A clinical-grade fat element preparation system and method

Abstract

The present invention discloses a system and method for preparing clinical-grade fat elements, comprising a fat processing device, connecting pipelines, a reagent chamber, a weight chamber, an incubation chamber, a collecting tube and a waste liquid bag; the fat processing device comprises a centrifugal motor and a centrifugal mechanism; the connecting pipeline comprises a first pipeline, a second pipeline, a third pipeline and a fourth pipeline respectively connected to the centrifugal mechanism with the reagent chamber, the collecting tube, the weight chamber and the first branch, the reagent chamber is connected to the first pipeline, a first peristaltic pump is installed between the reagent chamber and the centrifugal mechanism, the waste liquid bag is connected to the first pipeline through the first branch, the first branch is connected to the second peristaltic pump, the incubation chamber is connected to the first pipeline through the second branch, the second branch is connected to the third peristaltic pump; the second pipeline of the collecting tube is connected to the fourth peristaltic pump; the weight chamber and the third pipeline of the centrifugal mechanism are connected to the fifth peristaltic pump; the present application realizes highly automated preparation of fat elements in different devices under GMP conditions.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and in particular to a system and method for preparing clinical-grade fat elements. Background Art

[0002] Adipocytes are surrounded by adipose matrix, namely SVF (Stromal Vascular Fraction), which represents a group of heterogeneous cells, also known as adipocytes. SVF can be obtained by processing fat cells in the human abdomen, buttocks, breasts and internal organs. The method is as follows: obtain adipose tissue after liposuction, wash to remove blood cells, digest with collagenase to obtain a single cell suspension, filter through a 0.22um filter to remove undigested tissue residues, and the precipitate obtained after centrifugation due to its light specific gravity is SVF.

[0003] At present, the processing and separation of fat in the human body are carried out using different devices. The fat needs to be manually transferred between different devices, and the degree of automation is low, which cannot meet clinical needs. Summary of the invention

[0004] In view of the defects in the prior art, the object of the present invention is to provide a clinical-grade fat element preparation system integrating fat processing and separation.

[0005] Furthermore, the present application also provides a method for preparing clinical-grade fat elements.

[0006] The technical solution adopted by the present invention is:

[0007] A clinical-grade fat element preparation system, comprising a fat processing device, a connecting pipeline, a reagent chamber, a counterweight chamber, an incubation chamber, a collecting tube and a waste liquid bag; the fat processing device comprises a centrifugal motor and a centrifugal mechanism connected to the centrifugal motor; the connecting pipeline comprises a first pipeline, a second pipeline, a third pipeline and a fourth pipeline respectively connected to the centrifugal mechanism; the reagent chamber is connected to the first pipeline, and a first peristaltic pump is installed between the reagent chamber and the centrifugal mechanism; the waste liquid bag is connected to the first pipeline through a first branch; the incubation chamber is connected to the first pipeline through a second branch, and a third peristaltic pump is connected to the second branch; the collecting tube is connected to the second pipeline, and a fourth peristaltic pump is connected to the second pipeline; the counterweight chamber is connected to the third pipeline, and a fifth peristaltic pump is connected to the third pipeline; one end of the fourth pipeline is connected to the centrifugal tube, and the other end is connected to the first branch, and the fourth pipeline is connected to the second peristaltic pump.

[0008] Furthermore, the incubation chamber includes an incubation bag, a shaking table, a constant temperature device, and an imaging system; the shaking table is connected to the incubation bag, and the shaking frequency range is 160rpm-190rpm; the constant temperature device maintains the constant temperature of the incubation bag at 36°C±2.

[0009] Furthermore, the imaging system includes a camera and an AI software processing system.

[0010] Furthermore, the centrifugal mechanism includes a centrifugal tube, a counterweight tube and a mounting bracket, the output shaft of the centrifugal motor is connected to the center of the mounting bracket, the centrifugal tube and the counterweight tube are respectively installed at both ends of the mounting bracket, and the first pipeline and the second pipeline are respectively connected to the centrifugal tube.

[0011] Further, one end of the third pipeline is connected to the counterweight tube, the other end of the third pipeline is connected to the counterweight chamber, and the fifth peristaltic pump is located between the counterweight chamber and the counterweight tube.

[0012] Furthermore, the reagent chamber includes an enzyme bag, a protective liquid bag and a physiological saline bag, and a first pinch valve, a second pinch valve and a third pinch valve are respectively installed between the enzyme bag, the protective liquid bag and the physiological saline bag and the first pipeline; a fourth pinch valve is installed at the liquid inlet end of the second peristaltic pump; and a fifth pinch valve is installed at the liquid inlet end of the third peristaltic pump.

[0013] Further, the centrifugal mechanism also includes an anti-winding device, which includes a fixed part, a rotating part, a first liquid pipe, a second liquid pipe, a third liquid pipe and a fourth liquid pipe, the rotating part is fixedly mounted on the mounting bracket, the fixed part is rotatably mounted on the outer periphery of the rotating part, and a first annular cavity, a second annular cavity, a third annular cavity and a fourth annular cavity are provided between the fixed part and the rotating part and spaced apart in the vertical direction; the first pipeline, the second pipeline, the third pipeline and the fourth pipeline are all fixed to the side wall of the fixed part, and are respectively connected to the first annular cavity, the second annular cavity, the third annular cavity and the fourth annular cavity; the first ends of the first liquid pipe, the second liquid pipe, the third liquid pipe and the fourth liquid pipe are all vertically connected to the rotating part, and are respectively connected to the first annular cavity, the second annular cavity, the third annular cavity and the fourth annular cavity, the second ends of the first liquid pipe, the second liquid pipe and the fourth liquid pipe are connected to the centrifuge tube, and the second end of the third liquid pipe is connected to the counterweight pipe.

[0014] Furthermore, the height of the second end of the fourth liquid tube is 2 ml higher than the height of the second end of the first liquid tube.

[0015] Furthermore, a 100um filter membrane is installed between the collecting tube and the centrifugal mechanism, and a hydrophobic breathable membrane is installed between the collecting tube and the fourth peristaltic pump.

[0016] Furthermore, the enzyme bag and the protective liquid bag use a fat element extraction instrument kit, collagenase and physiological saline are added to the enzyme bag, 1-5 ml of physiological saline is added to 1200 units of injectable collagenase, and the mixture is fully mixed. 1200 units of injectable collagenase can dissociate 10-20 ml of fat.

[0017] Furthermore, the protective liquid bag is filled with protective liquid, which is composed of amino acids, glucose and physiological saline. Specifically, 800ul of amino acids and 10ml of 10% glucose are added to every 100ml of physiological saline.

[0018] Furthermore, the present application provides a method for preparing clinical-grade adipocytes, using the above-mentioned clinical-grade adipocyte preparation system, comprising the following steps:

[0019] S1. Inject the blood-containing fat into the centrifuge tube from the injection port;

[0020] S2, open the third pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, and pump the saline in the saline bag into the centrifuge tube through the first pipeline;

[0021] S3, the fifth peristaltic pump rotates forward, and the physiological saline in the weight-balancing chamber is pumped into the weight-balancing tube through the third pipeline, and the weight-balancing tube is weighted according to the weight of the centrifuge tube;

[0022] S4, turning on the centrifugal motor, and shaking the blood fat and saline in the centrifuge tube evenly by the action of the centrifugal motor; or turning on the second peristaltic pump, and continuously rotating the second peristaltic pump forward and reversely, so that the mixture of blood fat and saline is continuously moved between the centrifuge tube and the fourth pipeline in a "pumping-discharging" manner, and the two are evenly mixed;

[0023] S5. After mixing evenly, start the centrifugal motor and centrifuge at 64G for 1 minute to wash and layer the fat;

[0024] After cleaning, the fourth pinch valve is opened, the other pinch valves are closed, the second peristaltic pump is reversed, and the third and fourth layers of waste liquid, cell fragments, blood cells and other residues after cleaning are discharged into the waste liquid bag through the first pipeline and the first branch;

[0025] S6, opening the fifth pinch valve, closing the other pinch valves, and reversing the first and third peristaltic pumps to pump the fat particles of the second layer into the incubation chamber;

[0026] S7, opening the first pinch valve, closing the other pinch valves, reversing the third peristaltic pump, and adding the enzyme into the incubation chamber;

[0027] S8, the incubation chamber is kept at 37°C for 1 hour; at the same time, the third pinch valve is opened, the other pinch valves are closed, the first peristaltic pump is rotated forward, physiological saline is added into the centrifuge tube, and the centrifuge tube is cleaned; after the cleaning is completed, the fourth pinch valve is opened, the other pinch valves are closed, the first peristaltic pump and the second peristaltic pump are reversed, and the cleaned liquid is discharged into the waste liquid bag;

[0028] S9, open the fifth pinch valve, close the other pinch valves, and rotate the first peristaltic pump and the third peristaltic pump forward to pump the fat in the incubation chamber into the centrifuge tube;

[0029] S10, after the weight tube is weighted according to the method in step S3, the centrifugal motor is started, and the centrifugal treatment is performed at 1500G for 5 minutes to centrifugally separate the fat;

[0030] After centrifugal stratification, the fourth pinch valve is opened, the other pinch valves are closed, the second peristaltic pump is reversed, and the upper fatty oil of the first layer is discharged into the waste liquid bag through the fourth pipeline and the first branch pipe;

[0031] The fourth pinch valve is opened, the other pinch valves are closed, the first peristaltic pump and the second peristaltic pump are reversed, and the tissue fragments of the second layer and the waste liquid of the third layer are discharged into the waste liquid bag through the first pipeline and the first branch pipe;

[0032] S11, open the second pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, add the protective liquid into the centrifuge tube; and weight the weight tube according to the method in step S3;

[0033] S12, turning on the centrifugal motor to evenly shake the fat and protective solution in the centrifuge tube through the action of the centrifugal motor; or turning on the second peristaltic pump to continuously rotate forward and reversely through the second peristaltic pump to continuously move the mixture of the fat and protective solution between the centrifuge tube and the fourth pipeline in a "pumping-spitting" manner to evenly mix the two;

[0034] S13, after mixing evenly, start the centrifugal motor, centrifuge at 600-800G for 3-5 minutes to wash and layer the fat;

[0035] After centrifugation, the fourth pinch valve is opened, the other pinch valves are closed, the first peristaltic pump and the second peristaltic pump are reversed, and the first layer and the second layer after centrifugation are discharged into the waste liquid bag through the first pipeline and the first branch;

[0036] S14, the fourth peristaltic pump is reversed to extract the third layer of fat cell population in the centrifuge tube into the collection tube; after collection, the front and rear ends of the collection tube are cut off, and the tube is sealed for freezing or injection; or the third layer of fat cell population is extracted into a syringe through a manual interface for clinical application, or transferred to a collection tube through a syringe for freezing or storage.

[0037] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become apparent from the description below, or will be learned through the practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following is a brief introduction to the drawings required for the specific embodiments or the description of the prior art. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the elements or parts are not necessarily drawn according to the actual scale.

[0039] Figure 1 This is a schematic diagram of the structure of the clinical-grade adipocyte preparation system provided in Example 1 of the present application;

[0040] Figure 2 A partial top view of the clinical-grade fat element preparation system provided in Example 1 of the present application;

[0041] Figure 3 A cross-sectional view of a clinical-grade fat element preparation system provided in Example 1 of the present application;

[0042] Figure 4 This is a schematic diagram of the structure of the anti-winding device provided in Example 1 of the present application;

[0043] Figure 5 A control system diagram provided in Example 1 of the present application;

[0044] Figure 6 This is the pipeline diagram provided in Example 1 of the present application.

[0045] Figure 7 This is a schematic diagram of the collection tube structure provided in Example 1 of the present application.

[0046] Among them, the fat processing device 1, the centrifugal motor 11, the centrifugal tube 12, the counterweight tube 13, the mounting bracket 14, the anti-winding device 15, the rotating part 151, the fixing part 152, the first annular cavity 153, the second annular cavity 154, the third annular cavity 155, the fourth annular cavity 156, the collecting tube 2, the filter membrane 201, the hydrophobic breathable membrane 202, the first pipeline 31, the second pipeline 32, the third pipeline 33, the fourth pipeline 34, the first branch 35, the second branch 36, the waste liquid bag 4, the first peristaltic pump 51, the second peristaltic pump 52, the third peristaltic pump 53, the fourth peristaltic pump 54, the fifth peristaltic pump 55, the first pinch valve 61, the second pinch valve 62, the third pinch valve 63, the fourth pinch valve 64, the fifth pinch valve 65, the incubation chamber 7, the enzyme bag 81, the protective liquid bag 82, the physiological saline bag 83, and the counterweight chamber 9. DETAILED DESCRIPTION

[0047] The following embodiments of the technical solution of the present invention are described in detail in conjunction with specific embodiments. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and are therefore only used as examples, and cannot be used to limit the protection scope of the present invention.

[0048] It should be noted that, unless otherwise specified, the technical terms or scientific terms used in this application should have the common meanings understood by those skilled in the art to which the present invention belongs.

[0049] Example 1

[0050] See also Figure 1 to Figure 7 The present embodiment provides a clinical-grade fat element preparation system, including a fat processing device 1, a connecting pipeline, a reagent chamber, a weight chamber 9, an incubation chamber 7, a collection tube 2 and a waste liquid bag 4; the fat processing device 1 includes a centrifugal motor 11 and a centrifugal mechanism connected to the centrifugal motor 11; the connecting pipeline includes a first pipeline 31, a second pipeline 32, a third pipeline 33 and a fourth pipeline 34 respectively connected to the centrifugal mechanism; the reagent chamber is connected to the first pipeline 31, and a first peristaltic pump 51 is installed between the reagent chamber and the centrifugal mechanism; the waste liquid bag 4 is connected to the first pipeline 31 through the first branch 35; the incubation chamber 7 is connected to the first pipeline 31 through the second branch 36, and the second branch 36 is connected to the third peristaltic pump 53; the collecting tube 2 is connected to the second pipeline 32, and the second pipeline 32 is connected to the fourth peristaltic pump; the weight chamber 9 is connected to the third pipeline 33, and the third pipeline 33 is connected to the fifth peristaltic pump 55; one end of the fourth pipeline 34 is connected to the centrifuge tube 12, and the other end is connected to the first branch 35, and the fourth pipeline 34 is connected to the second peristaltic pump 52.

[0051] The present application provides a fat processing device 1, connecting pipelines, a reagent chamber, a counterweight chamber 9, an incubation chamber 7, a collecting tube 2, a waste liquid bag 4 and a peristaltic pump, so that fat can be injected into a centrifugal mechanism, and physiological saline is injected into the centrifugal mechanism through a first pipeline 31 by the action of the peristaltic pump for cleaning under the action of a centrifugal motor 11; clean fat and enzymes are injected into the incubation chamber 7 by the action of the peristaltic pump to digest the fat, and the digested fat particles are injected into the centrifugal mechanism by the action of the peristaltic pump for separation under the action of the centrifugal motor 11, and the obtained fat elements are collected in the collecting tube 2 or manually sucked out for clinical application; the waste liquid after cleaning or centrifugal treatment of the fat can be pumped into the waste liquid bag 4 by the peristaltic pump, and in the sterile pipeline, the fat does not need to be manually converted between different devices, and the degree of automation is high.

[0052] The incubation room 7 includes an incubation bag, a shaking table, a constant temperature device, and an imaging system.

[0053] In order to facilitate fat digestion, the incubation bag is selected to be flat and bag-shaped.

[0054] To facilitate fat digestion, a shaker was connected to the incubation bag with a parameter range of 160 rpm-190 rpm.

[0055] In order to facilitate fat digestion, a constant temperature device is installed in the incubation chamber 7 to maintain the temperature of the incubation bag at 36°C±2; the constant temperature device is not limited to resistance wire heating, but also includes a water bath pot and the like.

[0056] In order to facilitate the judgment of fat digestion completion, the camera in the imaging system continuously records the fat digestion status and identifies it through the AI ​​software processing system; the camera is not limited to continuously recording the fat digestion status, and can also record it once at a certain interval, for example: 5 minutes.

[0057] The centrifugal mechanism includes a centrifugal tube 12, a counterweight tube 13 and a mounting bracket 14. The output shaft of the centrifugal motor 11 is connected to the center of the mounting bracket 14. The centrifugal tube 12 and the counterweight tube 13 are respectively installed at both ends of the mounting bracket 14. The first pipeline 31, the second pipeline 32 and the fourth pipeline 34 are respectively connected to the centrifugal tube 12; the third pipeline 33 is connected to the counterweight tube 13; one end of the fourth pipeline 34 is connected to the centrifugal tube 12, and the other end is connected to the first branch 35.

[0058] The weight tube 13 is used for weighting. The top of the centrifuge tube 12 is provided with an injection port for injecting fat, and is also provided with an extraction port, so that the fat elements can be manually sucked out using a syringe.

[0059] In order to facilitate balancing, it also includes a third pipeline 33, a fifth peristaltic pump 55 and a balancing weight chamber 9. One end of the third pipeline 33 is connected to the balancing weight tube 13, and the other end of the third pipeline 33 is connected to the balancing weight chamber 9. The fifth peristaltic pump 55 is connected to the third pipeline 33 and is located between the balancing weight chamber 9 and the balancing weight tube 13; wherein, the balancing weight chamber 9 is used to contain the balancing weight liquid, which can be physiological saline.

[0060] In order to facilitate the discharge of the fatty oil, a fourth pipeline 34 is also included, one end of which is connected to the centrifuge tube 12, and the other end is connected to the first branch 35. Through the fourth pipeline 34, the treated upper fatty oil in the centrifuge tube 12 can be pumped into the waste liquid bag 4.

[0061] The reagent chamber includes an enzyme bag 81, a protective liquid bag 82 and a physiological saline bag 83. A first pinch valve 61, a second pinch valve 62 and a third pinch valve 63 are respectively installed between the enzyme bag 81, the protective liquid bag 82 and the physiological saline bag 83 and the first pipeline 31, and the three are connected to the first pipeline 31 through corresponding pinch valves.

[0062] In order to facilitate the control of the connection and disconnection between the first branch 35 and the second branch 36 and the first pipeline 31 , a fourth pinch valve 64 is installed at the liquid inlet end of the second peristaltic pump 52 ; a fifth pinch valve 65 is installed at the liquid inlet end of the third peristaltic pump 53 .

[0063] In order to facilitate the collection of processed fat cells, a collection tube 2 is installed at the liquid inlet end of the fourth peristaltic pump, a 100u filter membrane 201 is installed between the collection tube 2 and the centrifugal mechanism, and a hydrophobic breathable membrane 202 is installed between the collection tube 2 and the fourth peristaltic pump. The processed fat cells are pumped into the collection tube 2 under the action of the fourth peristaltic pump.

[0064] In order to prevent the pipeline from being entangled, the centrifugal mechanism also includes an anti-entanglement device 15, which includes a fixed part 152, a rotating part 151, a first liquid pipe, a second liquid pipe, a third liquid pipe and a fourth liquid pipe. The rotating part 151 is fixedly installed on the mounting bracket 14 and rotates with the mounting bracket 14 during centrifugal treatment. The fixed part 152 is rotatably installed on the outer periphery of the rotating part 151. A first annular cavity 153, a second annular cavity 154, a third annular cavity 155 and a fourth annular cavity 156 are provided between the fixed part 152 and the rotating part 151 and at intervals in the vertical direction; in this embodiment, the outer wall of the middle part of the rotating part 151 is slightly concave inward to form a circumferentially arranged groove, which cooperates with the inner wall of the outer fixed part 152 to form a cavity, and sealing rings are provided at the upper and lower ends and the middle interval of the cavity. , dividing the cavity into a first annular cavity 153, a second annular cavity 154, a third annular cavity 155 and a fourth annular cavity 156 which are arranged vertically; the first pipeline 31, the second pipeline 32, the third pipeline 33 and the fourth pipeline 34 are all fixed on the side wall of the fixed part 152, and are respectively connected with the first annular cavity 153, the second annular cavity 154, the third annular cavity 155 and the fourth annular cavity 156; the first ends of the first liquid pipe, the second liquid pipe, the third liquid pipe and the fourth liquid pipe are all vertically connected to the rotating part 151, and are respectively connected with the first annular cavity 153, the second annular cavity 154, the third annular cavity 155 and the fourth annular cavity 156, the second ends of the first liquid pipe, the second liquid pipe and the fourth liquid pipe are connected with the centrifuge tube 12, and the second end of the third liquid pipe is connected with the counterweight pipe 13.

[0065] In order to facilitate the discharge of different substances, the second end of the first liquid tube is close to the bottom of the centrifuge tube 12, and the height of the second end of the fourth liquid tube is 2 ml higher than the height of the second end of the first liquid tube.

[0066] The present application also includes an instrument kit for extracting fat cells, which includes tissue dissociation powder and protective solution.

[0067] In order to facilitate the dissociation of adipose tissue, collagenase is used for dissociation of adipose tissue. Collagenase and physiological saline are added to the enzyme bag 81. 1-5 ml of physiological saline is added to 1200 units of injectable collagenase, and the mixture is fully mixed. 1200 units of injectable collagenase can dissociate 10-20 mL of fat.

[0068] In order to reduce damage to fat cells during centrifugation, the fat cells after tissue dissociation are dissolved in protective solution for operation. The protective solution is filled in the protective solution bag 82, and the protective solution is composed of amino acids, glucose and normal saline. Specifically, 800ul amino acids and 10ml 10% glucose are added to every 100ml normal saline.

[0069] The present application also includes a cabinet, in which the fat processing device 1 is installed, and connecting pipes, reagent chambers, weight chambers 9, waste liquid bags 4, peristaltic pumps and incubation chambers 7 are all installed on the cabinet. A touch screen is also installed on the cabinet, which can realize human-computer interaction.

[0070] Example 2

[0071] This embodiment provides a method for preparing clinical-grade fat element, which uses the bed-level fat element preparation system provided in Example 1, and the preparation method is as follows:

[0072] S1. Inject the blood-containing fat into the centrifuge tube 12 from the injection port.

[0073] S2, the third pinch valve 63 is opened, the other pinch valves are closed, the first peristaltic pump 51 rotates forward, and the physiological saline in the physiological saline bag 83 is pumped into the centrifuge tube 12 through the first pipeline 31.

[0074] S3, the fifth peristaltic pump 55 rotates forward, and the physiological saline in the weight-balancing chamber 9 is pumped into the weight-balancing tube 13 through the third pipeline 33, and the weight-balancing tube 13 is weighted according to the weight of the centrifuge tube 12.

[0075] S4, turn on the centrifugal motor 11, and shake the blood fat and physiological saline in the centrifuge tube 12 evenly through the action of the centrifugal motor 11; or turn on the second peristaltic pump 52, and continuously rotate the second peristaltic pump 52 forward and reverse to continuously move the mixture of blood fat and physiological saline between the centrifuge tube 12 and the fourth pipeline 34 in a "pumping-discharging" manner to mix the two evenly.

[0076] S5. After mixing evenly, start the centrifugal motor 11 and centrifuge at 60-70G for 1-3 minutes to wash and layer the fat.

[0077] After stratification, stop and take pictures. The fat tissue from top to bottom is: the first layer: triglycerides; the second layer: fat particles arranged according to density gradient; the third layer: waste liquid; the fourth layer: cell fragments, blood cells and other residues; the waste liquid that needs to be discharged is the third layer: waste liquid and the fourth layer: cell fragments, blood cells and other residues. Multiple washings can be used to obtain the second layer of fat arranged according to density gradient, usually three times, until the bottom layer is clear.

[0078] After cleaning, the fourth pinch valve 64 is opened, the other pinch valves are closed, the second peristaltic pump 52 is reversed, and the third and fourth layers of waste liquid, cell fragments, blood cells and other residues after cleaning are discharged into the waste liquid bag 4 through the fourth pipeline 34 and the first branch 35.

[0079] S6, the fifth pinch valve 65 is opened, the other pinch valves are closed, the first peristaltic pump 51 and the third peristaltic pump 53 are reversed, and the fat particles of the second layer are pumped into the incubation chamber 7.

[0080] S7 , open the first pinch valve 61 , close the other pinch valves, reverse the third peristaltic pump 53 , and add the enzyme into the incubation chamber 7 .

[0081] S8, keep the incubator 7 at a constant temperature of 37°C for 1 hour; at the same time, open the third clamp valve 63, close the other clamp valves, rotate the first peristaltic pump 51 forward, add physiological saline into the centrifuge tube 12, and clean the centrifuge tube 12; after cleaning, open the fourth clamp valve 64, close the other clamp valves, and reverse the first peristaltic pump 51 and the second peristaltic pump 52 to discharge the cleaned liquid into the waste liquid bag 4.

[0082] S9, open the fifth pinch valve 65, close the other pinch valves, and rotate the first peristaltic pump 51 and the third peristaltic pump 53 forward to pump the fat in the incubation chamber 7 into the centrifuge tube 12.

[0083] S10, after weighting the weight tube 13 according to the method in step S3, start the centrifugal motor 11, and centrifuge at 1300-1600G for 5 minutes to centrifugally stratify the fat.

[0084] The adipose tissue after centrifugation is as follows from top to bottom: the first layer: the upper layer of fat oil; the second layer: tissue fragments; the third layer: waste liquid (saline solution); the fourth layer: cell precipitation; the waste liquid that needs to be discharged is the first layer: the upper layer of fat oil, the second layer: tissue fragments and the third layer: waste liquid (saline solution).

[0085] The fourth pinch valve 64 is opened, the other pinch valves are closed, and the second peristaltic pump 52 is reversed to discharge the upper fatty oil of the first layer into the waste liquid bag 4 through the fourth pipeline 34 and the first branch pipe.

[0086] The fourth pinch valve 64 is opened, the other pinch valves are closed, the first peristaltic pump 51 and the second peristaltic pump 52 are reversed, and the tissue fragments of the second layer and the waste liquid of the third layer are discharged into the waste liquid bag 4 through the first pipeline 31 and the first branch pipe.

[0087] S11, open the second pinch valve 62, close the other pinch valves, rotate the first peristaltic pump 51 forward, and add the protective liquid into the centrifuge tube 12; and weight the weight tube 13 according to the method in step S3.

[0088] S12, turning on the centrifugal motor 11, and shaking the fat and protective liquid in the centrifugal tube 12 evenly through the action of the centrifugal motor 11; or turning on the second peristaltic pump 52, and continuously rotating the second peristaltic pump 52 forward and reversely, in a "pumping-discharging" manner, so that the mixture of fat and protective liquid continuously moves between the centrifugal tube 12 and the fourth pipeline 34, and the two are evenly mixed.

[0089] S13, after uniform mixing, start the centrifugal motor 11, centrifuge at 600-800G for 3-5 minutes to wash and layer the fat.

[0090] After centrifugation, the fat tissue is divided into the following layers from top to bottom: first layer: upper layer of fat oil (very little, almost invisible); second layer: supernatant; third layer: adipocyte clusters. The waste liquid that needs to be discharged is the first layer: upper layer of fat oil (very little, almost invisible), second layer: supernatant; multiple centrifugation can be used to obtain the third layer: adipocyte clusters.

[0091] After centrifugation, the fourth pinch valve 64 is opened, the other pinch valves are closed, the first peristaltic pump 51 and the second peristaltic pump 52 are reversed, and the first layer and the second layer after centrifugation are discharged into the waste liquid bag 4 through the first pipeline 31 and the first branch 35.

[0092] S14, the fourth peristaltic pump is reversed to extract the third layer of fat cytogenes in the centrifuge tube 12 into the collection tube 2; after collection, the front and rear ends of the collection tube 2 are cut off, and the cells are sealed and frozen or injected; the third layer of fat cytogenes can also be extracted into a syringe through a manual interface for clinical application, or transferred to the collection tube 2 through a syringe for freezing or storage.

[0093] In this application, unless otherwise clearly specified and limited, the terms "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be an electrical connection; it can be a direct connection, or it can be an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0094] In the specification of the present invention, a large number of specific details are described. However, it is understood that embodiments of the present invention can be practiced without these specific details. In some instances, well-known methods, systems and techniques are not shown in detail so as not to obscure the understanding of this specification.

[0095] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, systems, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, systems, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

[0096] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. These modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be included in the scope of the claims and specification of the present invention.

Claims

1. A clinical-grade fat element preparation system, characterized in that: It comprises a fat processing device, connecting pipelines, a reagent chamber, a weight chamber, an incubation chamber, a collection tube and a waste liquid bag; the fat processing device comprises a centrifugal motor and a centrifugal mechanism connected to the centrifugal motor; the connecting pipelines comprise a first pipeline, a second pipeline, a third pipeline and a fourth pipeline respectively connected to the centrifugal mechanism; The reagent chamber is connected to the first pipeline, and a first peristaltic pump is installed between the reagent chamber and the centrifugal mechanism; the waste liquid bag is connected to the first pipeline through a first branch; the incubation chamber is connected to the first pipeline through a second branch, and a third peristaltic pump is connected to the second branch; the collecting tube is connected to the second pipeline, and a fourth peristaltic pump is connected to the second pipeline; the weight chamber is connected to the third pipeline, and a fifth peristaltic pump is connected to the third pipeline; one end of the fourth pipeline is connected to a centrifuge tube, and the other end is connected to the first branch, and the fourth pipeline is connected to a second peristaltic pump; The centrifugal mechanism comprises a centrifugal tube, a counterweight tube and a mounting bracket, the output shaft of the centrifugal motor is connected to the center of the mounting bracket, the centrifugal tube and the counterweight tube are respectively mounted at both ends of the mounting bracket, and the first pipeline and the second pipeline are respectively connected to the centrifugal tube; The centrifugal mechanism also includes an anti-winding device, which includes a fixed part, a rotating part, a first liquid pipe, a second liquid pipe, a third liquid pipe and a fourth liquid pipe, the rotating part is fixedly mounted on the mounting bracket, the fixed part is rotatably mounted on the outer periphery of the rotating part, and a first annular cavity, a second annular cavity, a third annular cavity and a fourth annular cavity are arranged between the fixed part and the rotating part and spaced apart in the vertical direction; the first pipeline, the second pipeline, the third pipeline and the fourth pipeline are all fixed to the side wall of the fixed part, and are respectively connected to the first annular cavity, the second annular cavity, the third annular cavity and the fourth annular cavity. The first ends of the first liquid pipe, the second liquid pipe, the third liquid pipe and the fourth liquid pipe are vertically connected to the rotating part, and are respectively connected to the first annular cavity, the second annular cavity, the third annular cavity and the fourth annular cavity. The second ends of the first liquid pipe, the second liquid pipe and the fourth liquid pipe are connected to the centrifuge tube, the second end of the third liquid pipe is connected to the counterweight tube, and the height of the second end of the fourth liquid pipe is 2ml higher than the height of the second end of the first liquid pipe; a 100um filter membrane is installed between the collecting tube and the centrifugal mechanism, and a hydrophobic breathable membrane is installed between the collecting tube and the fourth peristaltic pump; One end of the third pipeline is connected to the counterweight tube, the other end of the third pipeline is connected to the counterweight chamber, and the fifth peristaltic pump is located between the counterweight chamber and the counterweight tube; The reagent chamber includes an enzyme bag, a protective liquid bag and a physiological saline bag, and a first pinch valve, a second pinch valve and a third pinch valve are respectively installed between the enzyme bag, the protective liquid bag and the physiological saline bag and the first pipeline; a fourth pinch valve is installed at the liquid inlet end of the second peristaltic pump; and a fifth pinch valve is installed at the liquid inlet end of the third peristaltic pump; The preparation method of the clinical grade adipose tissue is as follows: S1. Inject the blood-containing fat into the centrifuge tube from the injection port; S2, open the third pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, and pump the saline in the saline bag into the centrifuge tube through the first pipeline; S3, the fifth peristaltic pump rotates forward, and the physiological saline in the weight-balancing chamber is pumped into the weight-balancing tube through the third pipeline, and the weight-balancing tube is weighted according to the weight of the centrifuge tube; S4, turning on the centrifugal motor, and shaking the blood fat and saline in the centrifuge tube evenly by the action of the centrifugal motor; or turning on the second peristaltic pump, and continuously rotating the second peristaltic pump forward and reversely, so that the mixture of blood fat and saline is continuously moved between the centrifuge tube and the fourth pipeline in a "pumping-discharging" manner, and the two are evenly mixed; S5. After mixing evenly, start the centrifugal motor and centrifuge at 60-70G for 1-3 minutes to wash and layer the fat; After cleaning, the fourth pinch valve is opened, the other pinch valves are closed, the second peristaltic pump is reversed, and the third and fourth layers of waste liquid, cell debris, and blood cells after cleaning are discharged into the waste liquid bag through the fourth pipeline and the first branch; S6, opening the fifth pinch valve, closing the other pinch valves, and reversing the first and third peristaltic pumps to pump the fat particles of the second layer into the incubation chamber; S7, opening the first pinch valve, closing the other pinch valves, reversing the third peristaltic pump, and adding collagenase into the incubation chamber; S8, incubate the chamber at 37°C for 1 hour at a constant temperature; at the same time, open the third pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, add physiological saline into the centrifuge tube, and clean the centrifuge tube; after cleaning, open the fourth pinch valve, close the other pinch valves, and rotate the first peristaltic pump and the second peristaltic pump in reverse to discharge the cleaned liquid into the waste liquid bag; S9, open the fifth pinch valve, close the other pinch valves, and rotate the first peristaltic pump and the third peristaltic pump forward to pump the fat in the incubation chamber into the centrifuge tube; S10, after the weight tube is weighted according to the method in step S3, the centrifugal motor is started, and the centrifugal treatment is performed at 1300-1600G for 5 minutes to centrifugally separate the fat; After centrifugal stratification, the fourth pinch valve is opened, the other pinch valves are closed, the second peristaltic pump is reversed, and the upper fatty oil of the first layer is discharged into the waste liquid bag through the fourth pipeline and the first branch pipe; The fourth pinch valve is opened, the other pinch valves are closed, the first peristaltic pump and the second peristaltic pump are reversed, and the tissue fragments of the second layer and the waste liquid of the third layer are discharged into the waste liquid bag through the first pipeline and the first branch pipe; S11, open the second pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, add the protective liquid into the centrifuge tube; and weight the weight tube according to the method in step S3; S12, turning on the centrifugal motor to evenly shake the fat and protective solution in the centrifuge tube through the action of the centrifugal motor; or turning on the second peristaltic pump to continuously rotate forward and reversely through the second peristaltic pump to continuously move the mixture of the fat and protective solution between the centrifuge tube and the fourth pipeline in a "pumping-spitting" manner to evenly mix the two; S13, after mixing evenly, start the centrifugal motor, centrifuge at 600-800G for 3-5 minutes to wash and layer the fat; After centrifugation, the fourth pinch valve is opened, the other pinch valves are closed, the first peristaltic pump and the second peristaltic pump are reversed, and the first layer and the second layer after centrifugation are discharged into the waste liquid bag through the first pipeline and the first branch; S14, the fourth peristaltic pump is reversed to extract the third layer of fat cell population in the centrifuge tube into the collection tube; after collection, the front and rear ends of the collection tube are cut off, and the tube is sealed for freezing or injection; or the third layer of fat cell population is extracted into a syringe through a manual interface for clinical application, or transferred to a collection tube through a syringe for freezing or storage.

2. The clinical-grade adipocyte preparation system according to claim 1, characterized in that: The incubation chamber includes an incubation bag, a shaking table, a constant temperature device, and an imaging system; the shaking table is connected to the incubation bag, and the shaking frequency range is 160rpm-190rpm; the constant temperature device maintains a constant temperature of the incubation bag at 36°C±2.

3. The clinical-grade adipocyte preparation system according to claim 2, characterized in that: The imaging system includes a camera and an AI software processing system.

4. The clinical-grade adipogen preparation system according to claim 1, characterized in that: The enzyme bag and the protective liquid bag use a fat element extraction instrument package, collagenase and physiological saline are added to the enzyme bag, 1200 units of collagenase for injection are added with 1-5 ml of physiological saline, and the mixture is fully mixed. 1200 units of collagenase for injection can dissociate 10-20 ml of fat.

5. The clinical-grade adipogen preparation system according to claim 4, characterized in that: The protective liquid bag is filled with protective liquid, which is composed of amino acids, glucose and physiological saline. Specifically, 800ul of amino acids and 10ml of 10% glucose are added to every 100ml of physiological saline.

6. A method for preparing clinical-grade adipogens, characterized in that: The clinical-grade fat element preparation system according to any one of claims 1 to 5 comprises the following steps: S1. Inject the blood-containing fat into the centrifuge tube from the injection port; S2, open the third pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, and pump the saline in the saline bag into the centrifuge tube through the first pipeline; S3, the fifth peristaltic pump rotates forward, and the physiological saline in the weight-balancing chamber is pumped into the weight-balancing tube through the third pipeline, and the weight-balancing tube is weighted according to the weight of the centrifuge tube; S4, turning on the centrifugal motor, and shaking the blood fat and saline in the centrifuge tube evenly by the action of the centrifugal motor; or turning on the second peristaltic pump, and continuously rotating the second peristaltic pump forward and reversely, so that the mixture of blood fat and saline is continuously moved between the centrifuge tube and the fourth pipeline in a "pumping-discharging" manner, and the two are evenly mixed; S5. After mixing evenly, start the centrifugal motor and centrifuge at 60-70G for 1-3 minutes to wash and layer the fat; After cleaning, the fourth pinch valve is opened, the other pinch valves are closed, the second peristaltic pump is reversed, and the third and fourth layers of waste liquid, cell debris, and blood cells after cleaning are discharged into the waste liquid bag through the fourth pipeline and the first branch; S6, opening the fifth pinch valve, closing the other pinch valves, and reversing the first and third peristaltic pumps to pump the fat particles of the second layer into the incubation chamber; S7, opening the first pinch valve, closing the other pinch valves, reversing the third peristaltic pump, and adding collagenase into the incubation chamber; S8, incubate the chamber at 37°C for 1 hour at a constant temperature; at the same time, open the third pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, add physiological saline into the centrifuge tube, and clean the centrifuge tube; after cleaning, open the fourth pinch valve, close the other pinch valves, and rotate the first peristaltic pump and the second peristaltic pump in reverse to discharge the cleaned liquid into the waste liquid bag; S9, open the fifth pinch valve, close the other pinch valves, and rotate the first peristaltic pump and the third peristaltic pump forward to pump the fat in the incubation chamber into the centrifuge tube; S10, after the weight tube is weighted according to the method in step S3, the centrifugal motor is started, and the centrifugal treatment is performed at 1300-1600G for 5 minutes to centrifugally separate the fat; After centrifugal stratification, the fourth pinch valve is opened, the other pinch valves are closed, the second peristaltic pump is reversed, and the upper fatty oil of the first layer is discharged into the waste liquid bag through the fourth pipeline and the first branch pipe; The fourth pinch valve is opened, the other pinch valves are closed, the first peristaltic pump and the second peristaltic pump are reversed, and the tissue fragments of the second layer and the waste liquid of the third layer are discharged into the waste liquid bag through the first pipeline and the first branch pipe; S11, open the second pinch valve, close the other pinch valves, rotate the first peristaltic pump forward, add the protective liquid into the centrifuge tube; and weight the weight tube according to the method in step S3; S12, turning on the centrifugal motor to evenly shake the fat and protective solution in the centrifuge tube through the action of the centrifugal motor; or turning on the second peristaltic pump to continuously rotate forward and reversely through the second peristaltic pump to continuously move the mixture of the fat and protective solution between the centrifuge tube and the fourth pipeline in a "pumping-spitting" manner to evenly mix the two; S13, after mixing evenly, start the centrifugal motor, centrifuge at 600-800G for 3-5 minutes to wash and layer the fat; After centrifugation, the fourth pinch valve is opened, the other pinch valves are closed, the first peristaltic pump and the second peristaltic pump are reversed, and the first layer and the second layer after centrifugation are discharged into the waste liquid bag through the first pipeline and the first branch; S14, the fourth peristaltic pump is reversed to extract the third layer of adipocytes in the centrifuge tube into the collection tube; after collection, the front and rear ends of the collection tube are cut off, and the tube is sealed and then frozen or injected; The third layer adipocyte population can be extracted into a syringe through a manual interface for clinical application, or transferred into a collection tube through a syringe for freezing or storage.

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