WO2010058898A1

WO2010058898A1 - Incubator for cell culture

Info

Publication number: WO2010058898A1
Application number: PCT/KR2009/004888
Authority: WO
Inventors: 박진오; 김현진; 홍성출
Original assignee: 전북대학교 산학협력단; 주식회사 지니스
Priority date: 2008-11-19
Filing date: 2009-08-31
Publication date: 2010-05-27
Also published as: KR20100056017A

Abstract

The present invention relates to an incubator for cell culture, comprising a culture chamber (10) for cell culture, a vacuum pump (20) for discharging air from the culture chamber (10) to the outside, an automatic control valve (30) for controlling a supply pipe (41) of a carbon dioxide tank (40) for supplying carbon dioxide into the culture chamber (10) and a supply pipe (51) of a nitrogen tank (50) for supplying nitrogen into the culture chamber (10) to control a supply of carbon dioxide and nitrogen, a carbon dioxide sensor (60) for sensing the concentration of carbon dioxide in the culture chamber (10), an oxygen sensor (70) for sensing the concentration of oxygen in the culture chamber (10), and a controller (80) for receiving signals from the vacuum pump (20) and sensors (60, 70), and controlling the automatic control valve (30) in accordance with the received signals, whereby the present invention with the aforementioned simple configuration can be selectively used as a carbon dioxide incubator in which a concentration of carbon dioxide in a culture chamber is controlled, or as a hypoxia incubator in which a concentration of oxygen in a culture chamber is controlled.

Description

Incubator for Cell Culture

The present invention relates to an incubator for culturing cells, and more particularly, to a carbon dioxide incubator using a higher carbon dioxide concentration in the culture chamber and a hypoxia incubator using a lower oxygen concentration in the culture chamber. It relates to a cell culture incubator that can be used.

Cell culture is the cultivation of cells isolated from tissues of living organisms. In plants, some of the callus, which is a tissue that occurs when the plant is injured, is also called oily tissue, callus, or callus. Incubate.

Incubators are generally used to culture cells. Cells must meet a variety of conditions for each type, the most commonly used being a carbon dioxide incubator for culturing most animal cells. Atmospheric air consists of 78% nitrogen, 21% oxygen, 0.03% carbon dioxide and other components, with a concentration of about 5% suitable for the cell culture in the carbon dioxide incubator.

Patent Publication No. 10-2004-27395 discloses a carbon dioxide incubator, the carbon dioxide incubator disclosed in the document is a detection means for detecting the carbon dioxide concentration in the incubator as shown in Figure 1, setting means for setting the carbon dioxide concentration, It consists of a supply means for supplying carbon dioxide and a control means for controlling the supply means is configured to adjust the carbon dioxide concentration in the culture space by pushing the air in the culture space through the discharge pipe to the outside of the culture space while supplying carbon dioxide from the supply means.

Although the carbon dioxide incubator is simple in configuration and relatively inexpensive, the carbon dioxide incubator has a disadvantage in that it can only cultivate cells satisfying the culture conditions through carbon dioxide concentration control.

Unlike cells cultured in the carbon dioxide incubator, there is a Hypoxia incubator for culturing cells of the same type as stem cells by adjusting oxygen concentration, and one example thereof is disclosed in US Patent Publication No. 2003-92178. have.

The oxygen concentration in the atmosphere is about 21%, and this state is called a nomoxia state, and a state where the oxygen concentration is lower than that in the air is called a hypoxia state. It is called a hypoxia incubator to be used while maintaining the state of this hypoxia. The oxygen concentration suitable for culturing stem cells or the like cultured by the oxygen concentration in the Hypoxia incubator is about 1 to 8% depending on the type thereof.

The HYPOXY incubator disclosed in the above document has an oxygen tank, a nitrogen tank, and a carbon dioxide tank connected to the culture chamber by a controller to control the supply of oxygen, nitrogen, and carbon dioxide to the culture chamber as shown in FIG. 2. It is configured to push out air while supplying the components to adjust the oxygen concentration in the culture chamber.

The Hypoxia incubator is considerably more expensive than the carbon dioxide incubator and requires considerable skill in mixing the components supplied from each of the three tanks, and even if there is a slight error, there are three sensors that detect each component. As the number of sensors is high, the probability of failure due to the sensors also increases.

On the other hand, it is necessary to selectively use the carbon dioxide incubator and the Hypoxia incubator, but there is currently no incubator that can use both the carbon dioxide incubator and the Hypoxia incubator as one incubator, and if purchased separately, There is a problem that increases the cost and maintenance costs.

The present invention has been made to solve the problems of the prior art, the carbon dioxide incubator mainly used for animal cell culture by adjusting the carbon dioxide concentration in the culture chamber and the hypoxia mainly used for stem cell culture by controlling the oxygen concentration in the culture chamber It is an object of the present invention to provide an incubator for cell culture that can selectively use an incubator.

In another aspect, the present invention is to provide a cell culture incubator that can be configured in combination with a carbon dioxide incubator and a Hypoxia incubator, but by simplifying the device can reduce the cost and facilitate the maintenance.

The present invention for solving the above problems is a culture chamber 10 for culturing cells, a vacuum pump 20 for discharging air in the culture chamber 10 to the outside, and a carbon dioxide tank for supplying carbon dioxide into the culture chamber 10 The supply pipe 41 of the 40 and the automatic control valve 30 for supplying carbon dioxide and nitrogen by controlling the supply pipe 51 of the nitrogen tank 50 for supplying nitrogen, and detecting the carbon dioxide concentration in the culture chamber 10 Controller for controlling the automatic control valve 30 by receiving the carbon dioxide detection sensor 60 and the oxygen detection sensor 70 for detecting the oxygen concentration, and the detection signal of the vacuum pump 20 and each

sensor

60, 70 It consists of 80, which has its technical features.

In addition, the present invention is characterized in that the automatic control valve 30 is connected to the carbon dioxide supply pipe 41 and the nitrogen supply pipe 51 is a three-way valve for selectively controlling and supplying carbon dioxide and nitrogen.

In addition, the present invention is characterized in that the automatic control valve 30 is connected to the carbon dioxide supply pipe 41 and the nitrogen supply pipe 51, respectively.

According to the present invention, the carbon dioxide incubator used to control the carbon dioxide concentration and the hypoxia incubator used to control the oxygen concentration are configured as one incubator, thereby reducing the cost of purchasing each incubator separately.

In addition, the carbon dioxide incubator and the Hypoxia incubator are configured in combination, but by reducing the number of tanks and sensors controlled in each incubator, the configuration is simple and easy to maintain accordingly, there is an effect of easy concentration control.

1 is a block diagram of a conventional carbon dioxide incubator,

2 is a block diagram of a conventional Hypoxia incubator,

3 is a schematic structural diagram of an incubator for cell culture according to one embodiment of the present invention,

4 is a schematic configuration diagram of an incubator for cell culture according to another embodiment of the present invention.

A: incubator 10 for cell culture: culture chamber

20: vacuum pump 30: automatic control valve

40: carbon dioxide tank 41, 51: supply pipe

50: nitrogen tank 60: carbon dioxide detection sensor

70: oxygen detection sensor 80: controller

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

3 is a schematic configuration diagram of a cell culture incubator according to an embodiment of the present invention, Figure 4 is a schematic configuration diagram of a cell culture incubator according to another embodiment of the present invention.

Referring to FIGS. 3 and 4, the cell culture incubator A according to the embodiment of the present invention includes a culture chamber 10 for culturing cells and a vacuum pump 20 for discharging air in the culture chamber 10 to the outside. And, the automatic control valve 30 for controlling and supplying carbon dioxide or nitrogen to the culture chamber 10, the carbon dioxide detection sensor 60 for detecting the carbon dioxide concentration in the culture chamber 10 and the oxygen detection sensor 70 for detecting the oxygen concentration (70) ) And a controller 80 for controlling the automatic control valve 30 according to the detection of the vacuum pump 20 and the

sensors

60 and 70.

The culture chamber 10 is a space for growing organisms (mainly microorganisms and embryos of living plants and animals) or parts of organisms (organs, tissues, cells, etc.) under environmental conditions in which artificial conditions are appropriately controlled. It is composed of a case shape (not shown) and puts a culture vessel (not shown) containing cells therein and is used to culture the cells while controlling the air composition ratio in the culture chamber 10 according to the stable growth conditions of the cells.

In the present invention, it is possible to selectively adjust the air composition ratio, in particular carbon dioxide concentration and oxygen concentration in the culture chamber (10), in which the use of the carbon dioxide concentration to control the carbon dioxide incubator (Carbon dioxide Incubator), to control the oxygen concentration It is referred to as a Hypoxia incubator.

The vacuum pump 20 is connected to the culture chamber 10 to forcibly discharge the air in the culture chamber 10 to the outside, and is also connected to the controller 80 and is controlled according to the situation in the culture chamber 10.

When the vacuum pump 20 discharges the air in the culture chamber 10 to the outside and uses it as a carbon dioxide incubator, the carbon dioxide concentration is 0.03% by supplying carbon dioxide from the carbon dioxide tank 40 to increase the amount of carbon dioxide in the culture chamber 10. Is controlled to about 5%, and when used as a HYPOXY incubator, by supplying nitrogen from the nitrogen tank 50 to increase the amount of nitrogen in the culture chamber 10 to reduce the concentration of oxygen of 21% to about 1 to 8% To control the oxygen concentration. In this case, when using as a HYPOXY incubator, it is usually not necessary to control the carbon dioxide, but when the carbon dioxide concentration is too low or high, it is necessary to adjust the carbon dioxide concentration from the carbon dioxide tank 40 or supply a vacuum pump 20 By discharging the air in the culture chamber 10 may be adjusted to the carbon dioxide concentration.

The automatic control valve 30 is connected to the supply pipe 41 of the carbon dioxide tank 40 for supplying carbon dioxide into the culture chamber 10 and to the supply pipe 51 of the nitrogen tank 50 for supplying nitrogen, and the vacuum pump ( 20 and carbon dioxide or oxygen in the culture chamber 10 by controlling the supply of carbon dioxide and / or nitrogen into the culture chamber 10 by the controller 80 receiving the detection signal from the carbon dioxide detection sensor 60 and the oxygen detection sensor 70. To adjust the concentration.

At this time, as shown in Figure 3, each supply pipe (41, 51) may be used by connecting with a three-way valve, or may be used separately as shown in FIG. In addition, although each

supply pipe

41 and 51 are separately installed in FIG. 3 and FIG. 4, when using as a carbon dioxide incubator, a carbon dioxide tank may be connected, and when using a hypoxia incubator, a carbon dioxide tank may be removed and a nitrogen tank may be connected. have. Of course, when it is necessary to control the carbon dioxide concentration in the Hypoxia incubator, both the carbon dioxide tank and the nitrogen tank are used.

When the carbon dioxide sensor 60 is used as a carbon dioxide incubator, the detection range is mainly about 5% of carbon dioxide concentration and can be freely adjusted to other desired concentrations. When the carbon dioxide is supplied from the carbon dioxide tank 40 into the culture chamber 10 and the carbon dioxide concentration in the culture chamber 10 is out of the carbon dioxide detection range input to the carbon dioxide sensor 60, the sensor 60 detects the signal from the controller 80. The controller 80 controls the carbon dioxide concentration in the culture chamber 10 by stopping the supply of carbon dioxide from the carbon dioxide tank 40 to the culture chamber 10 by operating the automatic control valve 30.

On the other hand, the oxygen detection sensor 70 is operated when used as a hypoxia, and the detection range is determined in a range that satisfies the conditions in which the cells are cultured, and is mainly set at an oxygen concentration of 1 to 8%. Free adjustment is possible in the range of% or less. When nitrogen is supplied from the nitrogen tank 50 into the culture chamber 10 and the oxygen concentration in the culture chamber 10 is out of the oxygen detection range input to the oxygen detection sensor 70, a signal is sent from the sensor 70 to the controller 80. The controller 80 operates the automatic control valve 30 to stop the nitrogen supply from the nitrogen tank 40 into the culture chamber 10 to adjust the oxygen concentration in the culture chamber 10.

The carbon dioxide detecting sensor 60 and the oxygen detecting sensor 70 are respectively connected to the controller 80 to transmit a detection signal to the controller 80.

The controller 80 is connected to the

sensors

60 and 70, the automatic control valve 30, and the vacuum pump 20, so that when a detection signal is transmitted from the

sensors

60 and 70 and the vacuum pump 20, the carbon dioxide tank ( 40) and the carbon dioxide concentration and the oxygen concentration in the culture chamber 10 by adjusting the automatic control valve 30 connected to the nitrogen tank 50.

The controller 80 is mainly composed of a PLC (automatic control circuit) is operated by a program configured to meet the conditions in the culture chamber 10, because the given conditions are not complicated or varied, it is possible to use a low-cost PCB circuit board or It can be composed of a variety of circuits.

Hereinafter, a method of using the cell culture incubator (A) of the present invention having the above structure will be described.

First, the carbon dioxide incubator used to culture most animal cells and used to control the concentration of carbon dioxide will be described.

After inserting the culture vessel containing the cells into the culture chamber 10, the vacuum pump 20 forcibly sucks the air in the culture chamber 10 and discharged to the outside, the controller 80 detects the signal to operate the automatic control valve 30 By operating the carbon dioxide tank 40 to supply as much carbon dioxide as the amount of air in the culture chamber 10.

As the carbon dioxide is supplied, the carbon dioxide concentration in the culture chamber 10 is improved, and when the carbon dioxide concentration suitable for cell culture is reached (mainly 5%), the carbon dioxide detection sensor 60 installed in the culture chamber 10 detects the signal and the controller 80 ), The controller 80 operates the automatic control valve 30 again to stop the supply of carbon dioxide.

When the cells are cultured in this state (carbon dioxide concentration of 5%) and the cells which have been cultured are pulled out of the culture chamber 10, the door of the culture chamber 10 is opened, and the air components in the culture chamber 10 again return to atmospheric carbon dioxide. It becomes equal to the concentration (0.03%). Inserting the culture vessel containing the cells in the culture chamber 10 for another cell culture and closing the door repeats the above operations in order starting with the operation of the vacuum pump 20, the carbon dioxide concentration in the culture chamber 10 during cell culture (Mainly 5%) is kept constant.

Next, we describe the Hypoxia incubator used to culture cells of the same type as stem cells and used to control the concentration of oxygen.

In the same manner as above, the culture vessel containing the cells is placed in the culture chamber 10, and the vacuum pump 20 forcibly sucks the air in the culture chamber 10 and discharges it to the outside. 30) is operated to supply nitrogen as much as the amount of air in the culture chamber 10 from the nitrogen tank 50.

When nitrogen is supplied, the oxygen concentration in the culture chamber 10 decreases, and when the oxygen concentration (mainly 1 to 8%) suitable for cell culture is reached, the signal is detected by the oxygen detecting sensor 70 installed in the culture chamber 10. 80, the controller 80 stops the supply of nitrogen by operating the automatic control valve 30 again.

When the cells are cultured in this state (oxygen concentration of 1 to 8%) and the cells which have been cultured are taken out of the culture chamber 10, the door of the culture chamber 10 is opened, and the air components in the culture chamber 10 are held back. It becomes equal to medium oxygen concentration (21%). Inserting the culture vessel containing the cells in the culture chamber 10 for another cell culture and closing the door, the operation is repeated in order starting with the operation of the vacuum pump 20 while the oxygen concentration in the culture chamber 10 during the cell culture (Mainly 1 to 8%) is kept constant.

In this case, the supply of nitrogen into the culture chamber 10 to adjust the oxygen concentration is lowered because the naturally occurring carbon dioxide is generated as the cells in the culture chamber 10 are cultured, but it is not necessary to supply carbon dioxide separately. If necessary, carbon dioxide is supplied from the carbon dioxide tank to control the carbon dioxide concentration.

By constructing the cell culture incubator (A) of the present invention as described above, the carbon dioxide incubator used while adjusting the carbon dioxide concentration according to the type of cells to be cultured, and the hypoxia incubator used while adjusting the oxygen concentration can be selectively used. Will be.

In addition, if the existing carbon dioxide incubator is modified and manufactured, the Hypoxia incubator can be used at the same time.

Claims

In an incubator for culturing cells,

A culture chamber 10 for culturing cells,

A vacuum pump 20 for discharging air in the culture chamber 10 to the outside,

An automatic control valve 30 for supplying carbon dioxide or nitrogen by controlling a supply pipe 41 of a carbon dioxide tank 40 supplying carbon dioxide into the culture chamber 10 and a supply pipe 51 of a nitrogen tank 50 supplying nitrogen; ,

A carbon dioxide detection sensor 60 for detecting the carbon dioxide concentration in the culture chamber 10 and an oxygen detection sensor 70 for detecting the oxygen concentration,

Cell vacuum incubator, characterized in that consisting of a controller (80) for receiving the detection signal of the vacuum pump 20 and each sensor (60, 70) to control the automatic control valve (30).
The method according to claim 1,

Automatic control valve 30 is a cell culture incubator, characterized in that the three-way valve is connected to the carbon dioxide supply pipe 41 and the nitrogen supply pipe 51 to selectively control the supply of carbon dioxide and nitrogen.
The method according to claim 1,

Automatic control valve 30 is a cell culture incubator, characterized in that for controlling the carbon dioxide and nitrogen connected to the carbon dioxide supply pipe 41 and the nitrogen supply pipe 51, respectively.