TWI720658B - Device for separating the adipose stem cells - Google Patents

Abstract

The present invention provides an adipose stem cell separation device including: a first sealing cover provided with an auxiliary cover and a lever extending from a lower surface; a first cylindrical housing formed on the outer peripheral surface of the upper end and formed in the lower end of the first sealing cover The internal threaded part of the peripheral surface is threadedly coupled with the external threaded part, which is selectively opened and closed by the upper part of the first sealing cover, and forms the first space; the second cylindrical shell will be formed in the first space where the joystick is arranged. The neck hole of the neck of the lower end of a cylindrical shell is used as a medium to communicate with the first cylindrical shell; the cover connecting part is assembled at the lower part of the second cylindrical shell so as to be formed between the second cylindrical shell The second space; the second sealing cover, which is provided with a corresponding closure member inserted through the discharge port formed in the cover connection portion, and assembled in the cover connection portion.

Description

Adipose stem cell separation device

The present invention relates to a device for separating adipose stem cells from adipose tissue. In more detail, it relates to a device for separating adipose stem cells (DEVICE FOR SEPARATING THE ADIPOSE STEM CELLS), which can be easily separated and extracted from adipose tissue by centrifugal separation with a centrifuge Adipose Stem Cells (ADIPOSE STEM CELLS).

Generally speaking, a stem cell is a cell that has both self-replication ability and the ability to differentiate into two or more cells. In recent years, in addition to research on obtaining stem cells from bone marrow, research on obtaining stem cells from adipose tissue is also ongoing.

Adipose-derived stem cells isolated from adipose tissue are pluripotent stem cells that have the ability to differentiate into adipocytes, muscle cells, chondrocytes, bone cells and other types of cells.

Compared with bone marrow-derived mesenchymal stem cells, this adipose stem cell has superior self-regeneration ability, is easy to be cultured in vitro, is rich in adipose tissue from which adipose stem cells can be separated, and the extraction method is simple and safe.

In addition, it is reported that adipose stem cells are the same as bone marrow-derived mesenchymal stem cells in almost all characteristics, such as differentiation ability, cytological characteristics, immunological characteristics, and tissue regeneration ability.

That is, adipose stem cells, as stem cells with a high possibility of being able to replace bone marrow-derived mesenchymal stem cells, have excellent medical utility.

Moreover, compared with mesenchymal stem cells, adipose stem cells are easier and safer to obtain tissues. They have the advantage of no restriction on tissue supply and demand. They are easy to be cultured in vitro. From the perspectives of tissue accessibility, safety, effectiveness and economy, it can be Considered to be on top of mesenchymal stem cells.

The commonly used adipose stem cell separation and culture method implements the following method: re-pulverize the inhaled or excised fat, and decompose the tissue into collagenase (collagenase), and then separate it by centrifugation. Cells are cultured in the stromal vascular fraction (SVF) separated from the sediment.

On the other hand, generally, all processes for separating adipose stem cells from adipose tissue are realized in one separation container. At this time, the process becomes complicated due to the discharge and disposal of products generated in the process. In order to solve the above problems, two separation containers are also used to realize the process of separating adipose stem cells from adipose tissue.

That is, when two separation containers are used, since the process of processing adipose tissue is realized in one separation container, and the process of processing separated stem cells is realized in the other separation container, the number of separation containers (consumables used) As the increase, the structural elements for rotating the separation container also increase, resulting in the problem of an increase in the separation time and the cost of the separation device.

Patent Document 1: KR10-1749932B1.

Therefore, the present invention is proposed to solve the above-mentioned technical problems, and its purpose is to provide an adipose stem cell separation device as follows, which can reduce the extraction time and cost by simplifying the process of extracting and separating adipose stem cells from fat, and improve the extraction and separation process. The filtration rate of adipose stem cells can increase the amount of extraction.

The technical problems to be solved by the present invention are not limited to the aforementioned technical problems, and those skilled in the art can clearly understand other technical problems that are not mentioned based on the following description.

A preferred embodiment of the present invention, as a specific technical means for achieving the above objective, provides an adipose stem cell separation device. The adipose stem cell separation device includes: a first sealing cover provided with an auxiliary cover for opening and closing the communication port formed through the upper surface , A joystick with a predetermined length extended from the lower surface; the first cylindrical housing, on the outer peripheral surface of the upper end, is formed with an external threaded portion that is threadedly coupled with the internal threaded portion formed on the inner peripheral surface of the lower end of the first sealing cap, by The upper part opened by the first sealing cover is selectively opened and closed, and forms a first space; a second cylindrical housing, which will form a neck hole at the neck of the lower end of the first cylindrical housing equipped with the above-mentioned joystick As a medium, it communicates with the above-mentioned first cylindrical shell; a cover connecting portion, which is assembled in the open lower part of the above-mentioned second cylindrical shell, so as to form a second space with the above-mentioned second cylindrical shell; a second seal The cover is provided with a corresponding closing member inserted into the discharge port formed through the cover connection portion, and is assembled to the cover connection portion.

Preferably, the above-mentioned joystick may be connected to the closed lower surface of the above-mentioned first sealing cover The female threaded part of the cap extension pipe extending straight down by a predetermined length is formed of a rod member of a predetermined length formed on the upper end of the threaded part, or is formed by a rod member of a predetermined length through which is formed on the closed upper surface of the first sealing cap. The through hole is formed in a rod member of a predetermined length in the middle of the length by screwing the female thread part of the cap extension pipe extended by a predetermined length toward the bottom of the through hole.

Preferably, the neck hole of the neck may include an upper guide tube, the upper guide tube is extended at a predetermined height to guide the lower end of the joystick to enter, and the upper guide tube may include at least one slit formed by an opening.

Preferably, the closing member may be formed in a cylindrical shape extending from the center of the upper surface of the second sealing cap by a predetermined height so as to enter and close the inner hole of the lower extension pipe extending downward from the discharge port.

More preferably, it is possible to include a connecting pipe with an internal thread formed on the inner peripheral surface on the outer side of the closing member so as to be screwed to the male thread formed on the outer peripheral surface of the lower extension pipe.

Preferably, it includes a filter portion for removing foreign matter contained in the object discharged to the outside by the cover connecting portion separated by the second sealing cover, and the filter portion may include: an upper cover provided in the center of the upper surface There is an inflow port that is in communication with the discharge port of the cap connecting portion; a filter body is provided at the end of a filter tube extended by a predetermined length from the lower surface of the upper cover corresponding to the inflow port, and is used to remove foreign matter; and The filter case is assembled with the upper cover and the open upper end, and has at least one outflow port so as to discharge the object filtered by the filter body.

More preferably, the filter body may include: an inner filter, the inlet end of which is fixed at the end of the filter tube; an outer filter, which has a relatively smaller mesh size than the inner filter, and is arranged in The outside of the above internal filter.

More preferably, the outlet end of the inner filter and the outlet end of the outer filter can be separated from each other at a predetermined interval.

The preferred embodiment of the present invention as described above has the following effects.

Through the precise and detailed rotating operation of the joystick, the neck hole connecting the neck of the first cylindrical shell and the second cylindrical shell can be easily opened and closed, so that the centrifugal separation and layered cell sediment can be separated Transfer to a different space, so it can improve the separation and extraction volume and purity of adipose stem cells.

The process of extracting only adipose stem cells from the adipose tissue obtained by liposuction is simplified, thereby reducing the extraction time and cost, thereby improving convenience in use and improving user satisfaction.

In the final process of separating and extracting from the cell pellets washed with the washing liquid, the filter section can remove foreign substances contained in the cell pellets. Therefore, the cleanliness of the finally obtained adipose stem cells can be improved, and the final extraction of the adipose stem cells can be improved. Cleanliness and purity.

100: Adipose stem cell separation device

10: The first sealing cover

11: Connecting port

11a: Hollow tube

12: auxiliary cover

13: Joystick

13a: O-ring

13b: Containment Department

14: Cover extension tube

15: Internal thread

20: The first cylinder shell

20a: neck

21: neck hole

22: Upper guide tube

22a: slit

23: O-ring parts

24: Lower guide tube

25: External thread

30: second cylinder shell

35: Internal thread

40: Cover connection part

41: Outlet

42: Lower extension tube

42a: External thread

43: upper face

45: External thread

50: The second sealing cover

51: closed parts

51a: Cut off area

52: connecting pipe

52a: Internal thread

53: O-ring parts

54: Outer skirt

60: Filter

61: upper cover

62: Inlet

62a: Internal thread

63: filter tube

64: filter body

64a: Internal filter

64b: External filter

65: filter housing

66: Outlet

66a: External thread

70, 71: Syringe

B: Junction layer

C: Enzyme reaction solution

C1: Enzyme solution

P: cell pellet

P1: Fat extract

S1: The first space

S2: second space

W1: One-time washing liquid

W2: secondary washing liquid

The above and/or other aspects and features of the present invention will become obvious and easy to understand from the following description of the embodiments in conjunction with the accompanying drawings, in which: FIG. 1a is an overall perspective view of an adipose stem cell separation device according to an embodiment of the present invention 1b is a sectional perspective view showing an adipose stem cell separation device according to an embodiment of the present invention; Figure 2 is an exploded perspective view showing an adipose stem cell separation device according to an embodiment of the present invention; Figures 3a and 3b show an implementation according to the present invention A cross-sectional perspective view and an exploded perspective view of other embodiments of the joystick used in the adipose stem cell separation device of the example; FIG. 4 shows the combined state of the second cylindrical housing and the lid connecting portion of the adipose stem cell separation device according to the embodiment of the present invention Figure 5 is a cross-sectional perspective view showing the filter portion of the adipose stem cell separation device according to an embodiment of the present invention; Figure 6 is a diagram of the use state of the filter portion in the adipose stem cell separation device according to an embodiment of the present invention; and 7a to 7h are diagrams of the use state of the adipose stem cell separation device according to an embodiment of the present invention.

Hereinafter, preferred embodiments that can be easily implemented by those skilled in the art will be described in detail with reference to the accompanying drawings. However, when the structural principle of the preferred embodiment of the present invention is described in detail, if it is considered that the specific description of related well-known functions or structures may unnecessarily obscure the gist of the present invention, the detailed description will be omitted.

In addition, throughout the drawings, the same reference numerals are used for parts having similar functions and effects.

In addition, throughout the specification, when a part is "connected" with another part, it includes not only the case of "direct connection", but also the case of "indirect connection" in which different elements are arranged in the middle. In addition, if there is no description to the contrary in particular, "including" a certain structural element means that other structural elements can also be included, rather than excluding other structural elements.

As shown in Figures 1a to 2, the adipose stem cell separation device 100 according to a preferred embodiment of the present invention may include a first sealing cover 10, a first cylindrical housing 20, a second cylindrical housing 30, a cover connecting portion 40, and a second The cover 50 is sealed so that a container can be used to more easily separate and obtain adipose stem cells from adipose tissue.

As shown in FIGS. 1a, 1b, and 2, the first sealing cover 10 includes an auxiliary cover 12 that opens and closes a communication port 11 formed through the upper surface, and a lever 13 extending from the lower part to the lower part by a predetermined length. The lid member that selectively opens or closes the open upper portion of the first cylindrical housing 20 described above.

The auxiliary cover 12 may be formed of a cover member having a female screw part screwed to a male screw part formed on the outer surface of the hollow tube 11a extending from the communication port 11 at a predetermined height.

The operating lever 13 may be formed of a rod member of a predetermined length that is integrally provided at the upper end on the lower surface of the first sealing cover 10 and moves together with the first sealing cover 10.

As shown in FIGS. 1b and 2, such a joystick 13 may be an external thread that is threadedly coupled with an internal thread formed on the inner peripheral surface of the cap extension tube 14 provided on the closed lower surface of the first sealing cap 10 The part is formed of a rod member of a predetermined length at the upper end, or is formed of a rod member of a predetermined length whose upper end is integrally provided on the closed lower surface of the first sealing cover 10.

Among them, for the joystick 13, the upper end is integrally coupled to the closed lower surface of the first sealing cap 10, and the first sealing cap 10 is rotated up and down to move up and down. However, It is not limited to this, and the above-mentioned first sealing cover 10 may be moved up and down by a single operation.

That is, as shown in FIGS. 3a and 3b, the joystick 13 is formed of a rod member having a predetermined length of an external thread portion screwed with an internal thread portion formed on the inner surface of the cover extension tube 14, wherein the cover extension The tube 14 extends downward from a through hole formed through the closed upper surface of the first sealing cover 10, and a part of the upper end of the joystick 13 passes through the cover extension tube 14 to make the first sealing cover The upper part of 10 is exposed to the outside.

Therefore, by rotating the first sealing cap 10 or the joystick 13 in the forward or reverse direction, the joystick 13 can move up and down to selectively open and close the neck hole 21 of the neck 20a described later.

As shown in Figures 1a, 1b, and 2, for the first cylindrical housing 20, a female thread 15 formed on the inner peripheral surface of the lower end of the first sealing cap 10 is formed on the outer peripheral surface of the open upper end. The coupled male screw portion 25 is selectively opened and closed by the upper portion opened by the first sealing cap 10, and the first cylindrical housing 20 is a hollow cylindrical member that forms a first space S1 of a predetermined size.

Preferably, the first cylindrical housing 20 uses an O-ring member 23 provided on the outer surface of the upper end as a medium, and is assembled with the first sealing cover 10 to perform a sealing process.

In addition, the first sealing cover 10 is provided with a silicon plate formed with a cross-shaped slit in the area corresponding to the communication port 11 opened and closed by the auxiliary cover 12, so that when injected, the communication opening is opened by the separation of the auxiliary cover 12 The port 11 and the injection port of the syringe can prevent outside air from flowing into the first space S1 when the adipose stem cell extract is injected into the object.

In addition, the first sealing cover 10 includes at least one one-way check valve that forms a channel for discharging from the first space S1 to the outside, so that when an injection target such as adipose stem cell extract is injected, the air in the first space S1 Since it is smoothly discharged to the outside, it is possible to prevent the internal pressure of the first space S1 from unnecessarily increasing excessively.

As shown in Figures 1a, 1b, 2 and 4, the second cylindrical housing 30 is formed with a neck hole 21 formed at the neck 20a of the lower end of the first cylindrical housing 20 on which the joystick 13 is arranged. The medium is a hollow cylindrical member communicating with the first space S1 of the first cylindrical housing 20 described above.

The neck portion 20a is roughly formed to extend so that the inner diameter gradually narrows from the first cylindrical housing 20 to the neck hole 21, so that the inner diameter increases from the neck hole 21 to the second cylindrical housing 30 The shape of the hourglass profile is extended in a way.

In addition, the neck hole 21 of the neck portion 20a includes an upper guide tube 22 extended at a predetermined height to guide the lower end of the joystick 13 to enter, and at least one slit 22a is formed on the outer surface of the upper guide tube 22 to facilitate The liquid filled in the inner space of the second cylindrical housing 30 can be guided and discharged to the first cylindrical housing 20 through the neck hole 21, and the neck hole 21 can be It may include a lower guide tube 24 extended to the lower part by a predetermined length.

In addition, the outer surface of the lower end of the joystick 13 may include at least one O-ring 13a that is in contact with the inner surface of the neck hole 21 to generate a sealing force.

Therefore, the first cylindrical housing 20 and the second cylindrical housing 30 use the neck portion 20a as a medium to be connected to each other as a single body. At the same time, the neck hole 21 is used as a medium to communicate with each other, which can be moved up and down through the rotating operation. The moving joystick 13 selectively opens and closes the above-mentioned neck hole 21.

That is, if the lower end of the joystick 13 is located in the neck hole 21 or the lower guide tube 24, the neck hole 21 is in a closed state. Unlike this, if the lower end of the joystick 13 is located in the slit of the upper guide tube 22 22a, a part of the slit is opened according to the position of the lower end of the joystick 13, therefore, the liquid filled in the first space S1 of the first cylindrical housing 20 can be guided and discharged through the slit 22a and the neck hole 21 To the second cylindrical shell 30.

As shown in Figures 1a, 1b, 2 and 4, the cover connecting portion 40 is detachably assembled in the open lower portion of the second cylindrical housing 30 so as to be formed inside the second cylindrical housing 30 The second space S2 has a volume that is relatively smaller than the above-mentioned first space S1.

For such a cover connecting portion 40, a discharge port 41 is formed in the center of the main body, and an assembly flange corresponding to the assembly groove formed on the outer edge of the lower end of the second cylindrical housing 30 may be included at the outer edge.

In addition, it is preferable that the upper surface 43 of the lid connection portion 40 corresponding to the second cylindrical housing 30 is formed in a downwardly inclined cross-sectional shape along the center where the discharge port 41 is formed through, so as to be able to increase the number of lid connections formed therethrough. The volume of the second space S2 between the portion 40 and the second cylindrical housing 30 described above.

Wherein, the connection between the open lower end of the second cylindrical housing 30 and the lid connecting portion 40 is connected to the lid through a female screw portion 35 formed on the inner peripheral surface of the open lower end of the second cylindrical housing. The threaded connection of the external threaded portion 45 on the outer peripheral surface of the portion 40 can be realized, so that the volume of the second space S2 can be adjusted to increase or decrease, but the present invention is not limited to this. The open lower end of the second cylindrical housing 30 The assembly with the lid connecting portion 40 can be realized by elastic coupling between an assembly groove formed at the lower end of the second cylindrical housing 30 and an assembly flange formed on the outer peripheral surface of the lid connecting portion 40.

Therefore, when the neck hole 21 and the slit 22a are communicated and connected to each other by the upward movement of the operating lever 13 whose lower end is located at the lower guide tube 24, a part of the liquid filled in the first cylindrical housing 20 passes through the neck The hole 21 guides and discharges to the second space S2 of the second cylindrical housing 30, thereby filling the second space S2.

In addition, when the neck hole 21 is in a closed state by the downward return movement of the joystick 13, only the liquid in the lower layer corresponding to the liquid layered by centrifugal separation is separated and filled in the second cylindrical housing 30 The second space S2.

As shown in FIGS. 1a, 1b, 2 and 4, the second sealing cover 50 is provided with a sealing member 51 that is inserted into the discharge port 41 formed through the center of the main body of the cover connecting portion 40 and is detachable. The cover member assembled to the lower end of the cover connecting portion 40 described above.

Such a closing member 51 may be formed in the shape of a column extending from the center of the upper surface of the second sealing cap 50 by a predetermined height so as to enter the lower extension pipe 42 extending downward by a predetermined length from the discharge port 41 of the cap connecting portion 40. The internal hole is closed.

It is possible to include a connecting pipe 52 having a female threaded portion 52a formed on the inner peripheral surface of the outer side of the closing member 51 so as to be screwed to the male threaded portion 42a formed on the outer peripheral surface of the lower extension pipe 42, wherein the connecting pipe 52 is from The center of the upper surface of the second sealing cap 50 is extended by a predetermined length.

At this time, preferably, the closing member 51 located at the inner center of the connecting pipe 52 is formed to be relatively higher than the height of the connecting pipe 52 so that the end of the lower extension pipe 42 enters first.

In addition, the outer edge of the second sealing cover 50 includes an upwardly curved outer skirt portion 54 so as to be combined with the outer edge of the lower end of the cover connecting portion 40, and there is at least one O-ring member 53 therebetween, thereby Can increase the sealing force.

Therefore, the second sealing cap 50 is coupled to the cap connecting portion 40 by the threaded coupling of the connecting pipe 52 and the lower extension pipe 42, and at the same time, the cap is closed by the closing member 51 that enters the inside of the lower extension pipe 42. The discharge port 41 of the connection part 40 is sealed.

On the other hand, it may include a filter unit 60 for removing foreign substances contained in an object (fat stem cell) discharged to the outside through the discharge port 41 of the cap connecting portion 40 separated by the second sealing cap 50 described above.

As shown in Figures 5 and 6, this filter portion 60 may include: an upper cover 61 on which In the center of the portion surface is provided an inflow port 62 communicating with the discharge port 41 of the cap connecting portion 40 separated from the second sealing cap; a filter body 64 is provided from the lower surface of the upper cover 61 corresponding to the inflow port 62 The end of the filter tube 63 extended by a predetermined length is used to remove foreign matter.

The filter unit 60 may include a filter housing 65 assembled with an open lower end and an open upper end of the upper cover 61, and the filter body 64 is arranged inside to discharge the fluid filtered by the filter body 64 There is at least one outlet 66.

The filter body 64 may include: an inner filter 64a, the inlet end of which is fixed to the end of the filter tube 63; an outer filter 64b, which has a relatively smaller mesh size than the inner filter 64a, and is arranged in The outer side of the internal filter 64a.

Preferably, the outlet end as the lower end of the inner filter 64a and the outlet end as the lower end of the outer filter 64b are separated from each other at a predetermined interval.

The inflow port 62 may be formed in the form of a hollow pipe with an internal threaded portion 62a formed on the inner surface so as to be screwed with an external threaded portion 42a formed on the outer surface of the lower extension pipe 42 extending from the discharge port 41.

At this time, when the inflow port 62 is formed as a hollow pipe with a male threaded portion formed on the outer surface, a separate adapter pipe can be used as a medium to communicate and assemble with the lower extension pipe 42 formed with the male threaded portion 42a.

The outflow port 66 may be formed in the form of a hollow tube formed on the outer surface with an external thread portion 66a threadedly coupled with an internal thread portion formed in the injection port of the syringe 70 so as to be able to collect and filter the filter body 64 passing through the filter portion 60 Object.

Therefore, the final separation object (fat stem cells) remaining in the second space S2 formed between the second cylindrical housing 30 and the lid connecting portion 40 can be filtered through the filtering portion 60 and then connected to the outflow port. The single syringe 70 connected with 66 is quickly separated and extracted without contact with the air in the atmosphere.

The method of separating adipose stem cells using the adipose stem cell separation device 100 having the above-mentioned structure extracts the final adipose stem cells by sequentially performing the preparation step, the adipose stem cell extraction step, the primary washing step, and the secondary washing step.

In the above preparation steps, first use a syringe with a luer lock to remove A predetermined amount of adipose tissue is extracted from the human body. At this time, the extracted adipose tissue is repeatedly extracted with a predetermined amount each time.

In addition, when the extracted adipose tissue is put into the inside of another syringe for storage, and the syringe is shaken to precipitate the adipose tissue, the inside of the syringe can be layered into a blood layer, a fat extraction liquid layer, and an oil layer due to the difference in specific gravity.

Next, the worker separates and extracts only the fat extraction liquid layer layered between the blood layer and the oil layer from the syringe through the piston operation of the syringe.

In the aforementioned adipose stem cell extraction step, the adipose stem cells are extracted from the fat extract obtained in the aforementioned preparation step.

That is, as shown in FIG. 7a, the auxiliary cover 12 (which closes the open upper portion of the first cylindrical housing 20) of the first sealing cover 10 is opened, and the neck hole 21 of the neck portion 20a is opened by the lower portion of the joystick 13 With the end closed, the fat extract P1 layered in the syringe is injected into the first space S1 of the first cylindrical housing 20 through the open communication port 11, and then injected into the first space S1 A predetermined amount of an enzyme solution (C1) such as a collagenase solution, wherein the collagenase is an enzyme that hydrolyzes collagen or gelatin components.

At this time, for the work of putting the above-mentioned fat extraction liquid P1 into the first space S1 of the first cylindrical housing 20, after the worker discharges the blood layer deposited on the lowermost layer of the syringe to the outside through the piston operation of the syringe, Only the fat extract liquid layer deposited on the intermediate layer can be injected into the first space S1 of the first cylindrical housing 20 through the opened communication port by the piston operation of the syringe described above.

Next, the adipose stem cell separation device 100 of the present invention with the auxiliary cover 12 closed to seal the communication port 11 is loaded into a shaking incubator and then stirred to perform the promotion of the enzyme reaction between the fat extract and the enzyme solution. The oscillation process.

In addition, if the adipose stem cell separation device 100 of the present invention taken out of the above-mentioned shaking incubator is placed in a centrifuge and centrifuged once with a predetermined intensity of centrifugal force within a predetermined time, the adipose tissue fluid of the enzyme reaction will be After performing centrifugal separation once, due to the difference in specific gravity, from the lower part of the first space S1 of the first cylindrical shell 20 to the upper part, the cell sediment P including the adipose stem cells and the enzyme reaction solution C of the enzyme reaction are sequentially layered.

Continuously, as shown in FIG. 7b, the worker raises the joystick 13 by rotating the first sealing cover 10 or the joystick 13 so that the lower end of the joystick 13 is located at the slit 22a of the upper guide tube 22.

In this state, if the secondary centrifugal separation process is performed in the centrifuge, the cell pellet P in the lowermost layer is transferred to the second cylindrical shell through the partially opened slit 22a and the completely opened neck hole 21. The second space S2 is thus filled in the above-mentioned second space S2.

And, if it is confirmed that the first and second cylindrical shells made of transparent materials that can be visually confirmed from the outside, and the boundary layer B between the cell sediment P layered through the neck 20a and the enzyme reaction solution C are located For the neck hole 21, the joystick 13 is lowered in such a manner that the lower end of the joystick 13 is located at the neck hole 21, thereby closing the opened neck hole 21.

Therefore, while the neck hole 21 is closed by the lower end of the joystick 13 in such a way that the enzyme reaction solution C of the first cylindrical housing flows into the second space S2 of the second cylindrical housing, the first circle Only the enzyme reaction solution C remains in the first space S1 of the cylinder shell. As the cell sediment P is filled in the second space S2 of the second cylindrical shell, the enzyme reaction solution and the cell sediment P are separated.

At this time, the joystick 13 that moves up and down in the neck hole 21 forms a receiving portion 13b with an open lower end inside the main body, so that a part of the cell sediment P separated by a volume larger than the internal volume of the second space S2 flows into the storage Part 13b, therefore, the separation efficiency can be improved.

In this state, as shown in FIG. 7c, the syringe 71 assembled to communicate with the lower extension tube 42 of the cap connecting portion 40 exposed to the outside by the separation of the second sealing cap 50 is used to extract and fill the outside. The cell pellet P in the second space S2 of the second cylindrical housing 30 described above.

In contrast, the enzyme reaction solution C remaining in the first space S1 of the first cylindrical casing 20 can be discharged to the outside through the opening of the auxiliary cover 12 or the opening of the first sealing cover 10.

Among them, as shown in FIG. 7c, for the process of washing the cell sediment P remaining in the second space S2 of the second cylindrical housing 30, the work of extracting to the outside may be carried out by separating the second sealing cover. The way in which the syringe 71 assembled in the lower extension tube 42 is connected in the state is shown and explained, but the present invention is not limited to this. As shown in FIG. 7b, the second sealing cap 50 can also be assembled by having a through hole. In the state of, the above-mentioned operation is performed corresponding to other syringes inserted into the injection needle of the closing member 51 of the lower extension tube 42 without contact with air.

That is, the sealing member 51 through which the injection needle penetrates is formed into a hollow tube column shape having a cut region 51a of a predetermined thickness cut in the middle of the length, and the cut region may be formed integrally with the hollow tube-shaped sealing member. Or formed as a separate press-in part, the material of the cut-off area may be composed of a rigid flexible material that can be penetrated by an injection needle.

As shown in FIGS. 7d and 7e, in the above-mentioned primary cleaning step, the first space S1 of the above-mentioned first cylindrical housing 20 is washed with the primary washing liquid W1 to clean the cell sediment P, and only the separation and extraction are performed to remove impurities by washing. The cell pellet P.

This washing solution can be used by selecting saline mixed with the extracted cell sediment in a predetermined ratio, but it is not limited to this, and other washing solutions can be selectively used.

The auxiliary cover 12 of the first sealing cover 10 (which closes the open upper part of the first cylindrical casing of the newly prepared separation device) is opened, and the open communication port 11 is used to extract the fat stem cells from the above-mentioned adipose stem cell extraction process. A certain amount of cell sediment P and a predetermined amount of primary washing liquid W1 such as saline are sequentially injected into the first space S1 of the first cylindrical housing, thereby filling the first space S1.

At this time, it is preferable that the above-mentioned primary washing liquid W1 is quantitatively input based on the input amount of the cell pellet P to be washed and a predetermined ratio.

In this state, if three centrifugal separations are performed by centrifugal force of a predetermined strength within a predetermined period of time in a centrifuge, the above-mentioned cell sediment P is washed by the primary washing liquid W1, and at the same time, it is removed and separated such as being adsorbed by the above-mentioned Contaminants of the enzyme reaction solution on the adipocytes, fat precursor cells and stem cells of the cell sediment P.

As shown in FIG. 7d, through this centrifugal separation, the washed cell sediment P and the primary washing liquid W1 in the first space S1 of the first cylindrical housing 20 are sequentially layered from the bottom to the top due to the difference in specific gravity. .

And, as shown in FIG. 7e, the primary washing liquid W1 stratified in the first space S1 of the first cylindrical casing 20 is sucked by the injection needle of the syringe that enters through the communication port 11 of the auxiliary cover 12, and is sucked by the injection needle of the syringe. By removing, only the cell sediment P remains in the first space S1.

At this time, the centrifugal separation process for sufficiently separating the cell sediment P and the primary washing liquid W1 in the first space S1 is closed by the lower end of the joystick 13 in the neck hole 21, keeping the second space S2 clean It is advisable to proceed in a low-degree state.

As shown in FIG. 7f, FIG. 7g and FIG. 7h, in the above-mentioned secondary cleaning step, the first space S1 of the first cylindrical housing 20 is used for the second washing of the cell sediment P with the secondary washing liquid W2. Washing separates only the cell sediment P from which impurities are removed by washing and extracts it to the outside.

That is, as shown in FIG. 7f, the auxiliary cover 12 is opened, and a predetermined amount of secondary washing liquid W2, such as salt water, is injected into the first space S1 of the first cylindrical housing through the opened communication port 11, and It is mixed with the cell sediment P remaining in the first space S1 at a predetermined ratio.

In addition, if the operating lever 13 is moved up and moved by the rotation operation of the first sealing cover 10 or the operating lever 13, as shown in FIG. 7g, the neck hole 21 will be completely opened, thereby the first space S1 and the second space S1 The space S2 is connected. In this state, if the centrifuge performs four centrifugal separations with a predetermined strength of centrifugal force within a predetermined period of time, the above-mentioned cell sediment P will be cleaned by the secondary washing solution W2, and at the same time, it will be removed and separated again. Such as contaminants adsorbed to the enzyme reaction solution of adipocytes, fat precursor cells, and stem cells included in the above-mentioned cell sediment P.

Through this centrifugal separation, the washed cell sediment P and the secondary washing liquid W2 in the first space S1, the second space S2 of the first cylindrical housing 20 and the second cylindrical housing 30 are separated from each other due to the difference in specific gravity. The lower part is layered successively to the upper part.

In addition, when the joystick 13 is moved down and the lower end of the joystick 13 is located at the neck hole 21 and the neck hole 21 is closed, the secondary washing liquid W2 remains in the first space S1 of the first cylindrical housing, and the second circle Washed cell sediments remain in the second space S2 of the cartridge shell.

At this time, a portion of the cell sediment that is relatively large compared to the internal volume flows from the second space S2 into the internal hole formed in the lower end of the joystick 13 in an open manner, thereby filling the internal hole. .

Therefore, the secondary washing liquid W2 remaining in the first space S1 of the first cylindrical housing 20 is discharged to the outside through the opening of the auxiliary cover 12 or the opening of the first sealing cover 10. For the cell sediment P separated and transferred in the second space S2 of the two-cylindrical casing 30, the filter unit 60 is assembled with the lower extension pipe 42 of the cover connection part 40 exposed to the outside by the separation of the second sealing cover 50. To filter the foreign matter contained in the cell sediment P, the cell sediment P is extracted to the outside.

That is, as shown in FIG. 6, the inflow port provided in the upper cover 61 of the filter unit 60 is 62 is threadedly connected with the extended lower extension pipe 42 of the discharge port 41 of the cap connection part exposed to the outside when separated from the second sealing cap to achieve communication, and the outlet port of the filter housing 65 provided in the filter part 60 and the extraction syringe 70 combined to achieve connectivity.

In the state where the upper part of the first cylindrical housing 20 has been opened by the opening of the first sealing cover 10 or the auxiliary cover 12, when the lower end of the joystick 13 is located at the slit 22a of the upper guide tube 22, the The first space S1 of the first cylindrical housing 20 from which the washing liquid is removed from the outside and the second space S2 of the second cylindrical housing 30 where the washed cell precipitate remains communicate with each other. Therefore, when the above-mentioned pulling of the extraction syringe is performed During operation, the cell pellet P is transferred to the inside of the filter unit 60 having an inflow port connected to the above-mentioned discharge port.

Continuously, the cell sediment P transferred to the inside of the filter unit 60 continuously passes through the filter body 64 composed of the internal filter 64a and the external filter 64b, thereby removing foreign matter, and the cell sediment P from which the foreign matter is removed is passed and installed The extraction syringe combined with the outflow port at the lower end of the filter housing 65 extracts the entire amount, thereby ending the fat stem cell separation work.

The present invention described above is not limited by the foregoing embodiments and drawings. It is obvious to those skilled in the art that various substitutions, modifications and changes can be made without departing from the technical idea of the present invention. .

100: Adipose stem cell separation device

10: The first sealing cover

11a: Hollow tube

12: auxiliary cover

13: Joystick

13a: O-ring

20: The first cylinder shell

20a: neck

25: External thread

30: second cylinder shell

40: Cover connection part

41: Outlet

43: upper face

50: The second sealing cover

51: closed parts

52: connecting pipe

54: Outer skirt

Claims (6)

An adipose stem cell separation device, comprising: a first sealing cover provided with an auxiliary cover for opening and closing a communication port formed through the upper surface, a joystick having a predetermined length extended from the lower surface; a first cylindrical housing, The outer peripheral surface of the upper end is formed with an external thread portion that is threadedly coupled with the female thread portion formed on the inner peripheral surface of the lower end of the first sealing cap, whereby the upper part opened by the first sealing cap is selectively opened and closed, and a first Space; a second cylindrical shell, which will be formed in the neck hole of the neck of the lower end of the first cylindrical shell configured with the joystick as a medium, communicating with the first cylindrical shell; a cover connecting portion , Which is assembled in the open lower part of the second cylindrical shell to form a second space with the second cylindrical shell; and a second sealing cover, which is provided with a corresponding insertion in the connecting portion formed through the cover The closure member of the discharge port is assembled in the cover connection part, wherein the closure member is formed in a column shape extending from the center of the upper surface of the second sealing cover by a predetermined height so as to enter the lower portion extending downward from the discharge port The inner hole of the extension tube is closed, the outer edge of the second sealing cover includes an outer skirt portion bent upward so as to be combined with the outer edge of the lower end of the cover connecting portion, and there is at least one O-ring component therebetween, Wherein, the outer side of the closing member includes a connecting pipe with an internal thread formed on the inner peripheral surface so as to be threadedly coupled with the external thread formed on the outer peripheral surface of the lower extension pipe, wherein the closing member is located at the inner center of the connecting pipe It is formed to be relatively higher than the height of the connecting pipe, so that when the cover connecting portion is combined with the second sealing cover, the closing component is first entered from the end of the lower extension pipe. The adipose stem cell separation device described in claim 1, wherein the joystick is composed of an outer threaded connection with an inner threaded portion of a cover extension tube extending from the closed lower part of the first sealing cover to a straight lower part by a predetermined length The threaded portion is formed by a rod member of a predetermined length at the upper end; or, it is formed by a threaded portion of a cap extension pipe extending by a predetermined length from a through hole formed through the closed upper surface of the first sealing cap to a straight lower part. The coupled male screw portion is formed in a rod member of a predetermined length in the middle of the length. The adipose stem cell separation device according to the first item of the scope of patent application, wherein the neck hole of the neck includes an upper guide tube, the upper guide tube is extended by a prescribed length to guide the lower end of the joystick to enter, and the upper guide The tube includes at least one slit formed by the opening. The adipose stem cell separation device described in claim 1 further includes: a filter part for removing the objects contained in the lid connection part separated by the second sealing lid and discharged to the outside Foreign matter, the filter portion includes: an upper cover, the center of the upper surface of which is provided with an inflow port communicating with the discharge port of the cover connecting portion; a filter body is provided on the lower surface of the upper cover corresponding to the inflow port The end of the filter tube extended by a predetermined length is used to remove foreign matter; and the filter housing, which is assembled with the upper cover and the open upper end, has at least one flow to discharge the object filtered by the filter body Exit. For the fat stem cell separation device described in item 4 of the scope of patent application, the filter body includes: an internal filter, the inlet end of which is fixed at the end of the filter tube; an external filter, which has a larger mesh than the internal filter The mesh size of the relatively small eye size is arranged on the outside of the inner filter. According to the adipose stem cell separation device described in item 5 of the scope of patent application, the outlet end of the inner filter and the outlet end of the outer filter are separated from each other by a predetermined distance.

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