CN117821214A - Device and method for separating and culturing adipose-derived stem cells - Google Patents

Abstract

The invention relates to a device and a method for separating and culturing adipose-derived stem cells, comprising a separation bin, wherein a protection bin is fixed at the center of the top surface of the separation bin, a feed inlet is fixed at the top of the front surface of the separation bin, a controller is fixedly arranged at the middle position of the front surface of the separation bin, a discharge pipe is fixed at the center of the bottom surface of the separation bin, a base is fixed at the bottom surface of the separation bin, supporting plates are fixed at the left and right sides of the inner wall of the base, fixing grooves are formed at the front and rear ends of the left and right sides of the inner wall of the separation bin, a crushing mechanism for crushing fat is arranged on the separation bin, a filtering mechanism for filtering adipose-derived cells is arranged on the separation bin, and a centrifugal mechanism for centrifuging the adipose-derived cells is arranged on the base. Through setting up twist grip and cam, when the mesh of filter screen is blockked up, accessible twist grip dwang rotates and utilizes cam and reset spring to make the filter screen shake from top to bottom, conveniently trembles away the adipose tissue that blocks.

Description

Device and method for separating and culturing adipose-derived stem cells

Technical Field

The invention relates to the technical field of stem cell separation and culture, in particular to a device and a method for separating and culturing adipose-derived stem cells.

Background

Stem cells are a class of cells with unlimited or immortalized self-renewing capacity, capable of producing at least one type of highly differentiated daughter cells, the definition of stem cells being constantly revised over the years and defined from different levels, and most biologists and medical practitioners currently consider stem cells to be a class of cells from embryonic, fetal or adult bodies with unlimited self-renewing and proliferative differentiation capacity under certain conditions, capable of producing daughter cells of the same phenotype as the genotype and themselves, and capable of producing specialized cells that make up tissues and organs of the body, while also differentiating into progenitor cells.

For example, chinese patent (publication No. CN 215440382U) discloses a separating device for stem cells, by arranging a sealing column, stem cells can not directly flow out of a discharging bin when the separating box is in operation, and by arranging an adjusting spring, a movable rod can be reset, layering liquid can flow out of discharging bins with different heights respectively, so that pollution of the stem cells is effectively avoided, and the aim of high practicability is achieved.

Be equipped with the filter plate among the above-mentioned technical scheme and filter adipose tissue, but the filter plate carries out filterable in-process filter plate on the adipose tissue the rate hole be liable to block up, above-mentioned technical scheme is inconvenient for handling the circumstances of blocking, and above-mentioned technical scheme is broken through the broken piece of single group only when carrying out the breakage to adipose tissue in addition, and its broken efficiency is lower.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a device and a method for separating and culturing adipose-derived stem cells, which have the advantages of high filtering efficiency, high crushing efficiency and the like, and solve the problems that the prior device is inconvenient to treat the blockage and has lower crushing efficiency of adipose tissues.

In order to achieve the above purpose, the present invention provides the following technical solutions: the utility model provides a device for separating and cultivateing fat stem cell, includes the separation storehouse, the center department of separation storehouse top surface is fixed with the protection storehouse, the positive top of separation storehouse is fixed with the feed inlet, the positive intermediate position fixed mounting of separation storehouse has the controller, the center department of separation storehouse bottom surface is fixed with the discharging pipe, the bottom surface of separation storehouse is fixed with the base, the left and right sides of base inner wall all is fixed with the backup pad, the fixed slot has all been seted up at the front and back both ends of separation storehouse inner wall left and right sides, be equipped with the crushing mechanism that is used for carrying out the breakage to fat on the separation storehouse, be equipped with the filtering mechanism that filters fat cell on the separation storehouse, be equipped with the centrifugation mechanism that is used for centrifuging fat cell on the base;

in addition, filtering mechanism is including fixing the dead lever in the fixed slot, the top movable sleeve of dead lever surface is equipped with reset spring, four the surface sliding connection of dead lever has the fixed frame, be fixed with the roof between the front and back both sides of the intermediate position of fixed frame inner peripheral wall, the left and right sides of fixed frame inner peripheral wall all is fixed with the filter screen, filtering mechanism still include through the bearing with the dwang that the back intermediate position of separation storehouse was rotated and is connected, the intermediate position of dwang surface is fixed with the cam.

Further, a fixed hole is formed in the bottom of the middle position of the front face of the separation bin, the rotating rod is rotationally connected with the fixed hole through a bearing, and a rotating handle is fixed on the front face of the rotating rod.

Further, the cam is located right below the top plate, and the back surface of the feed inlet extends to the inside of the separation bin.

Further, round holes are formed in the front end and the rear end of the left side and the right side of the top surface of the fixed frame, the fixed rods are round rods, and the fixed rods penetrate through the round holes and are in sliding connection with the round holes.

Further, the crushing mechanism comprises a crushing motor fixedly installed on the right side of the inner top wall of the protection bin, a first crushing part is fixed on an output shaft of the crushing motor, a driving gear is fixed on the top of the outer surface of the first crushing part, the crushing mechanism further comprises a second crushing part which is rotationally connected with the left side of the inner top wall of the protection bin through a bearing, and a driven gear is fixed on the top of the outer surface of the second crushing part.

Further, the driving gear is meshed with the driven gear, and through holes are formed in the left side and the right side of the top surface of the separation bin.

Further, the centrifugal mechanism comprises a centrifugal barrel which is rotationally connected with the bottom of the outer surface of the discharging pipe, a first belt pulley is fixed in the middle of the outer surface of the centrifugal barrel, a discharging pipe is fixed in the center of the bottom surface of the centrifugal barrel, the centrifugal mechanism further comprises a centrifugal motor which is fixedly installed on the left side of the top surface of the base, a driving shaft is fixed on an output shaft of the centrifugal motor, a second belt pulley is fixed in the middle of the outer surface of the driving shaft, and a driving belt is connected between the first belt pulley and the second belt pulley in a transmission mode.

Further, the top of drive shaft passes through the bearing with the left side the bottom surface of backup pad rotates to be connected, the bottom of centrifuge bowl surface passes through the bearing with the right side the backup pad rotates to be connected.

Further, the centrifugal barrel is cylindrical, a discharge valve is fixedly arranged on the discharge pipe, and a culture dish is placed right below the discharge pipe at the center of the top surface of the base.

A method of using a device for isolating and culturing adipose-derived stem cells, comprising the steps of:

s1, placing selected adipose tissues into a separation bin through a feed inlet, and then cutting and crushing the adipose tissues through a crushing mechanism;

s2, filtering the crushed adipose tissues through a filtering mechanism, continuously cutting and crushing the adipose tissues with larger particles, and allowing fluid cells to permeate from a filter screen to the bottom of a separation bin;

s3, fluid cells permeated to the bottom of the separation bin drop into the centrifugal barrel through the discharge pipe, and then the centrifugal barrel is driven to rotate at a high speed through the centrifugal motor, so that centrifugal treatment and standing are realized, and sediment A is obtained;

s4, placing the sterilized culture dish under a discharge pipe, opening a discharge valve on the discharge pipe, taking out the sediment A into the culture dish, and adding a nutrition additive for culturing to obtain fat stem cells;

s5, aliquoting the adipose-derived stem cells subjected to passage in the step S4, transferring the aliquotted adipose-derived stem cells into a new culture dish, carrying out timing culture, and periodically adding a nutrition additive with equal concentration;

s6, determining a culture result of the adipose-derived stem cells.

Compared with the prior art, the technical scheme of the application has the following beneficial effects:

1. the device and the method for separating and culturing the adipose-derived stem cells are provided with the filtering mechanism, and through arranging the rotating handle and the cam, when the meshes of the filtering net are blocked, the rotating rod can rotate through the rotating handle, and the cam and the reset spring can be utilized to enable the filtering net to vibrate up and down, so that the blocked adipose tissues can be conveniently shaken off.

2. The device and the method for separating and culturing the adipose-derived stem cells are provided with the crushing mechanism, and the first crushing part and the second crushing part are rotated in opposite directions by utilizing the driving gear and the driven gear through arranging the first crushing part and the second crushing part, so that the crushing efficiency of adipose tissues is improved.

3. The device and the method for separating and culturing the adipose-derived stem cells are provided with the centrifugal mechanism, and can drive the centrifugal barrel to rotate at high speed through the centrifugal motor and the transmission belt, so that adipose tissues can be subjected to centrifugal treatment in the process of rotating at high speed, and the adipose-derived stem cells are precipitated and separated from serum, thereby facilitating the subsequent stem cell culture treatment.

Drawings

FIG. 1 is a schematic diagram of the structure of the present invention;

FIG. 2 is a schematic view of the appearance structure of the present invention;

FIG. 3 is a schematic view of a filter mechanism according to the present invention;

FIG. 4 is a top view of a mounting frame of the present invention;

FIG. 5 is a schematic view of the crushing mechanism of the present invention;

fig. 6 is a schematic structural view of the centrifugal mechanism of the present invention.

In the figure: 1 separating bin, 2 protecting bin, 3 feed inlet, 4 controller, 5 discharging pipe, 6 base, 7 backup pad, 8 fixed slot, 9 crushing mechanism, 901 crushing motor, 902 first crushing portion, 903 driving gear, 904 second crushing portion, 905 driven gear, 10 filter mechanism, 1001 dead lever, 1002 reset spring, 1003 dead frame, 1004 roof, 1005 filter screen, 1006 dwang, 1007 cam, 11 centrifugal mechanism, 1101 centrifugal barrel, 1102 first belt pulley, 1103 discharging pipe, 1104 centrifugal motor, 1105 driving shaft, 1106 second belt pulley, 1107 driving belt.

Detailed Description

The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Referring to fig. 1-2, a device for separating and culturing adipose-derived stem cells in this embodiment includes a separation chamber 1, a protection chamber 2 is fixed at the center of the top surface of the separation chamber 1, a feed inlet 3 is fixed at the top of the front surface of the separation chamber 1, a controller 4 is fixedly installed at the middle position of the front surface of the separation chamber 1, a discharge pipe 5 is fixed at the center of the bottom surface of the separation chamber 1, a base 6 is fixed at the bottom surface of the separation chamber 1, supporting plates 7 are fixed at the left and right sides of the inner wall of the base 6, fixing grooves 8 are formed at the front and rear ends of the left and right sides of the inner wall of the separation chamber 1, a breaking mechanism 9 for breaking fat is arranged on the separation chamber 1, a filtering mechanism 10 for filtering adipose-derived cells is arranged on the separation chamber 1, and a centrifugal mechanism 11 for centrifuging the adipose-derived cells is arranged on the base 6.

Referring to fig. 1 to 4, in order to facilitate improvement of the penetration efficiency of the fluid adipose tissue, the filtering mechanism 10 includes a fixing rod 1001 fixed in a fixing groove 8, a return spring 1002 movably sleeved on the top of the outer surface of the fixing rod 1001, fixing frames 1003 slidably connected to the outer surfaces of the four fixing rods 1001, top plates 1004 fixed between the front and rear sides of the middle position of the inner peripheral wall of the fixing frames 1003, filter screens 1005 fixed to the left and right sides of the inner peripheral wall of the fixing frames 1003, and a rotating rod 1006 rotatably connected to the middle position of the back surface of the inner wall of the separation bin 1 through a bearing, wherein a cam 1007 is fixed to the middle position of the outer surface of the rotating rod 1006.

Wherein, the fixed orifices have been seted up to the bottom of separation storehouse 1 front intermediate position, and dwang 1006 passes through the bearing and rotates with the fixed orifices and be connected, and the front of dwang 1006 is fixed with the twist grip, can drive dwang 1006 rotation through rotating the twist grip, and dwang 1006 can drive the cam 1007 and rotate at pivoted in-process.

Meanwhile, the cam 1007 is located right under the top plate 1004, and the cam 1007 can push the fixing frame 1003 to move upward through the top plate 1004 in the rotating process, and the back surface of the feed port 3 extends to the inside of the separation bin 1, so that the adipose tissue can be conveniently added to the inside of the separation bin 1.

In addition, circular holes are formed in the front end and the rear end of the left side and the right side of the top surface of the fixed frame 1003, the fixed rod 1001 is a circular rod, the fixed rod 1001 penetrates through the circular holes and is in sliding connection with the circular holes, the fixed rod 1001 can play a role in supporting and limiting the fixed frame 1003, stability in the up-and-down moving process of the fixed frame 1003 can be improved, and meanwhile, the reset spring 1002 can realize automatic downward reset of the fixed frame 1003 and can realize the up-and-down vibration process of the filter screen 1005 in cooperation with the cam 1007.

Referring to fig. 1 and 5, in order to facilitate improvement of the crushing efficiency of adipose tissue, the crushing mechanism 9 includes a crushing motor 901 fixedly installed on the right side of the inner top wall of the guard bin 2, a first crushing portion 902 is fixed on an output shaft of the crushing motor 901, a driving gear 903 is fixed on the top of the outer surface of the first crushing portion 902, the crushing mechanism 9 further includes a second crushing portion 904 rotatably connected with the left side of the inner top wall of the guard bin 2 through a bearing, and a driven gear 905 is fixed on the top of the outer surface of the second crushing portion 904.

In addition, the driving gear 903 and the driven gear 905 are meshed, so that the first crushing portion 902 and the second crushing portion 904 can rotate in opposite directions, thereby improving the crushing efficiency of the adipose tissue, and through holes are formed in both the left and right sides of the top surface of the separation bin 1, so that the first crushing portion 902 and the second crushing portion 904 can extend into the separation bin 1.

Referring to fig. 1 and 6, in order to facilitate the centrifugation of adipose tissue, the centrifugal mechanism 11 comprises a centrifugal barrel 1101 rotatably connected to the bottom of the outer surface of the discharging pipe 5, a first belt pulley 1102 is fixed in the middle of the outer surface of the centrifugal barrel 1101, a discharging pipe 1103 is fixed in the center of the bottom surface of the centrifugal barrel 1101, the centrifugal mechanism 11 further comprises a centrifugal motor 1104 fixedly mounted on the left side of the top surface of the base 6, a driving shaft 1105 is fixed on the output shaft of the centrifugal motor 1104, a second belt pulley 1106 is fixed in the middle of the outer surface of the driving shaft 1105, and a driving belt 1107 is in driving connection with the first belt pulley 1102 and the second belt pulley 1106.

The top end of the driving shaft 1105 is rotatably connected to the bottom surface of the left support plate 7 through a bearing, and the bottom of the outer surface of the centrifugal barrel 1101 is rotatably connected to the right support plate 7 through a bearing, so that the supporting effect on the centrifugal barrel 1101 can be improved, and the stability of the centrifugal barrel 1101 in the rotation process can be improved.

It can be appreciated that the centrifugal barrel 1101 is cylindrical, the centrifugal barrel 1101 is conveniently driven to rotate by the driving belt 1107, the discharging pipe 1103 is fixedly provided with a discharging valve, the fat stem cells after centrifugation in the centrifugal barrel 1101 are conveniently taken out, and the culture dish is placed at the center of the top surface of the base 6 and under the discharging pipe 1103.

Finally, a method for using the device for separating and culturing the adipose-derived stem cells comprises the following steps:

s1, placing selected adipose tissues into a separation bin 1 through a feed inlet 3, and then cutting and crushing the adipose tissues through a crushing mechanism 9;

s2, filtering the crushed adipose tissues through a filtering mechanism 10, continuously cutting and crushing the adipose tissues with larger particles, and allowing fluid cells to permeate from a filter screen 1005 to the bottom of the separation bin 1;

s3, fluid cells permeated to the bottom of the separation bin 1 fall into the centrifugal barrel 1101 through the discharging pipe 5, and then the centrifugal barrel 1101 is driven to rotate at a high speed through the centrifugal motor 1104, so that centrifugal treatment is realized, and the sediment A is obtained;

s4, placing the sterilized culture dish under a discharge pipe 1103, opening a discharge valve on the discharge pipe 1103 to take out a precipitate A into the culture dish, and adding a nutrition additive for culturing to obtain fat stem cells;

s5, aliquoting the adipose-derived stem cells subjected to passage in the step S4, transferring the aliquotted adipose-derived stem cells into a new culture dish, carrying out timing culture, and periodically adding a nutrition additive with equal concentration;

s6, determining a culture result of the adipose-derived stem cells.

The working principle of the embodiment is as follows:

placing the selected adipose tissues into the separation bin 1 through the feed inlet 3, then driving the first crushing part 902 to rotate through the crushing motor 901 and cutting and crushing the adipose tissues, and simultaneously driving the second crushing part 904 to reversely rotate through the action of the driving gear 903 and the driven gear 905 in the rotating process of the first crushing part 902, so that the efficiency of cutting and crushing the adipose tissues is improved; the broken adipose tissue is filtered by the filter screen 1005, the adipose tissue with larger particles is cut and broken continuously, fluid cells permeate from the filter screen 1005 to the bottom of the separation bin 1, when the large-particle adipose tissue is blocked in the meshes of the filter screen 1005, and the fluid cell permeation efficiency is lower, the rotating rod 1006 is driven to rotate by the rotating handle, the rotating rod 1006 drives the cam 1007 to rotate in the rotating process, the cam 1007 can push the fixed frame 1003 to move upwards in the rotating process, and the filter screen 1005 is vibrated up and down continuously under the action of the elastic potential energy of the reset spring 1002, so that the large-particle adipose tissue blocked in the meshes of the filter screen 1005 is shaken off; the fluid cells permeated to the bottom of the separation bin 1 fall into the centrifugal barrel 1101 through the discharging pipe 5, then the centrifugal motor 1104 drives the second belt pulley 1106 to rotate, the first belt pulley 1102 is driven to rotate under the action of the driving belt 1107, meanwhile, the centrifugal barrel 1101 is driven to rotate at a high speed, the centrifugal barrel 1101 realizes centrifugal treatment on fat tissues in the process of rotating at the high speed, and substances after centrifugation are kept still to obtain sediment A; placing the sterilized culture dish under a discharge pipe 1103, opening a discharge valve on the discharge pipe 1103 to take out sediment A into the culture dish, and adding a nutrition additive for culturing to obtain fat stem cells; aliquoting the adipose-derived stem cells subjected to passage in the step S4, transferring the adipose-derived stem cells into a new culture dish, carrying out timing culture, and periodically adding a nutrition additive with equal concentration; and finally, determining the culture result of the adipose-derived stem cells.

It is noted that relational terms such as first and second, and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.

Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (10)

1. The device for separating and culturing the adipose-derived stem cells comprises a separation bin (1), and is characterized in that: the novel fat separator is characterized in that a protection bin (2) is fixed at the center of the top surface of the separation bin (1), a feed inlet (3) is fixed at the top of the front surface of the separation bin (1), a controller (4) is fixedly arranged at the middle position of the front surface of the separation bin (1), a discharge pipe (5) is fixed at the center of the bottom surface of the separation bin (1), a base (6) is fixed at the bottom surface of the separation bin (1), supporting plates (7) are fixed at the left and right sides of the inner wall of the base (6), fixing grooves (8) are formed in the front end and the rear end of the left and right sides of the inner wall of the separation bin (1), a crushing mechanism (9) for crushing fat is arranged on the separation bin (1), a filtering mechanism (10) for filtering fat cells is arranged on the separation bin (1), and a centrifugal mechanism (11) for centrifuging the fat cells is arranged on the base (6).

In addition, filtering mechanism (10) are including fixing dead lever (1001) in fixed slot (8), reset spring (1002) are equipped with to the top movable sleeve of dead lever (1001) surface, four the surface sliding connection of dead lever (1001) has fixed frame (1003), be fixed with roof (1004) between the front and back both sides of the intermediate position of fixed frame (1003) inner peripheral wall, the left and right sides of fixed frame (1003) inner peripheral wall all is fixed with filter screen (1005), filtering mechanism (10) still include through the bearing with dwang (1006) that the rotation of separation storehouse (1) inner wall back intermediate position is connected, the intermediate position of dwang (1006) surface is fixed with cam (1007).

2. The device for isolating and culturing adipose-derived stem cells according to claim 1, wherein: the bottom of the front middle position of the separation bin (1) is provided with a fixed hole, the rotating rod (1006) is rotationally connected with the fixed hole through a bearing, and the front of the rotating rod (1006) is fixedly provided with a rotating handle.

3. The device for isolating and culturing adipose-derived stem cells according to claim 1, wherein: the cam (1007) is located right below the top plate (1004), and the back surface of the feed inlet (3) extends to the inside of the separation bin (1).

4. The device for isolating and culturing adipose-derived stem cells according to claim 1, wherein: round holes are formed in the front end and the rear end of the left side and the right side of the top surface of the fixing frame (1003), the fixing rod (1001) is a round rod, and the fixing rod (1001) penetrates through the round holes and is connected with the round holes in a sliding mode.

5. The device for isolating and culturing adipose-derived stem cells according to claim 1, wherein: the crushing mechanism (9) comprises a crushing motor (901) fixedly mounted on the right side of the inner top wall of the protection bin (2), a first crushing part (902) is fixed on an output shaft of the crushing motor (901), a driving gear (903) is fixed on the top of the outer surface of the first crushing part (902), the crushing mechanism (9) further comprises a second crushing part (904) rotatably connected with the left side of the inner top wall of the protection bin (2) through a bearing, and a driven gear (905) is fixed on the top of the outer surface of the second crushing part (904).

6. The device for isolating and culturing adipose-derived stem cells according to claim 5, wherein: the driving gear (903) is meshed with the driven gear (905), and through holes are formed in the left side and the right side of the top surface of the separation bin (1).

7. The device for isolating and culturing adipose-derived stem cells according to claim 1, wherein: the centrifugal mechanism (11) comprises a centrifugal barrel (1101) which is rotationally connected with the bottom of the outer surface of the discharging pipe (5), a first belt pulley (1102) is fixed at the middle position of the outer surface of the centrifugal barrel (1101), a discharging pipe (1103) is fixed at the center of the bottom surface of the centrifugal barrel (1101), the centrifugal mechanism (11) further comprises a centrifugal motor (1104) fixedly installed on the left side of the top surface of the base (6), a driving shaft (1105) is fixed on an output shaft of the centrifugal motor (1104), a second belt pulley (1106) is fixed at the middle position of the outer surface of the driving shaft (1105), and a driving belt (1107) is connected between the first belt pulley (1102) and the second belt pulley (1106) in a transmission mode.

8. The apparatus for isolating and culturing adipose-derived stem cells according to claim 7, wherein: the top end of the driving shaft (1105) is rotationally connected with the bottom surface of the left supporting plate (7) through a bearing, and the bottom of the outer surface of the centrifugal barrel (1101) is rotationally connected with the right supporting plate (7) through a bearing.

9. The apparatus for isolating and culturing adipose-derived stem cells according to claim 7, wherein: the centrifugal barrel (1101) is cylindrical, a discharge valve is fixedly arranged on the discharge pipe (1103), and a culture dish is placed at the center of the top surface of the base (6) and under the discharge pipe (1103).

10. The device for separating and culturing adipose-derived stem cells according to any one of claims 1 to 9, which is a method for using the device for separating and culturing adipose-derived stem cells, comprising the following steps:

s1, placing selected adipose tissues into a separation bin (1) through a feed inlet (3), and then cutting and crushing the adipose tissues through a crushing mechanism (9);

s2, filtering the crushed adipose tissues through a filtering mechanism (10), continuously cutting and crushing the adipose tissues with larger particles, and allowing fluid cells to permeate from a filter screen (1005) to the bottom of a separation bin (1);

s3, fluid cells permeated to the bottom of the separation bin (1) drop into the centrifugal barrel (1101) through the discharging pipe (5), and then the centrifugal barrel (1101) is driven to rotate at a high speed through the centrifugal motor (1104), so that centrifugal treatment and standing are realized, and sediment A is obtained;

s4, placing the sterilized culture dish under a discharge pipe (1103), opening a discharge valve on the discharge pipe (1103) to take out the sediment A into the culture dish, and adding a nutrition additive for culturing to obtain fat stem cells;

s5, aliquoting the adipose-derived stem cells subjected to passage in the step S4, transferring the aliquotted adipose-derived stem cells into a new culture dish, carrying out timing culture, and periodically adding a nutrition additive with equal concentration;

s6, determining a culture result of the adipose-derived stem cells.