Stem cell extraction process

Abstract

The invention discloses a stem cell extraction process. The stem cell extraction process comprises the following steps: step 1, pretreating adipose tissues; step 2, dissolving fat particles, adding collagenase with the same amount into the fat tissue in the first centrifugal test tube, and carrying out water bath digestion; step 3, primary centrifugation: putting the plurality of first centrifugal test tubes into a centrifugal machine for centrifugation; step 4, primary extraction of supernate: placing the plurality of first centrifugal test tubes on a supernate extraction device to remove residues of the supernate; step 5, secondary centrifugation: placing a plurality of first centrifugal test tubes in an open state on a test tube fixing cover, connecting the test tube fixing cover with a second centrifugal test tube, and performing centrifugal processing on the second centrifugal test tube; and 6, secondary extraction of the supernatant: removing the supernatant in the second centrifugal test tube by using the supernatant extraction device to obtain the adipose-derived stem cells. The stem cell extraction process provided by the invention is convenient for accurate and efficient removal of the supernate during stem cell separation.

Description

Technical Field

[0001] This invention relates to the technical field of stem cell extraction, specifically a stem cell extraction process. Background Technology

[0002] Adipose-derived stem cells are a type of stem cell with multi-lineage differentiation potential isolated from adipose tissue. Centrifugation is an essential step in the extraction of adipose-derived stem cells, and the precision of removing the supernatant after centrifugation directly affects the extraction effect of stem cells.

[0003] According to patent application CN202210281736.5, a stem cell extraction device is provided. The product includes a base and a placement section comprising a turntable with several placement slots, the bottom of which is connected to a first drive shaft. The extraction section includes a second drive shaft with a third transmission component mounted on it. A second transmission component is rotatably mounted on the third transmission component. A first transmission component is mounted on the turntable. A first adjusting component is rotatably mounted on a mounting bracket, connected to a limit rod and fitted with a limit block. An extraction mechanism, comprising a reciprocating assembly and a collecting assembly, is also mounted on the mounting bracket. This product enables effective separation and automatic extraction of stem cells without manual operation. It also adaptively clamps test tubes of different sizes, resulting in good performance.

[0004] The products in the aforementioned patents can achieve effective separation and automatic extraction of stem cells without manual operation, but they are not convenient for the precise and efficient removal of the supernatant during stem cell separation. Summary of the Invention

[0005] This invention mainly provides a stem cell extraction process to solve the technical problems mentioned in the background section.

[0006] The technical solution adopted by the present invention to solve the above-mentioned technical problems is as follows:

[0007] A stem cell extraction process includes the following steps:

[0008] Step 1: Pretreatment of adipose tissue. The adipose tissue, after removing blood vessels and connective tissue, is minced and added to the first centrifuge tube.

[0009] Step 2: Dissolving fat particles. Add an equal amount of collagenase to the fat tissue in the first centrifuge tube, and after the collagenase is added, place the first centrifuge tube in a water bath at 37 degrees Celsius to digest until the fat particles disappear.

[0010] Step 3: Initial centrifugation. Place multiple first centrifuge tubes into a centrifuge for centrifugation.

[0011] Step 4: Initial extraction of supernatant. Place the multiple centrifuge tubes after centrifugation onto the supernatant extraction device to remove any residual supernatant.

[0012] Step 5: Secondary centrifugation. Place multiple open first centrifuge tubes on the tube fixing cap, connect the tube fixing cap to the second centrifuge tube, and place the second centrifuge tube in the centrifuge for centrifugation.

[0013] Step 6: Secondary extraction of supernatant. Remove the second centrifuge tube after centrifugation and remove the tube cap. Use a supernatant extraction device to completely remove the supernatant in the second centrifuge tube to obtain adipose stem cells.

[0014] Preferably, the top of the outer wall of the second centrifuge tube is threaded to the bottom of the inner wall of the test tube fixing cap. The top of the test tube fixing cap is provided with a sealing cap, and a test tube clamping plate is provided inside the test tube fixing cap. A rubber tube is provided on the inner wall of the second centrifuge tube, and multiple light-transmitting observation components and elastic squeezing components are provided through the side wall of the second centrifuge tube. The multiple light-transmitting observation components and elastic squeezing components are arranged in a ring array. In this preferred embodiment, the second centrifuge tube and the test tube fixing cap facilitate secondary centrifugation of the solutions in multiple first centrifuge tubes, and also facilitate the removal of the supernatant after centrifugation.

[0015] Preferably, the light-transmitting observation component includes a first observation port penetrating the side wall of the second centrifuge tube, a glass plate disposed within the first observation port, a second observation port penetrating the side wall of the rubber tube, and a transparent thin film disposed within the second observation port, wherein the first observation port and the second observation port are positioned correspondingly. In this preferred embodiment, the light-transmitting observation component facilitates the observation of the solution inside the second centrifuge tube.

[0016] Preferably, the elastic squeezing component includes a first groove embedded in the inner wall of the second centrifuge tube, a rectangular through hole passing through the side wall of the second centrifuge tube and communicating with the first groove, a slide rail disposed on the outer wall of the second centrifuge tube and covering the outside of the rectangular through hole, a slider slidably connected to the slide rail, a squeezing plate located in the first groove, a round-headed push rod with one end passing through the slider and connected to the squeezing plate, and a spring sleeved on the outer wall of the round-headed push rod and with one end abutting against the slider. In this preferred embodiment, the elastic squeezing component facilitates the squeezing of the interface between the upper clear liquid and the lower cell fluid, thereby reducing the area of ​​the interface and facilitating the precise removal of the supernatant.

[0017] Preferably, the supernatant extraction device includes a base plate, two positioning columns on the base plate, a flip-top plate connected at one end of the bottom to the top of the two positioning columns via a flip-drive component, a self-lifting suction component located on the top of the flip-top plate with its execution end penetrating the flip-top plate, a positioning tube located on the top of the base plate and on one side of the two positioning columns, a test tube placement tray rotatably connected to the top of the positioning tube, a rotation drive component located between the two positioning columns for driving the test tube placement tray to rotate, multiple first linear modules vertically arranged on the inner wall of the positioning tube and arranged in a circular array, a lifting ring connected to the execution ends of the multiple first linear modules on the outer wall, multiple magnetic suction squeezing components arranged in a circular array on the inner wall of the lifting ring, and a liquid level stratification observation component. In this preferred embodiment, the supernatant extraction device facilitates the removal of residual supernatant in the first centrifuge tube, ensuring the efficiency of supernatant removal, and facilitates the precise removal of supernatant in the second centrifuge tube.

[0018] Preferably, the flipping drive component includes a drive motor disposed on the outer wall of one of the positioning columns, a rotating shaft disposed at the actuating end of the drive motor and rotatably connected to the other positioning column at the other end, and an electromagnetic ring embedded in the positioning column and sleeved outside the rotating shaft. One bottom end of the flipping top plate is connected to the rotating shaft via a support plate. In this preferred embodiment, the flipping drive component facilitates the rotation of the flipping top plate, enabling the rapid insertion of the first and second centrifuge tubes.

[0019] Preferably, the self-lifting suction component includes an arc-shaped through hole passing through the top of the flip-top plate, a second linear module located on the top of the flip-top plate and on one side of the arc-shaped through hole, a telescopic cylinder vertically mounted on the actuating end of the second linear module, an L-shaped plate mounted on the actuating end of the telescopic cylinder, a third linear module mounted on the side wall of the L-shaped plate, and a suction tube mounted on the actuating end of the third linear module, with its bottom end passing through the arc-shaped through hole and extending to the lower part of the flip-top plate. In this preferred embodiment, the self-lifting suction component facilitates the removal of supernatant from the test tube.

[0020] Preferably, the rotation drive component includes a base plate with two positioning posts connected to its two ends, a stepper motor embedded in the base plate, a toothed ring disposed at the bottom of the test tube placement tray, and a drive gear disposed at the actuating end of the stepper motor and meshing with the toothed ring. In this preferred embodiment, the rotation drive component facilitates the rotation of the test tube placement tray.

[0021] Preferably, the magnetic suction extrusion component includes a herringbone-shaped positioning plate disposed on the inner ring of the lifting ring, an electric cylinder disposed on the side wall of the herringbone-shaped positioning plate, and a magnetic suction plate disposed on the actuating end of the electric cylinder. In this preferred embodiment, the magnetic suction extrusion component facilitates the movement of the elastic extrusion component to the interface between the upper clarified liquid and the lower cell fluid, and drives the elastic extrusion component to perform the extrusion operation.

[0022] Preferably, the liquid level observation component includes a photoelectric refractive liquid level sensor disposed within the inner ring of the lifting ring. In this preferred embodiment, the liquid level observation component facilitates the determination of the location of the liquid level interface.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0024] The extraction process in this invention facilitates the precise and efficient removal of supernatant during stem cell separation. After centrifugation and precipitation in the first centrifuge tube, residual supernatant is removed, ensuring removal efficiency while removing most of the supernatant. The second centrifuge tube performs a second centrifugation on the solution from the first centrifuge tube with multiple residual supernatant removed. After centrifugation, it is easier to reduce the liquid-liquid interface area to achieve precise removal of supernatant.

[0025] The second centrifuge tube and the tube fixing cap facilitate secondary centrifugation of the solutions in multiple first centrifuge tubes. At the same time, it facilitates the removal of supernatant after centrifugation. The light-transmitting observation component facilitates the observation of the solution in the second centrifuge tube. The elastic squeezing component facilitates the squeezing of the interface between the upper clear liquid and the lower cell liquid to reduce the interface area and facilitate the precise removal of supernatant.

[0026] The supernatant extraction device facilitates the removal of residual supernatant in the first centrifuge tube, ensuring efficient supernatant removal and enabling precise removal of supernatant from the second centrifuge tube. The device features a flip-drive mechanism to rotate the top plate, facilitating the rapid placement of the first and second centrifuge tubes. A self-lifting suction mechanism removes supernatant from the tubes. A rotation drive mechanism rotates the tube placement tray, moving each first centrifuge tube sequentially to the supernatant removal station. A magnetic squeezing mechanism moves the elastic squeezing mechanism to the interface between the upper clear liquid and the lower cell fluid, performing the squeezing action. A liquid surface layer observation mechanism helps determine the location of the liquid surface layer interface.

[0027] The present invention will be explained in detail below with reference to the accompanying drawings and specific embodiments. Attached Figure Description

[0028] Figure 1 This is a flowchart illustrating the overall process flow of the present invention.

[0029] Figure 2 This is an isometric view of the first centrifuge tube structure of the present invention;

[0030] Figure 3 This is an isometric view of the second centrifuge tube structure of the present invention;

[0031] Figure 4 This is an exploded view of the second centrifuge tube structure of the present invention;

[0032] Figure 5 This is an exploded view of the supernatant extraction device of the present invention;

[0033] Figure 6 This is an isometric view of the self-lifting suction component structure of the present invention;

[0034] Figure 7 This is an isometric view of the magnetic suction extrusion component structure of the present invention;

[0035] Figure 8 This is a cross-sectional view of the self-lifting suction component of the present invention;

[0036] Figure 9 This is a cross-sectional view of the second centrifuge tube structure of the present invention;

[0037] Figure 10 This is an enlarged view of the structure at point A of the present invention.

[0038] Figure Descriptions: 10. First centrifuge tube; 20. Second centrifuge tube; 21. Tube fixing cap; 211. Sealing cap; 212. Tube clamping plate; 22. Rubber tube; 23. Light-transmitting observation component; 231. First observation port; 232. Glass slide; 233. Second observation port; 234. Transparent film sheet; 24. Elastic extrusion component; 241. First groove; 242. Rectangular through hole; 243. Slide rail; 244. Slider; 245. Extrusion plate; 246. Round-headed push rod; 247. Spring; 30. Supernatant extraction device; 31. Base plate; 311. Positioning column; 312. Positioning tube; 313. Tilting top plate; 32. Tilting drive component 321. Drive motor; 322. Rotating shaft; 323. Electromagnetic ring; 33. Self-lifting suction component; 331. Arc-edge through hole; 332. Second linear module; 333. Telescopic cylinder; 334. L-shaped plate; 335. Third linear module; 336. Suction tube; 34. Test tube placement tray; 35. Rotation drive component; 351. Base plate; 352. Stepper motor; 353. Toothed wall ring; 354. Drive gear; 36. First linear module; 37. Lifting ring; 38. Magnetic suction extrusion component; 381. Herringbone positioning plate; 382. Electric cylinder; 383. Magnetic suction plate; 39. Liquid level stratification observation component; 391. Photoelectric refractive liquid level sensor. Detailed Implementation

[0039] To facilitate understanding of the present invention, a more comprehensive description of the present invention will be given below with reference to the accompanying drawings, which illustrate several embodiments of the present invention. However, the present invention can be implemented in different forms and is not limited to the embodiments described in the text. Rather, these embodiments are provided to make the disclosure of the present invention more thorough and complete.

[0040] It should be noted that when an element is referred to as being "fixed to" another element, it can be directly on the other element or there may be an intervening element. When an element is referred to as being "connected to" another element, it can be directly connected to the other element or there may be an intervening element. The terms "vertical," "horizontal," "left," "right," and similar expressions used in this document are for illustrative purposes only.

[0041] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0042] Please refer to the appendix carefully. Figure 1 , 2 As shown, in a preferred embodiment of the present invention, a stem cell extraction process includes the following steps: Step 1, pretreatment of adipose tissue, wherein the adipose tissue with blood vessels and connective tissue removed is shredded and added to a first centrifuge tube 10; Step 2, dissolution of fat particles, wherein an equal amount of collagenase is added to the adipose tissue in the first centrifuge tube 10, and after the collagenase is added, the first centrifuge tube 10 is placed in a water bath at 37 degrees Celsius for digestion until the fat particles disappear; Step 3, initial centrifugation, wherein multiple first centrifuge tubes 10 are placed in a centrifuge for centrifugation; Step 4, initial extraction of supernatant, wherein multiple centrifuged first centrifuge tubes 10 are placed on a supernatant extraction device 30 for residual removal of supernatant.

[0043] It should be noted that, in this embodiment, during the extraction of adipose stem cells, multiple first centrifuge tubes 10 are taken, the adipose tissue with blood vessels and connective tissue removed is cut into small pieces and added to the multiple first centrifuge tubes 10 respectively, an equal amount of collagenase is added to the adipose tissue in the first centrifuge tubes 10, and after the collagenase is added, the first centrifuge tubes 10 are placed in a water bath and digested in a shaking water bath at 37 degrees Celsius until the fat particles disappear. The multiple first centrifuge tubes 10 are placed in a centrifuge for centrifugation, and the centrifuged multiple first centrifuge tubes 10 are placed on a supernatant extraction device 30 to remove the supernatant residue.

[0044] Furthermore, when removing residual supernatant, the liquid level can be set to the unit distance upward from the interface between the supernatant and the lower cell fluid. This allows for rapid removal of the supernatant while preventing the cell fluid from being aspirated.

[0045] Please refer to the appendix carefully. Figure 1 , 3 As shown in Figures 4, 9, and 10, in another preferred embodiment of the present invention, in step five, secondary centrifugation, multiple open first centrifuge tubes 10 are placed on a tube fixing cap 21, and the tube fixing cap 21 is connected to a second centrifuge tube 20. The second centrifuge tube 20 is then placed in a centrifuge for centrifugation. The top of the outer wall of the second centrifuge tube 20 is threaded to the bottom of the inner wall of the tube fixing cap 21. The top of the tube fixing cap 21 is provided with a sealing cap 211, and a tube clamping plate 212 is provided inside the tube fixing cap 21. A rubber tube 22 is provided on the inner wall of the second centrifuge tube 20. Multiple light-transmitting observation components 23 and elastic compression components 24 are provided through the side wall of the second centrifuge tube 20. The multiple light-transmitting observation components 23 and elastic compression components 24 are arranged in a ring array. The light-transmitting observation component 23 includes a first observation port 231 that passes through the side wall of the second centrifuge tube 20. The first observation port 231 contains a glass plate 232, a second observation port 233 is inserted through the side wall of the rubber tube 22, and a transparent film 234 is disposed in the second observation port 233. The positions of the first observation port 231 and the second observation port 233 correspond. The elastic extrusion component 24 includes a first groove 241 embedded in the inner wall of the second centrifuge tube 20, a rectangular through hole 242 inserted through the side wall of the second centrifuge tube 20 and communicating with the first groove 241, a slide rail 243 disposed on the outer wall of the second centrifuge tube 20 and covering the outside of the rectangular through hole 242, a slider 244 slidably connected to the slide rail 243, an extrusion plate 245 located in the first groove 241, a round-headed push rod 246 with one end passing through the slider 244 and connected to the extrusion plate 245, and a spring 247 sleeved on the outer wall of the round-headed push rod 246 and with one end abutting against the slider 244.

[0046] It should be noted that, in this embodiment, when performing secondary centrifugation on multiple first centrifuge tubes 10, the cap 211 is first connected to the tube fixing cap 21, and the tube fixing cap 21 is placed upside down. The caps of multiple first centrifuge tubes 10 are removed and inserted into the tube retaining plate 212 inside the upside-down tube fixing cap 21. After the multiple first centrifuge tubes 10 are fixed, the body of the second centrifuge tube 20 is connected to the upside-down tube fixing cap 21. After the connection is completed, the second centrifuge tube 20 can be held and placed in the centrifuge for centrifugation. During centrifugation, the centrifugal force causes all the solution in the first centrifuge tube 10 to enter the second centrifuge tube 20, avoiding the residue of solution in the first centrifuge tube 10. After centrifugation, the tube fixing cap 21 is removed, and the supernatant can be finely removed. After the supernatant is removed, adipose stem cells can be obtained.

[0047] Furthermore, the light-transmitting observation component 23 facilitates the observation of the liquid layering by the liquid layering observation component 39, allowing the solution in the second centrifuge tube 20 to be observed directly through the glass slide 232 and the transparent film 234. The rubber tube 22 is connected to the inner wall of the second centrifuge tube 20 on both sides of the transparent film 234 by rubber strips, thus avoiding the impact on the observation effect of the light-transmitting observation component 23 when the elastic squeezing component 24 squeezes the rubber tube 22.

[0048] Furthermore, when the elastic extrusion component 24 is in use, the magnetic extrusion component 38 drives the round-headed push rod 246 to move up and down. When the round-headed push rod 246 moves up and down, the slider 244 slides in the slide rail 243. When the elastic extrusion component 24 extrudes, the round-headed push rod 246 drives the extrusion plate 245 to extrude the rubber tube 22, so as to reduce the interface area between the upper clear liquid and the lower cell liquid, so as to facilitate the precise removal of the supernatant. After the extrusion is canceled, the spring 247 drives the round-headed push rod 246 to reset.

[0049] Please refer to the appendix carefully. Figure 1 , 5As shown in Figures 6, 7, and 8, in another preferred embodiment of the present invention, in step six, the secondary extraction of the supernatant, the second centrifuge tube 20 after centrifugation is removed and the tube fixing cap 21 is removed. The supernatant in the second centrifuge tube 20 is completely removed using the supernatant extraction device 30 to obtain adipose stem cells. The supernatant extraction device 30 includes a base plate 31, two positioning columns 311 disposed on the base plate 31, a flipping top plate 313 connected at one end of the bottom of the two positioning columns 311 by a flipping drive component 32, a self-lifting suction component 33 disposed on the top of the flipping top plate 313 with its execution end penetrating through the flipping top plate 313, and a positioning... The positioning tube 312 includes a test tube placement tray 34 rotatably connected to the top of the positioning tube 312, a rotation drive component 35 located between the two positioning posts 311 for driving the test tube placement tray 34 to rotate, multiple first linear modules 36 vertically arranged in a circular array on the inner wall of the positioning tube 312, a lifting ring 37 connected to the actuating ends of the multiple first linear modules 36 on the outer wall, multiple magnetic suction and squeezing components 38 arranged in a circular array on the inner wall of the lifting ring 37, and a liquid level layer observation component 39. The flipping drive component 32 includes a drive motor 321 located on the outer wall of one of the positioning posts 311, a rotating shaft 322 located at the actuating end of the drive motor 321 and rotatably connected to the other positioning post 311, and... An electromagnetic ring 323 is embedded in the positioning post 311 and sleeved on the outside of the rotating shaft 322. The bottom of one end of the flip-top plate 313 is connected to the rotating shaft 322 through a support plate. The self-lifting suction component 33 includes an arc-shaped through hole 331 passing through the top of the flip-top plate 313, a second linear module 332 located on the top of the flip-top plate 313 and on one side of the arc-shaped through hole 331, a telescopic cylinder 333 vertically located at the execution end of the second linear module 332, an L-shaped plate 334 located at the execution end of the telescopic cylinder 333, a third linear module 335 located on the side wall of the L-shaped plate 334, and a third linear module 335 located at the execution end of the third linear module 335 with its bottom end passing through the arc-shaped through hole 331 and extending to the flip-top plate. The suction tube 336 at the lower part of 313; the rotation drive component 35 includes a base plate 351 with two positioning columns 311 connected to its two ends respectively; a stepper motor 352 embedded in the base plate 351; a toothed wall ring 353 located at the bottom of the test tube placement tray 34; and a drive gear 354 located at the execution end of the stepper motor 352 and meshing with the toothed wall ring 353; the magnetic suction squeezing component 38 includes a herringbone-shaped positioning plate 381 located in the inner ring of the lifting ring 37; an electric cylinder 382 located on the side wall of the herringbone-shaped positioning plate 381; and a magnetic suction plate 383 located at the execution end of the electric cylinder 382; the liquid level layer observation component 39 includes a photoelectric refractive liquid level sensor 391 located in the inner ring of the lifting ring 37.

[0050] It should be noted that, in this embodiment, when the supernatant extraction device 30 extracts the supernatant from the solution in the first centrifuge tube 10, the flipping drive component 32 drives the flipping top plate 313 to flip up, placing multiple first centrifuge tubes 10 into the tube placement holes on the tube placement tray 34 respectively. The flipping top plate 313 is then reset. At this time, the rotation drive component 35 can drive the tube placement tray 34 to rotate, so as to move each first centrifuge tube 10 to the supernatant removal station in sequence. The self-lifting suction component 33 can remove the supernatant residue in the tube. An optical sensor can be installed on the outer wall of the positioning tube 312 at the position corresponding to the supernatant removal station to facilitate understanding the location of the interface between the supernatant and the lower cell fluid.

[0051] When the supernatant extraction device 30 extracts the supernatant in the second centrifuge tube 20, the flipping drive component 32 drives the flipping top plate 313 to flip up, placing the second centrifuge tube 20 in the positioning tube 312. A snap-fit ​​groove can be provided at the bottom of the inner wall of the positioning tube 312 to facilitate the stable fixation of the second centrifuge tube 20. The flipping top plate 313 is reset. At this time, the execution end of the first linear module 36 drives the lifting ring 37 to rise and fall until the magnetic suction squeezing component 38 magnetically connects to the round-head push rod 246. The controller receives the boundary liquid level information from the photoelectric refractive liquid level sensor 391 and triggers the first linear module 36 until the round-head push rod 246 moves to the solution interface. The magnetic suction squeezing component 38 drives the round-head push rod 246 to move to produce a squeezing effect, so as to reduce the solution interface area. The self-lifting suction component 33 can then accurately suction the supernatant. The controller receives the liquid level information from the photoelectric refractive liquid level sensor 391 and stops the self-lifting suction component 33 after the supernatant is removed.

[0052] Furthermore, when the flipping drive component 32 is working, the drive motor 321 drives the rotating shaft 322 to rotate, and the rotating shaft 322 drives the flipping top plate 313 to rotate. After the flipping top plate 313 is reset, the electromagnetic ring 323 can magnetically fix the rotating shaft 322 to lock and fix the rotating shaft 322.

[0053] Furthermore, when the rotation drive component 35 is working, the stepper motor 352 drives the drive gear 354 to rotate, and the drive gear 354 drives the test tube placement tray 34 to rotate through the toothed ring 353.

[0054] Furthermore, when the self-lifting suction component 33 is working, the discharge end of the suction pipe 336 can be connected to the negative pressure system, the execution end of the second linear module 332 drives the suction pipe 336 to move linearly, the telescopic cylinder 333 can drive the suction pipe 336 to rise and fall quickly, and the third linear module 335 can drive the suction pipe 336 to rise and fall stably and accurately. The suction pipe 336 can suction and discharge the supernatant.

[0055] Furthermore, when the magnetic suction extrusion component 38 is working, the magnetic suction plate 383 is electromagnetically attracted to the end of the round-headed push rod 246, and the actuator of the electric cylinder 382 can drive the magnetic suction plate 383 to move, thereby driving the round-headed push rod 246 to extrude and push.

[0056] The specific process of this invention is as follows:

[0057] The controller model is “6ES7315-2EH14-0AB0”, and the photoelectric refractive liquid level sensor model 391 is “FS-IR02”.

[0058] During the extraction of adipose stem cells, multiple first centrifuge tubes 10 are taken, and the adipose tissue with blood vessels and connective tissue removed is cut into small pieces and added to the multiple first centrifuge tubes 10 respectively. An equal amount of collagenase is added to the adipose tissue in the first centrifuge tubes 10. After the collagenase is added, the first centrifuge tubes 10 are placed in a water bath and digested in a shaking water bath at 37 degrees Celsius until the fat particles disappear. The multiple first centrifuge tubes 10 are placed in a centrifuge for centrifugation. After centrifugation, the multiple first centrifuge tubes 10 are placed on a supernatant extraction device 30 to remove the supernatant residue.

[0059] When removing residual supernatant, the unit distance upward from the interface between the supernatant and the lower cell fluid can be set as the liquid level for liquid collection. This can achieve rapid removal of supernatant while avoiding the aspiration of cell fluid.

[0060] When performing secondary centrifugation on multiple first centrifuge tubes 10, first connect the cap 211 to the tube fixing cap 21 and place the tube fixing cap 21 upside down. Remove the caps of multiple first centrifuge tubes 10 and insert them into the tube clamping plate 212 inside the upside-down tube fixing cap 21. After the multiple first centrifuge tubes 10 are fixed, connect the body of the second centrifuge tube 20 to the upside-down tube fixing cap 21. After the connection is completed, the second centrifuge tube 20 can be held and placed in the centrifuge for centrifugation. During centrifugation, the centrifugal force causes all the solution in the first centrifuge tube 10 to enter the second centrifuge tube 20, avoiding the solution remaining in the first centrifuge tube 10. After centrifugation, remove the tube fixing cap 21 to perform fine removal of the supernatant. After the supernatant is removed, adipose stem cells can be obtained.

[0061] The light-transmitting observation component 23 facilitates the observation of the liquid layering by the liquid layering observation component 39. The solution in the second centrifuge tube 20 can be observed directly through the glass slide 232 and the transparent film 234. The rubber tube 22 is connected to the inner wall of the second centrifuge tube 20 on both sides of the transparent film 234 by rubber strips, which avoids the impact of the elastic squeezing component 24 on the observation effect of the light-transmitting observation component 23 when squeezing the rubber tube 22.

[0062] When the elastic extrusion component 24 is in use, the magnetic extrusion component 38 drives the round-headed push rod 246 to move up and down. When the round-headed push rod 246 moves up and down, the slider 244 slides in the slide rail 243. When the elastic extrusion component 24 extrudes, the round-headed push rod 246 drives the extrusion plate 245 to extrude the rubber tube 22, so as to reduce the interface area between the upper clear liquid and the lower cell liquid, so as to facilitate the precise removal of the supernatant. After the extrusion is canceled, the spring 247 drives the round-headed push rod 246 to return to its original position.

[0063] When the supernatant extraction device 30 extracts the supernatant from the solution in the first centrifuge tube 10, the flipping drive component 32 drives the flipping top plate 313 to flip up, placing multiple first centrifuge tubes 10 into the test tube placement holes on the test tube placement tray 34 respectively. The flipping top plate 313 is then reset. At this time, the rotation drive component 35 can drive the test tube placement tray 34 to rotate, so as to move each first centrifuge tube 10 to the supernatant removal station in sequence. The self-lifting suction component 33 can remove the supernatant residue in the test tube. An optical sensor can be installed on the outer wall of the positioning tube 312 at the position corresponding to the supernatant removal station to facilitate understanding the location of the interface between the supernatant and the lower cell fluid.

[0064] When the supernatant extraction device 30 extracts the supernatant in the second centrifuge tube 20, the flipping drive component 32 drives the flipping top plate 313 to flip up, placing the second centrifuge tube 20 in the positioning tube 312. A snap-fit ​​groove can be provided at the bottom of the inner wall of the positioning tube 312 to facilitate the stable fixation of the second centrifuge tube 20. The flipping top plate 313 is reset. At this time, the execution end of the first linear module 36 drives the lifting ring 37 to rise and fall until the magnetic suction squeezing component 38 magnetically connects to the round-head push rod 246. The controller receives the boundary liquid level information from the photoelectric refractive liquid level sensor 391 and triggers the first linear module 36 until the round-head push rod 246 moves to the solution interface. The magnetic suction squeezing component 38 drives the round-head push rod 246 to move to produce a squeezing effect, so as to reduce the solution interface area. The self-lifting suction component 33 can then accurately suction the supernatant. The controller receives the liquid level information from the photoelectric refractive liquid level sensor 391 and stops the self-lifting suction component 33 after the supernatant is removed.

[0065] When the flipping drive component 32 is working, the drive motor 321 drives the rotating shaft 322 to rotate, and the rotating shaft 322 drives the flipping top plate 313 to rotate. After the flipping top plate 313 is reset, the electromagnetic ring 323 can magnetically fix the rotating shaft 322 to lock and fix the rotating shaft 322.

[0066] When the rotation drive component 35 is working, the stepper motor 352 drives the drive gear 354 to rotate, and the drive gear 354 drives the test tube placement tray 34 to rotate through the toothed ring 353.

[0067] When the self-lifting suction component 33 is working, the discharge end of the suction pipe 336 can be connected to the negative pressure system. The execution end of the second linear module 332 drives the suction pipe 336 to move linearly. The telescopic cylinder 333 can drive the suction pipe 336 to rise and fall quickly. The third linear module 335 can drive the suction pipe 336 to rise and fall stably and accurately. The suction pipe 336 can suck out the supernatant.

[0068] When the magnetic suction extrusion component 38 is working, the magnetic suction plate 383 is electromagnetically attracted to the end of the round-headed push rod 246, and the actuator of the electric cylinder 382 can drive the magnetic suction plate 383 to move, thereby driving the round-headed push rod 246 to extrude and push.

[0069] The present invention has been described by way of example in conjunction with the accompanying drawings. Obviously, the specific implementation of the present invention is not limited to the above-described manner. Any non-substantial improvement made by adopting the inventive concept and technical solution of the present invention, or the direct application of the inventive concept and technical solution of the present invention to other occasions without modification, shall be within the protection scope of the present invention.

Claims

1. A stem cell extraction process, characterized in that... This includes the following steps: Step 1: Pretreatment of adipose tissue. The adipose tissue, after removing blood vessels and connective tissue, is shredded and added to the first centrifuge tube (10). Step 2: Dissolving fat particles. Add an equal amount of collagenase to the fat tissue in the first centrifuge tube (10), and after the collagenase is added, place the first centrifuge tube (10) in a water bath at 37 degrees Celsius to digest until the fat particles disappear. Step 3: Initial centrifugation. Place multiple first centrifuge tubes (10) into a centrifuge for centrifugation. Step 4: Initial extraction of supernatant. Place multiple first centrifuge tubes (10) after centrifugation on the supernatant extraction device (30) to remove residual supernatant. Step 5: Secondary centrifugation. Place multiple open first centrifuge tubes (10) on the tube fixing cap (21), connect the tube fixing cap (21) to the second centrifuge tubes (20), and place the second centrifuge tubes (20) in the centrifuge for centrifugation. Step 6: Secondary extraction of supernatant. Remove the second centrifuge tube (20) after centrifugation and remove the tube cap (21). Use the supernatant extraction device (30) to completely remove the supernatant in the second centrifuge tube (20) to obtain adipose stem cells.

2. The stem cell extraction process according to claim 1, characterized in that, The top of the outer wall of the second centrifuge tube (20) is threaded to the bottom of the inner wall of the tube fixing cap (21). The top of the tube fixing cap (21) is provided with a sealing cap (211). The tube fixing cap (21) is provided with a tube clamping plate (212). The inner wall of the second centrifuge tube (20) is provided with a rubber tube (22). The side wall of the second centrifuge tube (20) is provided with multiple light-transmitting observation components (23) and elastic squeezing components (24). The multiple light-transmitting observation components (23) and elastic squeezing components (24) are arranged in a ring array.

3. The stem cell extraction process according to claim 2, characterized in that, The light-transmitting observation component (23) includes a first observation port (231) passing through the side wall of the second centrifuge tube (20), a glass plate (232) disposed in the first observation port (231), a second observation port (233) passing through the side wall of the rubber tube (22), and a transparent film (234) disposed in the second observation port (233). The positions of the first observation port (231) and the second observation port (233) correspond.

4. The stem cell extraction process according to claim 2, characterized in that, The elastic extrusion component (24) includes a first groove (241) embedded in the inner wall of the second centrifuge tube (20), a rectangular through hole (242) passing through the side wall of the second centrifuge tube (20) and communicating with the first groove (241), a slide rail (243) disposed on the outer wall of the second centrifuge tube (20) and covering the outside of the rectangular through hole (242), a slider (244) slidably connected to the slide rail (243), an extrusion plate (245) located in the first groove (241), a round-headed push rod (246) with one end passing through the slider (244) and connecting to the extrusion plate (245), and a spring (247) sleeved on the outer wall of the round-headed push rod (246) and with one end abutting against the slider (244).

5. The stem cell extraction process according to claim 1, characterized in that, The supernatant extraction device (30) includes a base plate (31), two positioning columns (311) on the base plate (31), a flip-top plate (313) connected at one end of the bottom to the top of the two positioning columns (311) via a flip-top drive component (32), a self-lifting suction component (33) located on the top of the flip-top plate (313) with its execution end penetrating through the flip-top plate (313), and a positioning tube (312) located on the top of the base plate (31) and on one side of the two positioning columns (311), rotatably connected to the... The positioning tube (312) has a test tube placement tray (34) at the top, a rotation drive component (35) located between the two positioning columns (311) for driving the test tube placement tray (34) to rotate, a plurality of first linear modules (36) vertically arranged on the inner wall of the positioning tube (312) and arranged in a ring array, a lifting ring (37) connected to the execution end of the plurality of first linear modules (36) on the outer wall, a plurality of magnetic suction squeezing components (38) arranged in a ring array on the inner wall of the lifting ring (37), and a liquid level layer observation component (39).

6. The stem cell extraction process according to claim 5, characterized in that, The flipping drive component (32) includes a drive motor (321) disposed on the outer wall of one of the positioning posts (311), a rotating shaft (322) disposed on the execution end of the drive motor (321) and rotatably connected to the other positioning post (311) at the other end, and an electromagnetic ring (323) embedded in the positioning post (311) and sleeved on the outside of the rotating shaft (322). The bottom of one end of the flipping top plate (313) is connected to the rotating shaft (322) through a support plate.

7. The stem cell extraction process according to claim 5, characterized in that, The self-lifting suction component (33) includes an arc-edge through hole (331) passing through the top of the flip-top plate (313), a second linear module (332) located on the top of the flip-top plate (313) and on one side of the arc-edge through hole (331), a telescopic cylinder (333) vertically located at the execution end of the second linear module (332), an L-shaped plate (334) located at the execution end of the telescopic cylinder (333), a third linear module (335) located on the side wall of the L-shaped plate (334), and a suction pipe (336) located at the execution end of the third linear module (335) with its bottom end passing through the arc-edge through hole (331) and extending to the lower part of the flip-top plate (313).

8. The stem cell extraction process according to claim 5, characterized in that, The rotation drive component (35) includes a base plate (351) with two positioning columns (311) connected to its two ends respectively, a stepper motor (352) embedded in the base plate (351), a toothed ring (353) located at the bottom of the test tube placement tray (34), and a drive gear (354) located at the execution end of the stepper motor (352) and meshing with the toothed ring (353).

9. A stem cell extraction process according to claim 5, characterized in that, The magnetic suction and pressing component (38) includes a herringbone-shaped positioning plate (381) disposed in the inner ring of the lifting ring (37), an electric cylinder (382) disposed in the side wall of the herringbone-shaped positioning plate (381), and a magnetic suction plate (383) disposed in the actuating end of the electric cylinder (382).

10. A stem cell extraction process according to claim 5, characterized in that, The liquid level observation component (39) includes a photoelectric refractive liquid level sensor (391) located in the inner ring of the lifting ring (37).

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