KR101649031B1 - Hypoxia cell culture device comprising a lid for various gas concentration - Google Patents

Abstract

The present invention relates to a hypoxic cell incubator capable of realizing various concentrations of gas, including a cover configured to control the concentration of a gas in each of a plurality of culture spaces, wherein at least one gas inlet 110 and at least one gas A cover portion 100 including an outlet 120; And an intermediate layer (200) located under the cover part (100), wherein the intermediate layer (200) comprises at least one layer having a plurality of grooves. It is possible to construct various concentrations of gas through this, so that it is possible to secure a large amount of experimental data more effectively, thereby saving time for conducting the experiment. It is economical because it can be reused and continuous experiment for observation under a microscope can be performed have.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a hypoxia cell culture device comprising a cover for various concentrations of gas,

The present invention relates to a cell incubator, and more particularly to a cell incubator for hypoxic cells capable of realizing various concentrations of gas, including a cover configured to control the concentration of the gas in each of a plurality of culture spaces.

In general, cell culture has been widely used in various technical fields such as biology, medicine, pharmacy, or agriculture. The unit cells or microorganisms isolated from the living body are grown and proliferated, and studies on the cells and the microorganisms are actively carried out, so that information useful for the human body and the environment can be obtained. For example, microorganisms obtained through microbial cultivation include the food industry such as miso, cheese, and Yakult; Medical materials such as antibiotics and anticancer drugs; It is also used as an energy material for high value added chemical materials and methane, hydrogen, and electricity in the environmental industry. In recent years, its application to engineering fields such as wastewater treatment and waste oil treatment has been expanded.

In particular, in the field of biology, cell culture is available for study to observe biological changes occurring inside the human body. In order to more effectively observe the biological changes in the human body, the inside of the human body has a low oxygen concentration environment in comparison with the oxygen concentration in the atmosphere. In order to more effectively observe biological changes inside the human body, .

In order to meet these conditions, cell cultivators of various characteristics have been proposed in the past. For example, Korean Patent Registration No. 10-1294521 discloses an apparatus for controlling the concentration of dissolved oxygen in a cell, and includes an oxygen absorbent supply unit for supplying an oxygen absorbent. These devices are simple in terms of their ability to control the amount of dissolved oxygen by controlling the concentration of the oxygen absorbing agent. However, the reaction time and oxygen absorption degree depend on the type of oxygen absorbing agent used, the concentration of each type of oxygen absorbing agent And the response time of the absorbent depending on the temperature. Therefore, the procedure is complicated to acquire information for research, there are many items to be considered besides the cell culture, and time for accurate data analysis It is difficult to use it because it takes a lot of time.

Korean Patent No. 10-0815074 proposes a cell culture incubator including a heater for supplying heat energy and a sensor part for sensing a cell culture environment capable of automatically controlling the culture environment. It is easy to analyze data in a favorable environment in which real-time observation can be performed. However, the cell culture chip, which is a space where cell cultivation is performed, is formed differently from the culture chip used in the past, , The conventional culture chip can not be used, and thus there is a limit to the usability. In addition, since the culture chip is used for observing cells having minute dimensions, it is generally regarded as a one-time consumable, and therefore, it is unsuitable for disinfection and recycling, so that the unit price may increase as compared with a general culture chip.

Furthermore, there is a conventional hypoxic cell incubator having a structure capable of maintaining the oxygen concentration uniformly throughout the incubator or injecting and discharging the culture medium of oxygen concentration for controlling the amount of dissolved oxygen. However, in this case, it is troublesome to repeat the experiment repeatedly in order to observe the cells at various oxygen concentrations, and when the culture dish needs to be moved out of the cell incubator for observation during the cell culture experiment, There is a limit to accurate data collection because the concentration fluctuates.

KR 10-1294521 B1 KR 10-0815074 B1

Accordingly, the present invention has been made in order to solve the above problems.

Specifically, it is intended to provide a lid capable of effectively creating an environment with various gas concentrations and a cell incubator comprising the lid.

In addition, it is possible to obtain fast and accurate experimental results by using this, and at the same time, to provide a cost-effective lid and a cell incubator containing the lid.

Also, a cover for preventing hypoxic conditions from being destroyed when cells are observed under the microscope during the experiment, and a hypoxic cell incubator containing the same are proposed.

In order to solve the above-mentioned problems, an embodiment of the present invention includes a cover part 100 including at least one gas inlet 110 and at least one gas outlet 120; And an intermediate layer (200) located under the cover part (100), wherein the intermediate layer (200) comprises at least one layer having a plurality of grooves.

The cover part 100 and the intermediate layer 200 are preferably connected to each other by a detachable coupling part 130.

Further, it is preferable to further include a valve 140 detachable from the gas inlet 110 or the gas outlet 120.

The intermediate layer 200 may include a first induction layer 210 having a plurality of induction grooves 211 at positions corresponding to the gas inlet 110 and the gas outlet 120.

The intermediate layer 200 may include a secondary induction layer 220 formed below the primary induction layer 210 and formed with a slit guide groove 221 overlapping at least a part of the plurality of induction grooves 211 , And the gas inlet (110) and the gas outlet (120) are arranged in a row in parallel with each other, and the slit is disposed perpendicularly to the parallel row.

The intermediate layer 200 may include a tertiary induction layer 230 formed below the secondary induction layer 220 and having a plurality of induction grooves 231 overlapping at least a part of the slit- And the guide groove 231 is smaller than the guide groove 221.

The primary induction layer 210, the secondary induction layer 220 and the tertiary induction layer 230 are connected to each other and the induction grooves 211, 221, and 231 included in the respective layers are connected to each other, It is preferable to induce the flow of the gas passing through the gas inlet 110 through the action of the gas.

Further, there is provided a cell incubator including a lid according to an embodiment of the present invention.

In addition, the cell incubator may further include a culture unit 300 having a culture dish 310 at the lower end of the lid.

In addition, the cell incubator preferably further includes a sensor for measuring the concentration of the gas in the culture unit 300.

In the case of adopting the lid according to the present invention, various concentrations of gas can be formed by the structure of the lid, so that experimental data can be obtained more effectively.

In addition, since the lid includes a plurality of gas injection openings, an intermediate layer having a gas outlet and an induction groove, various concentrations of gas can be simultaneously realized in one cell incubator, and a large amount of experimental results can be obtained, .

In addition, unlike culture dishes, which are consumables, the cover is easy to be cleaned and disinfected, and is reusable and economical.

In addition, since a plurality of gas injection openings and gas discharge openings allowing a constant environment composition of the culture dish are provided, continuous experiments can be carried out by observing the cells under a microscope in a state where the hypoxic condition is not destroyed.

As such hypoxic conditions are established, various studies are possible. For example, studies on human embryonic stem cells, studies on infection in human body such as Mycobacterium tuberculosis, intestinal hemorrhagic Escherichia coli, effects of cancer drugs on cancer cells, And research on the relationship between hypoxic or specific gas concentrations for storage.

1 is a plan view of a cell incubator having a lid according to the present invention.
2 is a cross-sectional view taken along the line a-a 'in Fig.
3 is an exploded perspective view for explaining an intermediate layer according to the present invention.
4 is a perspective view for explaining an intermediate layer according to the present invention.
5 is a conceptual perspective view of a cell incubator for explaining the operation principle of a lid according to the present invention.
6A and 6B are conceptual diagrams for explaining the direction in which gas is guided in the lid according to the present invention, and show the flow direction of the gas in accordance with injection and discharge, respectively.

A cell incubator including a lid and a lid of the present invention will be described as an example. However, a cover to which the same principle is applied may be applied to devices other than the cell incubator. Accordingly, it is apparent that the scope of the present invention is narrowed to the scope of the claims appended to other apparatus to which the cover of the same principle is applied.

The constituent elements of the present invention can be used integrally or individually as needed. In addition, some components may be omitted depending on the usage form.

Preferred embodiments of the cover according to the present invention will be described with reference to Figs. 1 to 6B. In this process, the thicknesses of the lines and the sizes of the components shown in the drawings may be exaggerated for clarity and convenience of explanation. In addition, the terms described below are defined in consideration of the functions of the present invention, which may vary depending on the intention or custom of the user, the operator. Therefore, the definitions of these terms should be described based on the contents throughout this specification.

In the following description, 'gas' is used to mean a gas including all gaseous substances such as oxygen, mixed gas and the like.

1. Description of cover and configuration of cell incubator containing it

Hereinafter, a lid according to the present invention and a cell incubator including the same will be described with reference to the accompanying drawings.

First, the overall configuration of the lid and the cell incubator will be described with reference to Figs. 1 and 2. Fig.

The cell incubator includes a cover part 100; An intermediate layer 200 positioned below the cover part 100; And a culture unit 300 located below the intermediate layer 200. [ "Cover" means that it includes the cover part 100 and the intermediate layer 200.

An example of the means for connecting the lid and the culturing unit 300 is an engaging part 130. The coupling part 130 may be formed to be connected to the side surface of the cell incubator, and the induction action of the gas through the intermediate layer 200 may be more stably performed by the coupling part 130. It goes without saying that any other kind of coupling structure not shown may be applied.

The cover portion 100 includes at least one gas inlet 110 and at least one gas outlet 120. The intermediate layer 200 includes at least one or more layers having a plurality of grooves.

The gas inlet 110 and the gas outlet 120 are preferably formed in parallel in a row and formed in the cover 100. A gas such as oxygen or a mixed gas is supplied through the gas inlet 110 And is discharged to the outside of the cell incubator through the gas outlet 120.

The cover part 100 may further include a valve 140 including a coupling part formed in the gas injection port 110 so as to facilitate detachment and attachment. The valve 140 may be used in connection with an apparatus for setting, measuring, and controlling the flow of gas, for example, one or more mass flow controllers (MFCs) And maintains the mass flow rate of the gas at a predetermined set value. The mass flow controller is a known technique, and a detailed description thereof is omitted.

A plurality of mass flow controllers may be applied to the cell incubator so that the ratio of the components of the gas injected through the gas inlet 110 (for example, the ratio of nitrogen and oxygen) can be adjusted and included in the incubator 300 Various concentrations of gas may be provided to the cells 330 and the culture medium 340 accommodated in the respective culture wells 320 of the hollow formed in the culture dish 310. The cell incubator may further comprise a sensor capable of measuring the mass flow rate of the gas at various concentrations and / or a sensor capable of measuring the concentration of the gas.

The cover and the culturing unit 300 are not limited to the cells 330, and various kinds of microorganisms can also be tested. The culture dish 310 used in this case may be a widely used disposable cell culture dish, and the cover may be detachable from the culture dish 310, so that the culture dish 310 of various shapes and sizes can be used.

The culture well 320 should be disposed so that the culture liquid 340 can be influenced by the gas inlet 110 and the gas outlet 120 located in the cover 100, respectively. Further details related thereto will be described later with reference to Figs. 3 to 6A.

The structure of the intermediate layer 200 according to the present invention will be described in more detail with reference to FIGS. 3 and 4. FIG.

3 and 4 are an exploded perspective view for explaining the intermediate layer 200 according to the present invention and a perspective view of the connected intermediate layer 200, respectively. The individual inductive layers 210,220 and 230 forming the intermediate layer 200 may be connected in various ways and other embodiments may be added by adding different kinds of members between the inductive layers 210,220 and 230 shown. It should be understood. Also, since each of the inductive layers 210, 220, and 230 is relatively easily formed individually, the lid is economical because it is easy to clean and disinfect, and can be reused, unlike a culture dish as a consumable.

The intermediate layer 200 includes a primary induction layer 210 having a plurality of induction grooves 211 at positions corresponding to the gas inlet 110 and the gas outlet 120, respectively. It is preferable that the guide groove 211 is formed in the primary induction layer 210 by the number of the gas inlet 110 and the gas outlet 120.

A secondary induction layer 220 including a slit-shaped induction groove 221 formed to overlap with at least a part of a plurality of induction grooves 211 is located under the primary induction layer 210. [ The plurality of slit-shaped guide grooves 221 of the secondary inductive layer 220 may be disposed perpendicular to the guide grooves 211 of the primary inductive layer 210 formed in a row in parallel. The gap formed by the overlapping of the slit-shaped guiding grooves 221 and the guiding grooves 211 of the primary guiding layer 210 enables the gas injected or discharged into the cell incubator to pass therethrough.

The intermediate layer 200 is formed of a tertiary induction layer 220 having a plurality of induction grooves 231 which are located below the secondary induction layer 220 and overlap at least a part of the slit guide groove 221 of the secondary induction layer 220 230). The guide groove 231 of the tertiary guide layer 230 is formed to be smaller than the guide groove 221 of the primary guide layer 210 and overlaps with the slit guide groove 221 of the secondary guide layer 220 Gas can be injected or discharged in each culture well 320 through the formed gap.

The primary induction layer 210, the secondary induction layer 220 and the tertiary induction layer 230 are connected to each other through the induction grooves 211, 221, and 231 included in the respective layers, It is possible to induce the flow of the gas that has passed through the flow path 110. The gas injected into the gas inlet 110 may be passed through each of the plurality of guide grooves 211, 221 and 231 so that the gas can reach the culture dish 310 and the culture solution accommodated in the culture well 320 340 and cells 330 may be provided with various concentrations of gas.

2. Description of the lid and cell incubator containing it

5 to 6B, the operation of the lid and the cell incubator including the lid according to the present invention and the direction in which the gas lid is guided will be described in more detail.

5 is a conceptual perspective view of a cell incubator for explaining the principle of operation of a lid according to the present invention. The plurality of gas copper lines A, B, and C shown in the figure respectively include a primary induction layer 210, 6A and 6B are schematic representations of gas flow lines A, B, and C in the case where gas is injected and discharged, respectively. FIG. 2 is a conceptual diagram for explaining a direction in which gas is introduced through the gas.

First, the disinfected and sterilized lid is connected to the culture unit 300 by the coupling unit 130, and the valve 140 can be connected to the gas inlet 110. A predetermined concentration of gas (for example, oxygen or a mixed gas) may be injected into the gas inlet 110 by being connected to the mass flow controller through the valve 140.

At this time, the gas can be continuously injected, the concentration of the gas can be controlled by the valve 140, and the concentration can be measured through a sensor that can be further included in the cell incubator, It is easy. The apparatus further includes a viewing window or an imaging device formed so that a constant environment is created by continuous entry and exit of the gas through the plurality of gas inlet ports 110 and the gas outlet port 120 so that at least a part of the lid can be observed with a microscope, The continuous observation can be carried out by observing the cells without the hypoxic condition being discarded.

The injected gas passes through the induction groove 211 of the primary inductive layer 210 in the same flow as the gas copper wire (A). The gas passes through the overlapping portion of the guide groove 211 of the first induction layer 210 and the slit guide groove 221 of the second induction layer 220 so that the gas flows into the slit guide groove 221, As shown in FIG.

The gas flowing into the slit-shaped guide groove 221 of the secondary inductive layer 220 flows through the portion of the tertiary inductive layer 230 which overlaps with the guide groove 231 as the gas copper line C, And reaches the culture unit 300. The gas reaching the culture unit 300 is formed in an experimental environment in which different concentrations of the cells 330 and the culture liquid 340 accommodated by the respective culture wells 320 act.

At this time, the number of the plurality of guide grooves 231 formed in the tertiary inductive layer 230 is not limited, and if necessary, by changing the size and number of the guide grooves 231 of the tertiary inductive layer 230, The number of portions of the induction layer 220 that overlap the slit-shaped guide groove 221 can be varied. For example, the culture dish 310 may be made of 6 wells, 64 wells, or 96 wells, and each induction layer 210, 220, and 230 may have different sizes, numbers , Or having guide grooves 211, 221, and 231 having a shape.

In the course of the gas being injected, the gas flows along the gas copper lines A, B, and C from A to B and B to C in the direction as shown in Fig. 6A. On the other hand, the gas flows from C to B and from B to A in the direction as shown in FIG. 6B along the gas copper lines A, B, and C during the discharge, .

100:
110: gas inlet
120: gas outlet
130:
140: Valve
200: middle layer
210: primary induction layer
220: secondary induction layer
230: tertiary induction layer
211, 221, 231: guide groove
300: culture unit
310: Petri dish
320: culture well
330: cells
340: culture solution
A, B, C: Gas circulation line

Claims (10)

A cover part (100) comprising at least one gas inlet (110) and at least one gas outlet (120); And
And an intermediate layer (200) located below the cover part (100)
The intermediate layer (200)
A primary induction layer 210 having a plurality of induction grooves 211 at positions corresponding to the gas inlet 110 and the gas outlet 120, respectively;
A secondary induction layer 220 formed below the primary induction layer 210 and having a slit-shaped induction groove 221 overlapping at least a part of the induction recesses 211; And
And a tertiary induction layer (230) located below the secondary induction layer (220) and having a plurality of induction grooves (231) overlapping at least part of the slit - shaped induction recess (221)
cover.
The method according to claim 1,
The cover part 100 and the intermediate layer 200 are connected by a detachable coupling part 130,
cover.
The method according to claim 1,
Further comprising a valve (140) detachable to the gas inlet (110) or the gas outlet (120)
cover.
delete The method according to claim 1,
Wherein the gas inlet (110) and the gas outlet (120) are arranged in a row in parallel and the slits are arranged perpendicular to the parallel row,
cover.
The method according to claim 1,
The guide groove 231 is smaller than the slit guide groove 221,
cover.
The method according to claim 1,
The primary inductive layer 210, the secondary inductive layer 220 and the tertiary inductive layer 230 are connected to each other and the interaction of the induction grooves 211, 221, and 231 included in the respective layers (110) through the gas inlet (110)
cover.
10. A method of manufacturing a lid, comprising a lid according to any one of claims 1 to 3 and 5 to 7,
Cell incubator.
9. The method of claim 8,
Further comprising a culture unit (300) having a culture dish (310) at the lower end of the lid,
Cell incubator.
10. The method of claim 9,
Further comprising a sensor for measuring the concentration of the gas in said culture unit (300)
Cell incubator.

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