KR102313128B1 - Continuous cell culturing apparatus which be able to separate liquid - Google Patents

Abstract

The present invention is a bioreactor (bioreactor); An inlet port disposed on the upper side so that a mixture of cells and liquid of a first concentration is introduced from the bioreactor, and a mixture introduced from the inlet port causes a spiral-shaped vortex to form a descending path. a hydrocyclone formed at the lower end of the body and comprising a discharge port through which cells of a second concentration lower than the first concentration separated from the mixture (hereinafter, first separated cells) are discharged; and a filtration membrane mounted on the ceiling surface of the main body, extending toward the discharge port, and separating cells (hereinafter, second separated cells) and liquid (hereinafter, separated liquids) having a third concentration lower than the second concentration from the mixture, and the first separation The cells are returned to the bioreactor side, so that it is possible to obtain the advantages of the perfusion culture apparatus, as well as to continuously increase the production of cell culture, which is a desired product, a continuous cell culture apparatus capable of removing liquid. is about

Description

Continuous cell culture device capable of liquid removal {CONTINUOUS CELL CULTURING APPARATUS WHICH BE ABLE TO SEPARATE LIQUID}

The present invention relates to a continuous cell culture apparatus capable of removing liquid, and more particularly, a liquid capable of acquiring the advantages of a perfusion culture apparatus and continuously increasing the production of cell culture, a desired product. It relates to a continuous cell culture device capable of being removed.

The most important thing in a general perfusion culture medium including the "perfusion culture medium" (hereinafter referred to as the prior art) of Patent Publication No. 10-2003-0001391 is to seek a method of separating only the liquid from the mixture mixed with the cells and liquid in the incubator. can

That is, the perfusion culture adopts a method of continuously supplying fresh media through a pipe, and continuously discharging the cultured media after a certain period of time through another pipe.

In other words, perfusion culture is often used in experiments to examine whether a specific substance is secreted as a function of treatment versus time by sampling a substance or protein secreted by cells at each time period.

For this conventional perfusion culture, cells and liquids are separated by gravity difference using density differences, cells and liquids are separated using centrifugation technology, cells and liquids are separated using membranes, or cells are separated into a certain matrix. A method of separating the liquid by trapping it in the cell or aggregating and separating the cells is used.

However, the conventional perfusion culture method mentioned above has a clear limitation in that it lacks the technology for increasing the cell concentration and increasing the cell culture production by separating the liquid and recycling the egg cells, that is, returning them to a bioreactor and recultivating them. this existed.

Patent Publication No. 10-2003-0001391

The present invention was invented to improve the above problems, and it is possible to obtain the advantages of a perfusion-type culture apparatus, as well as a continuous method capable of removing a liquid so that the production of cell culture, a desired product, can be continuously increased. To provide a cell culture device.

And, an object of the present invention is to provide a continuous cell culture apparatus capable of removing a liquid to completely separate a liquid from a mixture of cells and liquid to increase cell culture efficiency.

In order to achieve the above object, the present invention is a bioreactor (bioreactor); An inlet port disposed on the upper side so that a mixture of cells and liquid of a first concentration is introduced from the bioreactor, and a path in which the mixture introduced from the inlet port causes a spiral-shaped vortex and descends toward the lower side. A hydrocyclone comprising a main body that is gradually narrowed, and an outlet port formed at the lower end of the main body and through which cells of a second concentration lower than the first concentration separated from the mixture (hereinafter, first separated cells) are discharged. ; and a filtration membrane mounted on the ceiling surface of the main body, extending toward the discharge port, and separating cells (hereinafter, second separated cells) and the liquid (hereinafter, separated liquids) having a third concentration lower than the second concentration from the mixture, , It will be possible to provide a continuous cell culture device capable of liquid removal, characterized in that the first separated cells are returned to the bioreactor side.

Here, an inlet pipe interconnecting the bioreactor and the inlet port, a fluid motor mounted on the inlet pipe to transport the mixture to the body side, and a return pipe interconnecting the discharge port and the bioreactor; It is characterized in that it further comprises.

At this time, the second separated cell and the separated liquid is characterized in that it is transferred to a harvest tank (harvest tank).

And, the hydrocyclone, it is characterized in that it further comprises a discharge pipe extending from the ceiling surface of the main body to form a flow path through which the second separated cells and the separated liquid are transferred to the harvest tank.

And, the filtration membrane is characterized in that it is formed in an inverted truncated cone shape.

And, the main body, the upper passage having a cylindrical shape having an outer circumferential surface on which the inlet port is formed, and a lower hopper in the shape of an inverted truncated cone whose diameter gradually decreases from the edge of the lower end of the upper tube toward the discharge port. .

And, as the mixture forms a descending path while generating the spiral vortex, the concentration value of the cells in the mixture descending along the inner peripheral surface of the main body is the concentration of cells in the mixture close to the filtration membrane side is greater than the value.

And, the main body is radially disposed along the outer circumferential surface of the upper cylinder, is mounted at an angle with respect to the outer circumferential surface to form a flow path communicating with the inner space of the main body, from the outside of the main body to the inside of the main body It further comprises a plurality of check valves for injecting compressed air into the space and regulating the flow from the inner space to the outer side of the body.

And, the main body is formed in a spiral shape along the inner circumferential surface of the upper tube and the lower hopper, and further comprises a vortex inducing rib for guiding the mixture flowing in from the inlet port to generate a spiral-shaped vortex to be transported. do.

In addition, the filtration membrane is a metal or resin material containing at least one of palladium (Pd), platinum (Pt), gold (Au), silver (Ag), iridium (lr), rhodium (Rh), or ruthenium (Ru). It is characterized in that it consists of any one or a combination of at least one or more.

In addition, the filtration membrane is characterized in that it is made of a porous material that forms pores having a diameter or width of 1 to 10 μm.

And, the angle formed by the inclined surface of the lower side of the filtration membrane having an inverted truncated cone shape is characterized in that I to II degrees.

And, the angle formed by the inclined surface of the lower side of the lower hopper having an inverted truncated cone shape is characterized in that i to ii degrees.

And, the ratio of the diameter (W2) of the top surface of the filtration membrane having an inverted truncated cone shape to the distance (H2) from the center of the top surface to the center of the lower end of the filtration membrane is (a) to (b).

In addition, the ratio of the diameter (W1) of the ceiling surface of the upper tube having an inverted truncated cone shape to the distance (H1) from the center of the ceiling surface of the upper tube to the center of the lower end of the lower hopper is (A) to (B) characterized in that

According to the present invention having the configuration as described above, the following effects can be achieved.

First, the present invention is expected to significantly increase the cell concentration inside the bioreactor while reducing the culture period by significantly lowering the concentration of cells in contact with the surface of the filtration membrane inside the hydrocyclone.

In other words, since the hourly production of the desired product is generally proportional to the cell concentration, cell culture at a much higher concentration than the conventional perfusion incubator in the bioreactor is possible, thereby increasing the productivity of the culture device. have

In addition, the present invention uses a hydrocyclone connected to a bioreactor to remove the liquid and return the cells, and at the same time to perform cell culture continuously by injecting new media, so that the production of the target product according to the cell culture can be increased. It has the advantage of being able to continuously pursue it.

In particular, the present invention allows to substantially completely separate the liquid from the mixture while adopting the conventional perfusion culture technology so that only pure cells can be re-cultured in the bioreactor, so that the quality of the target product according to cell culture can be improved. will be.

Above all, the present invention greatly improves the liquid separation rate by using the cell concentration gradient inside the hydrocyclone together with the filtration membrane by centrifugal force according to the generation of the spiral vortex, significantly reducing the time and cost required for cell culture It will have the effect of being able to do it.

In addition, according to the present invention, a wide range of applications will be possible to perfusion culture of animals, plants, insects, microorganisms, etc. that produce useful substances including biopharmaceuticals.

In particular, if the function of the hydrocyclone according to the present invention is utilized, it is determined that it will be particularly effective for culturing antibody-producing animal cells.

1 is a conceptual diagram showing the overall structure of a continuous cell culture apparatus capable of removing a liquid according to an embodiment of the present invention;
2 is a conceptual diagram illustrating the internal structure of a hydrocycle and a filtration membrane, which are main parts of a continuous cell culture device capable of removing liquid according to an embodiment of the present invention;

Advantages and features of the present invention, and methods for achieving them, will become apparent with reference to the embodiments described below in detail in conjunction with the accompanying drawings.

However, the present invention is not limited to the embodiments disclosed below, but will be implemented in various different forms.

In the present specification, the present embodiment is provided to complete the disclosure of the present invention, and to fully inform those of ordinary skill in the art to which the present invention pertains to the scope of the present invention.

And the invention is only defined by the scope of the claims.

Accordingly, in some embodiments, well-known components, well-known operations, and well-known techniques have not been specifically described to avoid obscuring the present invention.

In addition, like reference numerals refer to like elements throughout the specification, and terms used (referred to) herein are for the purpose of describing the embodiments and are not intended to limit the present invention.

In this specification, the singular also includes the plural unless specifically stated otherwise in the phrase, and elements and operations referred to as 'comprising (or having)' do not exclude the presence or addition of one or more other elements and operations. .

Unless otherwise defined, all terms (including technical and scientific terms) used herein may be used with the meaning commonly understood by those of ordinary skill in the art to which the present invention belongs.

Also, terms defined in commonly used dictionary are not to be interpreted ideally or excessively unless defined.

Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings.

First, FIG. 1 is a conceptual diagram illustrating the overall structure of a continuous cell culture apparatus capable of removing a liquid according to an embodiment of the present invention.

And, FIG. 2 is a conceptual diagram illustrating the internal structure of a hydrocycle and a filtration membrane, which are main parts of a continuous cell culture apparatus capable of removing a liquid according to an embodiment of the present invention.

For reference, the arrow indicated by the dashed-dotted line in the drawing indicates the transfer direction of the mixture flowing from the bioreactor (100, bioreactor) to the hydrocyclone (200, hydrocyclone), as a solid line from the inside of the filtration membrane 300 toward the discharge pipe 240. The indicated arrows indicate the transfer direction of the liquid separated from the above-described mixture, and the dotted spiral arrows indicate the transfer direction of the cells remaining after the liquid has been separated.

As shown in the present invention, a bioreactor 100 in which cell culture is performed, a hydrocyclone 200 connected to the above-described bioreactor 100, and a filtration membrane 300 embedded in the above-described hydrocyclone 200 are provided. It can be seen that the structure contains

First, the hydrocyclone 200 includes an inlet port 201 disposed on the upper side so that a mixture of cells and liquid of a first concentration flows from the bioreactor 100 .

In addition, the hydrocyclone 200 includes a body 210 that is gradually narrowed toward the lower side so that the mixture introduced from the inlet port 201 generates a spiral-shaped vortex and descends to form a descending path.

In addition, the hydrocyclone 200 is formed at the lower end of the body 210 and includes an outlet port 202 through which cells of a second concentration lower than the first concentration separated from the mixture (hereinafter, first separated cells) are discharged.

In addition, the filtration membrane 300 is mounted on the ceiling surface of the main body 210 and extends toward the discharge port 202, and from the mixture, cells (hereinafter referred to as second cells) and liquid (hereinafter referred to as separated liquid) having a third concentration lower than the second concentration. ) to separate

Accordingly, the first separated cells may be returned to the bioreactor 100 to be utilized for cell culture.

That is, by using the hydrocyclone 200 connected to the bioreactor 100 to remove the liquid, return the cells, and at the same time inject new media, so that the cell culture can be continuously performed, Production can be done continuously.

Here, the third concentration is negligibly low compared to the second concentration, that is, almost dilute, and refers to a cell concentration in which most of the cells are diluted in the above-described separation liquid.

That is, the third concentration may be considered to be a negligible concentration level close to zero.

It goes without saying that the present invention can be applied to the above-described embodiments, and can also be applied to the following various embodiments.

First, the bioreactor 100 includes a culture vessel 110 having a culture space 111 in which an influent and a mixture including fresh cells introduced from the outside and a returned first separated cell are accommodated therein. may include.

In addition, the bioreactor 100 may include a cover 120 covering the open upper surface of the culture vessel 110 .

In addition, the bioreactor 100 includes a rotation shaft 131 that penetrates through the cover 120 and is accommodated in the culture space 111 and is rotatably mounted, and is provided at the upper end of the rotation shaft 131 to apply a rotational force to the rotation shaft 131 . It may include a motor 132 for transmitting the, and a stirrer 130 including a blade 133 provided at the lower end of the rotation shaft 131 to stir the contents in the culture space 111 .

In addition, the bioreactor 100 may include a supply pipe 140 that forms a flow path through which the influent flows into the culture space 111 through the cover 120 .

In addition, the bioreactor 100 may include a fluid supply motor 150 mounted on the supply pipe 140 to transmit a transfer force for introducing an influent from the outside to the culture space 111 .

The bioreactor 100 is not particularly shown in the present invention, but in some cases, an aerator for supplying oxygen to the culture space 111 is added and applied and modified design is also possible, such as using it for culturing aerobic organisms. .

Therefore, the first separated cells discharged through the hydrocyclone 200 return to the bioreactor 100 again in a state in which the liquid is almost completely separated, so that the cells continue to grow in the bioreactor 100 through this. Accordingly, the cell concentration in the bioreactor 100 also increases, so that the cell culture yield can be increased.

On the other hand, the inlet pipe 220 interconnects the bioreactor 100 and the inlet port 201, and a fluid motor 221 is mounted on the inlet pipe 220 to transfer the mixture to the body 210 side, and return The pipe 230 interconnects the discharge port 202 and the bioreactor 100 so that the first separated cells return to the culture space 111 .

In addition, the second separated cells and the separated liquid may be transferred to a harvest tank (hereinafter not shown).

In addition, the hydrocyclone 200 may further include a discharge pipe 240 extending from the ceiling surface of the main body 210 to form a flow path through which the second separated cells and the separated liquid are transferred to the harvest tank.

On the other hand, the filtration membrane 300 is manufactured to be formed in an inverted truncated cone shape to correspond to the body 210 shape of the hydrocyclone 200 to be described later by using the liquid with a low cell concentration rushing to the center of the body 210 . desirable.

In addition, the lower end of the discharge pipe 240 is arranged in the center of the open upper surface of the filtration membrane 300, and although not shown in particular, on the discharge pipe 240, a pump for discharging fluid for smooth discharge of the separated and discharged liquid or Of course, it is also possible to add a fluid motor.

On the other hand, the main body 210, a cylindrical upper cylinder 211 having an outer circumferential surface on which the inlet port is formed, and an inverted truncated cone shape whose diameter is gradually reduced from the lower end edge of the upper cylinder 211 toward the discharge port 202 side. It may be manufactured in a structure including a lower hopper 212 .

As the shape of the body 210 described above forms a path for the mixture to descend while generating a spiral-shaped vortex, the concentration value of cells in the mixture descending along the inner circumferential side of the body 210 is on the filtration membrane 300 side. Since it becomes larger than the concentration value of the cells in the adjacent mixture, it can be said to be an optimal structure to increase the separation yield of the liquid from the mixture.

Meanwhile, the hydrocyclo 200 and the filtration membrane 300, which are main parts of a continuous cell culture apparatus capable of removing liquid according to another embodiment of the present invention, will be described with reference to FIG. 2 .

First, the main body 210 is radially disposed along the outer circumferential surface of the upper cylinder 211, is mounted at an angle with respect to the outer circumferential surface to form a flow path communicating with the inner space of the main body 210 check valve 213 may be provided with more.

The check valve 213 serves to inject compressed air from the outside of the main body 210 to the inner space side of the main body 210 and to regulate the flow from the inner space to the outer side of the main body 210 .

That is, the check valve 213 is provided to more strongly form a spiral-shaped vortex to increase the centrifugal force when the mixture is transferred to the discharge port 202 side, thereby further increasing the separation amount of the liquid from the cells.

In addition, the main body 210 is formed in a spiral shape along the inner circumferential surface of the upper cylinder 211 and the lower hopper 212 to induce a vortex to guide the mixture flowing in from the inlet port 201 to generate a spiral-shaped vortex and transport it. It may further include ribs 214 .

The vortex induction rib 214 is also strongly and constantly formed in a standardized spiral pattern with the check valve 213 together with the check valve 213 to increase the centrifugal force when the mixture is transferred to the discharge port 202 side. It is provided to further increase the separation amount of the liquid.

On the other hand, the filtration membrane 300 is a metal material containing at least one of palladium (Pd), platinum (Pt), gold (Au), silver (Ag), iridium (lr), rhodium (Rh), or ruthenium (Ru) or It may be made of any one of the resin materials or a combination of at least one or more.

The metal filtration membrane 300 is manufactured using, for example, lithography using X-rays or ultraviolet rays and electroforming.

The resin filtration membrane 300 is molded using, for example, a mold manufactured using X-ray or ultraviolet lithography and electroforming.

Although it is preferable that resin is transparent, it may be opaque.

The metal material contains, for example, at least one of palladium (Pd), platinum (Pt), gold (Au), silver (Ag), iridium (lr), rhodium (Rh), or ruthenium (Ru).

The material may be a single metal of palladium (Pd), platinum (Pt), gold (Au), silver (Ag), iridium (lr), rhodium (Rh), or ruthenium (Ru), for example, palladium (Pd) ) · a nickel (Ni) alloy, a platinum (Pt) · nickel (Ni) alloy, or a gold (Au) · nickel (Ni) alloy or the like.

In the case of an alloy, it is preferable that there are many ratios of said metal with respect to a counter metal, such as nickel.

On the other hand, the filtration membrane 300 is preferably made of a porous material that forms pores having a diameter or width of 1 to 10 μm so that only a liquid can pass through without passing cells from the mixture.

In addition, the angle θ2 formed by the lower inclined surface of the filtration membrane 300 having the inverted truncated cone shape is in the range of I to II degrees, and the angle θ1 formed by the lower inclined surface of the lower hopper 212 having the inverted truncated cone shape. can be appropriately designed in the range of i to ii.

The range of the aforementioned angles (θ1, θ2) is to secure an optimal structure for the mixture introduced from the inlet port 201 to smoothly separate only the liquid in the filtration membrane 300 while descending while smoothly generating a spiral-shaped vortex. will be.

In addition, the ratio of the diameter (W1) of the ceiling surface of the upper cylinder (211) having an inverted truncated cone shape and the distance (H1) from the center of the ceiling surface of the upper cylinder (211) to the center of the lower end of the lower hopper (212) 1 ratio) is preferably (A) to (B).

Here, when the value of the above-mentioned first ratio is very small, less than (A), the body 210 of the hydrocyclone 200 has a shape that is close to a cylinder or polygonal pillar having a small diameter or width, so that the mixture is spiral As it forms a vortex of shape and flows down in the direction of gravity before going down, you will face a problem that the liquid cannot be properly separated.

At this time, when the value of the above-described first ratio becomes very large exceeding (B), the diameter or width of the body 210 of the hydrocyclone 200 is a circular block or polygonal block in which the diameter or width corresponds to a height of approximately one-to-one. Since the shape is closer to , as the mixture goes straight to the discharge port 202 without a gap to form a spiral-shaped vortex, it will face a problem that the liquid cannot be properly separated.

In addition, the ratio of the diameter (W2) of the ceiling surface of the inverted truncated cone-shaped filtration membrane 300 and the distance (H2) from the center of the ceiling surface to the center of the lower end of the filtration membrane 300 (hereinafter the second ratio) is (a) to (b) is preferable.

Here, when the value of the above-mentioned second ratio is very small less than (a), the filtration membrane 300 has a shape that is close to a cylinder or polygonal pillar having a small diameter or width, and thus forms a spiral-shaped vortex to the lower side. The problem will be that the liquid will not be properly separated from the mixture going down along the flow path forming a narrowing diameter and width.

At this time, when the value of the above-described second ratio becomes very large exceeding (b), the filtration membrane 300 has a shape that is close to a circular block or a polygonal block in which the diameter or width corresponds to almost one-to-one correspondence with the height. , it will face a problem that the liquid cannot be properly separated from the mixture descending along the flow path forming a spiral-shaped vortex and narrowing in diameter and width to the lower side.

In other words, if the structure is out of the upper limit or the lower limit of the range of the value of the second ratio described above, it means that the lower end of the filtration membrane 300 has an area enough to form a surface in general and becomes a wider structure.

Therefore, due to the structural characteristics of the filtration membrane 300 formed in this way, the portion below the middle portion of the outer peripheral surface of the filtration membrane 300 collides with a vortex formed in a spiral shape that gradually narrows from the inside of the body 210 to the lower side, so that the liquid is not properly separated. You may face a problem that a significant amount of liquid will return to the discharge port 202 side as it is with the cells.

Hereinafter, the operation and effect of the continuous cell culture apparatus capable of removing liquid according to a preferred embodiment of the present invention will be looked at as follows.

First, the present invention is a bioreactor 100; The inlet port 201 disposed on the upper side so that a mixture of cells and liquid of a first concentration is introduced from the bioreactor 100, and a path in which the mixture introduced from the inlet port 201 descends while generating a spiral-shaped vortex Hydro comprising a main body 210 that is gradually narrowed toward the lower side to form, and an outlet port 202 that is formed at the lower end of the main body 210 and discharges the first separated cells of a second concentration separated from the mixture. cyclone 200; and a filtration membrane 300 mounted on the ceiling surface of the main body 210 and extending toward the discharge port 202 to separate the second separated cells and the separated liquid having a third concentration lower than the second concentration from the mixture, the first Since the separated cells are returned to the bioreactor 100 side, it will be possible to continuously increase the production according to cell culture as a desired target product while maintaining the advantages of the existing perfusion culture apparatus.

And, according to the present invention, an inlet pipe 220 interconnecting the bioreactor 100 and the inlet port 201, and a fluid motor mounted on the inlet pipe 220 to transfer the mixture to the body 210 side ( 221), and a return pipe 230 for interconnecting the discharge port 202 and the bioreactor 100, so that the pure first separated cells from which the separation liquid has been removed from the mixture are again cultured in the bioreactor 100 It will be possible to reduce the supply load of the influent and reduce unnecessary power and energy use.

That is, since the hourly output of the desired product is generally proportional to the cell concentration, cell culture at a much higher concentration than the conventional perfusion incubator in the bioreactor 100 is possible, thereby increasing the productivity of the culture apparatus. has the advantage of being

And, according to the present invention, the second separated cells and the separated liquid are transferred to the harvest tank, the hydrocyclone 200 according to the present invention is extended from the ceiling surface of the main body 210, the second separated cells and the separated liquid By further providing a discharge pipe 240 that forms a flow path through which is transferred to the harvest tank, it will be possible to significantly improve the recycling capacity of the cell culture.

And, the filtration membrane 300 according to the present invention is formed in an inverted truncated cone shape, and the main body 210 according to the present invention has a cylindrical upper cylinder 211 having an outer peripheral surface on which an inlet port is formed, and an upper cylinder 211 ) including a lower hopper 212 of an inverted truncated cone shape that gradually decreases in diameter from the lower edge of the lower end toward the discharge port 202 side, and as the mixture forms a descending path while generating a spiral-shaped vortex, the body 210 Since the concentration value of the cells in the mixture descending along the inner circumferential side of the filtration membrane 300 is greater than the concentration value of the cells in the mixture adjacent to the filtration membrane 300 side, the cell in contact with the surface of the filtration membrane 300 inside the hydrocyclone 200 is By significantly lowering the concentration, it will be possible to continuously increase the cell concentration in the bioreactor 100 while reducing the incubation period.

As described above, the present invention is to provide a continuous cell culture apparatus capable of liquid removal that can obtain the advantages of the perfusion culture apparatus and continuously increase the production of cell culture, which is a desired product, as a basic technical goal. It can be seen that he is doing it as a thought.

And, within the scope of the basic technical spirit of the present invention, many other modifications and applications are also possible for those of ordinary skill in the art.

100...Bioreactor
110...Incubation vessel
111...incubation space
120...cover
130...agitator
131... the axis of rotation
132...Motor
133...blade
140...supply pipe
150...Fluid supply motor
200...hydrocyclone
201...entry port
202...Exhaust port
210...Body
211... upper pain
212...lower hopper
213...Check valve
214...Vortex induction rib
220...inlet piping
221...fluid motor
230...return piping
240...exhaust piping
300...Filtration membrane

Claims (15)

bioreactors;
An inlet port disposed on the upper side so that a mixture of cells and liquid of a first concentration is introduced from the bioreactor, and a path in which the mixture introduced from the inlet port causes a spiral-shaped vortex and descends toward the lower side. A hydrocyclone comprising a main body that is gradually narrowed, and an outlet port formed at the lower end of the main body and through which cells of a second concentration lower than the first concentration separated from the mixture (hereinafter, first separated cells) are discharged. ; and
It is mounted on the ceiling surface of the main body and extends toward the discharge port and includes a filtration membrane that separates cells of a third concentration lower than a second concentration (hereinafter referred to as second separated cells) and the liquid (hereinafter referred to as separated liquid) from the mixture,
The first separated cells are returned to the bioreactor side,
The hydrocyclone further comprises a discharge pipe extending from the ceiling surface of the main body to form a flow path through which the second separated cells and the separated liquid are transported,
The second separated cell and the separated liquid are transferred to a harvest tank through the discharge pipe,
The filtration membrane is formed in an inverted truncated cone shape,
The main body is radially disposed along the outer circumferential surface of the upper tube of the main body, is mounted at an angle with respect to the outer circumferential surface to form a flow path communicating with the inner space of the main body, and from the outside of the main body to the inside of the main body A continuous cell culture apparatus capable of liquid removal, characterized in that it further comprises a plurality of check valves for injecting compressed air into the space and regulating the flow from the inner space to the outer side of the body.
The method according to claim 1,
an inlet pipe interconnecting the bioreactor and the inlet port;
a fluid motor mounted on the inlet pipe to transport the mixture to the body side;
A continuous cell culture apparatus capable of removing liquid further comprising a return pipe interconnecting the discharge port and the bioreactor.
delete delete delete The method according to claim 1,
The body is
A cylindrical upper passage having an outer peripheral surface on which the inlet port is formed;
A continuous cell culture apparatus capable of removing liquid, characterized in that it comprises a lower hopper in the shape of an inverted truncated cone, the diameter of which gradually decreases from the edge of the lower end of the upper cylinder toward the discharge port.
The method according to claim 1,
As the mixture forms a descending path while generating the spiral-shaped vortex, the concentration value of the cells in the mixture descending along the inner circumferential side of the body is,
A continuous cell culture apparatus capable of removing liquid, characterized in that it is larger than the concentration value of the cells in the mixture close to the side of the filtration membrane.
delete 7. The method of claim 6,
The body is
Continuous liquid removal possible, characterized in that it is formed in a spiral shape along the inner circumferential surface of the upper tube and the lower hopper and further comprises a vortex inducing rib for guiding the mixture flowing in from the inlet port to generate a spiral-shaped vortex to be transported. Phagocytic cell culture apparatus.
The method according to claim 1,
The filtration membrane is any one of a metal material or a resin material containing at least one of palladium (Pd), platinum (Pt), gold (Au), silver (Ag), iridium (lr), rhodium (Rh), or ruthenium (Ru) A continuous cell culture apparatus capable of removing liquid, characterized in that it consists of one or at least one combination.
The method according to claim 1,
The filtration membrane is a continuous cell culture device capable of removing liquid, characterized in that it is made of a porous material that forms pores having a diameter or width of 1 to 10 μm.
The method according to claim 1,
A continuous cell culture device capable of removing liquid, characterized in that the angle formed by the inclined surface of the lower side of the filtration membrane having an inverted truncated cone shape is 1 to 2 degrees.
7. The method of claim 6,
A continuous cell culture apparatus capable of removing liquid, characterized in that the angle formed by the inclined surface of the lower side of the lower hopper having an inverted truncated cone shape is 1 to 2 degrees. delete delete

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