KR20200084644A - Cell separator and method for cell separating using thereof - Google Patents

Abstract

The present invention relates to a cell separator and a cell separation method using the same. When a first body and a second body are separated after centrifugation, a supernatant provided in the first body can be easily separated, and the supernatant in the first body does not need to be discharged using a pipette or the like, thereby acquiring an effect that stem cells located in the second body can be safely extracted.

Description

Cell separator and cell separation method using the same{CELL SEPARATOR AND METHOD FOR CELL SEPARATING USING THEREOF}

The present invention relates to a cell separator and a method for separating cells using the same, and more specifically, to separate the first body and the second body after centrifugation, thereby allowing the cells located in the second body to be safely separated and It relates to a cell separation method using the same.

Recently, as the separation and use of stem cells from adipose tissue has become possible, adipose tissue has received much attention as a source of stem cells. Adipose-derived stem cells (ASCs) differentiate into fat cells as well as various cells such as bone cells, chondrocytes, neurons, muscle cells, endothelial cells, white blood cells, hepatocytes, epithelial cells, and pancreatic beta cells. There are characteristics that can be. In the past, most of the adult human stem cells were obtained from the bone marrow, but the process was very invasive and difficult to obtain large amounts at a time, whereas adipose tissue had the advantage of being able to collect large amounts of tissue more easily. have.

Adipose tissue is easy to collect a large amount of tissue and has good conditions for harvesting stem cells, and ASCs show stable growth and proliferation in culture and can differentiate into various cells when induced differentiation. Currently, ASCs are obtained by separating and culturing stromal vascular fraction (SVF) after removing mature fat cells and red blood cells of adipose tissue. However, this SVF contains various components according to each differentiation stage of adipocytes such as adipocytes and adipocytes. Therefore, a method for separating these components using antibodies specific to proteins present on the cell membrane surface has been developed, but has a disadvantage that it is difficult to use limitedly and publicly due to high cost.

Accordingly, methods based on the size and density of cells are used as a method for separating cells having high survival rate and purity from organs and tissues of humans and animals, and in detail, fat extraction step from adipose tissue, collagenase It includes an enzyme treatment step using Naz et al., a fat stem cell separation and washing step using centrifugation, and a filtration step to remove agglomerated (lump) fat stem cells. In addition, the step of separating and washing adipose stem cells using centrifugation is a process performed multiple times, and centrifugation is used to extract adipose stem cells from fat or to wash the extracted adipose stem cells with a washing solution.

However, in the process of performing the step of separating and washing the adipose stem cells using centrifugation, since the centrifugation operation proceeds, the cells are separated by a specific gravity difference, but because they are unstable, they can be mixed with each other by slight disturbance However, when separating only the desired components, there is a problem that additional extraction may be performed.

Korean Registered Patent Publication No. 10-1091052

An object of the present invention for solving the problems of the prior art as described above is to separate the first body and the second body after centrifugation, and a cell separator to safely extract stem cells located in the second body and using the same It is to provide a method of cell isolation.

In order to achieve the above object, the present invention includes a first body in which a first space is formed so that the mixture material is accommodated, but a first upper opening part is formed so that the mixture material is injected, and a first lower opening part is formed at the lower part; A first upper cap opening or closing the first upper opening; A second body in which a second space is formed, and a second upper opening is formed so that the upper portion can be coupled to the first lower open portion, and a second lower opening is formed at the lower portion; And a second lower cap opening or closing the second lower opening portion, wherein the first upper cap and the second lower cap respectively close the first upper opening portion and the second lower opening portion, and the second lower cap. 1 In the state in which the lower open portion and the second upper open portion are mutually coupled, when centrifugation is performed, supernatant is separated and stacked on the first body, and stem cells are separated and stacked on the second body, and the centrifugation is performed. When is completed, the second body is separated from the first body, the diameter of the first lower opening is characterized in that the supernatant located in the first body is formed to a size that can not be discharged downward by the surface tension To provide a cell separator.

In addition, the lower portion of the first body provides a cell separator characterized in that it is formed to be narrower as it gets closer to the second body.

In addition, it provides a cell separator characterized in that it further comprises a first upper opening and a first upper opening and closing portion for opening and closing the first upper opening formed on the upper portion of the first upper cap.

In addition, a first 1-1 upper opening is formed at an upper side of the first upper cap, a first 1-2 upper opening is formed at an upper other side of the first upper cap, and the first 1-1 upper opening is a first -1 provides a cell separator characterized in that it is opened and closed by the upper opening and closing part, and the first and second upper opening parts are opened and closed by the first and second upper opening and closing parts.

In addition, a cell separator is formed, wherein a male thread is formed along the outer periphery of the first lower opening portion, and a female thread is formed so as to be screwed in the state where the male thread is inserted along the inner periphery of the second upper opening portion. to provide.

In addition, the mixture material provides a cell separator characterized in that the stromal cells extracted from adipose tissue and the enzyme are mixed substances.

In addition, the lower side of the first body is formed such that the diameter becomes narrower toward the lower end, the first lower opening portion is formed to open downwardly at the lower end of the first body, and the lower side of the second body becomes smaller toward the lower end. It is formed to be narrowed, the second lower opening is formed to open downward to the lower end of the second body, a first lower cap is detachably coupled to the first lower opening, and the second lower opening is It provides a cell separator characterized in that the second lower cap is detachably coupled to.

In addition, the first lower cap, the first lower plate having a larger diameter than the first lower opening, and the first lower plate is integrally formed to open and close the first lower opening in the center of the first lower plate It includes a lower opening and closing portion, the second lower cap, the second lower plate having a larger diameter than the second lower opening, and the second lower opening integrally to open and close the second lower opening in the center of the second lower plate When the first lower opening and closing portion is coupled to the first lower opening portion in a state in which the second lower opening and closing portion is formed, and the first main body and the second main body are separated from each other, the first lower plate may be the first lower plate. When the second lower opening and closing portion is coupled to the second lower opening, the second lower plate provides a cell separator characterized in that the second main body is supported so as not to collapse.

In addition, the first lower plate and the second lower plate provide a cell separator, characterized in that formed in a circular or polygonal plate shape.

In addition, the upper side of the second body is formed so that the diameter becomes narrower toward the upper end, the second upper opening portion is formed to open upwardly at the upper end of the second body, and the agent separated from the first upper opening portion 2 A second upper cap is provided to open and close the upper open portion, and the second upper cap has a second upper plate having a larger diameter than the second upper open portion, and the second upper portion in the center of the second upper plate. It includes a second upper opening and closing portion integrally formed to open and close the opening, the second upper opening and closing portion is coupled to the second upper opening portion when the second upper plate is supported on the ground, the second upper The plate provides a cell separator characterized in that the second body is supported so as not to fall.

In addition, before the centrifugation is performed, the second lower cap is coupled to close the second lower opening, and when the centrifugation is completed and the second body is separated from the first body, the second The lower cap provides a cell separator characterized in that the second lower opening is opened so that the stem cells separated and stacked on the second body are discharged to the outside.

In addition, in the method of separating cells using a cell separator, the present invention includes the first lower opening part and the second upper opening part in a state where the second lower cap is coupled to the second lower opening part of the second body. A first step in which the additions are combined; A second step in which the mixture material is injected through the opened first upper opening; A third step in which the opened first upper opening is closed by the first upper cap; A fourth step of performing primary centrifugation to separate and stack supernatant on the first body, and to separate and stack stem cells on the second body; And a fifth step in which the first lower opening portion and the second upper opening portion are separated, and the supernatant located in the first body is not discharged downwardly through the first lower opening portion through the first lower opening portion by surface tension. It provides a cell separation method using a cell separator characterized in that it comprises a.

In addition, the sixth step of disposing of the first body; A seventh step of coupling the first lower opening of the new first body to the second upper opening of the second body; An eighth step of injecting physiological saline into the first upper open portion of the opened first body; A ninth step in which a first upper cap is coupled to the first upper open portion; A tenth step in which secondary centrifugation is performed to separate and stack supernatant on the first body, and to separate and stack stem cells on the second body; And an eleventh step in which the first lower opening portion and the second upper opening portion are separated, and the supernatant located in the first body is not discharged downwardly through the first lower opening portion through the first lower opening portion by surface tension. It provides a cell separation method using a cell separator characterized in that it further comprises.

In addition, a twelfth step of opening the first upper opening so that the supernatant located in the first body is discharged to the first lower opening; A thirteenth step of closing the first upper opening; A fourteenth step of coupling the first lower opening and the second upper opening; A fifteenth step of rotating the first body and the second body so that they are turned over; A sixteenth step of opening the second lower opening; A 17th step of injecting physiological saline into the opened second lower opening; 18th step of closing the second lower opening; A 19th step of rotating the first body and the second body so as to return to the original position; A 20th step of performing a third centrifugation to separate and stack supernatant on the first body, and to separate and stack stem cells on the second body; And the first lower opening portion and the second upper opening portion are separated, but the supernatant located in the first body is not discharged downwardly through the first lower opening portion through the first lower opening portion by the surface tension. ; And it provides a cell separation method using a cell separator, characterized in that it further comprises a step 22 to proceed to the 12th step to the 21st step n times.

In addition, a twelfth step of opening the first upper opening so that the supernatant located in the first body is discharged to the first lower opening; A thirteenth step of opening the second lower opening to extract stem cells located in the second body and then closing the second lower opening; A 14th step of combining the first body and the second body; A 15th step of injecting the stem cells and physiological saline extracted in the 13th step into the first body through the opened first upper opening; A 16th step in which the supernatant is separated and stacked on the first body and stem cells are separated and stacked on the second body by performing a third centrifugation; A 17th step in which the first lower opening portion and the second upper opening portion are separated, but the supernatant located in the first body is not discharged downward through the first lower opening portion through the first lower opening portion by a surface tension; And it provides a cell separation method using a cell separator, characterized in that it further comprises a step 18 to proceed to the 12th to 17th step n times.

In the present invention, when the first body and the second body are separated after centrifugation, the supernatant provided in the first body can be easily separated, so that the supernatant located in the first body does not need to be discharged using a pipette or the like. , It has the effect of safely extracting the stem cells located in the second body.

In addition, the upper cap or the lower cap is formed to support the body, there is an effect that does not need to use a separate mechanism to stand the body.

In addition, since the size of the first upper cap is variously configured, there is an effect of appropriately applying the first upper cap of various sizes according to the purpose.

1 is a view schematically showing a first body of a cell separator according to a preferred embodiment of the present invention.
2 is a view illustrating various examples of a first upper cap coupled to a first body of a cell separator according to a preferred embodiment of the present invention.
3 is a view schematically showing a second body of a cell separator according to a preferred embodiment of the present invention.
4 is a view schematically showing a state in which the first body and the second body of the cell separator according to a preferred embodiment of the present invention are mutually coupled.
5 is a view schematically showing a state in which the first body and the second body of the cell separator according to a preferred embodiment of the present invention are separated from each other.
6 is a view schematically illustrating another structure of a first upper cap coupled to a first body of a cell separator according to a preferred embodiment of the present invention.
7 is a diagram schematically showing a state in which the second body is coupled to the first body of FIG. 6.
8 is a view showing in detail a state in which the first lower opening portion and the second upper opening portion of FIG. 7 are combined.
9 is a view for explaining various examples of the first lower cap for upstanding the first body of the cell separator according to a preferred embodiment of the present invention.
10 is a view for explaining various examples of the second upper cap for upstanding the second body of the cell separator according to a preferred embodiment of the present invention.
11 is a view for explaining an example of a second lower cap for upstanding the second body of the cell separator according to a preferred embodiment of the present invention.

Hereinafter, a cell separator according to a preferred embodiment of the present invention and a cell separation method using the same will be described in more detail with reference to the accompanying drawings.

1 is a view schematically showing a first body of a cell separator according to a preferred embodiment of the present invention.

Referring to FIG. 1, the first body 100 is elongated in a cylindrical shape, for example, and a first space 102 is formed so that the mixture material is accommodated therein. The mixture may be, for example, a substance in which stromal cells extracted from adipose tissue and a collagenase enzyme are mixed, or a substance in which stromal vascular fraction (SVF) and physiological saline are mixed. The length of the first body 100 is composed of, for example, about 100 mm, the width of the inner diameter of the first body 100 is composed of about 35 mm, and the volume of the first body 100 is about 70 mm 2 to 80 mm. It may be ㏄, but it is natural that it is not limited. And the first upper opening portion 104 is formed so that the mixture material is injected into the first space 102 on the upper portion of the first body 100, the first lower opening portion on the lower portion of the first body 100 106 is formed to open. In addition, the upper portion of the first body 100 may be formed such that the width becomes narrower toward the upper portion to facilitate collection when liquid is discarded. The lower portion of the first body 100 is formed in a funnel shape so that the width becomes narrower toward the lower direction, so that the stem cells do not stagnate in the first body 100 during centrifugation and easily flow into the second body 200 direction. It is preferably configured to be.

In addition, the first upper cap 110 may be detachably coupled to the first upper open portion 104, and the first lower cap 140 may be detachably coupled to the first lower open portion 106. . The first upper cap 110 is formed in a lid shape to open or close the first upper opening portion 104, and the first lower cap 140 is shaped like a lid to open or close the first lower opening portion 106. Is formed into.

On the other hand, in the embodiment of the present invention, the cross sections of the first body 100 and the first upper and lower caps 110 and 140 are formed in a circular shape, but it is not limited to this, it is natural, and in some cases an ellipse, polygon, etc Of course, it may be formed in various ways.

2 is a view illustrating various examples of a first upper cap coupled to a first body of a cell separator according to a preferred embodiment of the present invention.

Referring to (a) of FIG. 2, the first upper opening portion 104 is formed to have a size large enough to easily inject the mixture, and the first upper cap 110 is the first upper opening portion 104 ) It is formed in a large size to open and close.

Referring to (b) of FIG. 2, the first upper opening portion 104a is formed relatively small, and the first upper cap 110a is also formed small to open and close the first upper opening portion 104a.

Referring to (c) of FIG. 2, the first upper open portion 104 is formed in an intermediate size, and the first upper cap 120 is formed in an intermediate size to open and close the first upper open portion 104. In addition, the first upper opening 120a is formed to be opened at the upper portion of the first upper cap 120, and the first upper opening 120a is opened and closed by the first upper opening and closing portion 122.

As described above, the present invention has a first upper cap which is variously formed, and thus has an effect of appropriately applying the first upper cap of various sizes according to the purpose.

3 is a view schematically showing a second body of a cell separator according to a preferred embodiment of the present invention.

Referring to FIG. 3, the second body 200 is formed in a cylindrical shape, for example, and a second space 202 is formed therein. In addition, the second upper opening 204 is formed on the upper portion of the second body 200, and the second lower opening portion 206 is formed on the lower portion of the second body 200. The inner body width of the second body 200 is composed of about 35 mm, the volume of the second body 200 may be about 7 mm 2 to 10 mm 2, but it is not limited.

In addition, the lower portion of the second body 200 is preferably formed in a funnel shape so that the width becomes narrower as it goes downward, so that the stem cells introduced in the direction of the second body 200 can be easily discharged to the lower portion. That is, before centrifugation is performed, the second lower cap 230 is coupled to close the second lower opening portion 206, and the centrifugation is completed, so that the second main body 200 is the first main body 100. When separated from, the second lower cap 230 opens the second lower opening portion 206 so that the stem cells separated and stacked on the second body 200 are easily discharged to the outside.

In addition, the second upper cap 210 may be detachably coupled to the second upper open portion 204, and the second lower cap 230 may be detachably coupled to the second lower open portion 206. . The second upper cap 210 is formed in a lid shape to open or close the second upper opening portion 204, and the second lower cap 230 is shaped like a lid to open or close the second lower opening portion 206. Is formed into.

4 is a view schematically showing a state in which the first body and the second body of the cell separator according to the preferred embodiment of the present invention are mutually coupled, and FIG. 5 is a first view of the cell separator according to the preferred embodiment of the present invention It is a diagram schematically showing a state in which the main body and the second main body are separated from each other.

First, referring to FIG. 4, when the first lower opening portion 106 is mutually coupled with the second upper opening portion 204 inserted therein, the first body 100 and the second body 200 are coupled. . At this time, a male thread is formed along the outer periphery of the first lower opening portion 106, and a female thread is formed to be screwed in a state where the male thread is inserted along the inner periphery of the second upper opening portion 204. The lower opening portion 106 and the second upper opening portion 204 are screwed together firmly. In addition, the mixture material is injected into the first body 100 through the first upper opening 104 while the second lower cap 230 is closed to the second lower opening 206. Subsequently, after the first upper cap 110 closes the first upper opening portion 104, and then proceeds to centrifugation, the supernatant is separated and stacked on the first body 100, and the second body 200 ), the stem cells are stacked separately.

Subsequently, referring to FIG. 5, after centrifugation is completed, when the first lower opening portion 106 and the second upper opening portion 204 are separated, the second body 200 is separated from the first body 100. do. The diameter of the first lower opening portion 106 is formed to be small in size such that the supernatant located in the first body 100 cannot be discharged downward by surface tension. The diameter of the first lower opening portion 106 may be, for example, about 5 mm. The surface tension is the tension acting on the surface of the liquid and corresponds to the scientific principle. That is, in a state in which the first main body 100 and the second main body 200 are separated, and the first upper open portion 104 is closed by the first upper cap 110, the first lower open portion 106 Even if the first lower cap 140 is not separately mounted, the supernatant liquid located in the first body 100 is not discharged to the lower portion arbitrarily by the surface tension. Accordingly, when the first body 100 is separated from the second body 200 after centrifugation is performed, the supernatant provided in the first body 100 can be easily separated. As described above, since the present invention does not need to discharge the supernatant located in the first body 100 using a pipette after centrifugation, the effect of safely extracting stem cells located in the second body 200 is obtained. have.

On the other hand, the opened first upper opening 104 is of sufficient size to allow the liquid contained in the first body 100 to be discharged to the outside, regardless of the surface tension of the liquid.

6 is a view schematically illustrating another structure of a first upper cap coupled to a first body of a cell separator according to a preferred embodiment of the present invention.

Referring to FIG. 6, the first upper cap 130 is formed in the shape of a lid to cover the first upper opening 104. In addition, a first 1-1 upper opening 130a is formed through an opening shape on an upper side of the first upper cap 130, and a first 1-2 upper opening 130b is formed on the other upper side of the first upper cap 130. It is formed through an opening shape. The diameter of the 1-1 upper opening 130a and the 1-2 upper opening 130b may be, for example, about 5 mm. And the 1-1 upper opening 130a is opened and closed by the 1-1 upper opening and closing portion 132, and the first 1-2 upper opening 130b is opened and closed by the first 1-2 upper opening and closing portion 134. The 1-1 upper opening 130a and the 1-2 1-2 upper opening 130b are combined with a separate medical syringe (not shown) to be used as an inlet for injecting fat, water, or other solutions, or after centrifugation. It can be used as an outlet to discard the supernatant.

7 is a diagram schematically showing a state in which the second body is coupled to the first body of FIG. 6.

Referring to FIG. 7, the second upper open portion 204 is coupled to the first lower open portion 106. At this time, an o-ring (r2) made of rubber is provided on the outer periphery of the first lower opening portion 106 to seal the watertightness between the first lower opening portion 106 and the second upper opening portion 204. Are combined. And the second lower cap 230 is coupled to the second lower opening portion 206. At this time, a rubber-shaped O-ring (r5) is provided on the outer periphery of the second lower opening 206, and is tightly coupled between the second lower opening 206 and the second lower cap 230. And the first upper cap 130 is coupled to the first upper opening 104. At this time, the O-ring (r1) is provided in the first upper opening portion 104, and is tightly coupled between the first upper opening portion 104 and the first upper cap 130.

8 is a view showing in detail a state in which the first lower opening portion and the second upper opening portion of FIG. 7 are combined.

Referring to FIG. 8, the distance between the inner circumference and the outer circumference of the first lower opening portion 106, that is, the thickness is configured to be thicker than the thickness of the first body 100. In addition, the distance between the inner peripheral edge and the outer peripheral edge of the second upper opening portion 204, that is, the thickness is configured to be thicker than the thickness of the second main body 200. As such, since the thicknesses of the first lower opening portion 106 and the second upper opening portion 204 are thick, the first lower opening portion 106 and the second upper opening portion 204 have a solid strength. 1, the coupling structure between the lower opening portion 106 and the second upper opening portion 204 is solidified.

9 is a view for explaining various examples of the first lower cap for upstanding the first body of the cell separator according to a preferred embodiment of the present invention.

Referring to (a) of FIG. 9, in a state where the first body 100 and the second body 200 are separated from each other, the first lower cap 150 has a larger diameter than the first lower opening portion 106. A first lower plate 152 having a first lower border portion 153 protruding upward along the rim of the first lower plate 152, and a first lower portion in the center of the first lower plate 152 It includes a first lower opening and closing portion 154 that is integrally formed to open and close the opening portion 106. The interior of the first lower plate 152 is configured to form an empty space. The O-ring r3 is interposed at the inner periphery of the first lower opening and closing portion 154. And in the state that the first body 100 and the second body 200 are separated from each other, when the first lower opening and closing portion 154 is coupled to the first lower opening portion 106, the first lower plate 152 is the first The body 100 is supported so as not to fall. Thus, since the first lower cap 140 is formed to support the first body 100, an effect of not having to use a separate mechanism to stand the first body 100 formed to be narrower in width toward the lower side There is. In addition, the first lower opening portion 106 and the first lower opening and closing portion 154 are interposed with O-rings r2 and r3, respectively, so that the first lower opening portion 106 and the first lower opening and closing portion 154 are watertight with each other. Combined.

Referring to (b) of FIG. 9, the first lower cap 150 has a first lower plate 152 having a larger diameter than the first lower opening portion 106 and a center of the first lower plate 152 It includes a first lower opening and closing portion 154 integrally formed to open and close the first lower opening portion (106). The inside of the first lower plate 152 is configured such that no empty space is formed, and the first lower plate 152 has a more rigid strength, so that the first main body 100 can be stably erected.

10 is a view for explaining various examples of the second upper cap for upstanding the second body of the cell separator according to a preferred embodiment of the present invention.

Referring to (a) of FIG. 10, the second upper cap 220 is in the center of the second upper plate 222 and the second upper plate 222 having a larger diameter than the second upper opening 204. It includes a second upper opening and closing portion 224 that is integrally formed to open and close the second upper opening portion (204). The interior of the second upper plate 222 is configured to form an empty space. Then, after the first main body 100 and the second main body 200 are separated from each other, after the second upper opening and closing part 224 is coupled to the second upper open part 204, the second main body 200 is followed. When pinched, the second upper plate 222 supports the second body 200 so that it does not collapse.

Referring to (b) of FIG. 10, the second upper cap 220 is in the center of the second upper plate 222 and the second upper plate 222 having a larger diameter than the second upper opening 204. It includes a second upper opening and closing portion 224 that is integrally formed to open and close the second upper opening portion (204). The inside of the second upper plate 222 is configured such that no empty space is formed, so that the second upper plate 222 has a more rigid strength, so that the inverted second body 200 can be stably erected.

11 is a view for explaining an example of a second lower cap for upstanding the second body of the cell separator according to a preferred embodiment of the present invention.

Referring to FIG. 11, the second lower cap 240 includes a second lower plate 242 having a larger diameter than the second lower opening 206, and a second lower portion in the center of the second lower plate 242. It includes a second lower opening and closing portion 244 is integrally formed to open and close the opening portion 206. When the second lower opening and closing portion 244 is coupled to the second lower opening portion 206, the second lower plate 242 can stably erect the second main body 200. In addition, an O-ring r4 is interposed in the second lower opening and closing portion 244, and the second lower opening portion 206 and the second lower opening and closing portion 244 are tightly coupled to each other.

Hereinafter, an example of a cell separation method using a cell separator according to a preferred embodiment of the present invention will be described.

Example 1-Stem cell extraction

In the state in which the second lower cap 230 is coupled to the second lower opening portion 206 of the second body 200, the opened first lower opening portion 106 and the second upper opening portion 204 are mutually mutually connected. After bonding, the mixture was injected through the opened first upper opening 104. The mixture is composed of a mixture of stromal cells extracted from adipose tissue and collagenase enzyme. Thereafter, the opened first upper opening 104 was closed using the first upper cap 110. Thereafter, the first and second bodies 100 and 200 are placed in a shaker, reacted at 37°C for 35 minutes, and then subjected to the first centrifugation to separate the supernatant from the first body 100. The cells were stacked, and the stem cells, that is, the stromal vascular fraction (SVF) were separated and stacked on the second body 200. Thereafter, the first lower opening portion 106 and the second upper opening portion 204 were separated. At this time, the supernatant located in the first body 100 is not discharged to the lower side of the first body 100 through the first lower opening portion 106 by the surface tension.

In the conventional enzyme treatment process, when removing unnecessary supernatant using centrifugation, the operator must elaborately remove the supernatant using a pipette, which takes a very long time and there is a risk of infection. On the other hand, according to the present invention, only the operation of separating the first main body 100 and the second main body 200 removes the supernatant, thereby effectively removing the supernatant in a simple process.

Example 2-Stem cell washing

The first main body 100 separated from the second main body 200 in Salsi Example 1 is disposed of, and the first first body 100 of the new first body 100 in the second upper opening 204 of the second main body 200 is disposed. After combining the lower opening portion 106, physiological saline was injected into the first upper opening portion 104 of the opened first body 100. Thereafter, after the first upper cap 110 is coupled to the first upper opening 104, the second centrifugation is performed to separate and stack the supernatant in the first body 100, and the second body 200 ) To separate and stack the stem cells. Thereafter, the first lower opening portion 106 and the second upper opening portion 204 were separated. At this time, the supernatant located in the first body 100 is not discharged to the lower side of the first body 100 through the first lower opening portion 106 by the surface tension.

In this way, the present invention does not use much of the first and second bodies 100 and 200 in the washing process, thus simplifying the operation and reducing the cost.

On the other hand, the stem cells separated in the second body 200 may be recovered and used as they are, but if higher purity stem cells are desired, the following reprocessing (1) or preparation (2) may be further performed. Hereinafter, the reprocessing (1) process will be described, and then the reprocessing (2) process will be described.

Example 3-Stem cell reprocessing (2)

In the example 2, the first upper opening 104 of the first main body 100 separated from the second main body 200 is opened, so that the supernatant located in the first main body 100 is the first lower opening 106. After being discharged to, the first upper opening 104 was closed. Thereafter, after combining the first lower opening portion 106 and the second upper opening portion 204, the first main body 100 and the second main body 200 were rotated 180 degrees to be turned over. Subsequently, the second lower opening 206 was opened, and after the physiological saline was injected into the opened second lower opening 206, the second lower opening 206 was closed. Thereafter, after the first main body 100 and the second main body 200 are reversely rotated to return to the original position, the third centrifugation is performed to separate the supernatant from the first main body 100 and stack the second body. The stem cells were separated and stacked at 200. Thereafter, the first lower opening portion 106 and the second upper opening portion 204 were separated. At this time, the supernatant located in the first body 100 is not discharged to the lower side of the first body 100 through the first lower opening portion 106 by the surface tension. If another reprocessing is desired, the above-described process can be repeated.

And after the reprocessing is completed, the stem cells located in the second body 200 are extracted to the outside or only the second body 200 is stored separately.

On the other hand, the stem cells separated in the second body 200 may be recovered and used as it is, but if more high-purity stem cells are desired, the following reprocessing process may be further performed. Hereinafter, the reprocessing process will be described.

Example 4-Stem cell reprocessing (2)

In the example 2, the first upper opening 104 of the first main body 100 separated from the second main body 200 is opened, so that the supernatant located in the first main body 100 is the first lower opening 106. After being discharged to, the second lower open portion 206 was opened to extract the stem cells located in the second body 200 and then the second lower open portion 206 was closed. Thereafter, after combining the first body 100 and the second body 200, the stem cells and physiological saline extracted above were injected into the first body 100 through the opened first upper opening 104. . Thereafter, the third centrifugation was performed to separate the supernatant from the first body 100 and stack the stem cells separately from the second body 200. Thereafter, the first lower opening portion 106 and the second upper opening portion 204 were separated. At this time, the supernatant located in the first body 100 is not discharged to the lower side of the first body 100 through the first lower opening portion 106 by the surface tension. If another reprocessing is desired, the above-described process can be repeated again.

Although the present invention has been described in detail in the above embodiments, it is obvious that the present invention is not limited thereto, and it is apparent to those skilled in the art that various modifications and modifications are possible within the technical scope of the present invention. If it falls within the scope of the claims, the technical idea should also be regarded as belonging to the present invention.

100: first body 102: first space
104: first upper opening 104a: first upper opening
106: first lower opening portion 110: first upper cap
110a: first top cap 120: first top cap
120a: first upper opening 122: first upper opening
130: first upper cap 130a: 1-1 upper opening
130b: first 1-2 upper opening 132: 1-1 upper opening
134: first 1-2 upper opening and closing part 140: first lower cap
150: first lower cap 152: first lower plate
153: first lower border portion 154: first lower opening and closing portion
200: second body 202: second space
204: second upper opening portion 206: second lower opening portion
210: second upper cap 220: second upper cap
222: second upper plate 224: second upper opening and closing portion
230: second lower cap 240: second lower cap
242: second lower plate 243: second lower border portion
244: second lower opening and closing portion r1, r2, r3, r4, r5: O-ring

Claims (15)

A first body in which a first space is formed to accommodate the mixture material, and a first upper opening part is formed so that the mixture material is injected, and a first lower opening part is formed at the lower part;
A first upper cap opening or closing the first upper opening;
A second body in which a second space is formed, and a second upper opening is formed so that the upper portion can be coupled to the first lower open portion, and a second lower opening is formed at the lower portion; And
And a second lower cap opening or closing the second lower opening portion,
The first upper cap and the second lower cap respectively close the first upper opening portion and the second lower opening portion, and in the state where the first lower opening portion and the second upper opening portion are mutually coupled, centrifugal separation When proceeding, the supernatant is separated and laminated on the first body, the stem cells are separated and laminated on the second body,
When the centrifugation is completed, the second body is separated from the first body, and the diameter of the first lower opening is formed to a size in which the supernatant located in the first body cannot be discharged downward by surface tension. Cell separator, characterized in that.
According to claim 1,
The lower portion of the first body is characterized in that the cell separator is formed so that the width becomes narrower as it approaches the second body.
According to claim 1,
A cell separator further comprising a first upper opening formed on an upper portion of the first upper cap, and a first upper opening and closing portion for opening and closing the first upper opening.
According to claim 1,
A 1-1 upper opening is formed on an upper side of the first upper cap, and a 1-2 upper opening is formed on an upper other side of the first upper cap, and the 1-1 upper opening is 1-1. A cell separator characterized in that it is opened and closed by the upper opening and closing part, and the first and second upper opening parts are opened and closed by the first and second upper opening and closing parts.
According to claim 1,
A cell separator characterized in that a female screw thread is formed along the outer periphery of the first lower opening portion, and a female screw thread is formed so as to be screwed with the male screw thread inserted along the inner periphery of the second upper opening portion.
According to claim 1,
The mixture material is a cell separator, characterized in that the material is a mixture of matrix cells and enzymes extracted from adipose tissue.
According to claim 1,
The lower side of the first body is formed to be narrower in diameter toward the lower end, and the first lower opening portion is formed to open downward to the lower end of the first body,
The lower side of the second body is formed so that the diameter becomes narrower toward the lower end, and the second lower opening portion is formed to open downwardly at the lower end of the second body,
A cell separator characterized in that a first lower cap is detachably coupled to the first lower opening, and a second lower cap is detachably coupled to the second lower opening.
The method of claim 7,
The first lower cap has a first lower plate having a larger diameter than the first lower opening, and a first lower opening and closing part integrally formed to open and close the first lower opening in the center of the first lower plate. Including,
The second lower cap has a second lower plate having a larger diameter than the second lower opening, and a second lower opening and closing part integrally formed to open and close the second lower opening in the center of the second lower plate. Including,
In a state in which the first body and the second body are separated from each other, when the first lower opening and closing portion is coupled to the first lower opening, the first lower plate supports the first body so that it does not collapse, and the second When the lower opening and closing portion is coupled to the second lower opening, the second lower plate is a cell separator, characterized in that to support the second body from falling.
The method of claim 8,
The first lower plate and the second lower plate is a cell separator, characterized in that formed in a circular or polygonal plate shape.
According to claim 1,
The upper side of the second body is formed such that the diameter becomes narrower toward the upper end, and the second upper opening part is formed to open upwardly to the upper end of the second body,
A second upper cap is provided to open and close the second upper open portion separated from the first upper open portion,
The second upper cap has a second upper plate having a larger diameter than the second upper open portion, and a second upper opening and closing portion integrally formed to open and close the second upper open portion in the center of the second upper plate. Including, but when the second upper plate is supported on the ground in a state where the second upper opening and closing portion is coupled to the second upper opening, the second upper plate is characterized in that it supports the second body so that it does not fall over. Cell separator.
According to claim 1,
Before the centrifugation proceeds, the second lower cap is coupled to close the second lower opening,
When the centrifugation is completed and the second body is separated from the first body, the second lower cap opens the second lower opening so that the stem cells separated and stacked on the second body are discharged to the outside. Cell separator characterized by.
In the cell separation method using the cell separator of claim 1,
A first step in which the first lower opening portion and the second upper opening portion are coupled while the second lower cap is coupled to the second lower opening portion of the second body;
A second step in which the mixture material is injected through the opened first upper opening;
A third step in which the opened first upper opening is closed by the first upper cap;
A fourth step of performing primary centrifugation to separate and stack supernatant on the first body, and to separate and stack stem cells on the second body; And
A fifth step in which the first lower opening portion and the second upper opening portion are separated, but the supernatant located in the first body is not discharged downwardly through the first lower opening portion through the first lower opening portion by surface tension. Cell separation method using a cell separator comprising a.
The method of claim 12,
A sixth step of disposing of the first body;
A seventh step of coupling the first lower opening of the new first body to the second upper opening of the second body;
An eighth step of injecting physiological saline into the first upper open portion of the opened first body;
A ninth step in which a first upper cap is coupled to the first upper open portion;
A tenth step in which secondary centrifugation is performed to separate and stack supernatant on the first body, and to separate and stack stem cells on the second body; And
The eleventh step of separating the first lower opening portion and the second upper opening portion, wherein the supernatant located in the first body is not discharged downward through the first lower opening portion through the first lower opening portion by surface tension. Cell separation method using a cell separator characterized in that it further comprises.
The method of claim 13,
A twelfth step of opening the first upper opening portion so that the supernatant located in the first body is discharged to the first lower opening portion;
A thirteenth step of closing the first upper opening;
A fourteenth step of coupling the first lower opening and the second upper opening;
A fifteenth step of rotating the first body and the second body so that they are turned over;
A sixteenth step of opening the second lower opening;
A 17th step of injecting physiological saline into the opened second lower opening;
18th step of closing the second lower opening;
A 19th step of rotating the first body and the second body so as to return to the original position;
A 20th step of performing a third centrifugation to separate and stack supernatant on the first body, and to separate and stack stem cells on the second body; And
A 21st step in which the first lower opening portion and the second upper opening portion are separated, and the supernatant located in the first body is not discharged downwardly through the first lower opening portion by the surface tension; And
Cell separation method using a cell separator, characterized in that it further comprises a step 22 to proceed to the 12th to 21st step n times.
The method of claim 14,
A twelfth step of opening the first upper opening portion so that the supernatant located in the first body is discharged to the first lower opening portion;
A thirteenth step of opening the second lower opening to extract stem cells located in the second body and then closing the second lower opening;
A 14th step of combining the first body and the second body;
A 15th step of injecting the stem cells and physiological saline extracted in the 13th step into the first body through the opened first upper opening;
A 16th step in which the supernatant is separated and stacked on the first body and stem cells are separated and stacked on the second body by performing a third centrifugation;
A 17th step in which the first lower opening portion and the second upper opening portion are separated, but the supernatant located in the first body is not discharged downward through the first lower opening portion through the first lower opening portion by a surface tension; And
Cell separation method using a cell separator, characterized in that it further comprises a step 18 to proceed to the 12th to 17th step n times.

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