KR101982766B1 - Device for separating the adipose stem cells - Google Patents

Abstract

Provided is an apparatus for isolating adipose stem cells. The apparatus of the present invention comprises: a first sealing cap having an auxiliary cap for opening or closing a communication hole formed by penetrating the upper surface, and an operation bar having predetermined length and extended from the lower surface; a first cylindrical housing forming a first space and having an opened upper part selectively opened or closed by the first sealing cap by forming a male screw portion on an external circumference of the upper end to be thread-engaged with a female screw portion formed on the inner circumference of the lower end of the first sealing cap; a second cylindrical housing communicating with the first cylindrical housing by using a neck hole of a neck portion formed on the lower end of the first cylindrical housing where the operation bar is disposed; a cap connector assembled into an opened lower part of the second cylindrical housing in order to form a second space between the same and the second cylindrical housing; and a second sealing cap having a sealing member correspondingly inserted into a discharge hole formed by penetrating the cap connector, thereby being assembled into the cap connector. Accordingly, isolation of adipose stem cells can be simplified to reduce time for extraction and costs.

Description

TECHNICAL FIELD [0001] The present invention relates to a stem cell separator,

The present invention relates to a device for separating adipose stem cells from adipose tissue, and more particularly to a device for separating adipose stem cells (SVF) from adipose tissue by centrifugation through a centrifuge And a cell separator.

In general, stem cells have the ability to self-replicate and to differentiate into two or more cells. In recent years, studies have been conducted to acquire stem cells in adipose tissue in addition to bone marrow.

Fat-derived stem cells isolated from adipose tissue are pluripotent stem cells having the ability to differentiate into various types of cells such as adipocytes, muscle cells, chondrocytes, and bone cells.

These adipocyte stem cells have excellent self-regenerating ability, easy in vitro culture, and are rich in adipose tissue that can separate adipose stem cells, and are simple and safe to collect, compared with mesenchymal stem cells of bone marrow origin.

In addition, it is known that almost all characteristics of adipose stem cells such as differentiation ability, cytologic characteristic, immunological characteristic, and tissue regeneration ability are the same as those of mesenchymal stem cells of bone marrow origin.

In other words, it is known that adipose stem cells are highly useful as stem cells having a high possibility of replacing mesenchymal stem cells of bone marrow origin.

In addition, the adipose stem cells have advantages in that the tissue acquisition process is easier and safer than the mesenchymal stem cells, and there is no limitation in the supply and demand of the tissues, and the in vitro culture is facilitated, which is superior in terms of organization accessibility, safety, efficacy and economics.

In general, the method of isolating and culturing the adipocyte stem cells is a method in which the inhaled or resected fat is shredded to decompose the tissue into collagenase, and the stromal vascular fraction (SVF ) Are cultured in a culture medium.

On the other hand, in general, all the processes for separating adipose stem cells from adipose tissue are performed in one separation vessel. In this case, the process becomes complicated due to the discharge and disposal of the products generated during the process. To solve this problem, two separate vessels are used to separate fat stem cells from adipose tissue.

That is, when two separation vessels are used, the process of treating adipose tissue is performed in one separation vessel, and the process of treating the separated stem cells is performed in the other one separation vessel. Therefore, The number of vessels increases and the number of components used to rotate the separation vessel also increases, resulting in an increase in the cost of the separation time and the apparatus for separation.

(Patent Document 1) KR10-1749932 B1

DISCLOSURE Technical Problem Accordingly, the present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to simplify the process of extracting and isolating adipose stem cells (SVF) from fat to reduce extraction time and cost, And to provide an apparatus for separating adipose stem cells that can improve the filtration rate of stem cells (SVF) and increase the extraction amount.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, unless further departing from the spirit and scope of the invention as defined by the appended claims. It will be possible.

As a specific means for achieving the above object, a preferred embodiment of the present invention is a sealing cap comprising: a first sealing cap having an auxiliary cap for opening and closing a communication hole formed in an upper surface thereof and having a predetermined length of operating bar extending from a lower surface; A first cylindrical housing having an upper outer circumferential surface formed with a male screw portion that is screwed with a female screw portion formed on a lower end inner circumferential surface of the first sealing cap, the upper portion being selectively opened and closed by the first sealing cap to form a first space; A second cylindrical housing communicating with the first cylindrical housing via a neck of a neck portion formed at a lower end of the first cylindrical housing in which the operation bar is disposed; A cap connecting portion assembled to an open lower portion of the second cylindrical housing to form a second space between the first cylindrical housing and the second cylindrical housing; And a second sealing cap having a sealing member corresponding to an outlet formed in the cap connecting portion and assembled to the cap connecting portion, wherein a center portion of the upper surface of the second sealing cap is provided with an inner And an outer skirt portion that is upwardly bent to be coupled to the outer edge of the lower end of the cap connection portion via at least one O-ring member is provided at an outer edge of the second sealing cap, Wherein the sealing member is located at an inner center of the coupling tube, and the sealing member is formed at the outer periphery of the sealing member so as to form a female threaded portion on an inner circumferential surface so as to be screwed with a male screw portion formed on an outer circumferential surface of the lower extension tube, When the second sealing cap is coupled with the first sealing cap, And the height of the connection tube is relatively higher than the height of the connection tube.

Preferably, the operation bar is formed of a bar member having a predetermined length, which is formed at an upper end thereof with a male threaded portion screwed with a female threaded portion of a cap extension pipe extending a predetermined length directly below the closed lower surface of the first sealing cap, 1 sealing cap, and a screw member having a predetermined length formed in the middle of the length of the threaded portion to be threadedly engaged with the female threaded portion of the cap extension pipe extending a certain length from the through hole formed directly through the sealed upper surface of the sealing cap.

Preferably, the neck portion of the neck portion includes an upper guide tube extending to a predetermined height so as to guide the lower end of the operation bar to be guided, and the upper guide tube may include at least one slit opening.

Preferably, the second sealing cap includes a filtering unit for removing foreign matter contained in an object discharged to the outside through the separated cap connection unit. The filtration unit includes an inlet port communicating with an outlet port of the cap connection unit, And an upper cover provided at an end of the filtration tube extending from the lower surface of the upper cover corresponding to the inflow port and having a predetermined length so as to remove foreign substances and an upper end opened with the upper cover, And a filter housing having at least one outlet to allow the filtered object to pass therethrough.

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More preferably, the filter body includes an internal filter network having an inlet end fixed to an end of the filtration tube, and an external filter network having a mesh size relatively smaller than the mesh size of the internal filter network and disposed outside the internal filter network .

More preferably, the outlet end of the inner filtration net may be spaced apart from the outlet end of the outer filtration net by a predetermined distance.

According to a preferred embodiment of the present invention as described above, the following effects can be obtained.

The opening and closing of the neck portion connecting the first cylindrical housing and the second cylindrical housing to the neck hole can be easily performed by a precise and delicate rotation operation of the operation bar to separate the centrifugally separated cell precipitate into another space Since the cells can be separated and transferred, the isolation and purity of adipose stem cells can be increased.

The extraction and extraction of lipid stem cells (SVF) from adipose tissue obtained from liposuction can be simplified to reduce the extraction time and cost, thus improving the usability and satisfaction of demand.

In the process of ultimately separating and extracting cell sediments washed with the washing solution, the foreign substances contained in the cell sediments can be removed by passing through the filtration section. Therefore, the cleanliness of the stem cells obtained at the highest level can be enhanced, It is possible to improve the cleanliness and purity of cells.

FIG. 1A is an overall perspective view showing an adipose stem cell separator according to an embodiment of the present invention. FIG.
1B is a cross-sectional perspective view illustrating an adipose stem cell separator according to an embodiment of the present invention.
2 is an exploded perspective view illustrating an adipose stem cell separator according to an embodiment of the present invention.
3A and 3B are a perspective view and an exploded perspective view showing another embodiment of the operation bar employed in the apparatus for separating adipose stem cells according to the embodiment of the present invention.
FIG. 4 is an enlarged view showing a state in which the second cylindrical housing and the cap connecting portion of the adipose stem cell separator according to the embodiment of the present invention are engaged with each other.
5 is a cross-sectional perspective view showing a filtering unit of an adipose stem cell separator according to an embodiment of the present invention.
FIG. 6 is a state in which a filtering unit is applied to an adipose stem cell separator according to an embodiment of the present invention.
7A to 7H are diagrams illustrating the state of use of an adipose stem cell separator according to an embodiment of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the following description, a detailed description of known functions and configurations incorporated herein will be omitted when it may obscure the subject matter of the present invention.

The same reference numerals are used for portions having similar functions and functions throughout the drawings.

In addition, in the entire specification, when a part is referred to as being 'connected' to another part, it may be referred to as 'indirectly connected' not only with 'directly connected' . Also, to include an element does not exclude other elements unless specifically stated otherwise, but may also include other elements.

As shown in FIGS. 1A and 2, the apparatus for separating adipose stem cells according to a preferred embodiment of the present invention includes a device for separating and obtaining adipose stem cells from adipose tissue more easily by using a single container, 1 sealing cap 10, a first cylindrical housing 20, a second cylindrical housing 30, a cap connection 40, and a second sealing cap 50.

As shown in Figs. 1A, 1B, and 2, the first sealing cap 10 has an auxiliary cap 12 for opening and closing a communication hole 11 formed in an upper surface thereof, And a cap member for selectively opening or closing an open upper portion of the first cylindrical housing 20 by having a long operating bar 13.

The auxiliary cap 12 may be formed of a cap member having a female screw portion that is screwed with a male screw portion formed on the outer surface of the hollow tube 11a extending from the communication hole 11 to a predetermined height.

The operation bar 13 may include a bar member having a predetermined length that is integrally formed with the lower surface of the first sealing cap 10 and moves together with the first sealing cap.

1B and FIG. 2, the operating bar 13 is provided with a male threaded portion 14a which is screwed with a female threaded portion formed on the inner circumferential surface of the cap extension tube 14 provided on the closed lower surface of the first sealing cap 10, A bar member having a predetermined length to form an upper end of the first sealing cap 10 or a bar member having a predetermined length with an upper end integrally formed on a closed lower surface of the first sealing cap 10.

Here, the operation bar 13 is shown as being vertically moved to the closed lower surface of the first sealing cap by the upper end integrally coupled and rotated by the rotation of the first sealing cap, but the present invention is not limited thereto, Can be independently operated with respect to the first sealing cap and can be moved up and down.

That is, as shown in FIGS. 3A and 3B, the operation bar 13 includes an inner surface of a cap extension pipe 14 extending downward from a through hole formed in a closed upper surface of the first sealing cap 10, And an upper end portion of the operating bar 13 is connected to the upper surface of the first sealing cap 10 through the cap extension pipe 14 Externally exposed.

The operating bar 13 selectively opens and closes the neck hole 21 of the neck portion 20a to be described later by operating the first sealing cap 10 or the operating bar 13 in the forward or reverse direction, As shown in FIG.

The first cylindrical housing 20 has a male screw portion 25 screwed with a female screw portion 15 formed on the inner peripheral surface of the lower end of the first sealing cap 10, as shown in FIGS. 1A, 1B and 2, Is a hollow cylindrical member which is formed on an opened upper peripheral surface and selectively opened and closed by an upper portion opened by the first sealing cap 10 to form a first space S1 of a predetermined size.

Preferably, the first cylindrical housing 20 is assembled with the first sealing cap 10 via an O-ring member 23 provided on the upper outer surface thereof, and then sealed.

The first sealing cap 10 is provided with a silicone pad which forms a cross-shaped slit in a region corresponding to a communicating port that is opened by the auxiliary cap, It is possible to prevent the outside air from being introduced into the first space when injecting an injection object such as an adipose stem cell extract through an injection port of the injection port.

The first sealing cap 10 may include at least one unidirectional check valve that forms a passage to be discharged to the outside in the first space S1. Accordingly, when the injection object is injected, such as an adipose stem cell extract, It is possible to prevent the air in the first space S1 from being smoothly discharged to the outside so that the internal pressure of the first space is unnecessarily raised and excessively increased.

The second cylindrical housing 30 includes a neck portion formed at the lower end of the first cylindrical housing 20 in which the operating bar 13 is disposed, as shown in FIGS. 1A, 1B, 2, 20a of the first cylindrical housing 20 via the neck hole 21 of the second cylindrical housing 20a.

The neck portion 20a is extended from the first cylindrical housing 20 toward the neck hole 21 to gradually narrow the inner diameter and then the inner diameter from the neck hole 21 toward the second cylindrical housing 30 And an approximately hourglass cross section extending to be enlarged.

The neck hole 21 of the neck portion 20a includes an upper guide pipe 22 extending to a predetermined height so that the lower end of the operating bar 13 is guided and guided. At least one slit 22a is provided on the outer surface of the second cylindrical housing 30 so as to guide and discharge the liquid filling the inner space of the second cylindrical housing 30 to the second cylindrical housing 30 through the neck hole 21 And the neck hole 21 may include a lower guide tube 24 extending a predetermined length downward.

The operation bar 13 may include at least one O-ring 13a that generates a sealing force in contact with the inner surface of the neck hole 21 on the outer surface of the lower end.

Accordingly, the first cylindrical housing 20 and the second cylindrical housing 30 are integrally connected to each other through the neck portion 20a, and at the same time, communicated with each other via the neck hole 21 , And the neck bar can be selectively opened and closed by the operation bar 13 which is moved up and down during the rotation operation.

That is, when the lower end of the operation bar 13 is positioned in the neck hole 21 or the lower guide pipe 24, the neck hole 21 is closed while the lower end of the operation bar is connected to the upper guide pipe 22 Since the slit is partially opened according to the position of the lower end of the operating bar, the liquid filled in the first space of the first cylindrical housing 20 flows through the slit 22a and the slit 22a, And can be guided and discharged to the second cylindrical housing 30 side through the neck hole 21.

As shown in FIGS. 1A, 1B, 2 and 4, the cap connecting portion 40 is formed by connecting a second space S2 having a relatively smaller volume than the first space S1 to the second cylindrical housing 2, (30) so as to be detachable from the open lower portion of the second cylindrical housing (30).

The cap connecting portion 40 may include an assembling jaw which is formed at the center of the body so as to penetrate the discharge port 41 and to be coupled to an assembling groove formed at the outer edge of the lower end of the second cylindrical housing 30 on the outer rim .

The upper surface 43 of the cap connecting portion 40 corresponding to the second cylindrical housing 30 may increase the volume of the second space S2 formed between the second cylindrical housing 30 and the upper surface 43 It is preferable that the discharge port 41 has a downwardly inclined cross-sectional shape at the center where the discharge port 41 is formed.

The coupling between the opened lower end of the second cylindrical housing 30 and the cap coupling portion 40 is achieved by a female screw portion 35 formed on the inner peripheral surface of the lower end of the second cylindrical housing, The second cylindrical housing 30 may be formed by screwing the male and the female threads into the second cylindrical housing 30 and the second cylindrical housing 30, Can be assembled to the outer circumferential surface of the housing 40 by elastic coupling between the assembling jaws.

 Accordingly, when the neck hole 21 and the slit 22a are communicated with each other by the upward movement of the operation bar 13 having the lower end in the lower guide tube 24, the first cylindrical housing 20 Is guided through the neck hole 21 to the second space S2 of the second cylindrical housing 30 to be filled with the liquid.

When the neck hole 21 is closed by the downward return movement of the operation bar 13, the second space S2 of the second cylindrical housing 30 is divided into a lower layer Can be separated and filled.

As shown in FIGS. 1A, 1B, 2 and 4, the second sealing cap 50 includes a sealing member 51 (see FIG. 1) inserted into a discharge port 41 formed in the center of the body of the cap connecting portion 40 The cap member 40 is detachably assembled to the lower end of the cap connection unit 40.

The sealing member 51 penetrates into the inner hole of the lower extension pipe 42 extending downward from the discharge port 41 of the cap connection portion 40 by a predetermined length and is provided at the center of the upper surface of the second sealing cap 50 And may be provided on the column extending in the height direction.

A female screw portion 52a is formed on the inner circumferential surface of the sealing member 51 so as to be screwed with a male screw portion 42a formed on the outer circumferential surface of the lower extension pipe 42, And a connection pipe 52 extending at a predetermined height in the center of the surface.

The end of the sealing member 51 located at the center of the connection pipe 52 may be first introduced into the lower extension pipe when the cap connection unit 40 and the second sealing cap 50 are coupled with each other. The height of the connecting pipe 52 is relatively higher than the height of the connecting pipe 52.

The second sealing cap 50 includes an outer skirt portion 54 that is upwardly bent to be engaged with a lower outer edge of the cap connecting portion 40. At least one o-ring member 53 So that the sealing force can be increased.

The second sealing cap 50 is engaged with the cap connecting portion 40 by the screw connection between the connection pipe 52 and the lower extension pipe 42 and enters the inside of the lower extension pipe 42 And the discharge port 41 of the cap connecting portion 40 is sealed by the sealing member 51.

The filtration unit 60 removes foreign matter contained in the adipose stem cells, which is an object to be discharged to the outside through the discharge port 41 of the cap connection unit 40 separated from the second sealing cap 50. have.

5 and 6, the filtration unit 60 has an inlet 62 communicating with the discharge port 41 of the separated cap connection unit 40 at the center of the upper surface thereof And a filtering member 64 provided at an end of the filtration pipe 63 extending from the lower surface of the upper cover 61 corresponding to the inlet 62 to a predetermined length to remove foreign matter, .

An open lower end of the upper cover 61 and an open upper end are assembled to arrange the filtration body 64 and have at least one outlet 66 for discharging the filtered fluid through the filtration body And a filtering housing 65.

The filtration body 64 includes an inner filtration network 64a having an inlet end fixed to an end of the filtration pipe 63 and has a mesh size relatively smaller than a mesh size of the inner filtration network 64a, And an external filtering network 64b disposed outside of the apparatus.

The lower end of the inner filtration net 64a may be spaced apart from the lower end of the outer filtration net 64b by a predetermined distance.

The inlet 62 may be provided in the form of a hollow tube that forms a female thread on the inner surface so as to be threadedly engaged with the male screw portion 42a formed on the outer surface of the lower extension pipe 42 extending from the discharge port 41 .

In this case, when the inlet 62 is formed of a hollow tube that forms a male screw portion 62a on the outer surface, the lower extension tube 42 forming the male screw portion 42a is connected to the communication assembly .

The outlet 66 is provided with a threaded portion 66a which is screwed with the female threaded portion formed at the injection port of the syringe 70 so as to collect the filtered object while passing through the filtration body 64 of the filtration unit 60 Or a hollow tube formed on the surface.

As a result, the adipose stem cells, which are the ultimate separation target remaining in the second space S2 formed between the second cylindrical housing 30 and the cap connecting part 40, pass through the filtration part 60, And then separated and extracted by the separate syringe 70 communicating with the outlet 66, without contact with air in the atmosphere.

The method for isolating adipose stem cells using the above-described apparatus for separating adipose stem cells comprises sequentially preparing the adipose stem cells by preparing the adipose stem cell extracting step, the first washing step and the second washing step, Respectively.

The preparation step is to extract a predetermined amount of adipose tissue from a human body using a luer lock syringe. At this time, the extracted adipose tissue is repeatedly extracted a predetermined number of times a predetermined number of times.

When the extracted fat tissue is put into another syringe and stored, and then the syringe is shaken to sediment the fat tissue, the syringe is divided into the blood layer, the fat extract layer and the oil layer by the specific gravity difference .

Subsequently, the operator separates and extracts only the layered fat extract layer between the blood layer and the oil layer in the syringe by piston operation of the syringe.

The lipid stem cell extracting step extracts adipose stem cells from the fat extract obtained in the preparation step.

7A, the auxiliary cap 12 of the first sealing cap 10, which hermetically seals the open upper portion of the first cylindrical housing 20, is opened, and the neck of the neck portion 20a is opened, The hole 21 is communicated with the first space of the first cylindrical housing through the communication hole 11 in which the fat extract P1 separated in the syringe is opened in a state of being closed by the lower end of the operating bar 13 S1), and then a predetermined amount of the enzyme solution (C1) such as collagen or a collagenase solution, which is an enzyme for hydrolyzing the gelatin component, is injected into the first space (S1).

At this time, in the operation of injecting the fat extract (P1) into the first space (S1) of the first cylindrical housing, the worker discharges the wetted liquid layer precipitated in the bottommost layer of the syringe to the outside by the piston operation of the syringe, Only the fat extract layer precipitated in the intermediate layer can be introduced into the first space S1 of the first cylindrical housing through the opened communication hole by the piston operation of the syringe.

Subsequently, the separation device 100 of the present invention in which the auxiliary cap 12 is closed and the communication port 11 is sealed is loaded on a shaking incubator, and then stirred to effect an enzyme reaction between the fat extract and the enzyme solution A shaking process is performed.

When the separating device 100 of the present invention taken out from the shaking incubator is subjected to primary centrifugation for a certain period of time under a centrifugal force of a constant centrifugal force, the enzyme-reacted fat extract is subjected to primary centrifugation, (P) containing the adipose stem cells to the upper side from the lower side of the first space (S1) of the first cylindrical housing (20) and the enzyme reaction solution (C) which is reacted with the enzyme.

7B, when the worker rotates the first sealing cap 10 or the operating bar 13, the lower end of the operating bar is moved to the slit 22a of the upper guide pipe 22, As shown in Fig.

When the centrifugal separator is subjected to a secondary centrifugation process in this state, the cell precipitate P in the lowermost layer is separated into the second space 22a of the second cylindrical housing through the slit 22a partially opened and the neck hole 21 being completely opened. (S2).

A boundary layer B between the cell precipitate (P) layered through the first and second cylindrical housings and the neck portion made of a transparent material and the enzyme reaction solution (C) is formed in the neck hole 121 The lower end of the operating bar 13 is lowered to be positioned at the neck hole 121 to seal the opened neck hole.

Accordingly, the neck hole is closed by the lower end of the operation bar to block the entry of the enzyme reaction liquid (C) of the first cylindrical housing into the second space (S2) of the second cylindrical housing, Only the enzyme reaction liquid C remains in the first space S1 of the cylindrical housing and the cell precipitate P is filled in the second space S2 of the second cylindrical housing so that the enzyme reaction solution and the cell precipitate are separated .

At this time, the operation bar (13) moved up and down in the neck hole forms a receiving portion (13b) having a lower end opened inside the body, so that a part of the cellular precipitate separated by the capacity larger than the inner volume of the second space So that the separation efficiency can be increased.

In this state, the cell precipitate P filled in the second space S2 of the second cylindrical housing 30 is exposed to the outside by the separation of the second sealing cap 50 as shown in Fig. 7C And is extracted to the outside by using a syringe 71 assembled to be connected to the lower extension pipe 42 of the cap connection portion 40.

On the other hand, the enzyme reaction liquid C remaining in the first space S1 of the first cylindrical housing 20 is discharged to the outside by opening of the auxiliary cap 12 or opening of the first sealing cap 10. [ Lt; / RTI >

Here, as shown in FIG. 7C, the operation for extracting the cell precipitate P remaining in the second space S2 of the second cylindrical housing 30 to the outside is performed as follows. That is, And a syringe 71 connected to the lower extension pipe 42 in a state where the cap is separated. However, the present invention is not limited to this. As shown in FIG. 7B, when the second sealing cap 50 is assembled The sealing member 51 corresponding to the lower extension pipe 42 may be performed without contact with air by another syringe passing through the injection needle.

That is, the sealing member 51 through which the injection needle penetrates is formed in a column shape of a hollow tube having a blocking region 51a of a certain thickness, the middle of which is interrupted, and the blocking region is integrally formed with a sealing member in the form of a hollow tube Or a separate pressing member. The material of the blocking region may be a soft material having rigidity through which the injection needle can penetrate.

7D and 7E, the cell pellet P is washed in the first space S1 of the first cylindrical housing 20 as the primary cleaning solution W1, And separating and extracting only the cell precipitate (P) which has been washed and the impurities have been removed.

The washing solution may be selected from a saline solution mixed at a certain ratio with respect to the extracted cell precipitate, but the washing solution is not limited thereto and other washing solutions can be selectively used.

The auxiliary cap 12 of the first sealing cap 10 which hermetically seals the open upper portion of the first cylindrical housing of the newly prepared separating device is opened and opened through the open communication hole 11 in the above- The first washing solution W1 such as a predetermined amount of cell precipitate P and a predetermined amount of saline solution is sequentially injected into the first space S1 of the first cylindrical housing.

At this time, it is preferable that the primary washing solution W1 is injected in a fixed amount in accordance with a charged amount of the cell precipitate (P) to be washed.

In this state, if the centrifugal separator is subjected to the third centrifugal force for a predetermined period of centrifugal force at a constant intensity, the cell precipitate (P) is washed by the primary washing solution (W1) , Lipid precursor cells, and enzyme reaction liquid attached to stem cells.

As shown in FIG. 7D, in the first space S1 of the first cylindrical housing 20, the cell pellets P and the primary washing solution W1, which have been washed from the lower side to the upper side, And are sequentially layered by difference.

As shown in FIG. 7E, the primary cleaning solution W1 layered in the first space S1 of the first cylindrical housing 20 passes through the communication port 11 in which the auxiliary cap is opened Only the cell precipitate (P) remains in the first space (S1) by being sucked and removed by the injection needle of the incoming syringe.

At this time, in the centrifugal separation process for separating the cell precipitate (P) and the primary washing solution (W1) in the first space, the neck hole (21) is sealed by the lower end of the operating bar (13) It is preferable to maintain the cleanliness of the space.

7f, 7g and 7h, the second washing step is a step of washing the cell pellets P washed first in the first space S1 of the first cylindrical housing 20 with the second washing solution (W2), and only the cell precipitate (P) washed and removed from the impurities is separated and extracted to the outside.

7f, the auxiliary cap 12 is opened and the secondary cleaning solution W2, such as a predetermined amount of saline solution, is supplied to the first space of the first cylindrical housing through the open communication hole 11. Then, (S1) and mixed with the cell precipitate (P) remaining in the first space at a predetermined ratio.

When the operation bar is moved upward by the rotation operation of the first sealing cap or the operation bar, as shown in Fig. 7G, the neck space is completely opened to allow the first space and the second space to communicate with each other, The cell pellets P are washed by the secondary washing solution W2 and the fat cells contained in the cell pellets are washed by the centrifugal separator and centrifuged at a constant centrifugal force for a predetermined period of time. The contaminants such as the enzyme reaction liquid adhering to the lipid precursor cells and the stem cells are once again removed and separated.

By the centrifugal separation, the cell precipitate (P) and the secondary washing solution (W2) washed from the lower side to the upper side in the first and second spaces S1 and S2 of the first and second cylindrical housings 20, And then sequentially separated by the difference.

When the operation bar 13 is moved downward and the lower end of the operation bar is positioned at the neck hole to close the neck hole, the secondary cleaning solution remains in the first space of the first cylindrical housing, The washed cell pellet remains in the second space of the cylindrical housing.

At this time, a relatively large amount of cell sediment in the second space may be filled into the inner hole formed in the lower end of the operation bar to fill the inner space.

The secondary cleaning solution W2 remaining in the first space S1 of the first cylindrical housing 20 is opened by the opening of the auxiliary cap 12 or the opening of the first sealing cap 10, The cell deposit P separated and transferred to the second space S2 of the second cylindrical housing 30 is discharged to the cap connecting portion 40 exposed to the outside by the separation of the second sealing cap 50 The foreign substance contained in the cell precipitate P is filtered out and extracted to the outside by the filtration unit 60 assembled with the lower extension pipe 42 of the filter.

6, the inlet port provided in the upper cover of the filtration unit is screwed to the lower extension pipe 42 extending from the discharge port 41 of the cap connection unit which is exposed when the second seal cap is separated And an outlet provided in the filtration housing 65 of the filtration unit is connected to the extraction syringe 70 for communication.

The lower end of the operation bar is slid in a state in which the upper end of the first cylindrical housing 20 is opened by the opening of the first sealing cap 10 or the auxiliary cap 12, The first space S1 of the first cylindrical housing 20 where the cleaning solution is removed to the outside and the second space S2 of the second cylindrical housing 30 where the washed cell precipitate remains remain So that the cell precipitate (P) is transferred to the inside of the filtration unit having the inlet connected to the discharge port during the pulling operation of the extraction syringe.

Subsequently, the cell pellets P transferred to the interior of the filtration unit 60 are continuously passed through the filtration body 64 composed of the internal filtration net 64a and the external filtration net 64b to remove foreign matter, The removed cell precipitate (P) is extracted by a syringe for extraction combined with an outlet provided at the lower end of the filtration housing to complete the lipid stem cell separation operation.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the inventions. It will be apparent to those of ordinary skill in the art.

10: first sealing cap
20: first cylindrical housing
30: second cylindrical housing
40: cap connection
50: second sealing cap
60:
70: Syringe

Claims (8)

A first sealing cap having an auxiliary cap opening and closing a communication port formed in an upper surface thereof and having a predetermined length of operating bar extending from a lower surface thereof;
A first cylindrical housing having an upper outer circumferential surface formed with a male screw portion that is screwed with a female screw portion formed on a lower end inner circumferential surface of the first sealing cap, the upper portion being selectively opened and closed by the first sealing cap to form a first space;
A second cylindrical housing communicating with the first cylindrical housing via a neck of a neck portion formed at a lower end of the first cylindrical housing in which the operation bar is disposed;
A cap connecting portion assembled to an open lower portion of the second cylindrical housing to form a second space between the first cylindrical housing and the second cylindrical housing;
And a second sealing cap having a sealing member corresponding to an outlet formed through the cap connecting portion and assembled to the cap connecting portion,
And a columnar sealing member extending into the inner hole of the lower extension pipe extending downward from the discharge port and extending to a predetermined height so as to close the inner hole of the lower extension pipe, And an outer skirt portion that is upwardly bent so as to be coupled with the lower outer rim through at least one O-ring member,
And a connection pipe formed on an outer side of the sealing member to form a female threaded portion on an inner circumferential surface to be screwed with a male screw portion formed on an outer circumferential surface of the lower extension pipe,
The sealing member located at the inner center of the connection pipe is formed to be relatively higher than the height of the connection pipe so as to be able to enter the end of the lower extension pipe extending from the outlet when the cap connection portion and the second seal cap are coupled Wherein the stem cell is a stem cell. The method according to claim 1,
The operation bar may be formed of a bar member having a predetermined length to form a male threaded portion screwed with the female threaded portion of the cap extension tube which extends a predetermined length directly below the sealed lower face of the first sealing cap at the upper end, And a bar portion having a predetermined length to form a male thread portion screwed with a female thread portion of a cap extension tube extending a certain length from a through hole formed in a closed top surface of the cap. The method according to claim 1,
Wherein the neck hole of the neck portion includes an upper guide tube extending to a predetermined height so that the lower end of the operation bar is guided and guided, and at least one slit opening in the upper guide tube Separating device. delete delete The method according to claim 1,
And a filtering unit for removing foreign matter contained in an object discharged to the outside through the separated cap connecting portion of the second sealing cap,
The filtration unit includes an upper cover having an inlet port communicating with an outlet port of the cap connection unit at the center of the upper surface and a filter cover provided at an end of the filtration pipe extending from the lower surface of the upper cover corresponding to the inlet port, And a filtration housing having at least one outlet so that the filtration body, the upper cover, and the open upper end are assembled and the filtered object is discharged while passing through the filtration body. The method according to claim 6,
Wherein the filtration body includes an inner filtration network having an inlet end fixed to an end of the filtration tube and an outer filtration network disposed outside the inner filtration network with a mesh size relatively smaller than a mesh size of the inner filtration network, Adipose stem cell separation device. 8. The method of claim 7,
Wherein the outlet end of the inner filtration mesh is spaced apart from the outlet end of the outer filtration mesh by a predetermined distance.

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