical rod (31). The front end of the pressing block (32) is hingedly connected to a hinge rod (33). The other end of the hinge rod (33) is hingedly connected to the rear end of the second rack (29) on the upper side. A pressing mechanism is provided at the bottom end of the first connecting rod (4). The pressing mechanism is used to continuously push the hinge rod (33) to a horizontal state when the installation box (3) moves downward. The driving mechanism comprises a connecting frame (36) on which two ends of the two mounting boxes (3) are fixedly mounted away from each other, electric slide rails (37) are arranged on both sides of the front end of the storage tray (2), driving columns (38) are installed at the top ends of the electric slide rails (37), the top ends of the driving columns (38) pass through the front end of the connecting frame (36) and are slidably connected thereto, and electric push rods (39) are fixedly mounted at the front ends of the driving columns (38), and the output ends of the electric push rods (39) are connected to the bottom of the front end of the connecting frame (36).

2. The stem cell extraction method according to claim 1, characterized in that: Two limit plates (15) are fixedly mounted on both left and right ends of the mounting rod (9), and the limit plates (15) are respectively fitted with the upper and lower ends of the mounting frame (6), and a certain gap exists between the bottom end of the mounting frame (6) and the top end of the clamping box (8).

3. The stem cell extraction method according to claim 1, characterized in that: The discharging mechanism comprises a discharging platform (16) arranged on the right side of the front end of the storage tray (2), a first conveyor (17) is installed in the middle of the discharging platform (16), and two first partitions (18) are arranged on the top of the discharging platform (16), and the left ends of the first partitions (18) are fixedly installed with first racks (19), and the top of the left end of the discharging platform (16) is rotatably connected with a first gear (20), and the two first racks (19) are respectively located on the left and right sides of the first gear (20) and meshed with it, and a plurality of limit blocks (21) are fixedly installed on the top of the discharging platform (16), and the middle parts of the limit blocks (21) are slidably connected to limit rods (22), and the ends of the limit rods (22) are fixedly connected to the side walls of the first partitions (18), and the first partitions (18) are connected to the plurality of limit blocks (21) through compression springs.

4. The stem cell extraction method according to claim 1, characterized in that: A material receiving platform (23) is provided on the left side of the front end of the storage tray (2), a second conveyor (24) is installed in the middle of the material receiving platform (23), and two second partitions (25) are fixedly installed on the top of the material receiving platform (23).

5. The stem cell extraction method according to claim 1, characterized in that: The pressing mechanism comprises two support plates (34) fixedly mounted on the rear end of the first connecting rod (4), and the rear end of the support plate (34) is slidably connected to the pressing rod (35) via a tension spring.

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