KR20140093821A - Device and System for extracting regenerative cells and Method for extracting regenerative cells using the same - Google Patents

Abstract

Disclosed is a regenerative cell extracting system. The regenerative cell extracting system comprises: a regenerative cell separating device which is rotated by receiving rotary power from the outside in order to separate a tissue; and a regenerative cell extracting device for extracting a regenerative cell through a secondary centrifugation by receiving a separated object which is discharged by being separated from the regenerative cell separating device. Especially, the regenerative cell extracting device has: a main container which forms an accommodating unit which accommodates a separation object; and a hollow access tube which is inserted into the main container. The present invention smoothly guarantees the tightness and relative rotation of the devices for separating and extracting a regenerative cell and also easily solves a problem of a heat generation which is caused by a contact rotation by using a compressible sealing member, a first friction reduction member, and a second friction reduction member which are formed in a ring shape and relatively rotate by a mutual contact. Multiple protruding accommodation units, which are formed to be convex to the outside in a radial direction around the rotary shaft of the regenerative cell extracting device in order to accommodate relatively heavy separated objects among separated objects which are separated from a separation object, is formed in a wave shape which continuously and alternatively arranges a convex part and a concave part in the outer circumference surface of the main container by being continuously arranged in the outer circumference of the main container.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a regenerative cell extracting apparatus, a regenerative cell extracting apparatus,

The present invention relates to a device for separating tissues and extracting cells, and more particularly, to a device for extracting regenerated cells such as stem cells by rotating tissues such as adipose tissue, bone marrow, cord blood and the like, .

The centrifugal separator is a device for separating a substance into constituent components by using a centrifugal force generated when an object rotates, and various forms have been developed depending on the object to be separated.

In the field of biotechnology, centrifuges have been used for the purpose of separating substances having specific gravity and adhesion higher than cells or liquids mixed with liquid by specific gravity. For example, the use of centrifuges is increasing in separating blood into components such as red blood cells and plasma or extracting stem cells from fats and bone marrow.

A centrifugal separator for extracting stem cells generally has a container in which an object to be separated is contained and an inlet and an outlet to be inserted into the container. The container is rotated by receiving a rotational force from the outside, but the inlet and outlet pipes are not rotated while being inserted into the container. In order to prevent foreign matter from entering into the container during the centrifugation process, the space between the container and the inlet and outlet must be sealed. However, the inlet pipe and the container are not physically coupled to each other but must be relatively rotated. As a result, in the centrifuge, the inlet pipe and the container are relatively rotated and must be sealed between them. In order to satisfy such a condition, an airtight bearing is employed, but it is not economical, so its use is limited, and development of a highly economical part having a substitute effect is insufficient.

In addition, when relative rotation is performed in a sealed state, heat is generated due to contact generated during rotation, and heat dissipation is not smoothly performed, so that a relative rotation site is melted or damaged.

Accordingly, it is required to develop a physical structure which satisfies both of the hermeticity and the relative rotation, can smoothly discharge heat generated by the contact during rotation, and satisfies economy.

In the meantime, an important factor in the centrifuge for extracting stem cells is the ratio of the impurity content to how much different components are incorporated into the isolated stem cells, the yield rate of how much stem cells can be extracted from the separated subject And the throughput per hour of how much can be extracted within a certain amount of time.

Conventional centrifuges have a cylindrical shape. When the body is rotated and then stopped, the constituents are layered and separated according to the specific gravity. The stem cells were extracted by inserting a pipette into the layer containing the stem cells and selectively pumping only the layer containing the stem cells. Since the stem cells are relatively heavy, they are arranged in the lower layer. Therefore, although pumping is easier than selectively pumping only the middle layer, the components of the other layers are flowed together in the process of selectively pumping the stem cell layer, There was a problem of incorporation.

In order to solve this problem, there has been an attempt to reduce the impurity content by forming a container that can accommodate only stem cells at the point where the centrifugal force is maximized by using the fact that the stem cells are the heaviest component. In this case, There is a problem that the amount is significantly reduced.

In the prior patent application of the present applicant, in order to solve such a problem, as disclosed in Patent Application No. 10-2010-0102987, the extraction volume of stem cells is increased by forming a receiving portion convexly along the circumferential direction on the body of the centrifugal separator, There have been attempts to reduce the amount of incorporation and satisfactory performance has been achieved. In the case of the centrifugal separator disclosed in the above-mentioned prior patent application, the maximum stem cell extraction rate and the lowest impurity content rate among the currently used stem cell centrifuges are shown.

However, the centrifugal separator of the prior patent application has a relatively weak point in terms of the throughput per hour as to how much fat tissue can be processed to extract stem cells within a predetermined time. Of course, the throughput per hour is superior to that of other conventional centrifuges, but it has a weak point in terms of throughput compared to the efficiency of stem cell extraction and impurity incorporation.

Particularly, when a centrifugal separator is connected to the human body and the stem cells are separated from the centrifuge immediately after extracting adipose tissue from the human body, and then the separated stem cells are injected into the human body, It is very important whether it can be processed.

From the viewpoint of increasing the throughput, simply increasing the size of the centrifuge is not economical. A rotating device is used to rotate the centrifuge, and the rotating device is provided with a mounting part that can be installed by inserting the centrifuge body in concave. That is, if the size of the centrifugal separator is simply increased, it is not preferable from the viewpoint of economical efficiency because the structure of the conventional rotating device, particularly the mounting portion, must be changed or the structure of the rotating device as a whole must be changed.

Therefore, it is required to improve the throughput of the centrifugal separator while maintaining the standard of the conventional rotary device.

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a regenerative cell extracting apparatus and system which can reliably discharge heat generated by contact during rotation while ensuring relative rotation and hermeticity, And an extraction method thereof.

It is another object of the present invention to provide a regenerative cell extracting apparatus having an improved structure in which the amount of fat tissue per hour can be increased while maintaining the structure and specification of a conventional rotating device.

A regenerative cell extracting apparatus according to the present invention includes: a main container rotatably provided with a rotational force from outside and having a receiving portion for receiving an object to be separated; A hollow type inlet / outlet pipe inserted into a through hole formed in the main container so that one end thereof is disposed inside the main container and the other end is disposed outside the main container and relatively rotated with the main container; A first friction reducing member formed in a ring shape so as to surround the inlet / outlet pipe; A main body rotatable together with the main container and formed in an annular shape corresponding to the first friction reducing member so as to surround the through hole and closely contacted with the first friction reducing member; A second frictional reducing member having a plurality of wings which are inclined with respect to the rotational surface of the body; And a sealing member provided between the main container and the outlet pipe so as to seal the gap between the inner circumferential surface of the through-hole of the main container and the outer circumferential surface of the outlet pipe.

According to the present invention, the sealing member is installed in the main container with a compressible rubber material and rotated together with the main container, and the second friction reducing member is coupled to the sealing member and rotated together with the main container. Particularly, the first frictional reducing member is made of a ceramic material, and the second frictional reducing member is made of a carbon material so that even when the first frictional reducing member and the second frictional reducing member are in close contact with each other, do. Above all, when the second friction reducing member rotates, the plurality of vanes may form a sudden flow (air) of air to easily release heat generated from the first friction reducing member and the second friction reducing member.

Meanwhile, the regenerative cell extracting apparatus according to an embodiment of the present invention may be configured to extract the regenerated cell from the separated object, the regenerated cell extractor being convex outward along the radial direction about the rotation axis of the main container, It is preferable that a plurality of protruding storage portions formed are continuously disposed on the outer peripheral surface of the main container and the outer peripheral surface of the main container is formed in a wave shape in which convex portions and concave portions are alternately arranged continuously.

The main container includes a main body portion in which the accommodating portion and the projecting portion are formed, and a lid portion which is coupled to the upper end of the main body portion to seal the accommodating portion. The lower end of the projecting accommodating portion is provided with a lower end And a blocking member for blocking a gap between the protrusion receiving portion and the protrusion receiving portion.

The regenerative cell extracting system according to the present invention is a regenerative cell extracting system for separating tissues, comprising: a sub container provided with a space portion rotatably received from the outside to receive tissue, and one end portion of the sub container, And a pump connected to the inlet pipe and providing a suction force, the regeneration cell separation device comprising: a regeneration cell separation device having a hollow inlet / outlet pipe inserted into a through hole formed in the sub container so that the other end thereof is disposed outside the sub container; And a regeneration cell extracting apparatus for extracting the regeneration cells from the regeneration cell separating apparatus through the secondary centrifugation by receiving the separated material discharged through the inlet and outlet tubes, have.

In one embodiment of the present invention, the sub-container is provided with a screen member installed in the space of the container so as to prevent a lump of a predetermined size or more formed in the process of separating the tissue from descending to the lower end of the container do.

Wherein the screen member has a plurality of engaging members formed outwardly from a central portion of the sub container, wherein the plurality of engaging members are spaced apart from each other along the circumferential direction, And is made of a flexible material so as to be bent toward the outer frame portion.

The regenerative cell separator includes a first frictional reducing member that is annularly formed to surround the inlet and outlet pipes, and a second frictional reducing member that is rotated together with the sub-container and is adapted to correspond to the first frictional reducing member And a plurality of wing parts disposed along the outer circumferential surface of the body part and inclined with respect to the rotation surface of the body part. 2 frictional reducing member and a sealing member provided between the sub container and the outlet pipe so as to seal the gap between the inner peripheral surface of the through hole of the sub container and the outer peripheral surface of the outlet pipe.

Meanwhile, the method for extracting regenerated cells according to the present invention is a method for extracting regenerated cells from tissue using an aliphatic cell extracting system having the above-mentioned constitution, Washing the tissue; And then discharging the heavy component including blood and body fluid continuously through the tube from the regenerative cell separating apparatus after the washing of the fatty tissue, if yellow fat tissue is discharged, stopping the discharge by sensing yellow using a photosensor; Decomposing the adipose tissue by rotating the regenerative cell separating device while the collagenase is put in the regenerative cell separating device; An aqueous solution containing regenerated cells separated from the adipose tissue and submerged in the lower part of the regenerative cell separator is discharged from the regenerative cell separator through the tube and transferred to the regenerative cell extractor, Stopping the discharge when mature adipocytes are detected; The regenerative cell extracting apparatus is rotated to centrifuge the aqueous solution containing the regenerative cells so that the regenerative cells adhere to the inner circumferential surface of the main container of the regenerative cell extracting apparatus and the remaining light components are accommodated in the lower portion of the main container step; And separating the regenerative cell from the inner circumferential surface of the main container by causing the remaining light component to be first discharged from the regenerative cell extractor and then injecting the cleaning liquid into the main container and rotating the detergent cell to fall to the lower portion of the main container, And discharging and recovering the sample from the cell extracting apparatus.

The regeneration cell extracting apparatus and the extraction system according to the present invention are advantageous in that they are relatively durable and economic as well as smoothly dissipating heat while ensuring relative rotation and hermeticity.

Particularly, there is an advantage that a flow of air can be formed as the wing portion formed on the second friction reducing member rotates to easily discharge the heat generated by the rotational contact between the first friction reducing member and the second friction reducing member. The first friction-reducing member and the second friction-reducing member which are in contact with each other have an advantage that heat generation itself is lowered because the number of friction is low, for example, ceramic and carbon are used as materials.

Meanwhile, the regenerative cell extracting system according to the present invention comprises two units of a separating device and an extracting device, and extracts the regenerating cells through a second separating process. The separating device of the regenerative cell extracting system is advantageous in that the screen member can be installed to filter large tissues such as collagen, thereby improving separation efficiency and yield of adipose tissue.

Particularly, the regenerative cell extracting apparatus has an advantage that the amount of the aqueous solution containing adipose tissue or regenerated cells can be treated per hour while maintaining the structure and specification of the conventional rotating apparatus, and the area where the stem cells can be attached And the yield of stem cells is improved.

In addition, the regenerative cell extracting method according to the present invention has an advantage in that it is possible to extract only pure regenerated cells in a state in which no impurities are incorporated, and extract as many regenerated cells as possible from adipose tissue.

1 is a schematic exploded perspective view of a regenerative cell separation apparatus employed in a regenerative cell extraction system according to an embodiment of the present invention.
2 is a schematic cross-sectional view taken along a line II-II in Fig.
3 is a schematic cross-sectional view taken along the line III-III in Fig.
4 is a schematic exploded perspective view of a regenerative cell extracting apparatus according to an embodiment of the present invention.
5 is a schematic perspective view of the regenerative cell extracting apparatus shown in FIG.
6 is a schematic sectional view taken along the line aa in Fig.
7 is a schematic cross-sectional view for explaining a process of collecting and discharging a separated material using a vaginal member and a guide member in a regenerative cell extracting apparatus.

Hereinafter, the regenerative cell extracting system according to the present invention will be described in more detail.

The regenerative cell extraction system according to the present invention includes a regenerative cell separation device and a regenerative cell extraction device. The regenerative cell separation device and regenerative cell extraction device are interconnected by pumps and tubes. In addition, the regenerative cell separator and the regenerative cell extractor are installed in the rotary jig and perform centrifugal separation while being rotated at a predetermined speed.

First, with reference to Figs. 1 and 2, a regenerative cell separator will be described.

FIG. 1 is a schematic exploded perspective view of a regenerative cell separation apparatus employed in a regenerative cell extraction system according to an embodiment of the present invention. FIG. 2 is a schematic sectional view taken along a line II-II in FIG. 2, III in Fig.

1 to 3, the regenerative cell extracting apparatus 100 is for separating the tissue containing regenerated cells such as fat tissue according to the specific gravity by rotating the tissue, And a tube (90).

The subcontainer 10 is for rotating and separating the adipose tissue and the subcontainer 10 has a bowl 11 and a cap 12 coupled to the top of the ball 11. When the ball 11 and the cap 12 are coupled, a space portion 13 in which the fat tissue can be accommodated is formed therein.

The body of the ball 11 is formed to have a smaller or constant diameter as it goes downward. However, the lower portion of the ball 11 has a shape in which the diameter decreases sharply toward the lower side, and the groove portion 14 is formed concavely at the lowermost end of the ball 11.

A through hole 15 is formed in a central portion of the cap 12 and a protruding wall portion 16 is formed to protrude upward from the outer periphery of the through hole 15. [

 Further, a stirring blade (17) is formed on the inner wall of the lower part of the ball (11). The stirring vanes 17 are formed in a plate shape and are formed so as to protrude from the inner wall toward the space portion 13. The stirring vanes 17 are arranged symmetrically with respect to the rotation center of the sub container 10 at four angular intervals of 90 degrees. Of course, there may be only one stirring vane 17 and it is not necessarily symmetrical, but it is preferable to be symmetrically arranged. The stirring vanes 17 are rotated together with the rotation of the sub-container 10, thereby playing a role of smoothly stirring the fat tissue.

The outlet pipe 90 is for transferring the material to the inside of the sub-container 10 or vice versa and discharging the material from the sub-container 10. In this embodiment, the center pipe 30 and the auxiliary pipe 40 are provided do.

The central pipe 30 functions as a flow path for transferring the washing liquid, enzyme, etc. to the space portion 13 of the sub container 10, or discharging the substance in the sub container 10 to the outside. The center pipe 30 is formed in a hollow shape and inserted into the sub container 10 through the through hole 15 of the sub container 10. The lower end of the central pipe 30 is disposed in the groove 14 of the sub container 10 in the state of being close to the bottom of the sub container 10 and the upper end of the center pipe 30 is disposed in the upper part of the sub container 10 .

Also, in the present embodiment, the auxiliary pipe 40 communicates the inside and the outside of the sub-container 10 with a path that is independent of the center pipe 30. Particularly, the auxiliary pipe 40 functions as a passage through which the air flows into and out of the sub-container 10. Therefore, the auxiliary pipe 40 does not need to be inserted into the lower end of the sub-container 10, and may be inserted only up to the upper end of the sub-container 10.

The center pipe 30 and the auxiliary pipe 40 may be separately formed, but they are integrally formed in the present embodiment. The auxiliary pipe 40 is formed to have a larger diameter than the central pipe 30 and is disposed to surround the central pipe 30. The auxiliary pipe 40 is disposed in the center pipe 30, The upper portion of the auxiliary pipe 40 is formed by being bent in a direction opposite to the direction in which the center pipe 30 is bent. A space between the inner surface of the auxiliary pipe 40 and, more precisely, the inner surface of the auxiliary pipe 40 and the outer peripheral surface of the central pipe 30 forms a flow path 45 through which air can flow. However, the flow path 45 is not necessarily used only for the inflow / outflow of air, and may be used as an inflow / outflow path for various materials such as a material for blood contaminants or the like, or a cleaning liquid can be injected.

An insertion port 35 is formed at the upper end of the central tube 30 and a stopper 36 is screwed into the insertion port 35. When a cap 36 is removed, a syringe needle (not shown) is inserted into the center tube 30 through the insertion port 35 to suck the liquid material placed in the groove 14 of the sub container 10 . When the syringe needle is not used, the insertion port 35 is closed by the cap 36 to prevent the space 13 inside the sub container 10 from being contaminated. In addition, a tube is inserted into the insertion port 35 in addition to the syringe needle so that the substance can be exchanged with the inside of the sub-container 10.

  At the lower side of the auxiliary pipe 40, a shielding member 47 is fitted to the center pipe 30 to be engaged. When the sub-container 10 is rotated for separation into adipose tissue, liquid substances such as blood or washing liquid can be abruptly raised to the upper portion of the sub-container 10, And can be drawn in between the auxiliary pipe 40 and the inner peripheral surface of the through hole 15 of the sub container 10. The shielding member 47 covers the through hole 15 and the auxiliary pipe 40 to prevent blood contamination or the like from entering the auxiliary pipe 40 or the like. However, since the auxiliary pipe 40 is in contact with the space 13, the wrapping is not completely closed but a gap is formed in the upper portion so as to communicate with the space 13.

Spiral auxiliary stirring vanes (not shown) are formed on the outer circumferential surface of the central tube 30 to stir the fatty tissue together with the stirring vanes 17 on the lower side of the sub vessel 10.

On the other hand, the sub container 10 has to be rotated relative to the outlet pipe 90. For example, as in the present embodiment, in the case of the access pipe 90 having the double pipe structure, the auxiliary pipe 40 is sealed between the auxiliary pipe 40 and the sub-container 10, The space portion 13 of the housing 10 must be sealed.

In this embodiment, the compressible sealing member 50, the first friction reducing member 61, and the second friction reducing member 65 are used to rotate the sub-container 10 and the outlet pipe 90 relative to each other, 13) is sealed. That is, the sealing member 50 employed in the present invention is interposed between the outlet pipe 90 and the sub-container 10 to seal the space inside the sub-container 10.

The sealing member 50 is formed in an annular shape and is fitted in the protruded wall portion 16 of the cap 12. The sealing member 50 is provided between the flange portion 48 formed on the outer peripheral surface of the auxiliary pipe 40 and the cap 12 . The sealing member 50 is made of an elastically compressible rubber material so that when the flange portion 48 of the auxiliary pipe 40 is compressed between the cap 12 and the through hole 15, Is fully sealable.

However, when the sealing member 50 is rotated together with the sub-container 10, the sealing member 50 of the rubber material has a high coefficient of friction, so that it is not smoothly rotated due to friction with the flange portion 48, Friction heat is generated. In the present invention, the first friction reducing member (61) and the second friction reducing member (65) are used to minimize heat generation and improve the structure so that the generated heat can be quickly released.

First, the first friction reducing member 61 is formed in an annular shape, is fitted to the auxiliary pipe 40, and is fixed to the lower surface of the flange portion 48. The second friction reducing member 65 includes a first friction reducing member 65 and a body portion 66 and a plurality of wing portions 67. The body portion 66 includes a first friction reducing member 61, And is disposed so as to surround the through-hole 15. As shown in Fig. And is physically coupled to the upper side of the sealing member 50. The upper surface of the body portion 66 is in surface contact with the lower surface of the first friction reducing member 61 and is in close contact with each other. The body portion 65 of the second friction reducing member 65 and the first friction reducing member 61 are relatively rotated in a state of being in close contact with each other. The wing portions 67 protrude from the outer circumferential surface of the body portion 66 along the radial direction and are disposed apart from each other along the circumferential direction of the body portion 66. The wing portion 67 is also inclined with respect to the rotation plane (generally horizontal plane) of the body portion 66. Accordingly, when the second friction reducing member 65 rotates, the wing portion 67 acts as a pan to form a sudden flow of air (i.e., wind). The wind is formed upward or downward according to the direction of inclination of the wing portion 67 and the direction of rotation of the second friction reducing member 65. Accordingly, the heat generated while the first friction reducing member 61 and the second friction reducing member 65 are rotated in contact with each other can be quickly discharged by the wind generated by the wing 67.

The first friction reducing member 61 and the second friction reducing member 65 are made of a ceramic or carbon material having a low number of friction and the frictional surface is flattened so that the generation of frictional heat itself is reduced. Which is advantageous in heat dissipation.

The flange portion 48 of the auxiliary pipe 40 and the subcontainer 10 are pressed toward each other in such a manner that the sealing member 50 of the rubber material is compressed and the through holes 15, The clearance between the auxiliary pipe 40 is closed and the first friction reducing member 61 and the second friction reducing member 65 are also brought into close contact with each other so that the sub container 10 can be sealed. When the sub container 10 rotates, relative rotation occurs between the first friction reducing member 61 and the second friction reducing member 65, but the first friction reducing member 61 and the second friction reducing member 65 generate little frictional heat and heat transfer is fast, and the wing portion 67 forms a sudden flow of air, so that heat can be smoothly discharged. Therefore, the centrifugal separator according to the present invention is also able to cope with the damage and durability of the centrifugal separator due to the generation of heat due to the contact rotation. In addition, there is an advantage in that it is possible to manufacture in a very economical manner compared with the conventional method using a hermetic bearing or a retainer ring.

The first frictional reducing member is installed and fixed to the outlet pipe 90 while the second frictional reducing member 65 is connected to the upper portion of the sealing member 50 and the sub container 10 ), But this configuration may be changed according to the embodiment. For example, the sealing member is provided below the flange portion 48 of the auxiliary pipe 40, the first friction reducing member is attached to the lower surface of the sealing member, and the second friction reducing member is engaged with the upper portion of the sub- . In this configuration, the sealing member does not rotate, unlike the previous case, but the same effect as that described above can be obtained. That is, the hermeticity of the sub-container is ensured by the first frictional reducing member and the second frictional reducing member which are in close contact with the compressible sealing member, and by the constitution of the material and the wing portion of the first frictional reducing member and the second frictional reducing member Heat dissipation can be ensured.

The central tube 30 is connected to a pump (not shown) by a connection tube (not shown) to supply a substance such as enzyme and cleaning solution to the sub-container 10 through the central tube 30, And discharges adipocytes or waste fluid from the container 10 to the outside. In this embodiment, since a substance such as a cleaning liquid must be introduced into and out of the sub-container 10, the pump must provide suction force in both directions. The pump uses a peristaltic pump that provides suction force while being rotated in both directions.

After the adipose tissue and washing solution are introduced into the regenerative cell separating apparatus 100 having the above-described structure, the subcontainer 10 is rotated to wash the adipose tissue, or the collagenase enzyme is added to decompose the adipose tissue, (10) is rotated, collagen is separated from adipose tissue. The discharged collagens can form a large lump with each other. When the collagen lumps are introduced into the grooves 14 of the sub container 10, the central tube 30 or the auxiliary tube 40 can be closed.

Accordingly, in the present invention, the screen member 70 is provided to prevent the collagen mass from closing the central tube 30 or the auxiliary tube 40.

In the present embodiment, the screen member 70 includes a support member 71 and a plurality of engagement members 72. [ The support member 71 is formed in an annular shape so as to surround the central tube 30 and the shielding member 47 and is coupled to the bottom surface of the cap 12. [ The plurality of engaging members 72 extend downward from the lower end of the support member 71 and are bent and bent outward of the sub-container 10 to be held in contact with the bottom or inner surface of the sub-container 10.

3, when the plurality of engaging members 72 are viewed from above, the engaging members 72 are formed to extend outward from the center of the sub-container 10, and are formed in the circumferential direction (more specifically, In the radial direction as a whole.

The spacing between the plurality of engaging members 72 is approximately 1 to 5 mm in order to prevent the collagen mass from being discharged through the interlocking members 72. However, the above interval may be variously changed depending on the kind of tissue and the like.

A slit 73 is formed at the lower end of the engaging member 72 disposed at a position corresponding to the plurality of stirring vanes 17 among the plurality of engaging members 72. The slit 73 is fitted with a plate-shaped stirring vane 17. The latching member 72 is made of a flexible material so that it can be bent in the vertical direction.

Hereinafter, a regenerative cell extracting apparatus according to the present invention will be described with reference to the accompanying drawings.

FIG. 4 is a schematic exploded perspective view of a regenerative cell extracting apparatus according to an embodiment of the present invention, FIG. 5 is a schematic perspective view of the regenerative cell extracting apparatus shown in FIG. Fig.

4 to 6, the regenerative cell extracting apparatus 200 according to the present embodiment receives centrifugal separation from the regenerative cell separating apparatus 100 to receive an aqueous solution containing stem cells, To extract the cells. However, when it is not used together with the regenerative cell separator 100 and used alone, it can be used for extracting stem cells directly from adipose tissue.

The regeneration cell extracting apparatus 200 performing the above function includes a main container 210 and an access pipe 290.

The main container 210 includes a main body 211 and a lid 212. The main body 210 has a receptacle 213 in which an object such as an aqueous solution containing adipose tissue or stem cells is received. The lower portion of the main body 211 is formed to be gradually narrowed from the upper side to the lower side, or formed to have a constant diameter. In particular, the recess 214 is formed at the lowermost end of the accommodating portion 213.

A through hole 215 is formed at the center of the top surface of the lid 210 so as to pass between the upper surface and the lower surface and an annular protruding wall portion 216 is formed to protrude upward from the outer periphery of the through hole 215.

In addition, a protruding receiving portion 217 is formed on the outside of the main container 210 in a convex manner. That is, the protrusion accommodating portion 217 is convex outwardly of the main container 210 along the radial direction around the rotation center axis of the main container 210, and is formed long along the height direction of the main container 210. The longer length means that the lower end portion and the upper end portion of the protrusion receiving portion 217 are elongated so as to be disposed at the lower portion and the upper portion of the receiving portion 213, respectively.

Also, the protrusion accommodating portions 217 are continuously disposed along the outer wall of the main container 210. [ Therefore, the outer circumferential surface of the main container 210 is formed in a wave form as a whole because the convex portion (protrusion receiving portion) and the concave portion are alternately and continuously arranged.

The protrusion accommodating portion 217 should be disposed symmetrically with respect to the main container 210 so as not to affect the rotation of the main container 210. In this embodiment, Are symmetrically arranged at eight angular intervals at an angle of 45 deg.

When the main container 210 is rotated, the separated objects are separated by weight, and the heavy components are disposed on the outer side in the main container 210 and the light components are disposed on the inner side. Since the protrusion accommodating portion 217 is disposed at the outermost periphery with respect to the center of rotation of the main container 210, the heaviest component separation is accommodated in the protrusion accommodating portion 217. Stem cells isolated from adipose tissue are heavier than other components and are accommodated in the protruding receptacle 217 during centrifugation.

As described in the present invention, the reason why the projecting accommodating portions 217 are continuously disposed on the outer peripheral surface of the main container 210 is to increase the throughput per hour. In the prior patent by the present applicant, the projecting accommodating portion was formed symmetrically in only four places in the main container, and the throughput per hour was small. In the present invention, the distance from the center of the main container to the vertex of the projecting accommodating portion is the same as that of the prior patent, that is, the diameter to the outermost portion of the main container is the same, but the projecting accommodating portion 217 is continuously arranged, The amount of the aqueous solution that can be accommodated in the main container 210 is greatly increased.

If the conventional centrifugal separator is filled with 15 cc, the centrifugal separator according to the present invention can be filled with 80 cc. In addition, the area of the protrusion accommodating portion 217 to which the stem cells can be attached is also increased more than two times, and the extraction rate of the stem cells is also increased.

This should be understood in terms of the process efficiency of maximizing the amount of aqueous solution that can be treated using the same rotating jig and the extraction rate of stem cells, rather than simply increasing the number of protrusion accommodating portions 217. Particularly, since the tissue separated from the human body is treated as an object to be treated, how many objects to be treated in the same time and how many target substances (stem cells) can be extracted therefrom is very important in terms of medical treatment . In addition, in the present invention, the maximum economic effect is achieved by changing the structure of the main vessel of the centrifuge at a level that does not supplement or change facilities by using the rotary jig as it is.

On the other hand, the lower end of the protrusion accommodating portion 217 is blocked by the blocking member 240. The blocking member 240 serves to prevent the stem cells from being lost during the process of increasing the rotation speed, which is the initial stage of centrifugation, and stopping the rotation by reducing the rotation speed which is the end of the centrifugation.

When the lower end of the projecting accommodating portion 217 is in communication with the lower end of the main container, stem cells are accommodated in the projecting accommodating portion 217 by centrifugal separation. In the process of increasing the rotation speed at the initial stage of centrifugal separation, The upper end of the main container 210 comes up to the projecting receiving portion. In addition, stem cells attached to the protrusion accommodating portion 217 may be lost as the aqueous solution rises. That is, the blocking member 240 is provided to stably accommodate stem cells attached to the protrusion accommodating portion 217 to prevent loss of stem cells. The blocking member 240 is formed with a hole (not shown) passing between the upper surface and the lower surface of the receiving member 213. The blocking member 240 receives centrifugal force from the lower portion of the receiving unit 213 and allows the stem cells coming along the inner wall of the main container to pass therethrough . That is, although the separation object is prevented from being abruptly lifted up by the protrusion receiving portion, a small amount of stem cells that are pushed up to the upper portion from the lower portion under centrifugal force can be introduced into the protrusion receiving portion.

The blocking member 240 also serves to preserve the stem cells attached to the protrusion receiving portion 217 during the termination of the centrifugation. That is, when the centrifugal force is reduced in the process of decelerating the main container after the centrifugal separation, the stem cells attached to the protrusion accommodating portion 217 are more affected by gravity, ) Along the inner wall of the bottom. The blocking member 240 also prevents the stem cells from being separated from the protrusion accommodating portion 217 in the course of stopping the main container after centrifugal separation.

The outlet pipe 290 has a first outlet pipe 220 and a second outlet pipe 230. The first discharge pipe 220 and the second discharge pipe 230 are for communicating the receiving portion 213 of the main container 210 with the outside of the main container 210. The first discharge pipe 220, The discharge pipes 230 form mutually independent paths. The first discharge pipe 220 and the second discharge pipe 230 are disposed coaxially with the main container 210. The first discharge pipe 220 is elongated so that the lower end thereof is connected to the lower end of the main container 210, And the second discharge pipe 230 is formed in a short shape so that the lower end of the second discharge pipe 230 is disposed at the upper end of the main container 210. [

The material flowing through the first discharge pipe 220 and the second discharge pipe 230 may be changed according to the use mode. In this embodiment, however, the first discharge pipe 220 is separated from the regenerative cell separator 100 An aqueous solution containing a stem cell, a washing solution, etc. may be introduced, and stem cells extracted from the main container 210, a washing solution and the like may be discharged. In addition, air can flow in and out through the second discharge pipe, and the cleaning liquid or the like after use can be discharged.

The constitution of the access tube 290 of the regenerative cell extracting apparatus 200 is completely the same as the constitution of the access tube 90 of the regenerative cell separator 100 described above. That is, the specific configurations of the first discharge pipe 220 and the second discharge pipe 230 of the regenerative cell extracting apparatus 200 are completely different from those of the central tube 30 and the auxiliary tube 40 in the regenerative cell separator 100, same. Also, the space between the inner circumferential surface of the second discharge pipe 230 and the outer circumferential surface of the first discharge pipe 220 forms a flow path 235 through which fluids such as air and cleaning fluid can flow. The configuration of the insertion hole 237 and the cap 236 is the same as that of the insertion port 35 and the cap 36 of the regenerative cell separation device. A detailed description thereof will be omitted.

Meanwhile, the main container 210 is relatively rotated with respect to the inlet / outlet pipe 290, and the space between the outlet pipe 290 and the main container 210 is sealed so that the receiving portion 213 of the main container 210 is sealed .

In the present invention, the problems of hermeticity, relative rotation, and heat generation are all solved by using the compressible sealing member 250, the first friction reducing member 261 and the second friction reducing member 265. This configuration is completely the same as that of the regenerative cell separation apparatus 100 described above, and therefore, the description will be briefly described only for the sake of clarity and redundancy.

That is, the compressible sealing member 250 is annularly fitted to the protruding wall portion 216 on the upper portion of the main container 210, and the second friction reducing member 265 in the form of an annular ring is formed on the compressible sealing member 250 . A flange portion 248 is formed in the inlet pipe (specifically, the second discharge pipe), and the first frictional reducing member 261 in the form of a ring surrounds the second outlet pipe 240 and is attached to the flange portion 248. The lower surface of the first friction reducing member 261 and the upper surface of the body portion 266 of the second friction reducing member 265 are mutually contacted and relatively rotated in close contact with each other. The wing portion 267 of the second frictional reducing member 265 forms a wind to allow the heat due to the rotational contact between the first frictional reducing member and the second frictional reducing member to be released easily. This configuration is completely the same as the sealing member 50, the first frictional reducing member 61 and the second frictional reducing member 65 of the regenerative cell separating apparatus described above. Accordingly, the problem of hermeticity, guarantee of relative rotation, and heat radiation due to heat generation can be completely solved.

Meanwhile, in this embodiment, during the rotation of the main container 210 for centrifugation, a cleaning liquid other than stem cells can be discharged to the outside of the regenerative cell extracting apparatus 100 through the second discharge pipe 230 to provide. That is, in this embodiment, the rotation of the main container 210 may be stopped and the cleaning liquid may be discharged from the lower portion of the main container 210 through the first discharge pipe 220. However, during the rotation of the main container 210 The cleaning liquid or the like may be discharged to the outside by the vowel member 270 and the guide member 280 to be described later.

The vowel member 270 is fitted to the first discharge pipe 220 and is formed with a collecting portion 271 concavely so as to temporarily receive the separated product guided by the guide member 280 to be described later. In the present embodiment, the vowel member 270 is formed in a concave disc shape. The vowel member 270 is disposed at the upper portion of the main container 210, more precisely, at the lower end of the second discharge pipe 230. The diameter of the vowel member 270 is set to a size such that the outer peripheral surface of the vowel member 270 can be close to the inner peripheral surface (concave portion connecting surface) of the main container 210.

The guide member 280 is disposed on the vowel member 270 to guide the separated product that is being centrifugally rotated in the main container 210 to the collection portion of the vowel member 270. In this embodiment, the guide member 280 is formed so as to protrude upward from the upper surface of the vowel member 270, and is disposed symmetrically with respect to the vowel member 270 at 90 degree angular intervals. The plurality of guide members 280 are formed so as to be curved toward the center from the outer peripheral surface of the vowel member 270. Accordingly, the plurality of guide members 280 disposed on the vowel member 270 are formed in a whirling manner as a whole.

Of course, the guide member 280 does not necessarily have to be curved, but may be straight, and may be formed in a short section at a portion that is not formed over the entire radius of the vowel member 270, It can be in various forms such as water supply. Also, the number of guide members may be variously selected from one to a plurality.

As described above, the separation object accommodated in the main container 210 is rotated together with the centrifugal force by the centrifugal force as the main container 210 is rotated. As described above, the stem cells and the like, which are heavy components, And a light component such as a cleaning solution maintains a close contact with the inner circumferential surface of the main container 210. When the main container starts to rotate, the separation object is rotated together to raise the water level from the lower part of the main container 210 to the upper part. As shown in FIG. 7, the separated water having a raised water level to the upper part of the vowel member 270 And is guided along the guide member 280 to the center side of the vowel member 270 while being in contact with and interfering with the guide member 280. When the suction force is applied to the second discharge pipe 230 from the pump, the separated substance guided to the center of the vowel member 270 is discharged to the outside through the second discharge pipe 230 by the suction force.

A plurality of flow holes 272 are formed in a central portion of the vowel member 270. The flow holes 272 lower the pressure applied to the vowel member 270 so that a large amount of separated water Thereby preventing the overflow of the separated product.

Meanwhile, the regenerative cell extracting apparatus 200 according to the present embodiment includes a suction device. In this embodiment, a first pump (not shown) and a second pump (not shown) are provided. The first pump is connected to the first discharge pipe 220 by the connecting tube and the second pump 292 is also connected to the second discharge pipe 230 by the connecting tube so that the first discharge pipe 220 and the second discharge pipe 220, (Not shown).

In this embodiment, since the fluid must be introduced and discharged through the first discharge pipe and the second discharge pipe, respectively, the pump uses a peristaltic pump that provides suction force in both directions.

Meanwhile, a photo sensor (not shown) is installed in the connection tube connected to the central tube 30 of the regenerative cell separator 100. The photosensor can sense the color of the fluid flowing along the connection tube. In particular, when the yellow adipose fat cells are discharged from the regenerative cell separator 100 through the central tube 30, Or a signal that the adipose tissue is being discharged to a controller (not shown) to be described later.

The connection tube connected to the central tube 30 of the regenerative cell separation apparatus 100 may be provided with a multi-filter capable of filtering large tissues such as collagen in a lump state.

The connection tube connected to the first discharge pipe 220 and the connection tube connected to the second discharge pipe 230 of the regeneration cell extractor 200 are provided with a sensor for sensing the flow of air through the connection tubes, It senses that the air is discharged from the cell extracting apparatus 200 through the first discharge pipe 220 and the second discharge pipe 230, and sends a signal to the controller.

The regenerative cell extracting system includes a regenerative cell separator 100, a regenerative cell extractor 200, a rotary jig (not shown), and a controller (not shown) for controlling the operation of components such as various valves , All operations are controlled by this controller.

Hereinafter, a method for extracting regenerated cells using the regenerative cell extracting system having the above-described constitution will be described.

As described above, the regeneration cell separation apparatus 100 and the regeneration cell extraction apparatus 200 are connected to each other by a plurality of connection tubes. The connection tubes are also connected to a storage bag in which a washing solution is stored, , A storage bag in which washed fat tissue is stored, a storage bag in which the used medicine is stored, and a storage bag in which stem cells are stored. In addition to the above, the storage bag may be connected to a storage bag in which adipose cells are stored or a storage bag for storing the extracted stem cells in a cryopreservation manner.

First, the adipose tissue to be separated is introduced into the regenerative cell separating apparatus 100 through the central tube 30. For example, the fat tissue may be preliminarily extracted from a human body through a syringe, and then the fat tissue may be introduced into the separation apparatus 100 again. However, in order to prevent contamination during the transferring process, a syringe needle is directly inserted into a human body, a tube connected to the syringe injector is connected to the central tube 30, The fat tissue can be directly introduced into the sub-container 10 from the human body.

As described above, when the adipose tissue is introduced into the sub-container 10, the valve is opened and the pump is operated to transfer the washing solution to the regenerative cell separation device 100. The rotation jig is operated to repeat the rotation and stop of the regenerative cell separator 100. When the rotation and the stopping are repeated at regular intervals, the fat tissue and the washing liquid are centrifuged while being stirred with each other, and the adipose tissues flowing along with the blood-derived contaminants are washed.

When the sub container 10 is left stationary for about one minute, heavy components such as a cleaning liquid, blood, body fluids, and the like are disposed in the lower part of the sub container 10, The ingredients are placed on the top layer.

The pump is operated again to discharge the blood, the washing liquid, etc. in the lower part of the sub container 10 through the central pipe 30. When the washing liquid is exhausted, the yellow fatty tissue is discharged through the central tube 30. The photosensor installed in the connecting tube senses the color of the fatty tissue and sends a signal to the controller. In the controller, the operation of the pump is stopped, and the above procedure is repeated three to four times to allow the fat tissue to be completely cleaned.

As described above, blood contaminants, body fluids, and oil components are separated from the fat tissue extracted from the human body, and after the fat tissue has been completely washed, the washed fat tissue is transferred to the storage bag, if necessary, Can be stored.

Thereafter, the valve is opened while operating the pump to transfer the collagenase from the enzyme storage bag to the regenerative cell separator 100 through the central tube 30. When the collagenase enters the sub-container 10, the operation of the pump is stopped and the sub-container 10 is rotated and agitated for approximately 30 minutes. At this time, the temperature of the sub container 10 is maintained at about 37 캜 through a rotating means such as a rotary jig. The collagenase degrades adipose tissue, and the degraded components are centrifuged by rotation of the sub-container 10. When the rotation of the sub-container 10 is stopped and left to stand for 1 minute, the aqueous solution containing the adipose-derived stem cells is disposed at the lower part of the sub-container 10 because it is a relatively heavy component, and matured fat cells ) And the oil component.

The pump is operated again to transfer the aqueous solution containing the stem cells to the regenerative cell extracting apparatus 200 through the central tube 30. During transfer, the pump and the second pump work together. When the yellow sensor is detected again by the photosensor, the aqueous solution containing the stem cells is exhausted and then the fat cells are discharged. And, if necessary, mature adipocytes can be stored in separate storage bags.

When the aqueous solution including the stem cells is introduced into the main container 210 of the regenerative cell extracting apparatus 200 as described above, the stem cells are extracted. The vowel member 270 and the guide member 280 It may or may not be used.

First, when the vowel member 270 and the guide member 280 are not used, all the valves are closed and the main container 210 is rotated for about five minutes in a state in which the pumps are also stopped. A relatively heavy stem cell is pressurized by the centrifugal force and attached to the inner wall of the receiving protrusion 217, and a body fluid, a washing liquid, an enzyme, and the like are attached to the lower part of the main container 210 . An important point is to naturally decelerate the main container 210 without artificially decelerating the rotation of the main container 210. The stem cells attached to the inner wall of the receiving protrusion 217 may fall to the lower portion of the main container 210 when the brake is artificially rotated.

When the centrifugal separation is completed, the first pump is operated to transfer the aqueous solution such as body fluid to the waste liquid storage bag through the first discharge pipe 220. When air is detected in the sensor installed in the connection tube connected to the first discharge pipe 220, the operation of the first pump is stopped and the process proceeds to the next step since the aqueous solution is discharged.

When the vowel member 270 and the guide member 280 are used, the second pump is operated together with the rotation of the main container 210 to provide a suction force through the second discharge pipe 230. As the main container 210 is rotated, the aqueous solution containing the stem cells is also rotated, and the stem cells are attached to the inner wall of the protrusion accommodating portion 217. The rest of the aqueous solution except the stem cells continues to rise in water level. When the aqueous solution is moved to the upper portion of the vowel member 270, the rotating aqueous solution is guided along the guide member 280 toward the center of the vowel member 270 while interfering with the guide member 280 do. The second supply pipe 230 is disposed above the central portion of the vowel member 270 so that the demanded liquid is supplied to the flow path 235 between the first discharge pipe 220 and the second discharge pipe 230 by the suction force of the second pump 292, And is transported to the waste liquid storage bag. At this time, when air is sensed by a sensor installed in the connection tube connected to the second discharge pipe 230, it is determined that the aqueous solution has been discharged, and the operation of the pump is stopped. Thereafter, when the main container 210 is naturally decelerated and stopped, only the stem cells are attached to the inner wall of the protrusion accommodating portion 217 in the main container 210. If some aqueous solution is not discharged and remains in the lower part of the main container 210, the first pump may be operated and transferred to the waste liquid storage bag through the first discharge pipe.

As described above, after centrifugation of the aqueous solution containing the stem cells is completed, a small amount of washing solution is put into the main container 210 to wash the stem cells attached to the inner wall of the protrusion receiving part 217, After rotating and stopping the vessel about 5 times, the main container 210 is decelerated again by centrifugal separation. When the main container is accelerated and decelerated while repeating the rotation and stopping, the stem cells attached to the inner wall of the protrusion accommodating portion 217 fall down to the lower portion of the main container 210. If the above-described stem cell extraction process is repeated several times, the stem cells finally obtained may contain at least the foreign material.

Finally, the stem cells existing in the lower part of the main container 210 are transferred to the stem cell storage bag through the first discharge tube 220, thereby completing the stem cell extraction process.

Since the entire process is automatically performed by the regeneration cell extraction system according to the present invention, it is not only accurate and efficient, but also does not cause contamination problems because the entire system is performed under a closed condition.

First of all, a primary centrifugation is performed in the regenerative cell separator 100 to form a concentrate containing stem cells. In the regenerative cell extractor 200, secondary centrifugation is performed on the concentrate, so that the extraction operation is efficient There is an advantage that the extraction yield is improved.

Although the present invention has been described and illustrated as separating adipose tissue, the regenerative cell extracting apparatus and the regenerative cell extracting system according to the present invention can be used for separating adipose tissue, Cells can also be extracted.

Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims. Of course.

Therefore, the scope of the present invention should not be limited to the described embodiments, but should be determined by the scope of the appended claims, as well as the appended claims.

100 ... regenerative cell separator 200 ... regenerative cell extractor
210 ... main container 211 ... main body portion
212 ... cover 213 ... accommodating portion
217 ... protrusion accommodating portion

Claims (11)

A main container rotatably provided with a rotational force from the outside and having a receiving portion for receiving the object to be separated;
A hollow type inlet / outlet pipe inserted into a through hole formed in the main container so that one end thereof is disposed inside the main container and the other end is disposed outside the main container and relatively rotated with the main container;
A first friction reducing member formed in a ring shape so as to surround the inlet / outlet pipe;
A main body rotatable together with the main container and formed in an annular shape corresponding to the first friction reducing member so as to surround the through hole and closely contacted with the first friction reducing member; A second frictional reducing member having a plurality of wings which are inclined with respect to the rotational surface of the body; And
And a sealing member provided between the main container and the inlet and outlet tube to seal the gap between the inner circumferential surface of the through-hole of the main container and the outer circumferential surface of the outlet tube. The method according to claim 1,
Wherein the sealing member is provided in the main container with a compressible rubber material and rotated together with the main container,
And the second friction reducing member is coupled to the sealing member and rotated together with the main container. The method according to claim 1,
Wherein the first friction reducing member is made of a ceramic material,
Wherein the second friction reducing member is made of a carbon material. The method according to claim 1,
A plurality of projecting portions formed to be convex outward along the radial direction around the rotation axis of the main container are continuously disposed on the outer circumferential surface of the main container so as to accommodate relatively heavy separates among the separated objects separated from the object to be separated , And the outer circumferential surface of the main container is formed in a wavy shape in which convex portions and concave portions are alternately arranged in succession. 6. The method of claim 5,
Wherein the main container includes a main body portion having the receiving portion and the protruding portion and a lid portion coupled to an upper end of the main body portion to seal the receiving portion,
And a blocking member is formed at a lower end of the protrusion receiving portion to block the lower end of the main container receiving portion and the protrusion receiving portion. 6. The method of claim 5,
Wherein the blocking member is provided with a hole for communicating the lower end of the main container accommodating portion with the projecting accommodating portion. A subcontainer for separating tissue, comprising: a subcontainer which is rotatably received by a rotating force from the outside and in which a space for receiving a tissue is formed; and one end is disposed inside the subcontainer and the other end is disposed outside the subcontainer A regeneration cell separation device having a hollow type entrance and exit tube inserted into a through hole formed in the sub container so as to be disposed, and a pump connected to the entrance and exit and providing a suction force; And
The regenerated cell according to any one of claims 1 to 6 for extracting regenerated cells from the regenerative cell separator through a secondary centrifugation, And an extracting device for extracting the regenerated cells from the regenerated cells. 8. The method of claim 7,
And a screen member installed in the space of the container so as to prevent a lump of a predetermined size or larger formed in the process of separating the tissue from falling down to the lower end of the container, system. 9. The method of claim 8,
Wherein the screen member has a plurality of engagement members formed outwardly from a central portion of the sub container,
Wherein the plurality of engaging members are spaced apart from each other along the circumferential direction and are made of a flexible material so as to be bent from an upper center portion of the sub container toward a lower outer frame portion of the container. 8. The method of claim 7,
The regenerative cell separation apparatus may further comprise:
A first friction reducing member formed in a ring shape so as to surround the inlet / outlet pipe;
A body portion which is rotated together with the sub-container and is formed in a ring shape corresponding to the first friction reducing member so as to surround the through-hole and is brought into close contact with the first friction reducing member; A second frictional reducing member having a plurality of wings that are inclined with respect to the rotational surface of the body,
Further comprising a sealing member provided between the sub-container and the outlet pipe so as to seal a gap between the inner circumferential surface of the through-hole of the sub-container and the outer circumferential surface of the outlet pipe. A subcontainer for separating fat tissue, comprising: a subcontainer provided with a space for receiving a rotating tissue and receiving a rotating force from the outside; A regeneration cell separation device having an entrance and exit tube of a mold and a pump connected to the entrance tube to provide a suction force;
The regenerated cell according to any one of claims 1 to 6 for extracting regenerated cells from the regenerative cell separator through a secondary centrifugation, A method for extracting regenerative cells from a tissue using an aliphatic cell extraction system comprising an extraction device,
Washing the fat tissue by injecting the fat tissue and the washing liquid into the regenerative cell separating apparatus;
And then discharging the heavy component including blood and body fluid continuously through the tube from the regenerative cell separating apparatus after the washing of the fatty tissue, if yellow fat tissue is discharged, stopping the discharge by sensing yellow using a photosensor;
Decomposing the adipose tissue by rotating the regenerative cell separating device while the collagenase is put in the regenerative cell separating device;
An aqueous solution containing regenerated cells separated from the adipose tissue and submerged in the lower part of the regenerative cell separator is discharged from the regenerative cell separator through the tube and transferred to the regenerative cell extractor, Stopping the discharge when mature adipocytes are detected;
The regenerative cell extracting apparatus is rotated to centrifuge the aqueous solution containing the regenerative cells so that the regenerative cells adhere to the inner circumferential surface of the main container of the regenerative cell extracting apparatus and the remaining light components are accommodated in the lower portion of the main container step; And
Separating the regenerative cell from the inner circumferential surface of the main container by allowing the remaining light component to be discharged first from the regenerative cell extracting device and then injecting the washing liquid into the main container and rotating the regenerative cell to fall down to the bottom of the main container, And extracting and recovering the regenerated cell from the extracting apparatus.

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