VESSEL FOR CENTRIFUGE, AND SVF ISOLATION METHOD USING SAME

Abstract

The present invention provides a vessel used for a centrifuge, a centrifugation method using same, and a method for stromal vascular fraction (SVF) isolation using same. In particular, the centrifuge vessel according to the present invention has a rotatable body unit (300) provided with a side accommodation part (310) which is on the side thereof and formed by extending at least a part thereof radially outwards and on which is positioned a plate (150) having a radial end positioned therein. As a result, the present invention substantially eliminates a stirring effect, has high isolation efficiency, and obtains a superior SVF yield.

Claims

1. A centrifuge vessel comprising: a body unit 300 that is rotatable; a cover part 200 that covers the body unit 300; and a connection part 100 provided in the cover part 200 and comprising a plurality of connection pipes 111 and 112 in fluid communication with an inside of the body unit 300, wherein the body unit 300 comprises a side accommodation part 310 formed on a side surface of the body unit 300 by extending at least a part thereof radially outwards, and a plate 150 having a radial end positioned on the side accommodation part 310.

2. The centrifuge vessel according to claim 1, wherein the side accommodation part 310 extends radially in all directions.

3. The centrifuge vessel according to claim 2, wherein the plate 150 is a body of revolution such that the stirring effect does not occur even when the plate 150 is not rotated during rotation of the body unit 300.

4. The centrifuge vessel according to claim 3, wherein a side flow path 152 in fluid communication with one connection pipe 112 of the plurality of connection pipes 111 and 112 is provided inside the plate 150.

5. The centrifuge vessel according to claim 4, wherein an end of the other connection pipe 111 of the connection pipes 111 and 112 reaches the lower body 303 of the body unit 300.

6. The centrifuge vessel according to claim 5, wherein the connection part 100 further comprises a connection pipe housing 115, the other connection pipe 111 of the plurality of connection pipes 111 and 112 reaches the lower body 303 of the body unit 300 through a central flow path 120, a part of the central flow path 120 is provided inside the connection pipe housing 115, the one connection pipe 112 is in fluid communication with the side flow path 152 through a space between the connection pipe housing 115 and the central flow path 120, and a side flow path opening 151 connecting a space between the connection pipe housing 115 and the central flow path 120 to the side flow path 152 is disposed at a center of the plate 150.

7. The centrifuge vessel according to claim 6, wherein outside of the centrifuge vessel and inside of the side accommodation part 310 are in fluid communication through the connection pipe 112 and the side flow path 152.

8. The centrifuge vessel according to claim 7, wherein a substance accommodated in the side accommodation part 310 is discharged to the outside of the centrifuge vessel through the one connection pipe 112 and the side flow path 152, and a substance outside the centrifuge vessel is introduced into the side accommodating part 310.

9. The centrifuge vessel according to claim 8, wherein a diameter of an upper portion of the connection pipe housing 115 is smaller than a diameter of a lower portion of the connection pipe housing 115, the central flow path 120 is inserted into and coupled with the upper portion of the connection pipe housing 115, and the side flow path opening 151 is inserted into and coupled with the lower portion of the connection pipe housing 115.

10. The centrifuge vessel according to claim 1, wherein the side accommodation part 310 extends radially outwards of an upper portion, a middle portion, or a lower portion of the body unit 300.

11. The centrifuge vessel according to claim 10, wherein the body unit 300 comprises an upper body 301, a middle body 302, and a lower body 303, a diameter of the upper body 301 gradually increases in an upward direction, and the side accommodating part 310 is a radially outer part of the upper body 301 with an increased diameter.

12. The centrifuge vessel according to claim 11, wherein the diameter of the lower body 303 gradually decreases in a downward direction, a plurality of stirring members 360 are positioned on an inner surface of the lower body 303, and the plurality of stirring members 360 are arranged to form a plurality of rows oriented toward a center of rotation axis of the body unit 300.

13. A centrifugation method using the centrifuge vessel according to claim 3, comprising steps of: separating a substance inside the centrifuge vessel as the body unit 300 is rotated by a rotation of the centrifuge vessel, which is accumulated in the side accommodating part 310, during which the plate 150 is not rotated when the body unit 300 is rotated, thus preventing a stirring effect of the substance that is separated and accumulated in the side accommodating part 310.

14. An stromal vascular fraction (SVF) isolation method using the centrifugation method according to claim 13, comprising steps of: (a) introducing adipose tissue into the body unit 300 through a first connection pipe 111 of the plurality of connection pipes 111 and 112; (b) supplying the washing solution through the first connection pipe 111 and centrifuging by the centrifugation method to separate the adipose tissue into a washed adipose tissue and a contaminant; (c) removing contaminants from the body unit 300 through the first connection pipe 111; (d) supplying an enzyme through the first connection pipe 111 and centrifuging by the centrifugation method to separate the washed adipose tissue into a digested adipose tissue and a primary aqueous solution; (e) removing the digested adipose tissue from the body unit 300 through the second connection pipe 112 of the plurality of connection pipes 111 and 112; (f) separating a primary aqueous solution into SVF and a secondary aqueous solution by supplying a washing solution through the first connection pipe 111, stirring, and centrifuging by the centrifugal separation method, wherein the SVF is adhered to the side accommodation part 310; (g) removing the secondary aqueous solution from the body unit 300 through the second connection pipe 112; (h) precipitating the SVF adhered to the side accommodating part 310 by inertia to the lower body 303 by supplying a small amount of washing solution through the first connection pipe 111 and then repeating rotating and stopping the centrifuge vessel; and (i) extracting the SVF precipitated in the lower body 303 through the first connection pipe 111.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0041] The accompanying drawings illustrate a preferred embodiment of the present disclosure and together with the foregoing disclosure, serve to provide further understanding of the technical features of the present disclosure, and thus, the present disclosure is not construed as being limited to the drawing.

[0042] FIG. 1 is a perspective view of a centrifuge vessel according to the present disclosure.

[0043] FIG. 2 is an exploded perspective view of a centrifuge vessel according to the present disclosure.

[0044] FIG. 3 is a cross-sectional view of a centrifuge vessel taken along line A-A of FIG. 1 according to the present disclosure.

[0045] FIG. 4 is a perspective view of a connection part 100 of a centrifuge vessel according to the present disclosure.

[0046] FIG. 5 is a perspective view of a part of a connection part 100 of the centrifuge vessel according to the present disclosure.

[0047] FIG. 6 is a cross-sectional perspective view of a part of a connection part 100 of a centrifuge vessel according to the present disclosure.

[0048] FIGS. 7 and 8 are perspective views of a centrifuge vessel of another embodiment of the present disclosure.

[0049] FIGS. 9 and 10 are conceptual views for explaining the characteristics for eliminating the stirring effect of a centrifuge vessel according to the present disclosure.

[0050] FIG. 11 is a flowchart for explaining an isolation method of SVF using a centrifuge vessel according to the present disclosure.

[0051] FIG. 12 is a schematic view for explaining an isolation method of SVF using a centrifuge vessel according to the present disclosure.

[0052] FIGS. 13 to 15 are the results of a verification test of comparing between SVF isolation using a centrifuge vessel according to the present disclosure and SVF isolation using conventional instruments.

BEST MODE

[0053] Hereinafter, a centrifuge vessel according to the present disclosure will be described with reference to the drawings.

1. Description of Centrifuge Vessel

[0054] The centrifuge vessel according to the present disclosure is mounted in a centrifuge. The centrifuge generally includes a power member that provides power for rotation, a plurality of bags connected to the centrifuge vessel to accommodate the substances to be separated and the additives, and the separated substances or contaminants therein, a control unit for controlling the operation, a display device for outputting the status of operation, and so on. The centrifuge vessel according to the present disclosure can be applied to any kind of centrifuge, and therefore, the centrifuge will not be redundantly described herein.

[0055] Hereinafter, a centrifuge vessel according to the present disclosure will be described in detail with reference to FIGS. 1 to 6.

[0056] 1.1. Connection Part 100

[0057] The centrifuge vessel according to the present disclosure includes a connection part 100, a cover part 200, and a body unit 300.

[0058] A first connection pipe 111, a second connection pipe 112, a sealing member housing 113, a connection pipe housing 115, and a latching jaw 119 are positioned on an upper portion of the connection part 100. Although the connection part 100 is shown in the drawing as including two connection pipes 111 and 112, two or more connection pipes may be provided as well.

[0059] The plurality of connection pipes 111 and 112 provided on the connection part 100 are connected to the bags of the centrifuge by separate tubes or the like. With this configuration, the user may be able to feed the analyte or additive into the centrifuge vessel, or remove the separated substance or contaminants out of the centrifuge vessel.

[0060] A plurality of connection pipes 111 and 112 are in fluid communication with a space within the body unit 300, and the flow path configuration will be specifically described below.

[0061] The sealing member housing 113 has a sealing member 130 positioned therein to protect the sealing member 130 from the outside. The sealing member 130 may include a first sealing member 131, a second sealing member 132, and a third sealing member 133 (see FIG. 2), and may be made of an elastic member, a ceramic member, and a magnetic member, respectively, to increase tightness.

[0062] The connection pipe housing 115 includes a flow path through which a plurality of connection pipes 111 and 112 are in fluid communication with a space inside the body unit 300, and also protects the flow path from the outside.

[0063] The connection pipe housing 115 is configured such that an upper portion and a lower portion are stepped, and the first connection pipe 111 is connected to the upper portion and the second connection pipe 112 is connected to the lower portion. When the upper and lower portions are stepped, this means that the upper and lower inner diameters of the connection pipe housing 115 are configured differently from each other, which allows the center flow path 120 to be easily inserted into the upper portion, and also allows a protruding portion of the upper plate 150 having the side flow path opening 151 positioned therein to be inserted into the lower portion, and thus allows easy assembly, while maintaining the tightness (see FIG. 3).

[0064] The latching jaw 119 refers to a portion where the centrifuge is latched when the centrifuge vessel is mounted on the centrifuge, and it serves to help the centrifuge vessel to be set in position in the centrifuge. When the centrifuge vessel is mounted for use, user can easily mount the centrifuge by latching the centrifuge to the latching jaw 119, and can also easily separate the centrifuge by releasing the latching jaw 119.

[0065] The lower portion of the connection part 100 is inserted into the space within the body unit 300 and is sealed from the outside by the cover part 200 and the sealing member 130. A circular plate 150 is positioned under the connection part 100. A radial end of the plate 150 is positioned in the side accommodation part 310. In the drawing, the plate 150 is shown as having a circular plane, that is, the plate 150 itself is in a flat cylindrical shape, but any body of revolution may be used. This will be described in detail below.

[0066] A side flow path 152 is provided inside the plate 150 to provide a fluid communication through the second connection pipe 112.

[0067] One of the advantages of the plate 150 according to the present disclosure is that the side flow path 152 provided in the plate 150 can be directly connected to the side accommodation part 310. Accordingly, another advantages are obtained such that it is possible to discharge some of the substances sequentially precipitated in the side accommodation part 310 by the centrifugation process through the second connection pipe 112, and in the discharge process and the centrifugation process, the plate 150 as a body of revolution does not cause a stirring effect on the solution collected in the side accommodation part 310, so that the outermost precipitate layer not in contact with the discharge port is not stirred. This will be discussed in detail below with reference to FIGS. 9 and 10.

[0068] 1.2. Cover Part 200

[0069] The cover part 200 refers to a portion that is seated on the upper portion of the body unit 300.

[0070] An opening 210 is positioned at an upper center of the cover part 200. The opening 210 is configured such that the protrusion of the upper plate 150 having the side flow path positioned therein is inserted into and coupled with the opening 210, and the central flow path 120 is passed through the opening 210.

[0071] A sealing member seating part 230 is positioned radially outward of the opening 210, and the sealing member 130 is seated thereon.

[0072] A body connection part 240 protruding upward is positioned radially outward of the cover part 200. A groove is provided on a lower portion of the body connection part 240 and a cover connection part 340 of the body unit 300 is inserted therein, such that the durability of the centrifuge vessel is increased and separation of the cover part 200 is prevented during rotation.

[0073] 1.3. Body Unit 300

[0074] The body unit 300 may be divided into an upper body 301, a middle body 302, and a lower body 303 (see FIG. 3).

[0075] The side accommodation part 310 is positioned radially outward of the upper body 301. The side accommodation part 310 refers to a portion that protrudes at least radially outward of the body unit 300. During an operation of the centrifuge, substances separated in the centrifuge vessel begin to be accumulated naturally in the side accommodation part 310 by centrifugal force. When the rotation of the centrifuge is completed, substances of low viscosity will descend by gravity and flow to a lower accommodating part 330 of the lower body 303, while substance of high viscosity (e.g., SVF) remain in the side accommodating part 310.

[0076] Preferably, the wall of the side accommodation part 310 is radially and outwardly protruded and smoothly connected to the wall of the body unit 300. That is, it is preferable that the upper body 301 and the middle body 302 are connected smoothly. In this case, substances separated in the middle body 302 or the lower body 303 can be naturally delivered to the side accommodation part 310 of the upper body 301. While the diameter of the middle body 302 is shown as being constant in FIG. 3, the diameter may be different as long as the middle body 302 is smoothly connected to the upper body 301.

[0077] Further, it is preferable that the side accommodation part 310 extends radially outwards in all directions (that is, by 360 degrees). When only a part of the lobe is extended, it will take the form of the conventional lobe where the SVF is accommodated to some extent. However, when the lobe extends radially outwards in all directions, SVF can be accommodated regardless of directions where the SVF is separated by centrifugation, and as a result, superior yield rate can be expected.

[0078] The lower accommodating part 330 is positioned at an end of the lower body 303 and the central flow path 120 is connected to the inside of the lower accommodating part 330. With this configuration, the substances accumulated in the lower accommodating part 330 can be discharged to the outside of the centrifuge vessel (see FIGS. 12c and 12j).

[0079] A plurality of stirring members 360 are positioned on an inner surface of the lower body 303, i.e., on a bottom surface of the lower body 303. It is preferable that the plurality of stirring members 360 are arranged so as to form a plurality of rows toward the center of rotation axis of the body unit 300. FIG. 2 illustrates five stirring members 360 in each of three rows, but the number and arrangement may be varied.

[0080] Meanwhile, in the illustrated embodiment, the side accommodation part 310 extending radially outward is positioned in the upper body 301, but another embodiment is also possible in which the side accommodation part is positioned in the middle body or the lower body. In the above case, the plate should also be positioned in the middle body or the lower body where the side accommodation part is positioned.

[0081] 1.4. Flow Path

[0082] The flow path of the centrifuge vessel according to the present disclosure will be described with further reference to FIG. 3. As used herein, flow channel refers to a fluid communication path between the outside of the centrifuge vessel and the inside of the body unit 300.

[0083] The present disclosure adopts two independent channels.

[0084] The first flow path includes the first connection pipe 111 and the central flow path 120, and the end thereof is positioned at the lower accommodating part 330. The second flow path includes the second connection pipe 112, a radially outer region of the connection pipe housing 115 (that is, an outer region of the central flow path 120), a side flow path opening 151, and a side flow path 152. An end of the second flow path is positioned in the side accommodation part 310. In this example, the side flow path opening 151 is in a shape that is bent at 90 degrees to connect the second connection pipe 112 and the side flow path 152 (see FIGS. 5 and 6).

[0085] The first flow path and the second flow path are independent of each other. In particular, when viewed from the inside of the connection pipe housing 115, the central flow path 120 is positioned nearest to the center, thus constituting the first flow path, and the second flow path is formed on the outside of the wall of the central flow path 120 that serves as a boundary. Accordingly, the two flow paths are independent of each other.

2. Description of Assembling Method of Centrifuge Vessel

[0086] The centrifuge vessel according to the present disclosure is advantageous in that the vessel is designed to be easily assembled while having an excellent tightness, thus providing low difficulty of manufacturing and low manufacturing cost.

[0087] The assembling method will be described with reference to FIG. 2.

[0088] The upper portion of the connection part 100, which includes the plurality of connection pipes 111 and 112, the connection pipe housing 115, the latching jaw 119, and the sealing member housing 113, is integrally molded.

[0089] The first sealing member 131, the second sealing member 132, and the third sealing member 133 are separately molded due to different substances thereof, and then an upper surface of the second sealing member 132 is inserted into a lower surface of the first sealing member 131 and an upper surface of the third sealing member 133 is inserted into a lower surface of the second sealing member 132 to form an integrated sealing member 130.

[0090] The upper surface of the sealing member 130 (that is, the upper surface of the first sealing member 131) is inserted into the inner surface of the sealing member housing 113 and the lower surface of the sealing member 130 (that is, the lower surface of the third sealing member 133) is seated on the sealing member seating part 230 of the cover part 200. In this example, the upper surface of the sealing member 130 is sized and shaped to fit the inner surface of the sealing member housing 113, and the lower surface of the sealing member 130 is sized and shaped to fit the sealing member seating part 230, such that the sealing is maintained.

[0091] Next, the lower portion of the connection part 100 including the central flow path 120 and the plate 150 is inserted into and coupled with the assembly of the upper portion of the connection part 100, the sealing member 130 and the cover part 200 from below. Specifically, the central flow path 120 is inserted into the upper portion of the connection pipe housing 115, and the protruding portion of the upper plate 150 where the side flow path opening 151 is positioned is inserted into the lower portion of the connection pipe housing 115 (see FIG. 3).

[0092] Next, the body unit 300 is mounted on the cover part 200. That is, the cover connection part 340 of the body unit 300 is inserted into a lower portion of the body connection part 240 of the cover part 200 and fixed in position. A separate adhering or fusing may be further provided for the inserting and fixing.

3. Description of Other Embodiments

[0093] As shown in FIGS. 1 to 6, the centrifuge vessel according to the embodiment of the present disclosure described above has the side accommodation part 310 positioned in the upper portion of the main body 301.

[0094] As shown in FIG. 7, a centrifuge vessel according to another embodiment of the present disclosure has the side accommodation part 310 positioned in the middle portion of the main body 302.

[0095] As shown in FIG. 8, a centrifuge vessel according to still another embodiment of the present disclosure has the side accommodation part 310 positioned in the lower portion of the main body 303.

[0096] In all of the three embodiments shown in FIGS. 1, 7 and 8, the side accommodation part 310 has at least a part extending radially outwards, or preferably, the side accommodation part 310 extends radially outwards in all directions. Further, in all three embodiments, an end of the plate 150 is positioned in the side accommodation part 310.

4. Substantial Elimination of the Stirring Effect

[0097] The characteristic of a centrifuge vessel of eliminating the stirring effect according to the present disclosure will be described below with reference to FIGS. 9 and 10. FIGS. 9 and 10a illustrate the stirring effect shown in the related art (or in particular, in Korean Patent Publication No. 10-2016-0006076), and FIG. 10b illustrates an example of the present disclosure.

[0098] In a centrifuge vessel according to the present disclosure, the substance to be separated in the cover part 200, the body unit 300 and the body unit 300 are rotated as the centrifuge is operated. In this situation, the components included in the connection part 100 are not rotated. That is, the central flow path 120, the plate 150, and the side flow path 152 inside the plate 150 included in the connection part 100 are not rotated. This is because the connection part 100 is connected to a substance accommodating bag or the like of the centrifuge that supplies the substance through a tube (not shown) or the like.

[0099] This is also the case of the related art in which the plate 150 is not provided, and accordingly, as shown in FIG. 9a, the body unit 300 and the substance to be separated included therein are rotated, but the side flow path 152 is not rotated when the centrifuge is operated. When the process proceeds to some extent, the substances to be separated are separated into SVF or the like to form a multi-layered structure, and the substances in the multi-layered structure are rotated, but the side flow path 152 is not rotated.

[0100] As shown in FIG. 10a, in a relative observation of this from the perspective of the multiple layers separated, it means that only the side flow path 152 is rotated, while the multiple layers are in a stationary state. Accordingly, the side flow path 152 stirs and disturbs the multiple layers as it is repeatedly dipped into and removed from the multiple separated layers. This causes some of the separated substances including SVF to be re-mixed, which lowers the isolation efficiency.

[0101] However, in the embodiment of the present disclosure provided with the circular plate 150, different aspects are provided.

[0102] As shown in FIG. 10b, in a relative observation from the side of the multiple layers separated, only the plate 150 is rotated, while the multiple layers are in a stationary state. However, only the plate 150 is rotated due to its circular shape, in the same three-dimensional position. That is, the plate is not newly dipped or removed. Accordingly, the multiple layers are not stirred or disturbed, and the stirring effect in which the separated substances including the SVF are re-mixed again is substantially eliminated.

[0103] Meanwhile, the plate 150 for preventing the stirring effect does not necessarily have to be circular, and any shape may be used as long as it is a shape of a body of revolution. As used herein, the body of revolution means a figure that always has a constant shape on the cross-section including the rotation axis.

[0104] For example, while the plate 150 is circular in FIG. 10b, the cross-section from the side of the multiple layers is constant as a square.

[0105] The plate 150 may be a body of revolution having a triangular cross-section rather than a rectangular shape. In this case, the plate 150 itself may have a flat conical shape, and the stirring effect is still not occurred, which is preferable.

[0106] The plate 150 may be a body of revolution having a parallelepipedal cross-section. In this case, the plate 150 itself will be of an umbrella shape, and again, the stirring effect is not occurred, which is preferable.

5. Description of SVF Isolation Method Using Centrifuge Vessel

[0107] An SVF isolation method using a centrifuge vessel according to the present disclosure will be described below with reference to FIGS. 11 and 12.

[0108] FIGS. 12a to 12j show the respective steps described with reference to FIG. 11, in which corresponding reference numerals (S100 to S800) are added at the bottom of each drawing.

[0109] The adipose tissue is supplied into the body unit 300 through the first connection pipe 111 (S100).

[0110] Next, washing solution is supplied into the body unit 300 through the first connection pipe 111, and then the centrifuge is operated, rotating and stirring for a predetermined time (S200). Upon completion of the rotating and stirring, the washed adipose tissue and contaminants including blood or the like are separated into upper and lower layers, respectively. The lower layer of the separated contaminants are discharged to the exterior through the central flow path 120 whose end is positioned in the lower accommodating part 330, and removed (S300).

[0111] Next, an enzyme such as collagenase for example is supplied and the centrifuge is operated to rotate and stir for a predetermined time (S400). Upon completion of rotating and stirring, the adipose tissue and primary aqueous solution digested by the enzyme are separated radially inwards and outwards, respectively. The digested adipose tissues that are separated radially inwards are discharged to the outside through the side flow path 152 whose end is positioned in the side accommodation part 310, and removed (S500).

[0112] Then the washing solution is additionally supplied to the remaining primary aqueous solution, after which rotating and stirring are performed for a predetermined time (S600). Thus, the secondary aqueous solution and SVF are separated radially inwards and outwards, respectively. The secondary aqueous solution that is separated radially inwards is discharged to the outside through the side flow path 152 whose end is positioned in the side accommodation part 310, and removed (S700).

[0113] Accordingly, only the SVF remains in the side accommodation part 310, and it is remained positioned in the side accommodation part 310 due to the viscosity of the SVF, that is, the SVF does not descend downward even with its own weight. Accordingly, after a small amount of washing solution is supplied through the first connection pipe 111, the centrifuge vessel repeats rotating and stopping. Because the centrifugal force from the rotation causes the washing solution to be spread widely in the side accommodation part 310, by the repeated rotation and stopping, the SVF is precipitated in the lower body 303 by the inertia imparted to the washing solution. Next, the SVF precipitated in the lower body 303 is extracted and collected through the first connection pipe 111 (S800).

6. Verification Test

[0114] Tests were conducted to verify the performance of the centrifuge vessel according to the present disclosure.

[0115] The fat which was aspirated from the abdomen and thigh of 10 male and female adults between the ages of 20 and 40 was introduced into a centrifuge vessel according to the present disclosure, and then SVF was separated according to the method described with reference to FIGS. 11 and 12. The tests were conducted with first group introduced with 50 ml and the second group introduced with 100 ml. As an enzyme, 50 mg of collagenase was diluted in 50 ml of Hartmann's solution, compared to 50 ml of fat.

[0116] The number of obtained cells per 1 g of introduced fat (Cells/g), the cell viability (%), and the collagenase activity (PZU/ml), that is, the residual amount of collagenase, were calculated based on the final product obtained according to the method described in FIGS. 11 and 12. Cell counting for calculating the number of obtained cells and viability was performed using an automated cell counter.

[0117] Among the conventional instruments currently available in the market, Multi Station, Cha-Station, Lipokit, and Celution were selected. The number of obtained cells per 1 g of introduced fat (Cells/g), the cell viability (%), and the collagenase activity (PZU/ml) of each of the instruments are already well known, and accordingly, the corresponding information was compared with the results according to the present disclosure.

[0118] (1) Number of Obtained Cells

[0119] As shown in FIG. 13, it was observed that when the centrifuge vessel according to the present disclosure was used, 2 to 50 times or more cells were obtained as compared to the results obtained from Multi Station, Cha-Station, and Lipokit. However, the number of obtained cells was similar to the results obtained from Celution.

[0120] (2) Cell Viability

[0121] As shown in FIG. 14, the viability of the centrifuge vessel according to the present disclosure was very excellent as compared to the results obtained from Lipokit and Celution. However, the cell viability of the centrifuge vessel according to the present disclosure was similar or somewhat lower when compared to the results obtained from Multi Station and Cha-Station.

[0122] Then the number of obtained cells was compared with the following results.

[0123] Compared with the present disclosure, Multi Station of the related art exhibited half of the number of obtained cells or less, but the cell viability was somewhat higher. Compared with the present disclosure, Lipokit and Celution exhibited similar number of obtained cells, but lower cell viabilitys. That is, the related art exhibited a low viability when a large amount of cells are obtained, and the number of obtained cells was low when the viability was high.

[0124] Therefore, it was confirmed that the present disclosure has excellent cell viability as well as the number of cells obtained, and thus has very excellent final yield rate as compared with the related art.

[0125] (3) Enzyme Residue

[0126] As a result of the collagenase activity as shown in FIG. 15, when the centrifuge vessel according to the present disclosure was used, the amount of collagenase residue was about 20% or less of the average value in Multi Station, Cha-Station, Lipokit, and Celution, thus it was confirmed that it is more stable.

[0127] It will be apparent to those skilled in the art that various modifications and variations can be made to the present disclosure without departing from the spirit and scope of the present disclosure as defined by the following claims and their equivalents. Therefore, the scope of protection of present disclosure should be determined by the claims.