TWI579030B - Cell-separating container, cell-separating system and cell-separating method - Google Patents

Description

Container, system and method for separating cells

The present invention relates to a cell separation container, a cell separation system using the same, and a cell separation method, and more particularly to a cell separation container, a cell separation system, and a cell separation method for separating stem cells from adipose tissue and culturing stem cells.

Stem cells are cells that are capable of replicating themselves and dividing into two or more cells. In recent years, studies on obtaining stem cells from adipose tissue and bone marrow have been conducted. Adipose stem cells isolated from adipose tissue are pluripotent stem cells that can differentiate into various cells such as fat cells, muscle cells, chondrocytes, and bone cells.

Adipose-derived stem cells have better self-regeneration ability than mesenchymal stem cells isolated from bone marrow and are easily used for in vitro culture. The amount of adipose tissue that can isolate adipose stem cells is large, easy to collect, and safe to collect. Furthermore, it is generally known that adipose stem cells have the same characteristics (differentiation ability, cytological property, immunological property, tissue regeneration ability, etc.) as that of stem cells isolated from bone marrow. That is, adipose stem cells have a high probability of replacing mesenchymal stem cells from the bone marrow and are highly medically usable.

Compared to mesenchymal stem cells, the collection process of adipose stem cells is easier and safer, and the supply and demand of the organization is unlimited. Since adipose stem cells are easily cultured in vitro, they are superior to mesenchymal stem cells in terms of tissue availability, safety, efficacy, and economic viability.

The conventional method for separating and culturing the adipose stem cells is to finely break the extracted or cut fat, decompose it with collagenase, centrifuge the stromal vessel fraction (SVF), and culture the cells in SVF.

Various devices for collecting stem cells from adipose tissue have been developed, but stem cells are usually detached from adipose tissue in a separate container. Therefore, it is complicated to discharge the waste material from the produced material and discard it. To solve these problems, two separate containers were used to separate stem cells from adipose tissue. In this case, the adipose tissue is treated in a separate container and the isolated stem cells are treated in another separate container. Therefore, since the number of separation containers for consumables increases, the structure for rotating the separation containers increases, so the time and cost for separating stem cells increases.

[Object of the Invention]

An object of the present invention is to provide a cell separation container, a cell separation system, and a cell separation method which can improve the efficiency of isolating stem cells from adipose tissue.

Another object of the present invention is to provide a cell separation container, a cell separation system, and a cell separation method which can easily separate stem cells from adipose tissue with a simple structure.

Another object of the present invention is to provide a reduction in fat A cell separation vessel, a cell separation system, and a cell separation method for organizing the time and cost of isolating stem cells.

[Structure of the invention]

A cell separation container according to a first embodiment of the present invention includes: a housing rotatable on a rotating shaft and having a centrifugal space in the housing, wherein the centrifugal space protrudes from a side surface of the housing in a peripheral direction ( Convexly protruding; the first pipe is embedded in the casing in the direction of the rotating shaft and connects the inner space of the casing with the outer space of the casing; the second pipe is embedded in the casing in the direction of the rotating shaft and is connected into the casing a space between the space and the outer space of the casing, wherein the second pipe is bent such that the end of the second pipe is disposed in the centrifugal space; and a plurality of agitating blades project from the inner surface of the casing.

Preferably, the agitating blades are disposed at a lower portion of the casing and arranged around the first duct.

Preferably, the agitating blade has a plurality of through holes.

Preferably, the cell separation container further comprises an air discharge device disposed on the upper portion of the housing to connect the inner space of the housing with the outer space of the housing, thereby controlling the internal pressure of the housing and having a function of filtering air.

Preferably, when the housing is rotated on the rotating shaft, the first duct and the second duct do not rotate.

Preferably, the first conduit extends to approximate the bottom surface of the housing.

Preferably, the housing is rotated by a motor disposed at a lower portion of the housing.

A cell separation method using the above cell separation container according to a second embodiment of the present invention comprises the steps of: (a) providing the shell adipose tissue through the first conduit; (b) providing the shell cleaning through the first conduit Liquid, rotating the shell to stir the adipose tissue and the washing liquid, so that after the shell stops rotating, it is separated into the washed adipose tissue and blood contaminants respectively in the upper layer and the lower layer; (c) through the first a conduit removes the blood contaminant from the housing; (d) providing a shell enzyme through the first conduit, rotating the housing to agitate the washed adipose tissue and the enzyme, thereby separating the shell after it stops rotating Forming the decomposed adipose tissue and the first aqueous solution in the upper layer and the lower layer respectively; (e) in the centrifugal space, by rotating the shell, centrifuging the decomposed adipose tissue and the first aqueous solution into inner and outer layers, respectively Removing the decomposed adipose tissue from the housing through the second conduit; (f) after the housing stops rotating, generating a second aqueous solution by mixing the first aqueous solution with the cleaning liquid supplied through the first conduit (g) in the centrifugal space, by rotating the housing, centrifuging the second aqueous solution into outer and inner layer stem cells and a second aqueous solution extracted by stem cells, and passing the shell during stem cell centrifugation The second conduit removes the second aqueous solution extracted by the stem cells; and (h) after the housing stops rotating, the target aqueous solution is generated by using the mixed stem cells and the cleaning liquid supplied through the second conduit, and passes through the first conduit The target aqueous solution is discharged from the casing.

A cell separation system according to a third embodiment of the present invention comprises: a storage device that separately stores adipose tissue, a washing solution and an enzyme; and a cell separation container connected to the storage device and supplied with adipose tissue and washing therefrom Cleansing liquid and enzyme, wherein the cell separation container is stirred Adipose tissue, a washing solution and an enzyme supplied from the storage device, and centrifuging the stem cells; a rotating unit connected to the cell separation container and rotating the cell separation container; and a waste storage device connected to the cell separation container and stored by the cell separation container a waste provided by a cell separation container, wherein the waste is produced in a cell separation container for agitating adipose tissue, a washing solution and an enzyme, and centrifuging the stem cells; a cell storage unit connected to the cell separation container and stored from the cell Separating the stem cells provided by the container, wherein the stem cell is obtained by stirring adipose tissue, a washing solution and an enzyme, and centrifuging the adipose tissue; and connecting a unit connecting the cell separation container to the storage device, the waste storage device and the cell storage unit; and transporting a unit disposed on the connecting unit, wherein the transport unit transports adipose tissue, a washing solution and an enzyme from the storage device to the cell separation container through the connecting unit, transports the waste from the cell separation container to the waste storage device, and separates from the cell Container transporting stem cells to cell storage unit

Preferably, the cell separation system further comprises an inductor disposed on the connection unit between the cell separation container and the transport unit, and sensing the concentration of the waste discharged from the cell separation container via the transport unit.

Preferably, the cell separation container comprises: a housing that is rotated on a rotating shaft by a rotating unit disposed at a lower portion of the housing and has a centrifugal space in the housing, wherein the centrifugal space is from the housing in a peripheral direction Projecting a convex surface in the body side surface; the first pipe is embedded in the casing in the direction of the rotating shaft and connects the inner space of the casing and the outer space of the casing; the second pipe is embedded in the direction of the rotating shaft The housing is connected to the housing a space between the portion and the outer space of the casing, wherein the second pipe is bent such that an end of the second pipe is disposed in the centrifugal space; and a plurality of agitating blades projecting from an inner surface of the casing.

Preferably, the agitating blades are disposed at a lower portion of the casing and arranged around the first duct.

Preferably, the agitating blade has a plurality of through holes.

Preferably, the cell separation container further comprises: an air discharge device disposed on the upper portion of the housing to connect the inner space of the housing and the outer space of the housing, thereby controlling the internal pressure of the housing and having the function of filtering air.

Preferably, when the housing is rotated on the rotating shaft, the first duct and the second duct do not rotate.

Preferably, the first conduit extends to approximate the bottom surface of the housing.

Preferably, the storage device comprises: an adipose tissue storage device that stores adipose tissue and is connected to the cell separation container via the connection unit; a cleaning liquid storage device that stores the washing liquid and is connected to the cell via the connection unit a separation container; and an enzyme storage device that stores the enzyme and is connected to the cell separation container via the connection unit.

Preferably, the connecting unit comprises: a common pipe connected to the transport unit; an adipose tissue pipe connecting the fat tissue storage device and the shared pipe; and a cleaning liquid pipe connected to the cleaning liquid storage device and the a shared conduit; an enzyme conduit connecting the enzyme storage device and the shared conduit; a waste conduit connecting the waste storage device and the shared conduit; and a separation conduit connecting the cell separation container and the transport unit; And a cell transport conduit connecting the cell storage unit to the shared conduit.

Preferably, the cell separation system further comprises an adjustment unit disposed on the connection unit and opening or closing the connection unit.

Preferably, the adjusting unit comprises: an adipose tissue regulator disposed on the adipose tissue duct to open or close the adipose tissue duct; and a detergent regulator disposed on the washing liquid pipeline to be opened or closed The cleaning fluid conduit; an enzyme regulator disposed on the enzyme conduit to open or close the enzyme conduit; a waste regulator disposed on the waste conduit to open or close the waste conduit; a separation regulator configured Opening or closing the separation conduit on the separation conduit; and transporting a cell regulator disposed on the cell delivery conduit to open or close the cell delivery conduit.

Preferably, the cell separation system further comprises a control unit coupled to the rotation unit, the transport unit and the adjustment unit, wherein the control unit controls the transport unit and the adjustment unit to transport adipose tissue, a cleaning solution, an enzyme via the connection unit , waste and stem cells, and control the rotating unit to rotate the cell separation container.

Preferably, the enzyme storage device and the cell separation container are connected to a temperature adjustment unit that maintains the temperature of the enzyme storage device and the cell separation container at 35 ° C to 40 ° C.

Preferably, the fat tissue storage device, the enzyme storage device and the cell storage unit are syringes.

Preferably, the connecting unit comprises: a first separating pipe connecting the conveying unit to the first pipe of the cell separation container and being openable or closed by the first separating regulator; and a second separating pipe connecting the conveying The second conduit of the unit to the cell separation vessel can be opened or closed by a second separation regulator.

Preferably, the cell storage unit comprises a cell filtration unit disposed on the connection unit between the cell storage unit and the transport unit, and filtering the stem cells transported to the cell storage unit via the connection unit.

The cell separation method according to the fourth embodiment of the present invention comprises the steps of: (a) transporting the adipose tissue from the adipose tissue storage device to the cell separation container via the transport unit; (b) transporting the wash from the detergent solution storage device via the transport unit Purging the liquid to the cell separation container, and rotating the cell separation container through the rotating unit to stir the adipose tissue and the washing liquid; thereby, after the cell separation container stops rotating, it is separated into the washed adipose tissue of the upper layer and the lower layer, respectively. (c) transporting blood contaminants from the cell separation container to the waste storage device via the transport unit; (d) transporting the enzyme from the enzyme storage device to the cell separation container via the transport unit, and rotating the cell separation via the rotating unit The container is stirred with the enzyme and the washed adipose tissue; thereby, after the cell separation container stops rotating, it is separated into the decomposed adipose tissue and the first aqueous solution which are respectively the upper layer and the lower layer; (e) when the rotating unit rotates the cell separation container When the decomposed adipose tissue and the first aqueous solution are separated into an inner layer and an outer layer by centrifugation, Transferring the decomposed fat tissue from the cell separation container to the waste storage device by the transport unit; (f) after the cell separation container stops rotating, transporting the washing liquid to the cell separation container via the transport unit, and passing through the cell separation container, stirring An aqueous solution and a washing solution to produce a second aqueous solution; (g) rotating the cell separation vessel via a rotating unit to centrifuge the second aqueous solution into stem cells of outer and inner layers, respectively And a second aqueous solution extracted by the stem cells; during the centrifugation of the stem cells, the second aqueous solution extracted by the stem cells is transported from the cell separation container to the waste storage device via the transport unit; (h) after the cell separation container stops rotating, via the transport unit, The washing liquid is transported to the cell separation container, and the centrifuged stem cells are dripped by using the washing liquid to produce a target aqueous solution; and (i) the target aqueous solution is transported from the cell separation container to the cell storage unit via the transport unit.

Preferably, step (c) is repeated until the sensor senses the cleaned adipose tissue.

Preferably, steps (b) and (c) are repeated until the sensor senses that the blood contaminant of step (c) has a concentration below a predetermined value.

Preferably, the cell separation container comprises: a housing that is rotated on a rotating shaft by a rotating unit and has a centrifugal space in the housing, wherein the centrifugal space protrudes from the side surface of the housing in a peripheral direction a first pipe, which is embedded in the casing in a direction of a rotating shaft and connects the inner space of the casing and the outer space of the casing; the second pipe is embedded in the casing and connected in the direction of the rotating shaft The inner space of the casing and the outer space of the casing, wherein the second pipe is bent such that the end of the second pipe is disposed in the centrifugal space; and a plurality of agitating blades project from the inner surface of the casing.

Preferably, the first conduit is used in steps (a), (b), (c), (d), (f) and (i), and the second conduit is used in step (e), ( g) and (h).

Preferably, in step (e), the decomposed adipose tissue and the first aqueous solution are separately centrifuged into an inner layer in the centrifugal space and In the outer layer; and in the step (g), the stem cells and the second aqueous solution extracted by the stem cells are separately centrifuged into the outer layer and the inner layer in the centrifugal space.

[Effects of the invention]

The cell separation container according to the present invention has an effect of efficiently separating stem cells from adipose tissue, washing the separated stem cells, storing the washed stem cells, and since the cell separation container according to the present invention has only a housing containing a centrifugal space, It has the same effect as two housings, thus reducing equipment costs.

The cell separation system and the cell separation method according to the present invention have the advantages of reducing the cell separation container used as a consumable, extracting the stem cells in a simple structure, and separating the stem cells from the adipose tissue only by manipulating the control unit, thereby reducing the time for extracting the stem cells. With cost.

100‧‧‧ cell separation system

101‧‧‧Storage device

102‧‧‧ cell separation container

102a‧‧‧Temperature adjustment unit

103‧‧‧Waste storage device

104‧‧‧ cell storage unit

104a‧‧‧cell filtration unit

105‧‧‧Shared pipeline

106‧‧‧Transportation unit

107‧‧‧ sensor

108‧‧‧Adjustment unit

109‧‧‧Control unit

111‧‧‧Adipose tissue storage device

113‧‧‧cleaning liquid storage device

115‧‧‧Enzyme storage device

115a‧‧‧Temperature adjustment unit

120‧‧‧Rotating unit

121‧‧‧Shell

121a‧‧‧Centrifugal space

122‧‧‧First pipeline

123‧‧‧Second pipeline

End of 123a‧‧

124‧‧‧Agitating blades

124a‧‧‧through holes

125‧‧‧Air discharge device

151‧‧‧Adipose tissue pipeline

152‧‧‧Clean liquid pipeline

153‧‧‧Enzyme Pipeline

154‧‧‧ scrap pipeline

155‧‧‧Separate pipeline

155a‧‧‧The first separation pipeline

155b‧‧‧Second separation pipeline

156‧‧‧cell transport pipeline

181‧‧‧Adipose tissue regulator

182‧‧•washing fluid regulator

183‧‧‧Enzyme regulator

184‧‧‧Waste regulator

185‧‧‧Separation regulator

185a‧‧‧First separation regulator

185b‧‧‧Second separation regulator

186‧‧‧cell transport regulator

A‧‧‧Rotary axis

a‧‧‧Adipose tissue

b‧‧‧Cleaned adipose tissue

c‧‧‧Blood contaminants

d‧‧‧First aqueous solution

E‧‧‧Decomposed adipose tissue

F‧‧‧second aqueous solution

G‧‧‧stem cells

H‧‧‧ target aqueous solution

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a schematic illustration of a cell separation system in accordance with a preferred embodiment of the present invention.

Figure 2 is a cross-sectional view of a cell separation vessel in accordance with a preferred embodiment of the present invention.

Figure 3 is a cross-sectional view of the cell separation container of Figure 2.

Figure 4 is a flow chart of a cell separation method in accordance with a preferred embodiment of the present invention.

Figures 5 through 13 are schematic views of the state of the cell separation system corresponding to the steps of the cell separation method of Figure 4.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to the accompanying drawings. Related Functions or Structures of the Related Art, if an understanding of the gist of the present invention is not necessary, it will not be described in detail herein.

1 is a schematic illustration of a cell separation system 100 in accordance with a preferred embodiment of the present invention. Figure 2 is a cross-sectional view of a cell separation vessel 102 in accordance with a preferred embodiment of the present invention. Fig. 3 is a cross-sectional view showing the cell separation container 102 of Fig. 2.

As shown in Figures 1 to 3, a cell separation system 100 according to a preferred embodiment of the present invention comprises a storage device 101, a cell separation container 102, a rotating unit 120, a waste storage device 103, a cell storage unit 104, a connection unit, and a transport. Unit 106 is for separating and extracting stem cells from adipose tissue. Adipose tissue means adipose tissue extracted by liposuction, and the washing liquid means physiological saline, phosphate buffered saline, Hartmann's solution, and the like. The enzyme means collagenase, trypsin, etc., and is used by being dissolved in the washing solution. The cell separation system 100 is completely sealed and sterile. Therefore, stem cells can be stably separated and extracted from adipose tissue.

The storage device 101 stores adipose tissue, a washing solution, and an enzyme for extracting stem cells. The storage device 101 includes an adipose tissue storage device 111 for storing adipose tissue, a cleaning solution storage device 113 for storing the cleaning solution, and an enzyme storage device 115 for storing the enzyme. The adipose tissue storage device 111 and the enzyme storage device 115 can be syringes. Therefore, the fat tissue supply and leaven can be controlled via the adipose tissue storage device 111 and the enzyme storage device 115. Supply.

The enzyme storage device 115 can be connected to the temperature adjustment unit 115a which is a separation unit, and the temperature adjustment unit 115a maintains the temperature of the enzyme storage device 115 at a specific temperature, for example, 35 ° C to 40 ° C. Therefore, the enzyme can be maintained at a specific temperature; at this temperature range, the enzyme is reactive.

The cell separation container 102 is connected to the storage device 101. Specifically, the fat tissue storage device 111, the cleaning solution storage device 113, and the enzyme storage device 115 are connected to the cell separation container 102. The cell separation container 102 obtains adipose tissue, a washing solution, and an enzyme from the storage device 101. The cell separation container 102 is rotatable on the rotation axis A. When the cell separation container 102 is rotated, the fat tissue, the washing solution and the enzyme are stirred in the cell separation container 102 and centrifuged.

The cell separation container 102 includes a housing 121, a first conduit 122, a second conduit 123, a stirring blade 124, and an air discharge device 125 for separating adipose tissue, a washing solution and an enzyme from adipose tissue by stirring and centrifuging. Isolation of stem cells. The cell separation vessel 102 is completely sealed and sterile, except for the portion that is attached to the external device. The isolated stem cells are cultured and used.

The housing 121 is coupled to the rotating unit 120 disposed at a lower portion of the housing 121, and is rotated by the rotating unit 120 on the rotating shaft A. Adipose tissue, a washing solution, and an enzyme are provided in the housing 121. The lower portion of the housing 121 has a curved surface. The cross-sectional area of the housing 121 becomes smaller toward the lower direction. Therefore, the fat tissue, the washing liquid and the enzyme supplied to the housing 121 can be easily aggregated. In the lower part of the housing 121.

The housing 121 has a centrifugal space 121a therein, and the centrifugal space 121a protrudes convexly from the side surface of the housing 121 along the peripheral direction.

The first duct 122 has a duct shape and is embedded in the casing 121 in the direction of the rotational axis A. Preferably, the first conduit 122 extends to approximate the bottom surface of the housing 121. The first duct 122 connects the internal space of the housing 121 with the outer space of the housing 121. The adipose tissue, the cleaning solution and the enzyme are transported into the housing 121 through the first conduit 122. When the housing 121 is rotated on the rotational axis A, the first duct 122 does not rotate. The adipose tissue, the washing solution and the enzyme are stirred in the housing 121, and the agitated material is discharged from the inside of the housing 121 through the first conduit 122. The first conduit 122 is connected to the first separation conduit 155a of the separation conduit 155.

The second duct 123 has a duct shape like the first duct 122 and is embedded in the casing 121 in the direction of the rotational axis A. The second duct 123 connects the internal space of the casing 121 and the outer space of the casing 121. The second duct 123 is bent so that its end 123a is disposed in the centrifugal space 121a. Therefore, when the housing 121 is rotated, the agitated material can be collected in the centrifugal space 121a by centrifugal force and discharged from the inside of the housing 121 through the second conduit 123. The second conduit 123 is connected to the second separation conduit 155b of the separation conduit 155.

When the housing 121 rotates, the second duct 123 does not rotate in addition to the first duct 122. Therefore, the hoses connected to the first pipe 122 and the second pipe 123 can be independent of the rotation of the casing 121. The connection with the first duct 122 and the second duct 123 is maintained to stably transport the fat tissue, the washing liquid, the enzyme, the waste, and the stem cells through the first duct 122 and the second duct 123.

The agitating blades 124 protrude from the inner surface of the housing 121. When the housing 121 rotates, the stirring blade 124 rotates around the rotation axis A. Therefore, the adipose tissue, the washing solution and the enzyme can be efficiently stirred and reacted with each other in the casing 121.

As shown in FIG. 3, the agitating blades 124 may be disposed at a lower portion of the housing 121 and arranged around the first duct 122. Therefore, even when a small amount of fat tissue, washing liquid, and enzyme are transported to the casing 121, the fat tissue, the washing liquid, and the enzyme can be stably stirred via the stirring blade 124.

The agitating blade 124 has a plurality of through holes 124a. Therefore, the rotational resistance of the agitating blade 124 is reduced, and since the adipose tissue, the washing liquid, and the enzyme can flow through the through hole 124a, the stirring efficiency is increased.

The air discharge device 125 is disposed on an upper portion of the casing 121 and connects an inner space of the casing 121 and an outer space of the casing 121. Therefore, the air discharge device 125 can control the internal pressure of the housing 121 to be a constant pressure. The air discharge device 125 has a function of filtering air. Therefore, the air discharge device 125 can prevent foreign materials from entering the housing 121, thereby maintaining the internal space of the housing 121 in a sterile state.

The size of the cell separation vessel 102 can correspond to the amount of stem cells to be obtained. Providing the cell separation container 102 with the amount of adipose tissue corresponding to the amount of the stem cells to be obtained, providing the cell separation container 102 washing solution and the enzyme in an amount corresponding to the amount of the adipose tissue, and then separating the fine tissue from the adipose tissue Cell. Therefore, the cell separation container 102 according to this embodiment can obtain a desired amount of stem cells at one time.

The cell separation vessel 102, and in particular the casing 121, can be coupled to a temperature conditioning unit 102a, which is a separation unit that maintains the temperature of the housing 121 at a particular temperature, such as 35 °C to 40 °C. Therefore, the fat tissue, the cleaning solution and the enzyme supplied from the storage device 101 can be maintained at a specific temperature; in the temperature range, the adipose tissue, the cleaning solution and the enzyme can react reactively.

The rotation unit 120 is connected to the lower portion of the cell separation container 102, and rotates the cell separation container 102 on the rotation axis A. The rotating unit 120 can be a motor that controls the rotational speed of the cell separation vessel 102 to provide agitation and centrifugation in the cell separation vessel 102.

The waste storage device 103 is connected to the cell separation container 102. When the adipose tissue, the washing solution, and the enzyme are stirred and centrifuged in the cell separation vessel 102, a stirred material is produced. In the agitated material, the material that is not used during the separation of stem cells from adipose tissue is called waste. The waste storage device 103 stores the waste removed from the cell separation container 102.

The cell storage unit 104 is connected to the cell separation container 102. When the adipose tissue, the washing solution, and the enzyme are stirred and centrifuged in the cell separation container 102, the stem cells separated from the adipose tissue are finally obtained in the cell separation container 102. The stem cells are aggregated in the cell storage unit 104 and cultured in the cell storage unit 104. Cell storage unit 104 can be a syringe.

The cell filtration unit 104a is disposed near the cell storage unit 104. After the stem cells pass through the cell filtration unit 104a, they are accumulated in the cell storage unit 104. Therefore, stem cells purified to high purity can be stored in the cell storage unit 104 and cultured.

The connecting unit can be a hose. The cell separation container 102 is connected to the storage device 101, the waste storage device 103, and the cell storage unit 104 via the connection unit. The connecting unit performs the function of connecting the cell separation container 102 to the storage device 101, the cell separation container 102 to the waste storage device 103, and the cell separation container 102 to the cell storage unit 104. Therefore, the adipose tissue, the abrad liquid and the enzyme can be transported from the storage device 101 to the cell separation container 102 along the connection unit; the waste material in the cell separation container 102 can be transported from the cell separation container 102 to the waste storage device along the connection unit. 103; and stem cells isolated in the cell separation vessel 102 can be transported from the cell separation vessel 102 to the cell storage unit 104 along the connection unit.

The transport unit 106 can be a pump connected to the connection unit. The transport unit 106 transports the fat tissue, the washing liquid and the enzyme to the cell separation container 102 from the storage device 101, specifically, the fat tissue storage device 111, the cleaning liquid storage device 113, and the enzyme storage device 115, respectively, along the connection unit. . The transport unit 106 transports the waste from the cell separation container 102 to the waste storage device 103 along the connection unit, and transports the stem cells from the cell separation container 102 to the cell storage unit 104 along the connection unit.

As explained above, the transport unit 106 is operated to control the direction of transport of fat tissue, washing liquid, enzymes, waste, and stem cells, and Aspiration of adipose tissue, detergent, enzymes, waste and stem cells.

The connecting unit includes a common duct 105, an adipose tissue duct 151, a washing liquid duct 152, an enzyme duct 153, a waste duct 154, a separating duct 155, and a cell transport duct 156. The shared conduit 105 is always open. The common pipe 105 is connected to the adipose tissue pipe 151, and connects the adipose tissue storage device 111 to the transport unit 106 and the cleaning liquid pipe 152, so that the connected cleaning liquid storage device 113 to the transport unit 106 and the enzyme conduit 153 are connected to the enzyme storage. The device 115 to the transport unit 106, the waste pipe 154, and the connection waste storage device 103 to the transport unit 106, and the cell transport conduit 156 are connected to the cell storage unit 104 to the transport unit 106.

The separation duct 155 connects the cell separation container 102 and the transport unit 106. The separation duct 155 includes a first separation duct 155a that connects the transport unit 106 with the first duct 122 of the cell separation container 102, and a second separation duct 155b that connects the transport unit 106 with the second duct 123 of the cell separation container 102.

The fat tissue stored in the fat tissue storage device 111 is transported to the cell separation container 102 through the fat tissue conduit 151, the common conduit 105, the transport unit 106, and the separation conduit 155 according to the structure of the connection unit. The cleaning liquid stored in the cleaning liquid storage device 113 is transported to the cell separation container 102 through the cleaning liquid pipe 152, the common pipe 105, the conveying unit 106, and the separation pipe 155. The enzyme stored in the enzyme storage device 115 is transported to the cell separation container 102 through the enzyme conduit 153, the common conduit 105, the transport unit 106, and the separation conduit 155. The waste generated in the cell separation container 102 passes through the separation pipe 155, the transport unit 106, The shared conduit 105 and the waste conduit 154 are transported to the waste storage device 103 and stored in the waste storage device 103. In the cell separation container 102, the stem cell line separated from the adipose tissue is transported to the cell storage unit 104 through the separation line 155, the transport unit 106, the common line 105, and the cell transport line 156, and cultured in the cell storage unit 104.

By using this structure, the adipose tissue is washed with a washing solution and decomposed by the enzyme. Then, only the stem cells are separated and extracted. The stem cells are aggregated and cultured in the cell storage unit 104.

The cell separation system 100 can further include an inductor 107, an adjustment unit 108, and a control unit 109.

The sensor 107 is disposed on the separation duct 155 between the cell separation container 102 and the transport unit 106, and senses the inside of the separation duct 155. The sensor 107 can be an optical sensor for checking whether fat tissue, washing liquid, enzymes, waste, and stem cells transported by the transport unit 106 are properly transported during separation and extraction of stem cells from adipose tissue. The sensor 107 senses the waste conveyed by the transport unit 106 from the cell separation container 102, thereby checking whether the fat tissue is completely washed, or whether the waste is transported. For example, by sensing the color change of the waste (ie, blood contaminants), the sensor 107 can sense whether the concentration of the blood contaminant is below a predetermined amount. Blood contaminants are wastes produced when cleaning adipose tissue with a cleaning solution. If the blood pollutant concentration is lower than the predetermined amount, it means that the adipose tissue is sufficiently washed by the washing liquid. Therefore, the washed adipose tissue can be used in the next step.

The adjusting unit 108 is disposed on the connecting unit, and is turned on or Close the connection unit. When the fat tissue, the washing liquid, the enzyme, the waste, and the stem cells are transported along the connection unit via the transport unit 106, the connection unit that does not need to transport the fat tissue, the washing solution, the enzyme, the waste, and the stem cells is closed by the adjustment unit 108. The regulating unit 108 prevents the fat tissue, the washing liquid, the enzyme, the waste, and the stem cells from being erroneously transported by the transport unit 106 when transporting the fat tissue, the washing solution, the enzyme, the agitated material, and the stem cells.

The adjustment unit 108 includes a fat tissue regulator 181 disposed on the fat tissue conduit 151, a detergent regulator 182 disposed on the cleaning liquid conduit 152, an enzyme regulator 183 disposed on the enzyme conduit 153, and a waste pipe disposed in the waste pipe A waste conditioner 184 on the 154, a separation regulator 185 disposed on the separation duct 155, and a cell transport regulator 186 disposed on the cell transport conduit 156. The separation regulator 185 includes a first separation regulator 185a disposed on the first separation conduit 155a and a second separation regulator 185b disposed on the second separation conduit 155b. The first separation regulator 185a operates in opposition to the second separation regulator 185b.

The cell filtration unit 104a is disposed on the cell transport conduit 156 between the cell transport regulator 186 and the cell storage unit 104.

The adipose tissue regulator 181, the detergent regulator 182, the enzyme regulator 183, the waste conditioner 184, the separation regulator 185, and the cell transport regulator 186 can be of the cassette type. The adjusters 181 to 186 are disposed at fixed positions. The cassette adjusters 181 to 186 are disposed on the cell separation system 100, and the connection unit is connected to the regulators 181 to 186. Therefore, the connecting units are not entangled and are easily connected to the storage device 101, the cell separation container 102, the waste storage device 103, and the cell storage unit 104. Regulator 181 The 186 can be valve or cam type.

For example, the adipose tissue regulator 181 opens the adipose tissue conduit 151, and the separation regulator 185 opens the separation conduit 155 (eg, the first separation regulator 185a opens the first separation conduit 155a and the second separation regulator 185b closes the second separation The conduit 155b), and the detergent regulator 182 closes the purge conduit 152, the enzyme regulator 183 closes the enzyme conduit 153, the waste regulator 184 closes the waste conduit 154, and the cell transport regulator 186 closes the cell delivery conduit 156. When the transport unit 106 is in operation, the adipose tissue stored in the adipose tissue storage device 111 is transported to the cell separation container 102. That is, the adipose tissue storage device 111 is connected to the cell separation container 102 through the opened adipose tissue duct 151, the common duct 105, and the separation duct 155 (for example, the first separation duct 155a), and the fat tissue can pass through the opened fat tissue duct. 151. The shared duct 105 and the separation duct 155 are transported by the transport unit 106.

The cleaning liquid regulator 182 opens the cleaning liquid pipe 152, and the separation regulator 185 opens the separation pipe 155 (for example, the first separation regulator 185a opens the first separation pipe 155a, and the second separation regulator 185b closes the second separation pipe 155b) And the adipose tissue regulator 181 closes the adipose tissue conduit 151, the enzyme regulator 183 closes the enzyme conduit 153, the waste regulator 184 closes the waste conduit 154, and the cell transport regulator 186 closes the cell delivery conduit 156. When the transport unit 106 is operated, the washing liquid stored in the detergent storage device 113 is transported to the cell separation container 102. That is, the cleaning liquid storage device 113 is connected by the opened cleaning liquid pipe 152, the common pipe 105, and the separation pipe 155 (for example, the first separation pipe 155a). The cell separation container 102, and the cleaning liquid can be transported by the transport unit 106 through the opened cleaning liquid pipe 152, the common pipe 105, and the separation pipe 155.

The enzyme regulator 183 opens the enzyme conduit 153, and the separation regulator 185 opens the separation conduit 155 (for example, the first separation regulator 185a opens the first separation conduit 155a, and the second separation regulator 185b closes the second separation conduit 155b). The adipose tissue regulator 181 closes the adipose tissue conduit 151, the detergent regulator 182 closes the detergent fluid conduit 152, the waste regulator 184 closes the waste conduit 154, and the cell transport regulator 186 closes the cell delivery conduit 156. When the transport unit 106 is in operation, the enzyme stored in the enzyme storage device 115 is transported to the cell separation container 102. That is, the enzyme storage device 115 is connected to the cell separation container 102 through the open enzyme tube 153, the common conduit 105, and the separation conduit 155 (for example, the first separation conduit 155a), and the enzyme can pass through the open enzyme tube 153, the shared conduit. The separation duct 155 is transported by the transport unit 106.

The waste regulator 184 opens the waste conduit 154, and the separation regulator 185 opens the separation conduit 155 (eg, one of the first separation regulator 185a and the second separation regulator 185b opens the first separation conduit 155a and the second separation conduit 155b Correspondingly, and the other of the first separation regulator 185a and the second separation regulator 185b closes the corresponding one of the first separation duct 155a and the second separation duct 155b), and the fat tissue regulator 181 closes the fat The tissue conduit 151, the detergent regulator 182 closes the cleaning fluid conduit 152, the enzyme regulator 183 closes the enzyme conduit 153, and the cell transport regulator 186 closes the cell delivery conduit 156. When the transport unit 106 is in operation, the waste generated in the cell separation container 102 is transported to Waste storage device 103. That is, the cell separation container 102 is connected to the waste storage device 103 through the open separation pipe 155, the common pipe 105, and the waste pipe 154, and the waste material can pass through the open separation pipe 155, the common pipe 105, and the waste pipe 154, by the transportation unit. 106 shipping.

The cell transport regulator 186 opens the cell transport conduit 156, and the separation regulator 185 opens the separation conduit 155, while the adipose tissue regulator 181 closes the adipose tissue conduit 151, the detergent regulator 182 closes the wash fluid conduit 152, and the enzyme regulator 183 The enzyme tube 153 is closed, and the waste regulator 184 closes the waste line 154. When the transport unit 106 is in operation, the stem cells separated from the adipose tissue are transported to the cell storage unit 104 in the cell separation container 102. That is, the cell separation container 102 is connected to the cell storage unit 104 through the open separation pipe 155, the common pipe 105, and the cell delivery pipe 156, and the stem cells can be transported by opening the separation pipe 155, the common pipe 105, and the cell transportation pipe 156. Unit 106 is shipped.

The separation regulator 185 maintains the separation duct 155 in an open state. That is, one of the first separation duct 155a and the second separation duct 155b is kept open. When the material that is not transported from the cell separation container 102 to the waste storage device 103 is transported to the separation conduit 155, the separation regulator 185 closes the separation conduit 155, thereby preventing the material from being discharged from the cell separation container 102.

The control unit 109 is connected to the rotation unit 120, the transport unit 106, and the adjustment unit 108. The control unit 109 drives the transport unit 106 and controls the adjustment unit 108 to open or close the connection unit. Therefore, fat Tissue, washings, enzymes, waste and stem cells can be transported through the connection unit. The control unit 109 operates the rotation unit 120 to rotate the cell separation container 102, thereby agitating and centrifuging the adipose tissue, the washing solution and the enzyme in the cell separation container 102. The stem cells can be separated from the adipose tissue via the simple operation control unit 109.

The control unit 109 can have a display and a plurality of buttons. The user checks the status of the cell separation system 100 on the display and manipulates the buttons to control the cell separation system 100.

Figure 4 is a flow chart of a cell separation method in accordance with a preferred embodiment of the present invention. Figures 5 to 13 are schematic views of the state of the cell separation system corresponding to the steps of the cell separation method of Figure 4.

As shown in Figs. 4 to 13, the cell separation method according to a preferred embodiment of the present invention is a method of separating and extracting stem cells from adipose tissue, and using the cell separation system 100.

First, the adipose tissue a is transported from the adipose tissue storage device 111 to the cell separation container 102 (S101) (see Fig. 5). In step S101, the fat tissue regulator 181 opens the fat tissue conduit 151, the separation regulator 185 opens the separation conduit 155, the cleaning fluid regulator 182 closes the cleaning fluid conduit 152, the enzyme regulator 183 closes the enzyme conduit 153, and the waste regulator The waste conduit 154 is closed 184, and the cell transport regulator 186 closes the cell transport conduit 156. In the separation regulator 185, only the first separation regulator 185a opens the first separation conduit 155a connected to the first conduit 122 of the cell separation vessel 102, and the second separation regulator 185b opens the second connection to the cell separation vessel 102. The second separation conduit 155b of the conduit 123.

The transport unit 106 transports the fat tissue a stored in the fat tissue storage device 111 of the storage device 101 through the fat tissue conduit 151, the common conduit 105, and the first separation conduit 155a to the cell separation container 102. The adipose tissue a is placed in the housing 121 through the first conduit 122 of the cell separation container 102.

Then, the cleaning liquid is transported from the cleaning liquid storage device 113 to the cell separation container 102, and the cell separation container 102 is rotated via the rotation unit 120, and the cleaning liquid is stirred together with the fat tissue a (S102) (see Fig. 6). . Step S102 is similar to step S101 except that the detergent regulator 182 opens the detergent liquid conduit 152 and the fat tissue regulator 181 to close the fat tissue conduit 151. The transport unit 106 transports the washing liquid stored in the washing liquid storage device 113 of the storage device 101 through the washing liquid pipe 152, the common pipe 105, and the first separating pipe 155a to the cell separating container 102. The washing liquid, which has a larger amount than the adipose tissue a, passes through the first conduit 122 of the cell separation vessel 102.

The rotation unit 120 repeats the rotation of the cell separation container 102 and the stop rotation of the cell separation container 102. The washing solution and the adipose tissue a are stirred in the cell separation container 102 by the stirring blade 124. The adipose tissue a is washed with the washing liquid, and thus the washed adipose tissue b and the blood contaminant c are produced in the cell separation container 102. After the adipose tissue a is sufficiently washed, the rotating unit 120 stops rotating the cell separation container 102, and then the two are separated by gravity due to the difference in density of the washed adipose tissue b and the blood contaminant c. Therefore, the blood contaminant c sinks in the cell separation container 102 to form a lower layer, and the washed adipose tissue b floats above the blood contaminant c to form an upper layer. The blood pollutant c contains a washing liquid, blood and body fluid.

Next, the transport unit 106 transports the waste (i.e., blood contaminant c) from the cell separation container 102 to the waste storage device 103 (S103) (see Fig. 7). Step S103 is similar to step S102 except that the cleaning liquid regulator 182 closes the cleaning liquid pipe 152, and the waste conditioner 184 opens the waste pipe 154. The transport unit 106 transports the blood contaminants c of the cell separation vessel 102 through the first separation conduit 155a, the common conduit 105 and the waste conduit 154 to the waste storage device 103. Blood contaminants c pass through the first conduit 122 of the cell separation vessel 102. That is, step S103 removes the waste (i.e., blood contaminant c) generated in step S102.

In step S103, the sensor 107 disposed on the separation duct 155 senses the concentration of the blood pollutant c discharged and removed from the cell separation container 102 via the transport unit 106. The removed blood contaminant c is red due to body fluid or blood, and the washed adipose tissue b is yellow. Therefore, in step S103, if the sensor 107 senses the yellow washed adipose tissue b, the transport unit 106 is stopped, and the first separation adjuster 185a closes the first separation duct 155a. Step S103 is repeated until the sensor 107 senses the yellow washed adipose tissue b. Therefore, the washed adipose tissue b can be prevented from being discharged from the cell separation container 102.

When blood contaminants c are transported via the transport unit 106, the sensor 107 senses the concentration of the waste (ie, blood contaminants c). If the sensor 107 senses that the blood contaminant c concentration is below a predetermined amount, the next step is performed. However, if not, step S102 is repeated after completion of step S103. Steps S102 and S103 are repeated until the sensor 107 senses that the blood pollutant c concentration is lower than a predetermined amount. Therefore, the adipose tissue a can be sufficiently washed Cleaning.

Next, the enzyme is transported from the enzyme storage device 115 to the cell separation container 102, and the cell separation container 102 is rotated by the rotation unit 120 to stir the enzyme together with the washed fat tissue b (S104) (see Fig. 8). Step S104 is similar to step S102, except that the cleaning liquid regulator 182 turns off the cleaning liquid conduit 152 and the enzyme regulator 183 to open the enzyme conduit 153. The transport unit 106 transports the enzyme stored in the enzyme storage device 115 of the storage device 101 through the enzyme conduit 153, the common conduit 105, and the first separation conduit 155a to the cell separation vessel 102. The enzyme is transported to the housing 121 of the cell separation vessel 102 through the first conduit 122 of the cell separation vessel 102.

The rotation unit 120 repeats the rotation of the cell separation container 102 and the stop rotation of the cell separation container 102. The enzyme and the washed adipose tissue b are stirred in the cell separation container 102 by the stirring blade 124, and the washed adipose tissue b is decomposed by the enzyme. Therefore, the decomposed fat tissue e and the first aqueous solution d are generated in the casing 121. After the washed adipose tissue b is completely decomposed, the rotating unit 120 stops rotating the cell separation container 102, and then separates the two by gravity based on the density of the decomposed fat tissue e and the first aqueous solution d. The first aqueous solution d is sunk in the cell separation container 102 to form a lower layer, and the decomposed adipose tissue e floats on the first aqueous solution d to form an upper layer. The first aqueous solution d contains stem cells g and an enzyme.

Next, the rotating unit 120 rotates the cell separation container 102, the decomposed fat tissue e and the first aqueous solution d are centrifuged, and the transport unit 106 transports the decomposed adipose tissue from the cell separation container 102. e to the waste storage device 103 (S105) (see Figure 9). In step S105, the cell separation container 102 is centrifuged. Due to the centrifugal force generated by the rotating unit 120, the decomposed adipose tissue e and the first aqueous solution d move to the centrifugal space 121a of the housing 121. The decomposed adipose tissue e and the first aqueous solution d are separated by centrifugal force due to their density. The first aqueous solution d is disposed on the inner surface of the centrifugal space 121a to form an outer layer, and the decomposed aliphatic tissue e is disposed inside the first aqueous solution d to form an inner layer.

Since the end 123a of the second duct 123 contacts the decomposed fat tissue e, the decomposed fat tissue e can be transported from the cell separation container 102 to the waste storage device 103 through the second duct 123. Step S105 is similar to step S104, except that the enzyme regulator 183 closes the enzyme conduit 153, the second separation regulator 185b opens the second separation conduit 155b, the first separation regulator 185a closes the first separation conduit 155a, and the waste regulator 184 The waste pipe 154 is opened. The transport unit 106 transports the decomposed fat tissue e of the cell separation vessel 102 through the second separation conduit 155b, the common conduit 105 and the waste conduit 154 to the waste storage device 103. The decomposed adipose tissue e is discharged from the casing 121 through the second conduit 123 of the cell separation vessel 102.

In step S105, the sensor 107 disposed on the separation duct 155 senses the decomposed fat tissue e separated and removed by the transport unit 106 from the cell separation container 102. The decomposed adipose tissue that was removed was yellow. Therefore, in step S105, if the sensor 107 does not sense the yellow decomposed fat tissue e, the transport unit 106 stops operating, and the second separation adjuster 185b closes the second separation duct 155b. When the sensor 107 does not sense the yellow broken fat tissue e, the step is completed. Step S105.

Next, the cleaning liquid is transported from the cleaning liquid storage device 113 to the cell separation container 102, and then the cell separation container 102 is rotated via the rotation unit 120 to stir the cleaning liquid together with the first aqueous solution d (S106) (see FIG. 10). ). Step S106 is similar to step S105 except that the cleaning liquid regulator 182 opens the cleaning liquid pipe 152, the second separation regulator 185b closes the second separation pipe 155b, and the first separation regulator 185a opens the first separation pipe 155a. The transport unit 106 transports the washing liquid stored in the washing liquid storage device 113 of the storage device 101 through the washing liquid pipe 152, the common pipe 105 and the separating pipe 155 (in particular, the first separating pipe 155a) to the cell separating container 102. . The cleaning liquid is transported to the housing 121 through the first conduit 122 of the cell separation vessel 102.

The rotation unit 120 repeats the rotation of the cell separation container 102 and the stop rotation of the cell separation container 102. The stem cells g in the first aqueous solution d are washed with a washing solution. The second aqueous solution f is produced by mixing the first aqueous solution d with the cleaning solution. The second aqueous solution f is an aqueous solution in which the washing liquid is mixed in the first aqueous solution d.

Next, the rotating unit 120 rotates the cell separation container 102, the stem cells g are centrifuged from the second aqueous solution f, and the transport unit 106 transports the second aqueous solution f extracted by the stem cells from the cell separation container 102 to the waste storage device 103 (S107) ( See Figure 11). In step S107, the cell separation container 102 is centrifuged. Due to the centrifugal force generated via the rotating unit 120, the stem cells g and the second aqueous solution f extracted by the stem cells (hereinafter, "the remaining second aqueous solution f") are moved to the centrifugal space 121a of the housing 121. The stem cell g and the remaining second aqueous solution f are different in density It is separated by centrifugal force. The stem cells g are disposed on the inner surface of the centrifugal space 121a to form an outer layer, and the remaining second aqueous solution f is disposed inside the stem cells g to form an inner layer.

Since the end 123a of the second conduit 123 contacts the remaining second aqueous solution f, the remaining second aqueous solution f can be transported from the cell separation vessel 102 to the waste storage device 103 through the second conduit 123. Step S107 is similar to step S106, except that the cleaning liquid regulator 182 closes the cleaning liquid pipe 152, the first separation regulator 185a closes the first separation pipe 155a, the second separation regulator 185b opens the second separation pipe 155b, and The waste regulator 184 opens the waste conduit 154. The transport unit 106 transports the second aqueous solution f of the cell separation vessel 102 through the second separation conduit 155b, the common conduit 105 and the waste conduit 154 to the waste storage device 103. The second aqueous solution f is discharged through the second conduit 123 of the cell separation vessel 102.

Next, the washing liquid is transported from the washing liquid storage device 113 to the cell separation container 102, and the centrifuged stem cells g are dropped to produce the target aqueous solution h (S108) (see Fig. 12). Step S108 is similar to step S107 except that the cleaning liquid regulator 182 opens the cleaning liquid pipe 152, and the waste conditioner 184 closes the waste pipe 154. The transport unit 106 transports the washing liquid stored in the cleaning liquid storage device 113 of the storage device 101 through the cleaning liquid pipe 152, the common pipe 105, and the second separation pipe 155b to the cell separation container 102. The washing liquid passes through the second conduit 123 of the cell separation container 102 and is ejected to the centrifugal space 121a of the housing 121 of the cell separation container 102. The stem cells g are washed by the drops to the lower portion of the cell separation container 102 to produce the target aqueous solution h. The target aqueous solution h contains a washing liquid, And the stem cells g float on the washing liquid.

Next, the transport unit 106 transports the target aqueous solution h from the cell separation container 102 to the cell storage unit 104 (S109) (see Fig. 13). In step S109, the first separation regulator 185a and the cell transport regulator 186 open the first separation conduit 155a and the cell delivery conduit 156, respectively, and the adipose tissue conduit 151, the cleaning fluid conduit 152, the enzyme conduit 153, and the waste conduit 154 are shut down. The transport unit 106 transports the target aqueous solution h in the cell separation container 102 from the cell separation container 102 through the first separation conduit 155a, the common conduit 105, and the cell delivery conduit 156 to the cell storage unit 104. The target aqueous solution h is transported to the cell storage unit 104 through the cell filtration unit 104a. Therefore, the stem cells g are purified and stored and cultured in the cell storage unit 104.

According to the system and method described above, in the cell separation container 102, the stem cells g are separated from the adipose tissue a, and the waste is discharged and removed, thereby obtaining only the high-purity stem cells g. In particular, since only one cell separation container 102 is used, the time and cost for separating and extracting stem cells g can be saved.

100‧‧‧ cell separation system

101‧‧‧Storage device

102‧‧‧ cell separation container

102a‧‧‧Temperature adjustment unit

103‧‧‧Waste storage device

104‧‧‧ cell storage unit

104a‧‧‧cell filtration unit

105‧‧‧Shared pipeline

106‧‧‧Transportation unit

107‧‧‧ sensor

108‧‧‧Adjustment unit

111‧‧‧Adipose tissue storage device

113‧‧‧cleaning liquid storage device

115‧‧‧Enzyme storage device

115a‧‧‧Temperature adjustment unit

120‧‧‧Rotating unit

151‧‧‧Adipose tissue pipeline

152‧‧‧Clean liquid pipeline

153‧‧‧Enzyme Pipeline

154‧‧‧ scrap pipeline

155‧‧‧Separate pipeline

155a‧‧‧The first separation pipeline

155b‧‧‧Second separation pipeline

156‧‧‧cell transport pipeline

181‧‧‧Adipose tissue regulator

182‧‧•washing fluid regulator

183‧‧‧Enzyme regulator

184‧‧‧Waste regulator

185‧‧‧Separation regulator

185a‧‧‧First separation regulator

185b‧‧‧Second separation regulator

186‧‧‧cell transport regulator

109‧‧‧Control unit

Claims (31)

A cell separation container comprising: a housing rotatable on a rotating shaft and having a centrifugal space in the housing, wherein the centrifugal space protrudes from the side surface of the housing in a peripheral direction; the first conduit Inserting into the housing in the direction of the rotating shaft and connecting the inner space of the housing with the outer space of the housing; the second duct is embedded in the housing in the direction of the rotating shaft and connects the shell The inner space of the body and the outer space of the casing, wherein the second pipe is bent such that the end of the second pipe is disposed in the centrifugal space; and the plurality of agitating blades are from the inner surface of the casing protruding. The cell separation container of claim 1, wherein the agitating blade is disposed at a lower portion of the housing and arranged around the first conduit. The cell separation container of claim 1, wherein the agitating blade has a plurality of through holes. The cell separation container according to claim 1, further comprising: an air discharge device disposed on the upper portion of the housing and the inner space connecting the housing and the outer space of the housing, thereby controlling The internal pressure of the housing and the function of filtering air. The cell separation container of claim 1, wherein the first pipe and the body are rotated when the housing is rotated on the rotating shaft The second pipe does not rotate. The cell separation container of claim 1, wherein the first conduit extends to a bottom surface of the housing. The cell separation container of claim 1, wherein the housing is rotated by a motor disposed at a lower portion of the housing. A cell separation method using the cell separation container according to claim 1, wherein the cell separation method comprises the steps of: (a) providing the shell adipose tissue through the first conduit; (b) passing the a pipe provides the casing cleaning liquid, and rotates the casing to stir the fat tissue and the washing liquid, so that after the casing stops rotating, it is separated into the washed adipose tissue of the upper layer and the lower layer respectively. And blood contaminants; (c) removing the blood contaminant from the housing through the first conduit; (d) providing the shell enzyme through the first conduit, and rotating the housing to agitate the washed fat Organizing the enzyme with the enzyme, so that after the shell stops rotating, it is separated into the decomposed adipose tissue and the first aqueous solution which are respectively the upper layer and the lower layer; (e) in the centrifugal space, by rotating the shell, The decomposed adipose tissue and the first aqueous solution are separated into the inner layer and the outer layer respectively, and the decomposed adipose tissue is removed from the shell through the second tube; (f) after the shell stops rotating By mixing the first An aqueous solution and a cleaning solution provided through the first conduit to produce a second aqueous solution; (g) in the centrifugal space, by rotating the housing, centrifuging the second aqueous solution into outer and inner layer stem cells and a second aqueous solution extracted by stem cells, and during centrifugation of the stem cells from the shell Removing the second aqueous solution extracted by the stem cells through the second conduit; and (h) generating a target aqueous solution by mixing the stem cells with the cleaning liquid supplied through the second conduit after the housing stops rotating, and The target aqueous solution is discharged from the housing through the first conduit. A cell separation system comprising: a storage device for separately storing adipose tissue, a washing solution and an enzyme; a cell separation container connected to the storage device, and the fat tissue is supplied from the storage device, the cleaning solution And the enzyme, wherein the cell separation container agitates the adipose tissue supplied from the storage device, the washing solution and the enzyme, and centrifuges the stem cells; the rotating unit is connected to the cell separation container and rotates the cell to separate a container; a waste storage device connected to the cell separation container and storing waste provided by the cell separation container, wherein the waste is in the cell separation container, the adipose tissue is stirred, the cleaning solution and the enzyme and centrifugation Produced during the process of isolating the stem cells; the cell storage unit is connected to the cell separation container and stores the stem cells provided from the cell separation container, wherein the stem cell is agitated by the adipose tissue, the washing solution and the stem cell Enzyme and centrifugation of the adipose tissue; a connecting unit connecting the cell separation container to the storage device, the waste storage device and the cell storage unit; and a transport unit disposed on the connection unit, wherein the transport unit is transported from the storage device via the connection unit The adipose tissue, the washing solution and the enzyme are transferred to the cell separation container, the waste container is transported from the cell separation container to the waste storage device, and the stem cell is transported from the cell separation container to the cell storage unit. The cell separation system of claim 9, further comprising: an inductor disposed on the connecting unit between the cell separation container and the transport unit, and sensing the separation container from the cell via the transport unit The concentration of the waste discharged. The cell separation system of claim 9, wherein the cell separation container comprises: a housing that is rotated on a rotating shaft by the rotating unit disposed at a lower portion of the housing and the housing a centrifugal space in which the centrifugal space protrudes from the side surface of the casing in a peripheral direction; the first pipe is embedded in the casing in the direction of the rotating shaft and connects the internal space of the casing with An outer space of the casing; the second pipe is embedded in the casing in the direction of the rotating shaft and the inner space connecting the casing and the outer space of the casing, wherein the second pipe is bent So that the end of the second pipe is disposed in the centrifugal space; and A plurality of agitating blades project from the inner surface of the housing. The cell separation system of claim 11, wherein the agitating blade is disposed at a lower portion of the housing and arranged around the first conduit. The cell separation system of claim 11, wherein the agitating blade has a plurality of through holes. The cell separation system of claim 11, wherein the cell separation container further comprises: an air discharge device disposed on the upper portion of the housing and the inner space connecting the housing and the housing The external space, thereby controlling the internal pressure of the housing and the function of filtering air. The cell separation system of claim 11, wherein the first conduit and the second conduit do not rotate when the housing is rotated on the rotating shaft. The cell separation system of claim 11, wherein the first conduit extends to a bottom surface of the housing. The cell separation system according to claim 9, wherein the storage device comprises: an adipose tissue storage device for storing the adipose tissue and being connected to the cell separation container via the connection unit; the cleaning liquid storage device The cleaning solution is stored and connected to the cell separation container via the connection unit; and an enzyme storage device stores the enzyme and is connected to the cell separation container via the connection unit. The cell separation system according to claim 17, wherein the connection unit comprises: a common pipe connected to the transport unit; a fat tissue pipe connecting the fat tissue storage device and the shared pipe; a pipe connecting the cleaning liquid storage device and the shared pipe; an enzyme pipe connecting the enzyme storage device and the shared pipe; a waste pipe connecting the waste storage device and the shared pipe; and a separation pipe connecting the cells a separation container and the transport unit; and a cell transport conduit connecting the cell storage unit and the shared conduit. The cell separation system of claim 18, further comprising an adjustment unit disposed on the connection unit and opening or closing the connection unit. The cell separation system of claim 19, wherein the conditioning unit comprises: an adipose tissue regulator disposed on the adipose tissue conduit, and opening or closing the adipose tissue conduit; a cleaning fluid regulator, Is disposed on the cleaning liquid pipeline, and opens or closes the cleaning liquid pipeline; an enzyme regulator is disposed on the enzyme pipeline, and is opened Or shutting down the enzyme conduit; a waste regulator disposed on the waste conduit and opening or closing the waste conduit; a separation regulator disposed on the separation conduit and opening or closing the separation conduit; and cell transport conditioning The device is disposed on the cell transport conduit and opens or closes the cell transport conduit. The cell separation system of claim 19, further comprising: a control unit connected to the rotating unit, the transport unit and the adjusting unit, wherein the control unit controls the transport unit and the adjusting unit, The adipose tissue, the washing solution, the enzyme, the waste material and the stem cells are transported through the connection unit, and the rotation unit is controlled to rotate the cell separation container. The cell separation system according to claim 17, wherein the enzyme storage device and the cell separation container are connected to a temperature adjustment unit, and the temperature adjustment unit maintains the temperature of the enzyme storage device and the cell separation container at 35 °C to 40 °C. The cell separation system of claim 17, wherein the adipose tissue storage device, the enzyme storage device, and the cell storage unit are syringes. The cell separation system of claim 11, wherein the connection unit comprises: a first separation conduit connecting the transport unit to the cell separation The first conduit of the container is opened or closed via a first separation regulator; the second separation conduit connects the delivery unit to the second conduit of the cell separation vessel and is opened via a second separation regulator or shut down. The cell separation system of claim 9, wherein the cell storage unit comprises: a cell filtration unit disposed on the connection unit between the cell storage unit and the transport unit, and filtering through the connection unit The stem cells delivered to the cell storage unit. A cell separation method comprising the steps of: (a) transporting adipose tissue from a fat tissue storage device to a cell separation container via a transport unit; (b) transporting the cleaning liquid from the detergent solution storage device via the transport unit to the a cell separation container, and rotating the cell separation container via a rotating unit to agitate the adipose tissue and the washing liquid, thereby separating the separated cells into upper and lower layers of washed adipose tissue after the cell separation container stops rotating. And (c) transporting the blood contaminant from the cell separation container to the waste storage device via the transport unit; (d) transporting the enzyme from the enzyme storage device to the cell separation container via the transport unit, and via The rotating unit rotates the cell separation container to stir the enzyme and the washed adipose tissue, thereby separating the upper layer into upper layers after the cell separation container stops rotating And the lower layer of the decomposed adipose tissue and the first aqueous solution; (e) when the rotating unit rotates the cell separation container to centrifugally separate the decomposed adipose tissue and the first aqueous solution into an inner layer and an outer layer, respectively a transport unit that transports the decomposed fat tissue from the cell separation container to the waste storage device; (f) after the cell separation container stops rotating, transports the cleaning liquid to the cell separation container via the transport unit, and Dissolving the first aqueous solution and the cleaning solution to produce a second aqueous solution; (g) rotating the cell separation container via the rotating unit to centrifuge the second aqueous solution into stem cells which are outer and inner layers, respectively And the second aqueous solution extracted by the stem cells, and during the centrifugation of the stem cells, the second aqueous solution extracted by the stem cells is transported from the cell separation container to the waste storage device via the transport unit; (h) the cell separation container After the rotation is stopped, the washing liquid is transported to the cell separation container via the transport unit, and the centrifugal separation is performed by using the washing liquid. Stem cells dripping an aqueous solution to produce the target; and (I), an aqueous solution of the target cell separation container conveyed to the reservoir from the cell through the cell carrier unit. The cell separation method of claim 26, wherein the step (c) is repeated until the sensor senses the washed adipose tissue. The cell separation method according to claim 26, wherein Steps (b) and (c) are repeated until the sensor senses that the blood contaminant in step (c) has a concentration below a predetermined value. The cell separation method of claim 26, wherein the cell separation container comprises: a housing that is rotatable on a rotating shaft by a rotating unit, and a centrifugal space in the housing, wherein The centrifugal space protrudes from the side surface of the casing in a peripheral direction; the first pipe is embedded in the casing in the direction of the rotating shaft, and the inner space connecting the casing and the outer space of the casing a second pipe embedded in the casing in the direction of the rotating shaft, and the inner space connecting the casing and the outer space of the casing, wherein the second pipe is bent so that the second pipe The end is disposed in the centrifugal space; and a plurality of agitating blades project from the inner surface of the housing. The cell separation method according to claim 29, wherein the first conduit is used in steps (a), (b), (c), (d), (f) and (i), and the The second pipe is used in steps (e), (g) and (h). The cell separation method according to claim 29, wherein in the step (e), the decomposed adipose tissue and the first aqueous solution are separately centrifuged into the inner layer and the outer layer in the centrifugal space; and the step (g) And the stem cells and the second aqueous solution extracted by the stem cells are separately centrifuged into the outer layer and the inner layer in the centrifugal space.