KR101489264B1 - Kit for separating stem cell and method for separating stem cell - Google Patents

Abstract

The present invention relates to a kit for separating a stem cell from various kinds of tissue of a human body, and to a method for separating a stem cell. The kit includes an upper main body which has an upper containing space and a lower containing space; a closed lower main body that has a first containing space in fluid-communication with the lower containing space, and a second containing space, which has a cross-sectional area smaller than that of the first containing space, includes a lower end portion in the shape of a cone, and is detachably joined with a neck part of the upper main body; a separation film rod which moves up through the upper main body, moves down to close the top of the neck, and thus prevents fluid-communication between the upper containing space and the lower containing space; and a sieve which is detachably mounted in the first containing space of the closed lower main body. Materials contained in the upper containing space are sieved by the sieve. Therefore, a material in the predetermined size or bigger size can be separated. According to the present invention, a separated stem cell is not transferred from one container to another container; and separation and cleaning of a stem cell and separation of impurities are performed by using one kit for separating a stem cell. Therefore, when the kit is used, the stem cell is prevented from being polluted and can be rapidly separated.

Description

[0001] The present invention relates to a stem cell isolation kit and a method for separating stem cells,

The present invention relates to a stem cell isolation kit and a stem cell isolation method for separating stem cells from various tissues of the human body.

Stem cells are cells that have the ability to differentiate into all kinds of cells that constitute organisms such as nerves, blood, cartilage, etc., if they are maintained without progressing to specific cells.

These stem cells can be divided into embryonic stem cells isolated from the embryo and adult stem cells isolated from the adult body.

Another class can be divided into pluripotency stem cells, multipotency stem cells, and unipotent stem cells depending on how many kinds of differentiated cells can be made from one stem cell. In general, embryonic stem cells are classified into pluripotent stem cells, and adult stem cells can be classified into multipotential stem cells and monodomain stem cells.

Adult stem cells, which are comparable to the performance of embryonic stem cells, have been found in various organs of the human body, and mesenchymal stem cells have been reported to be isolated from mesenchymal stem cells, fats and umbilical cord blood.

Among these, adipose stem cells isolated from adipose tissue have the ability to differentiate into various types of cells such as adipocytes, muscle cells, chondrocytes, and bone cells. These adipocyte stem cells have an advantage in that they have excellent self-regenerative ability, easy in vitro culturing, and are rich in adipose tissue that can separate adipose stem cells, are simple and safe to collect, compared with stem cell of bone marrow origin . In terms of function, adipocyte stem cells have almost the same characteristics as stem cell of bone marrow origin, such as differentiation ability, cytologic characteristic, immunological characteristic, tissue regeneration ability.

As a method for obtaining adipose stem cells from adipose tissue, a method has been generally proposed in which the inhaled or resected fat is finely crushed to dissolve the tissue with collagenase, and the stem cells that are precipitated through centrifugation are separated .

1 is a view showing a container for separating adipose stem cells from the prior art.

In order to separate the adipose stem cells from the fat, first the inhaled fat from the human body is injected into the container 10. Next, collagenase is injected into the container 10 as a fat dissolving agent to shake the container 10, followed by centrifugation. By centrifugation, the fat in the container 10 is separated into fat stem cells 11, collagenase and foreign matter 12, pure fat 13 and oil 14 in turn according to the specific gravity.

Next, only the adipose stem cells 11 are sucked in the container 10 with a syringe or the like, and then injected into the container 20 having a narrow width and a long length. Next, when the stem cell detergent 15 is injected into the container 20 and centrifugally separated, only the pure adipose stem cells 11 are located in the lower part of the container 10. The resulting adipose stem cells (11) are mixed with pure fat and injected into the patient.

In the conventional containers 10 and 20, only the adipose stem cells are difficult to accurately inhale during inhalation of the adipose stem cells located in the lower part of the container with a syringe or the like, and are sucked together with other foreign substances. Therefore, there is a problem in that the purity of the adipose stem cells is low or the adipose stem cells are separated separately. In addition, since the adipose stem cells must be transferred from one container to another, there is a risk that the adipose stem cells are contaminated during the transfer and the separation process of adipose stem cells becomes complicated.

Korean Patent No. 10-1049201 discloses a container for separating hematopoietic stem cells in blood.

Such a container includes a body portion having a bottleneck portion separating the upper accommodation space and the lower accommodation space. In addition, a syringe guide having a tubular structure that extends through the upper accommodation space from the upper lid coupled to the main body and blocks the space at the upper and lower ends of the bottleneck is formed. In the process of inhaling hematopoietic stem cells with a syringe, Stem cells are prevented from being inhaled together with blood plasma and the like at the top of the bottleneck.

However, such a container is limited to use for separating hematopoietic stem cells in blood, and there is a problem that hematopoietic stem cells can not be prevented from being inhaled together with red blood cells or the like at the bottom of the bottleneck. In addition, since the stem cells inhaled by the syringe need to be transferred to another container to obtain only pure stem cells, there is a problem that the stem cells are contaminated or the separation process becomes complicated.

Korean Patent No. 10-1049201

DISCLOSURE OF THE INVENTION Accordingly, the present invention has been made in view of the above circumstances, and it is an object of the present invention to provide a method for separating stem cells from bone marrow, fat, umbilical cord blood and the like, A cell separation kit, and a method for separating stem cells.

According to an aspect of the present invention, there is provided a stem cell separation kit including a neck portion having a narrow width at a lower portion thereof and having an upper upper accommodating space and a lower lower accommodating space with respect to an upper end of the neck, ; A second accommodation space in fluid communication with the lower accommodation space and a second accommodation space in fluid communication with the first accommodation space and having a conical shape with a lower end portion reduced in cross section than the first accommodation space, A closed lower body detachably coupled to the neck; A separating bar capable of being raised and lowered through the upper main body and closing the upper end of the neck portion when the lower portion is lowered to block fluid communication between the upper accommodating space and the lower accommodating space; A squeezing network detachably mounted in the first receiving space of the closed lower body; Wherein the closed lower body is detachable from the upper body in a state of blocking fluid communication between the upper body and the closed lower body by the separating rod, And a material having a predetermined size or more can be separated through the filtering net.

In addition, a stepped portion is formed at a boundary between the first accommodation space and the second accommodation space in the closed lower body so that the sieve can be seated.

An open lower body having a lower accommodating space in fluid communication with the lower accommodating space and detachably coupled to the neck of the upper body; A lower stopper movably coupled to a lower portion of the open lower body to seal the accommodation space of the open lower body; And the volume of the accommodating space of the openable lower body is varied by raising and lowering the lower stopper.

An upper cover coupled to the upper body to cover the upper body and having at least one opening; And the separator plate is coupled to the upper cover and extends into the upper body through the opening.

According to another aspect of the present invention, there is provided a method for separating adipose stem cells from fat by using a stem cell isolation kit, the stem cell separation kit comprising: An upper body having an upper upper accommodating space and a lower lower accommodating space with respect to an upper end of the neck portion; An open lower main body having a lower accommodating space in fluid communication with the lower accommodating space and detachably coupled to the neck of the upper main body; A lower stopper movably coupled to a lower portion of the open lower body to seal the accommodation space of the open lower body; A second accommodation space in fluid communication with the lower accommodation space and a second accommodation space in fluid communication with the first accommodation space and having a conical shape with a lower end portion reduced in cross section than the first accommodation space, A closed lower body detachably coupled to the neck; A separating bar capable of being raised and lowered through the upper main body and closing the upper end of the neck portion when the lower portion is lowered to block fluid communication between the upper accommodating space and the lower accommodating space; A squeezing network detachably mounted in the first receiving space of the closed lower body; / RTI >

The stem cell separation method comprises:

The open bottom main body is coupled to the upper main body, the lower stopper is coupled to the open bottom main body, and the separating bar is inserted into the upper main body so as not to close the upper end of the neck, And centrifuging in a centrifuge; A step of raising the lower stopper to place the level of the centrifugally separated pure fat at a boundary point between the upper accommodation space and the lower accommodation space; Separating the open lower body from the upper body in a closed state in which the separating bar closes the upper end of the neck; Coupling the closed lower body to the upper body; Injecting a fat dissolving agent into the upper body in an open state of the separating bar to shink; Centrifuging the upper body and the closed lower body in a centrifugal separator to place the adipocyte stem cells having a larger specific gravity at the lower end of the closed lower body; Discarding the contents in the upper body in a closed state of the separating bar; Injecting saline into the upper body; Moving the contents in the upper body and the closed lower body to the upper receiving space in an open state of the separating bar; Separating the closed bottom body from the upper body; Coupling the closed bottom body with the filtering net to the upper body; Centrifuging in a centrifugal separator, passing the contents moved to the upper accommodation space through the sieve, thereby obtaining pure adipose stem cells in the closed lower body; .

According to another aspect of the present invention, there is provided a method for separating adipose stem cells from fat by using a stem cell isolation kit, the stem cell separation kit comprising: An upper body having an upper upper accommodating space and a lower lower accommodating space with respect to an upper end of the neck portion; A second accommodation space in fluid communication with the lower accommodation space and a second accommodation space in fluid communication with the first accommodation space and having a conical shape with a lower end portion reduced in cross section than the first accommodation space, A closed lower body detachably coupled to the neck; A separating bar capable of being raised and lowered through the upper main body and closing the upper end of the neck portion when the lower portion is lowered to block fluid communication between the upper accommodating space and the lower accommodating space; A squeezing network detachably mounted in the first receiving space of the closed lower body; / RTI >

The stem cell separation method comprises:

Centrifuging the fat to obtain pure fat containing adipose stem cells; The pure water and the fat dissolving agent are injected into the upper main body in an open state inserted into the upper main body so as not to close the upper end of the neck, step; Centrifuging the upper body and the closed lower body in a centrifugal separator to place the adipocyte stem cells having a larger specific gravity at the lower end of the closed lower body; Discarding the contents in the upper body in a closed state in which the separating bar closes the upper end of the neck; Injecting saline into the upper body; Moving the contents in the upper body and the closed lower body to the upper receiving space in an open state of the separating bar; Separating the closed bottom body from the upper body; Coupling the closed bottom body with the filtering net to the upper body; Centrifuging in a centrifugal separator, passing the contents moved to the upper accommodation space through the sieve, thereby obtaining pure adipose stem cells in the closed lower body; .

According to the present invention, since isolated stem cells are isolated from one container to another, a single stem cell separation kit is used to separate and wash stem cells, and to separate foreign substances, thereby preventing contamination of stem cells A stem cell separation kit and a stem cell separation method in which a separation operation can be performed.

1 is a view showing a container for separating adipose stem cells from the prior art.
2 is a cross-sectional view illustrating a stem cell separation kit according to an embodiment of the present invention.
FIG. 3 is an exploded view of the stem cell separation kit shown in FIG. 2; FIG.
FIGS. 4 to 15 are diagrams sequentially illustrating embodiments for separating adipose stem cells using the stem cell isolation kit of the present invention. FIG.

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 drawings, like reference numerals refer to like elements throughout. The same reference numerals in the drawings denote like elements throughout the drawings.

2 is a cross-sectional view illustrating a stem cell separation kit according to an embodiment of the present invention. FIG. 3 is an exploded view of the stem cell separation kit shown in FIG. 2; FIG.

The stem cell separation kit 100 includes an upper body 110, an open lower body 120, a lower stopper 130, a separating bar 140, an upper cover 150, and the like.

The upper body 110 is formed in a substantially cylindrical shape and has an accommodation space for accommodating a fluid or the like therein. The upper body 110 has a narrow neck portion 110a at a lower portion thereof. The neck portion 110a has a screw portion 110a 'formed on the inner side and a downward protruding portion 110a "which is connected to the upper end of the screw portion 110a' and is spaced a predetermined distance inward from the screw portion 110a ' The upper body 110 may have a wide upper portion, a connecting portion where the width gradually decreases in a cone shape, and a neck portion 110a having a narrow width at the lower portion of the connecting portion. A scale 110b may be formed in the main body 110 to indicate the capacity of the fluid contained therein.

The upper main body 110 can be divided into an upper upper accommodating space S1 and a lower lower accommodating space S2 on the basis of the upper end of the neck 110a (indicated by the dotted line l1 in Fig. 2) .

The open lower body 120 has a substantially cylindrical shape and a receiving space in fluid communication with the lower receiving space S2. The open lower body 120 is opened at the bottom and is detachably connected to the neck 110a of the upper body 110. [

The open lower body 120 is, for example, threadably engaged with the upper body 110 and can be easily separated from the upper body 110 by rotation of the open lower body 120. For example, a female screw is formed on the inner surface of the neck portion 110a, and a male screw is formed on the upper outer side of the open lower body 120 so that the open lower body 120 is inserted into the neck portion 110a to be engaged . Accordingly, the receiving space of the open lower body 120 and the lower receiving space S2 of the upper main body 110 can be partially overlapped.

A lower stopper 130 for sealing the receiving space of the open lower body 120 is provided at the lower portion of the open lower body 120.

The lower stopper 130 is movably coupled to the open lower body 120. The fluid stored in the accommodating space of the openable lower body 120 can be raised and lowered together with the rising and falling of the lower stopper 130. The volume of the accommodating space of the openable lower body 120 is varied by raising and lowering the lower stopper 130.

The lower stopper 130 can be screwed with the open lower body 120 and can be easily raised and lowered along the inner surface of the open lower body 120 by rotation of the lower stopper 130.

For example, a female screw may be formed on the inner bottom surface of the open lower body 120, and a male screw may be formed on the outer side of the lower end of the lower stopper 130. The length of the portion where the female screw is formed on the open lower body 120 is longer than the length of the portion where the male screw is formed on the lower stopper 130. Accordingly, by rotating the lower stopper 130, the lower stopper 130 can move up and down along the female screw formed on the inner surface of the openable lower body 120.

A wrench groove 131 may be formed at a lower end of the lower stopper 130 to move up and down the lower stopper 130. The lower stopper 130 can be lifted and lowered along the inner surface of the openable lower body 120 by rotating a hexagonal wrench or the like into the wrench groove 131.

The upper cover 150 is engaged with the upper end of the upper body 110 to shield the upper end of the upper body 110. For example, the upper cover 150 may be screwed with the upper body 110.

An opening is formed in the middle of the upper cover 150, and the separating bar 140 can be inserted into the upper body 110 through the opening.

In addition, the top cover 150 may further include one or more openings 151. And fat or the like to be separated into the upper main body 110 may be introduced through the opening 151. The opening 151 may be provided with a lid 155 for sealing the opening 151. Although only one opening 151 is formed in the drawing, a plurality of such openings 151 may be formed in the upper cover 150.

The separating bar 140 is coupled to the upper cover 150 and extends into the upper body 110 through an opening formed in the center. The separating bar 140 is threadably engaged with the inner surface of the extension 152 extending vertically along the opening formed in the center of the upper lid 150, 110). The separating bar 140 can be raised and lowered while being kept closed with the inner surface of the extending portion 152.

The separating bar 140 includes a longitudinally long bar portion 141, a head portion 142 integrally connected to the upper end of the bar portion 141, And may include a cap 143. By rotating the head portion 142, the separating bar 140 can be raised and lowered in the upper body 110.

The detachable cap 143 may be formed of a material having elasticity. The separating stopper 143 is formed to be slightly larger than the inner width of the neck 110a of the upper body 110. When the separating bar 140 is lowered, 110a can be closed. To this end, the side surface of the separating stopper 143 may be tapered.

When the separating bar 140 is lowered and the separating stopper 143 closes the upper end of the neck portion 110a, the fluid communication between the upper accommodating space S1 on the upper side of the separating bar 140 and the lower accommodating space S2 on the lower side Lt; / RTI > The separating bar 140 blocks fluid communication between the upper body 110 and the open lower body 120 in a state where the upper body 110 and the open lower body 120 are engaged.

The stem cell separation kit 100 further includes a closed lower body 160 and a sieve 170.

The closed lower body 160 can be detachably coupled to the neck 110a of the upper body 110 while the upper body 110 is separated from the open lower body 120. [ For example, a screw thread 162 may be formed on the outer surface of the closed lower body 160 to be screwed into the upper body 110.

The closed lower body 160 has a cylindrical first accommodation space S3 in fluid communication with the lower accommodation space S2 of the upper body 110 and a second accommodation space S3 in fluid communication with the first accommodation space S3, And the lower end portion has a second accommodating space S4 having a cone shape while the cross section is smaller than the space S3. Due to the shape of the closed bottom body 160, a small amount of fluid can be gathered well in the closed bottom body 160.

The filter screen 170 is detachably mounted in the first receiving space S3 of the closed bottom body 160. [ The mesh screen 170 has a cylindrical portion 172 to be mounted in the first accommodation space S3 and a mesh mesh 171 is formed below the cylindrical mesh portion 172. The mesh mesh 171 A material having a predetermined size or larger can be separated. The size of the material that can be separated through the mesh network 171 can be controlled by the mesh size of the mesh network 171. The mesh size (mesh size) of the mesh network 171 may be several tens to several hundreds of 탆.

The stepped portion 161 may be formed at the boundary between the first accommodation space S3 and the second accommodation space S4 in the closed lower body 160 so that the sieve 170 may be seated. When the filter screen 170 is attached to the closed bottom body 160, it can be seated and fixed to the step portion 161. The closed lower body 160 can be divided into the first accommodation space S3 and the second accommodation space S4 on the basis of the line l2 passing through the step portion 161. [

The stem cell separation kit 100 having the above-described shape and structure can accurately separate only the stem cells when the stem cells are separated from the bone marrow, fat, umbilical cord blood, etc. of the human body, All the processes for separating stem cells can be performed and it is not necessary to transfer the separated stem cells to another container, thereby preventing contamination of the stem cells in the separation process.

Hereinafter, an embodiment of separating adipose stem cells using the stem cell isolation kit of the present invention will be described with reference to FIG. 4 to FIG. FIGS. 4 to 15 sequentially illustrate a method for separating adipose stem cells using the stem cell separation kit 100 of the present invention.

The human bone marrow, fat, umbilical cord blood, and blood contain stem cells, from which stem cells can be obtained. In particular, adipocyte stem cells isolated from adipose tissue are advantageous in that they have excellent self-regenerating ability, are easy to culture in vitro, have abundant adipose tissue capable of separating adipose stem cells, have a simple and safe method of harvesting, have. Recently, for the purpose of molding, a method of injecting fat stem cells into human body by mixing with pure fat has been developed. When the fat stem cells are mixed with the pure fat taken from the patient's body and transplanted into the patient, there is a good advantage of being able to engage the fat, which is widely utilized.

Hereinafter, a process for separating adipose stem cells using the stem cell isolation kit 100 of the present invention will be described. However, the stem cell separation kit 100 of the present invention is not limited to the use for isolating adipose stem cells, Can be used to isolate stem cells from other human tissues.

Referring to FIGS. 2 and 3, first, after collecting fat from the patient's body, the lid 155 is opened and the fat is injected into the upper body 110 through the opening 151 using a syringe or the like. Next, the lid 155 is closed and centrifuged at a suitable rpm in a centrifuge. In this case, the stem cell separation kit 100 includes an open lower body 120 coupled to the upper body 110, a lower stopper 130 coupled to the open lower body 120, Is inserted into the upper main body 110 so as not to close the upper end of the neck 110a of the main body 110.

Referring to FIG. 4, blood and foreign matter 101 are contained in the stem cell separation kit 100 by centrifugation, pure fat 102 (including adipose stem cells) .

5, the water level (i.e., the lower water level) of the centrifugally separated pure fat 102 rises in the lower stopper 130 to the boundary point between the upper accommodation space S1 and the lower accommodation space S2 . The lower stopper 130 can be lifted and lowered along the inner surface of the openable lower body 120 by rotating a hexagonal wrench or the like into the wrench groove 131 of the lower stopper 130. [

6, when the head portion 142 of the separating bar 140 is rotated so that the separating bar 140 closes the upper end of the neck portion 110a of the upper body 110, The open lower body 120 is removed. At this time, the pure fat 102 is held in the upper receiving space S1 of the upper main body 110 by the separating rod 140. [

Next, referring to Fig. 7, the closed bottom body 160 is engaged with the upper body 110. Fig.

Next, referring to FIG. 8, the head portion 142 of the separating bar 140 is rotated to bring the separating bar 140 into an open state. Thereby, the upper accommodation space S1 and the lower accommodation space S2 (or the first accommodation space S3) are in fluid communication with each other, so that the pure fat 102 is filled from the closed lower body 160. Next, the lid 155 is opened and the fat dissolving agent 104 is introduced into the upper main body 110. The amount of fat dissolving agent 104 may be similar to the amount of pure fat 102. Next, the lid 155 is closed and the stem cell separation kit 100 is shaken to shake so that the pure fat 103 and the fat dissolving agent are mixed well. Such a shaking operation can use a shaking device. The shaking work makes the pure fat (102) mush so that the fat stem cells attached between the pure fat (102) can be well separated. The fat dissolving agent 104 can be used, for example, by diluting collagenase in saline.

Next, referring to Fig. 9, the stem cell separation kit 100 (i.e., the upper body 110 and the closed lower body 160) is centrifuged in a centrifuge. Accordingly, the adipose stem cells 102 'having a large specific gravity are positioned at the lower end of the closed lower body 160. At this time, the adipose stem cells 102 'may contain some foreign matter and may be in a slightly contaminated state.

Next, the upper body 110 is detached from the closed bottom body 160 in the closed state of the separating bar 140, and the other upper body 110 (for example, 110) (preliminary upper main body 1) to the closed lower main body 160. Next, the separating rod 140 is shaken well in an open state to allow the saline solution and the adipose stem cells 102 'to be mixed and washed, followed by centrifugation in a centrifuge (washing process 1).

Next, the upper body 110 is detached from the closed bottom body 160 in the closed state of the separating bar 140, and another preliminarily prepared upper body 110 (preliminary upper body 2) filled with saline To the closed bottom body (160). Next, the separating rod 140 is shaken well in the open state to allow the saline solution and the adipose stem cells 102 'to be mixed and washed, followed by centrifugation in a centrifuge (washing process 2).

The cleaning processes 1 and 2 may be further performed or omitted if necessary.

10, in the closed state of the separating bar 140, the lid 155 is opened and the contents in the upper main body 110 are discarded.

11, a stem cell cleanser (for example, saline solution) 105 is introduced into the upper body 110 and shaken well in the open state of the separating bar 140 to remove saline and adipose stem cells 102 ').

12, the upper body 110 and the closed lower body 160 are tilted in the open state of the separating bar 140 so that the contents 106 in the upper body 110 and the closed lower body 160 To the upper accommodation space S1. At this time, the contents 106 are mixed with the saline solution and the adipose stem cells 102 '.

13, the closed lower body 160 is separated from the upper body 110 and the closed lower body 160 equipped with the filtering net 170 is coupled to the upper body 110 again . At this time, it is preferable that the closed lower body 160 equipped with the filtering net 170 uses another closed lower body 160 prepared in advance. The upper end of the cylindrical portion 172 of the sieve 170 is connected to the inner side surface of the closed lower body 160 and the neck portion 110a of the upper body 110. [ And the lower end of the cylindrical portion 172 may be seated in the step portion 161. The mesh size of the mesh net 171 of the mesh 170 may be several tens to several hundreds Mu] m, for example, 90 to 150 [mu] m.

Next, referring to Fig. 14, the upper body 110 and the closed lower body 160 are centrifuged in a centrifuge in an open state of the separating bar 140. Then, as shown in Fig. At this time, in a state in which the filter screen 170 is seated on the step portion 161 of the closed type lower body 160, the cylindrical portion 172 of the filter screen 170 is pressed against the inner side surface of the closed type lower body 160 and the neck portion 110a of the upper housing space S1 so that the contents moved into the upper housing space S1 can be passed through the filtering net 170. [ The cleaned adipose stem cells 106 'are obtained in the lower part of the closed type lower body 160 and the foreign substances 106' 'such as fat agglomerates having a relatively large size are filtered in the filtering net 170.

15, after separating the closed lower body 160 from the upper body 110 and separating the filtered mesh 170 from the closed lower body 160, the pure adipose stem cells 106 ' ) Is sucked by a syringe or the like and used for a necessary purpose.

According to the above-described stem cell separation method, the stem cell separation kit 100 of the present invention shown in FIG. 2 is used. The stem cell separation kit 100 includes an upper body 110, an open lower body 120, a separating rod 140, a lower stop 130, a closed lower body 160, a sieve 170, .

With this configuration, the lower level of the centrifugally separated pure fat in the stem cell separation kit 100 is positioned at the boundary portion between the upper housing space S1 and the lower housing space S2, After the main body 120 is separated, the closed type lower body 160 is coupled to the upper body 110, and further purified and filtered using the adipose stem cell washing and filtering system 170 to obtain pure adipose stem cells .

As described above, in the stem cell separation method of the present invention, operations such as separation, washing, and foreign matter separation of adipose stem cells are performed using a single stem cell separation kit 100 (here, The preliminary stem cell separation kit 100 having the same shape can be provided and used). Accordingly, the separation of the adipose stem cells can be performed more quickly than the conventional method, since the adipocyte stem cells are transferred from one vessel to another vessel as in the prior art, while the additional operation is carried out.

Although the method of separating stem cells in which the fat is put into the upper body 110 and centrifuged in the centrifugal separator to obtain pure fat and then the operation such as washing is carried out has been described, If so, such pure fat (including adipose stem cells) can be injected into the upper body 110 to proceed with the operation.

In this case, the stem cell separation kit 100 includes the upper body 110, the separating bar 140, the closed lower body 160, the filtering net 170, and the like without the open lower body 120 and the lower stopper 130 . In the opened state inserted into the upper main body 110 so that the separating bar 140 does not close the upper end of the neck 110a of the upper main body 110, The pure fat and the fat dissolving agent obtained in the upper body 110 are injected to perform shaking.

Next, the stem cell sorting operation is performed in the same manner as described with reference to Figs. 9 to 15.

The process of separating adipose stem cells from the stem cell separation kit 100 of the present invention is not limited to the above embodiment. Those skilled in the art can appropriately change the working order, add the stem cells, and apply them to other apparatuses in order to accurately separate the adipose stem cells using the stem cell separation kit 100.

For example, even when hematopoietic stem cells in peripheral blood are located in the middle layer between plasma and erythrocytes by centrifugation, the stem cell separation kit 100 of the present invention can be used to separate stem cells have. When the upper body 110 and the open lower body 120 are combined with each other after the red blood cells are separated and removed in the same manner as the method for separating the adipose stem cells of the above embodiment, the stem cells are positioned at the bottom, It is possible to carry out the separation in the same manner as the separation of the cells.

The stem cell separation kit 100 of the present invention can be used not only for separating adipose stem cells from fat but also for separating stem cells from bone marrow, fat, umbilical cord blood and the like.

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 and scope of the invention will be.

100: Stem Cell Separation Kit
110: upper body
110a: neck
120: open bottom body
130: Lower stopper
140: Split bar
141:
142:
143: Separation plug
150: upper cover
151: opening
155: Lid
160: closed type lower body
170: Filter
171: mesh network
S1: upper accommodation space
S2: Lower accommodating space
S3: First accommodation space
S4: the second accommodation space

Claims (6)

A stem cell separation kit comprising:
An upper body 110 having a narrow neck portion 110a at the lower portion and having an upper receiving space S1 and a lower receiving space S2 at an upper portion with respect to an upper end of the neck portion 110a;
A first receiving space S3 in fluid communication with the lower receiving space S2 and a second receiving space S3 in fluid communication with the first receiving space S3 and having a section smaller than the first receiving space S3, A closed bottom body 160 detachably coupled to the neck portion 110a of the upper body 110,
A separating bar 140 which is movable up and down through the upper body 110 and closes an upper end of the neck 110a when the lower part 110 is lowered to block fluid communication between the upper housing space S1 and the lower housing space S2, ;
And a mesh network 171 formed at a lower portion of the cylindrical portion 172 and detachably mounted in the first accommodation space S3 of the closed lower body 160 170);
/ RTI >
The closed lower body 160 is separated from the upper body 110 in a state where fluid communication between the upper body 110 and the closed lower body 160 is blocked by the separation bar 140 Can,
The material contained in the upper accommodation space S1 can be filtered through the sieve 170 to separate a material having a predetermined size or larger,
The neck portion 110a includes a screw portion 110a 'formed on the inner side and a downward protrusion 110a' connected to the upper end of the screw portion 110a 'and spaced apart from the screw portion 110a'"),
A step portion 161 is formed at a boundary between the first accommodation space S3 and the second accommodation space S4 in the closed lower body 160 so that the sieve 170 can be seated,
The upper end of the cylindrical portion 172 of the sieve 170 is connected to the upper end of the closed lower body 160 in a state where the closed lower body 160 is coupled to the neck 110a of the upper body 110. [ And the lower end of the cylindrical portion 172 is seated in the stepped portion 161. The stem cell separation kit according to claim 1, delete The method according to claim 1,
An open lower main body having a lower accommodating space in fluid communication with the lower accommodating space and detachably coupled to the neck of the upper main body;
A lower stopper movably coupled to a lower portion of the open lower body to seal the accommodation space of the open lower body;
Further comprising:
Wherein the volume of the accommodating space of the open lower body is varied by raising and lowering the lower stopper. The method according to claim 1,
An upper cover coupled to the upper body to cover the upper body and having at least one opening;
Further comprising:
Wherein the separator plate is coupled to the upper lid and extends into the upper body through the opening. A stem cell isolation method for separating adipose stem cells from fat using a stem cell isolation kit,
The stem cell separation kit includes: an upper body having a lower neck portion at a lower portion and having an upper upper accommodating space and a lower lower accommodating space based on an upper end of the neck portion; An open lower main body having a lower accommodating space in fluid communication with the lower accommodating space and detachably coupled to the neck of the upper main body; A lower stopper movably coupled to a lower portion of the open lower body to seal the accommodation space of the open lower body; A second accommodation space in fluid communication with the lower accommodation space and a second accommodation space in fluid communication with the first accommodation space and having a conical shape with a lower end portion reduced in cross section than the first accommodation space, A closed lower body detachably coupled to the neck; A separating bar capable of being raised and lowered through the upper main body and closing the upper end of the neck portion when the lower portion is lowered to block fluid communication between the upper accommodating space and the lower accommodating space; A squeezing network detachably mounted in the first receiving space of the closed lower body; / RTI >
The stem cell separation method comprises:
The open bottom main body is coupled to the upper main body, the lower stopper is coupled to the open bottom main body, and the separating bar is inserted into the upper main body so as not to close the upper end of the neck, And centrifuging in a centrifuge;
A step of raising the lower stopper to place the level of the centrifugally separated pure fat at a boundary point between the upper accommodation space and the lower accommodation space;
Separating the open lower body from the upper body in a closed state in which the separating bar closes the upper end of the neck;
Coupling the closed lower body to the upper body;
Injecting a fat dissolving agent into the upper body in an open state of the separating bar to shink;
Centrifuging the upper body and the closed lower body in a centrifugal separator to place the adipocyte stem cells having a larger specific gravity at the lower end of the closed lower body;
Discarding the contents in the upper body in a closed state of the separating bar;
Injecting saline into the upper body;
Moving the contents in the upper body and the closed lower body to the upper receiving space in an open state of the separating bar;
Separating the closed bottom body from the upper body;
Coupling the closed bottom body with the filtering net to the upper body;
Centrifuging in a centrifugal separator, passing the contents moved to the upper accommodation space through the sieve, thereby obtaining pure adipose stem cells in the closed lower body;
≪ / RTI > A stem cell isolation method for separating adipose stem cells from fat using a stem cell isolation kit,
The stem cell separation kit includes: an upper body having a lower neck portion at a lower portion and having an upper upper accommodating space and a lower lower accommodating space based on an upper end of the neck portion; A second accommodation space in fluid communication with the lower accommodation space and a second accommodation space in fluid communication with the first accommodation space and having a conical shape with a lower end portion reduced in cross section than the first accommodation space, A closed lower body detachably coupled to the neck; A separating bar capable of being raised and lowered through the upper main body and closing the upper end of the neck portion when the lower portion is lowered to block fluid communication between the upper accommodating space and the lower accommodating space; A squeezing network detachably mounted in the first receiving space of the closed lower body; / RTI >
The stem cell separation method comprises:
Centrifuging the fat to obtain pure fat containing adipose stem cells;
The pure water and the fat dissolving agent are injected into the upper main body in an open state inserted into the upper main body so as not to close the upper end of the neck, step;
Centrifuging the upper body and the closed lower body in a centrifugal separator to place the adipocyte stem cells having a larger specific gravity at the lower end of the closed lower body;
Discarding the contents in the upper body in a closed state in which the separating bar closes the upper end of the neck;
Injecting saline into the upper body;
Moving the contents in the upper body and the closed lower body to the upper receiving space in an open state of the separating bar;
Separating the closed bottom body from the upper body;
Coupling the closed bottom body with the filtering net to the upper body;
Centrifuging in a centrifugal separator, passing the contents moved to the upper accommodation space through the sieve, thereby obtaining pure adipose stem cells in the closed lower body;
≪ / RTI >

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