KR101412083B1 - Cell culture unit, cell culture device and cell culture system - Google Patents

Abstract

TECHNICAL FIELD The present invention relates to a cell culture unit, and more particularly, to a cell culture unit, a cell culture device, and a cell culture system capable of easily discharging a lung culture fluid. According to an aspect of the present invention, there is provided a cell culture system comprising a plurality of cell culture units, a plate for receiving cell culture units, and a cell culture unit An incubator, a culture solution storage container in which the culture solution is stored in a sealed state, and a waste culture solution storage container in which the waste solution is stored in a sealed state. Wherein the cell culture unit comprises: a culture unit which is an inner space in which cells are grown; an inlet for injecting a culture solution into the culture unit; an outlet for discharging a pulmonary culture solution of the culture unit; and an outlet for connecting the culture unit and the outlet Channel. The resistance of the outflow channel is preferably larger than the resistance between the inlet and the culture unit. The cell culture unit according to the present invention can easily discharge the pulmonary culture solution and improve the growth condition of the cells.

Description

[0001] The present invention relates to a cell culture unit, a cell culture unit, and a cell culture system,

TECHNICAL FIELD The present invention relates to a cell culture unit, and more particularly, to a cell culture unit, a cell culture device, and a cell culture system capable of easily discharging a lung culture fluid.

Cultivation of animal cells means that cells are grown by forming conditions similar to in vivo artificially in vitro. The cells grow in suspension or monolayer attachment. The cultivation of animal cells can be roughly divided into two parts: the production of medical therapeutic proteins such as enzymes, hormones, vaccines, immunomodulators, anticancer drugs, and the production of organs and tissues. The anti-cancer immune cell therapy for the treatment of tumor cells of cancer patients can be carried out by culturing the cells in vitro and then by increasing the number of immune cells selectively capable of selectively killing tumor cells or by functionally enhancing them, It is an anticancer therapy that is injected and treated.

As an example of a device for in vitro cell culture, US Pat. No. 5,026,649 discloses an apparatus for growing tissue cultures in vitro. The technology of this patent document consists of a tissue culture cluster dish, a membrane support assembly, a membrane and a lid. Tissue Culture The collecting dish contains a nutrient solution for the growth of tissue culture samples (Tissue culture sample). The bottom of the membrane support assembly is contained in a nutrient solution. The membrane that permeates the nutrient solution is attached to the bottom of the membrane support assembly. The tissue culture sample lies on the top surface of the membrane. The drug solution passes through the membrane and flows into the membrane support assembly to grow a tissue culture sample.

Since the cells grow under artificial growth conditions that are similar to the growth conditions of the human body, unlike the case where the cells are grown into the tissues of the human body, the in vitro culture growth apparatus described above exhibits various physiological and biochemical changes none. Therefore, there is a problem that the cell culture is very difficult and hard to improve the viability of the cells. In addition, waste products such as lactate and ammonia, which are generated during cell growth, are left in the nutrient solution and deteriorate the growth conditions.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above problems occurring in the prior art, and it is an object of the present invention to provide a cell culture unit capable of easily removing waste products generated during cell growth in a culture solution, and a cell culture apparatus and a cell culture system do.

It is another object of the present invention to provide a cell culture unit capable of culturing cells safely by minimizing unnecessary contact between cells and the atmosphere by increasing the degree of sealing and a cell culture apparatus and a cell culture system including the same.

According to an aspect of the present invention, there is provided a cell culture system comprising a plurality of cell culture units, a plate for receiving cell culture units, and a cell culture unit An incubator, a culture solution storage container in which the culture solution is stored in a sealed state, and a waste culture solution storage container in which the waste solution is stored in a sealed state.

Wherein the cell culture unit comprises: a culture unit which is an inner space in which cells are grown; an inlet for injecting the culture solution into the culture unit; an outlet for discharging the pulmonary culture solution of the culture unit; and an outlet for connecting the culture unit and the outlet Channel. The resistance of the outflow channel is preferably larger than the resistance between the inlet and the culture unit.

Wherein the plate includes partition walls for partitioning a space surrounded by the outer wall and the outer wall into a plurality of receiving portions capable of accommodating the plurality of cell culture units, The pathway is formed so that the pulmonary culture fluid stored in the pathway can flow in one direction.

The incubator includes a plate accommodating portion capable of accommodating the plate, a heater for heating the plate accommodating portion to a temperature according to the culturing condition, and a gas for maintaining the atmosphere according to the culturing condition to the plate accommodating portion A ditch formed around the plate accommodating portion for accommodating a pulmonary culture fluid flowing through an outer wall of the plate, and a trough formed in the trench so as to discharge the pulmonary culture fluid stored in the trench to the outside, And a culture liquid delivery pipe for supplying a culture liquid to an inlet of the cell culture units.

The cell culture system may further include a first connection pipe connecting the culture solution storage container and the culture solution distribution pipe, a first pump installed in the first connection pipe, and a second pump connected to the waste culture solution discharge pipe, A second connection pipe, and a second pump installed in the second connection pipe.

Preferably, the cell culture system further includes a pressure compensating pipe connecting the culture medium storage container and the waste culture medium storage container to each other.

The culture liquid distributor includes an inlet through which the culture liquid flows, a plurality of outlets for discharging the culture liquid and connected to the culture liquid distribution tubes, and a distributor partition wall disposed inside to divide the culture liquid distributor into the inlet space and the outlet space. .

According to an aspect of the present invention, there is provided a cell culture apparatus comprising: a culture unit which is an inner space in which cells are grown; an inlet for injecting a culture solution into the culture unit; an outlet for discharging a lung culture solution of the culture unit; A plurality of cell culture units each having a resistance larger than a resistance between the inlet and the culture unit, and a plurality of cell culture units each having an outer wall and a space surrounded by the outer wall, And a partition wall for partitioning the plurality of cell culture units into a plurality of receptacles capable of accommodating a plurality of cell culture units, wherein the outer wall and the partition walls are provided with passages for discharging the pulmonary culture fluid discharged from the outlets and flowing in one direction Lt; / RTI >

In order to accomplish the above object, a cell culture unit according to the present invention comprises a culture unit which is an inner space where cells are grown, an inlet for injecting a culture solution into the culture unit, an outlet for discharging a lung culture solution of the culture unit, And an outlet channel connecting the culture unit and the outlet. The resistance of the outflow channel is preferably larger than the resistance between the inlet and the culture unit.

In addition, it is preferable that one side of the bottom surface of the cell culture unit is formed to be stepped, and the outlet is formed so as to discharge the waste culture liquid toward a space formed by a step on the bottom surface.

The outlet channel may have at least one bend.

The cell culture unit may further include a culture medium reservoir for primarily storing the culture fluid introduced from the inlet, an inlet channel connecting the culture medium reservoir to the culture unit, and a resistance lower than that of the outlet channel.

The cell culture unit according to the present invention can easily discharge the pulmonary culture solution and improve the growth condition of the cells. In addition, it is possible to minimize contact with the atmosphere, which hinders cell growth during cell culture.

The cell culture apparatus according to the present invention is advantageous in that only a number of cell culture units required for the test are mounted on a plate, and when the test is completed, only a used cell culture unit is discarded and a new cell culture unit is mounted on a plate have.

1 is a conceptual view briefly showing a cell culture system according to the present invention.
2 is an exploded perspective view showing a part of the cell culture system shown in Fig.
3 is a perspective view and an exploded perspective view of the cell culture unit shown in Fig.
Figure 4 is a perspective view of the plate shown in Figure 1;
5 is a perspective view of the incubator shown in Fig.
6 is a perspective view of the incubator cover and the culture liquid distributor shown in Fig.
7 is a perspective view of the culture liquid distributor shown in Fig.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

The following embodiments are provided by way of example so that those skilled in the art can fully understand the spirit of the present invention. Therefore, the present invention is not limited to the embodiments described below, but may be embodied in other forms. In the drawings, the width, length, thickness, etc. of components may be exaggerated for convenience. Like reference numerals designate like elements throughout the specification.

FIG. 1 is a conceptual view briefly showing a cell culture system according to the present invention, and FIG. 2 is an exploded perspective view showing a part of the cell culture system shown in FIG.

1 and 2, the cell culture system according to the present invention comprises a plurality of cell culture units 10, a plate 20 for accommodating a plurality of cell culture units 10, And an incubator 30 for providing a temperature and an atmosphere suitable for the culture medium and the culture medium storage container 40 and 41. The culture medium and the culture medium storage containers 40 and 41 and the first and second culture media 40 and 41, First and second pumps 44 and 45 provided in the first and second connection pipes 42 and 43 for circulating the culture liquid and the pulmonary culture liquid and the culture liquid storage container 40 And a pressure-compensating pipe 46 connecting the waste-culture-solution storage container 41 with the waste-culture-solution storage container 41.

3 is a perspective view and an exploded perspective view of the cell culture unit shown in Fig.

Referring to Fig. 3, the cell culture unit 10 is generally a rectangular parallelepiped. The cell culture unit 10 is made of a transparent polymer material. For example, a polymer material such as PDMS (polydimethylsiloxane), PS (polystyrene), or PE (polyethylene).

At the center of the cell culture unit 10, a culture unit 11, which is an inner space in which cells grow, is formed. The cross section of the culture section 11 is generally circular or elliptical.

On the upstream side (left side in the drawing) of the culture unit 11, an inlet 12 for injecting the culture liquid into the culture unit 11 is formed long in the thickness direction of the cell culture unit 10. The end of the inlet port 12 on the side of the culture section 11 is connected to a reservoir 13 which can receive the culture liquid. The culture medium reservoir 13 has a smaller cross-sectional area than the culture unit 11. The culture medium reservoir 13 and the culture section 11 are connected to each other by an inlet channel 14. [ A steel pipe 15 is inserted into the inlet 12 (see Fig. 2). By inserting the steel pipe, the culture liquid can be stably flown regardless of whether the surface tension of the culture liquid and the material of the cell culture unit are hydrophobic or hydrophilic.

On the downstream side (right side in the drawing) of the cultivation part 11, an outflow port 16 for discharging the pulmonary culture fluid generated in the cultivation part 11 is formed at the lower side. When the cell culture unit 10 is placed on a flat surface, the bottom of the cell culture unit 10 on which the outlet 16 is formed has a height difference, and a waste culture liquid is discharged between the surface of the cell culture unit 10 and the bottom surface of the cell culture unit 10 A space 17 to be formed is formed. The outlet (16) is formed toward the space (17) so that the waste culture liquid can be discharged into the space.

The outlet 16 and the culture unit 11 are connected to each other by an outlet channel 18. The outlet channel 18 is curved in S or U shape to increase resistance to the pulmonary fluid flow. The resistance of the outlet channel 18 must be greater than the resistance of the inlet channel 14 so that the cells growing in the culture section 11 can flow through the outlet channel 18 It can be prevented that it is easily leaked.

Since the upstream side of the culture unit 11 is closed by the culture liquid and the downstream side is closed by the lung culture liquid, the culture unit 11 has an advantage that unnecessary contact with the atmosphere is minimized.

The cell culture unit 10 can be manufactured by using a photolithography technique or an injection molding technique. The cell culture unit 10 may be manufactured by a method in which upper and lower portions are respectively manufactured as shown in FIG. 3 (b), and then the upper portion and the lower portion are thermally joined to each other.

Figure 4 is a perspective view of the plate shown in Figure 1; 4, the plate 20 includes a plate 20 that divides the space surrounded by the outer wall 21 and the outer wall 21 forming the outer periphery of the plate 20 into a plurality of receiving portions 22, And barrier ribs 23 and 24 in the width direction. The upper and lower surfaces of the plate 20 are open.

Passages 26 are formed on one side (right side in the drawing) of the lower ends of the width direction partition walls 24. Also, a passage 25 is formed on one side of the lower end of one of the lateral walls 21 in the width direction. The passages 25 and 26 formed in the width direction partition walls 24 and the outer wall 21 become the passages 25 and 26 through which the waste culture liquid passes.

One edge 27 of the plate 20 is chamfered as a mark for indicating the direction in which the cell culture unit 10 is disposed.

5 is a perspective view of the incubator shown in Fig. Referring to FIG. 5, the bottom surface 31 of the incubator 30 has a lattice shape, and a heater (not shown) is installed inside the lattice. The cell culture unit 10 is disposed on the open portion of the bottom surface 31 of the incubator 30 and the intervals of the gratings are adjusted so that the partition walls 23 and 24 of the plate 20 are disposed on the grating . In order to prevent the lattice from covering the cell culture unit 10 when observing the cells growing in the cell culture unit 10 through the bottom surface 31 of the incubator 30 with a phase contrast microscope. A glass plate 28 having a thickness of 0.1 mm is disposed between the bottom surface 31 of the incubator 30 and the plate 20 to support the cell culture unit 10.

The incubator 30 has a plate receiving portion 32 which is a space for accommodating the plate 20 in the center portion thereof. A ditch 33 is formed around the plate receiving portion 32 to receive the waste culture liquid discharged through the passages 25 and 26 of the plate 20.

On the side surface of the incubator 30, a waste culture liquid discharge pipe 34 is installed to discharge the waste culture liquid stored in the ditch 33 to the outside. The trench 33 is inclined so that the pulp culture fluid can be collected at a position where the pulmonary fluid discharge pipe 34 is installed.

6 is a perspective view of the incubator cover and the culture liquid distributor shown in Fig. An incubator cover will be described with reference to Fig.

The incubator cover (35) covers the open upper surface of the incubator (30). A window 36 is provided at the center of the incubator cover 35 so that the cell culture units 10 inside the incubator 30 can be observed. A plurality of through holes 37 are formed in the window 36 at positions corresponding to the inlets 12 of the cell culture unit 10. On the window 36, a plurality of culture liquid distributors 50 are installed side by side.

A gas piping 38 for supplying a gas for controlling the atmosphere is provided inside the incubator 30 on the side surface of the incubator cover 35. As shown in FIG. 1, the gas pipe 38 is connected to a bomb 39 in which compressed gas is stored.

7 is a perspective view of the culture liquid distributor shown in Fig. 6 and 7, the culture liquid distributor 50 includes an inlet 51 into which a culture liquid is introduced, a plurality of outlets 52 through which the culture liquid is discharged, and a culture liquid distributor 50 into the inlet side space 53 and the outlet side space And a culture liquid distributing pipe 56 connected to the plurality of outlets 52 and a distributor partition wall 55 disposed inside the culture liquid distributor 50 so as to be distinguished from each other. The distributor partition wall 55 serves to discharge the culture liquid supplied to the inlet 51 to the plurality of outlets 52 evenly.

The culture liquid distributing pipes 56 are inserted into the plurality of through holes 37 of the window 36 and are connected to the ends of the culture liquid distributing pipe 56 and the steel pipe inserted into the inlets 12 of the cell culture unit 10 15 are connected to each other by a connection pipe 58 made of a plastic material.

The inlet (51) of the culture liquid distributor (50) is connected to the culture liquid storage container (40) through the first connection pipe (42). The culture medium storage container 40 maintains a closed state so that the culture medium is not in contact with air. The first connection pipe 42 is provided with a first pump 44 for supplying the culture solution of the culture solution storage container 40 to the culture solution distributor 50.

The pulmonary culture liquid discharge pipe (34) is connected to the pulmonary fluid storage container (41) through the second connection pipe (43). The waste culture medium storage container 41 is also kept in a sealed state. The second connection pipe (43) is provided with a second pump (45) for supplying a pulmonary fluid to the lung fluid storage container (41).

The culture medium storage container 40 and the waste culture medium storage container 41 are connected to each other by the pressure compensation pipe 46. Since the culture liquid is discharged in a closed state in the culture liquid storage vessel 40, negative pressure is generated in the culture liquid storage vessel 40. Since the pulmonary fluid is supplied in a closed state to the pulmonary fluid storage container 41, a positive pressure is generated in the pulmonary fluid storage container 41. Therefore, unless a separate gas is supplied or gas is discharged, the supply of the culture liquid and the discharge of the liquid medium may be stopped. This problem can be solved at once by connecting the culture medium storage container 40 and the waste culture medium storage container 41 with the pressure compensating pipe 46.

Hereinafter, the operation of the above-described cell culture system will be described.

A glass plate 28 is disposed on the bottom surface 31 of the incubator 30 and a plate 20 is disposed on the plate receiving portion 32 thereon.

Next, the cell culture unit 10 is arranged so that the chamfered edge 19 of the cell culture unit 10 and the chamfered edge 27 of the plate 20 are located in the same direction, and the cell culture unit 10 Is directed toward the passage (26) of the partition wall (24) of the plate (20).

The cells to be cultured and the culture medium are injected through the inlet 12 of each cell culture unit 10 to allow the cells to be placed in the culture unit 11. [

The culture liquid distributing pipes 56 of the culture liquid distributor 50 are connected to the steel pipes 15 inserted in the inlet 12 of the cell culture unit 10 through the connection pipe 58 while covering the incubator cover 35 .

Next, the culture solution storage container 40 and the inlet 51 of the culture solution distributor 50 are connected through the first connection pipe 42. Then, the waste culture medium storage container 41 and the waste culture medium discharge pipe 34 on the side of the incubator 30 are connected through the second connection pipe 43.

Power is supplied to the heater on the bottom surface 31 of the incubator 30 and necessary gas is supplied through the gas piping 38 on the side of the incubator 30 so that a temperature suitable for cell growth The atmosphere is formed.

The first pump 44 installed in the first connection pipe 42 and the second pump 45 installed in the second connection pipe 43 are operated.

When the pumps operate, the culture solution stored in the culture solution storage container 40 is supplied to the culture solution distributor 50 through the first connection pipe 42. The culture liquid supplied to the culture liquid distributor 50 first fills the inlet side space 53 of the culture liquid distributor 50 by the distributor barrier 55. When the inlet side space 53 becomes full, the culture liquid begins to fill the outlet side space 54 beyond the distributor partition wall 55. When the outlet side space 54 is filled to some extent and the culture liquid reaches or exceeds the height of the outlet 52 of the culture liquid distributor 50, the culture liquid is supplied to the outlets 52 of the culture liquid distributor 50, the culture liquid distributing pipes 56, Through the inlets 12 of the cell culture unit 10 of the cell culture unit 10 and supplied to the culture unit 11 of the cell culture unit 10. [

The pulmonary culture fluid generated during the cell growth process is pushed by the outflow channel 18 and discharged to the outlet 16 of the cell culture unit 10 as a new culture fluid is supplied to the culture unit 11. The discharged pulmonary culture liquid passes through the passage 25 formed in the outer wall 21 after sequentially passing through the passage 26 formed in the widthwise partition wall 24 of the plate 20. And is stored in the ditch 33 formed around the plate receiving portion 32 of the incubator 30. [ The pulp culture fluid stored in the trench 33 is discharged to the outside of the incubator 30 through the pulmonary fluid discharge pipe 34 and stored in the pulmonary fluid storage container 41 through the second connection pipe 43.

When the pressure of the pulmonary fluid storage container 41 increases with the inflow of the pulmonary fluid, the gas in the pulmonary fluid storage container 41 is transferred to the culture fluid storage container 40 through the pressure compensation pipe 46, (41) is lowered. At the same time, the pressure of the culture medium storage container 40 in which the pressure is lowered by the outflow of the culture liquid is increased.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the invention as defined by the appended claims. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention.

For example, although it has been described that the cells are injected through the inlet 12, it is also possible to separately inject the cells and the culture solution by forming a separate cell injection port connected to the culture unit. Further, in addition to the outlet formed below the cell culture unit 10, a separate outlet formed elongated in the thickness direction of the cell culture unit 10 may be additionally formed.

Although a passage is formed in the outer wall and the partition in the width direction of the plate, a passage may be formed in the outer wall and the partition in the longitudinal direction of the plate.

10: cell culture unit 11: culture unit
12: inlet 13: culture fluid reservoir
14: Inflow channel 15: Steel pipe
16: outlet 18: outlet channel
20: plate 21: outer wall
22: accommodating portion 23: longitudinal barrier rib
24: width direction bulkhead 25, 26: passage
27: edge 28: glass plate
30: Incubator 31: Bottom surface
32: plate receiving portion 33: ditch
34: Lung culture fluid discharge pipe 35: Incubator cover
36: window 37: through hole
38: gas piping 39:
40: Culture solution storage container 41: Lung culture solution storage container
42: first connection pipe 43: second connection pipe
44: first pump 45: second pump
46: pressure compensating tube 50: culture liquid distributor
51: inlet 52: outlet
53: inlet side space 54: outlet side space
55: Dispenser bulkhead 56:
57: Culture liquid distributor cover 58: Connector

Claims (8)

And an outflow channel for connecting the culture unit and the outflow port, wherein the outflow channel is for discharging the culture liquid of the culture unit, and the outflow channel for connecting the culture unit and the outflow port, Wherein a resistance of the outflow channel is larger than a resistance between the inlet and the culture unit,
And a partition wall for partitioning a space surrounded by the outer wall and the outer wall into a plurality of accommodating portions capable of accommodating the plurality of cell culture units, wherein the outer wall and the partition walls are provided with a waste A plate having passages for allowing the culture solution to flow in one direction,
A heater for heating the plate containing portion to a temperature according to a culture condition; a gas pipe for supplying a gas for maintaining the atmosphere according to the culture condition to the plate containing portion; A trench formed around the plate receiving portion to receive a pulmonary culture fluid flowing through the outer wall of the plate and a pulmonary culture fluid having one end disposed in the trench and the other end extending to the outside so as to discharge the pulmonary culture fluid stored in the trench to the outside An incubator including a discharge tube and a culture fluid distribution tube for supplying a culture fluid to an inlet of the cell culture units;
A culture medium storage container in which the culture medium is stored in a sealed state, a waste culture medium storage container in which the waste culture medium is stored in a sealed state,
A first connection pipe for connecting the culture liquid storage container and the culture liquid distribution pipe, a first pump installed on the first connection pipe, a second connection pipe for connecting the waste liquid discharge pipe and the waste liquid storage container, 2 cell culture system comprising a second pump installed in a connector. The method according to claim 1,
And a pressure compensating tube for connecting the culture medium storage container and the waste culture medium storage container to each other. The method according to claim 1,
And a distributor bulkhead disposed inside to divide the culture liquid distributor into an inlet side space and an outlet side space, and an inlet for introducing the culture liquid, a plurality of outlets for discharging the culture liquid and connected to the culture liquid distribution tubes, and a distributor partition wall Lt; / RTI > An inlet port for injecting the culture liquid into the culture section, an outlet port for discharging the lung culture liquid of the culture section, and at least one bend section connecting the culture section and the outlet port, Wherein the resistance of the outlet channel is larger than the resistance between the inlet and the culture unit,
And a partition wall for partitioning a space surrounded by the outer wall and the outer wall into a plurality of accommodating portions capable of accommodating the plurality of cell culture units, wherein the outer wall and the partition walls are provided with a waste And a plate in which passages are formed so that the culture can flow in one direction. A culture unit which is an inner space where cells grow,
An inlet for injecting the culture liquid into the culture unit,
An outlet for discharging the waste culture liquid of the culture unit,
And an outlet channel connecting the culture section and the outlet and having at least one bend,
Wherein a resistance of the outflow channel is larger than a resistance between the inlet and the culture section. A culture unit which is an inner space where cells grow,
An inlet for injecting the culture liquid into the culture unit,
An outlet for discharging the waste culture liquid of the culture unit,
An outlet channel connecting the culture unit and the outlet, the resistance of which is larger than the resistance between the inlet and the culture unit,
And a bottom surface formed so as to be stepped on one side,
And the outlet is configured to discharge the waste culture liquid toward a space formed by the step of the bottom surface. delete A culture unit which is an inner space where cells grow,
An inlet for injecting the culture liquid into the culture unit,
An outlet for discharging the waste culture liquid of the culture unit,
An outlet channel connecting the culture unit and the outlet, the resistance of which is larger than the resistance between the inlet and the culture unit,
A culture liquid reservoir in which the culture liquid introduced from the inlet is primarily stored,
And an inlet channel connecting the culture medium reservoir to the culture unit and having a resistance lower than that of the outlet channel.

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