Culture apparatus and culture method

Abstract

To provide novel culture apparatus and culture method that enable precise cell culture under conditions free from the effects of temperature and environmental disturbances.SOLUTION: The present invention provides a culture apparatus for culturing cells by housing a culture vessel containing at least cells and a culture medium within a culture chamber. The apparatus includes: an auxiliary device that is freely movable into and out of the culture chamber to support cell growth; an auxiliary device control unit for controlling the auxiliary device; and a temperature control device for controlling the temperature inside the culture chamber. The temperature control device is configured to preemptively correct temperature changes arising from the on / off operation of the auxiliary device.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a culture apparatus and a culture method. [Background technology]

[0002] A cell is the smallest unit of life, and the human body is made up of 37 trillion cells. From this perspective, it is important to stably culture cells generated by fertilization, for example, so that they can be used in various research projects, and culture devices play an important role in this regard.

[0003] Meanwhile, regenerative medicine aims to restore lost bodily functions when organs and tissues made of cells lose their functions due to illness or accident. The market size of regenerative medicine is expected to continue to expand. Cells that are important in regenerative medicine include stem cells, which have the ability to differentiate into various cells, and mesenchymal stem cells in particular are most widely used in clinical practice. The role of culture devices will also be important in responding to the regenerative medicine market, which is expanding year by year.

[0004] When culturing cells, it may be necessary to observe the culture state by some means, for example, by using a camera and to stir the cells with a shaker (Patent Document 1). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent No. 7072310 Summary of the Invention [Problem to be solved by the invention]

[0006] However, the method described in Patent Document 1 does not allow precise observation of cells because the cells being cultured are observed from outside the culture chamber. For this reason, attempts have been made to directly introduce a camera into the culture chamber to observe the culture state of the cells. In addition, a shaker is used by directly introducing it into the culture device.

[0007] For this reason, the camera and shaker generate heat during use, and the temperature rise caused by this heat causes a shift in the culture conditions related to the temperature of the cells, making it difficult to perform the culture accurately. In addition, changes in temperature conditions cause oxygen gas or carbon dioxide gas, as well as impurity gases caused by resins, etc., to be generated from the cells, culture medium, and other parts of the device, causing a problem that the culture conditions of the cells change not only in temperature but also in the environment.

[0008] An object of the present invention is to provide a novel culture device and culture method that can accurately culture cells while eliminating the influences of temperature, gas concentration, and other environmental disturbances. [Means for solving the problem]

[0009] In order to achieve the above object, the present invention provides a culture apparatus for culturing cells by installing a culture vessel containing at least cells and a culture medium in a culture chamber, comprising: An auxiliary device that can be freely inserted into and removed from the culture chamber to assist in culturing the cells; an auxiliary device control unit for controlling the auxiliary device; A temperature control device for controlling the temperature inside the culture chamber, The present invention relates to an incubation apparatus characterized in that the temperature inside the incubation chamber is kept constant by on / off control of the auxiliary device. The present invention also provides a culture apparatus for culturing cells by installing a culture vessel containing at least cells and a culture medium in a culture chamber, comprising: An auxiliary device that can be freely inserted into and removed from the culture chamber to assist in culturing the cells; an auxiliary device control unit for controlling the auxiliary device; A temperature control device for controlling the temperature inside the culture chamber, The present invention relates to an incubation apparatus, characterized in that the temperature inside the incubation chamber is kept constant by the temperature control device.

[0010] Furthermore, the present invention provides a culture method for culturing a culture vessel containing at least cells and a medium in a culture chamber, comprising: Heating the culture chamber to a predetermined temperature by a temperature control device to culture cells; A step of assisting the culture of the cells with an auxiliary device that can be freely inserted into and removed from the culture chamber; and maintaining a constant temperature in the culture chamber by on / off control of the auxiliary device. The present invention also provides a culture method for culturing a culture vessel containing at least cells and a medium in a culture chamber, comprising: Heating the culture chamber to a predetermined temperature by a temperature control device to culture cells; A step of assisting the culture of the cells with an auxiliary device that can be freely inserted into and removed from the culture chamber; and maintaining the temperature in the culture chamber constant using the temperature control device. Effect of the Invention

[0011] According to the present invention, the temperature in the culture chamber is always kept uniform by controlling the temperature rise and fall caused by the on / off operation of the auxiliary device disposed in the culture chamber using the on / off control of the auxiliary device or the temperature control device. Therefore, the environment in the culture chamber can be kept uniform by maintaining a uniform temperature. In other words, it is possible to provide a novel culture device and culture method that can accurately culture cells while eliminating the influence of disturbances in temperature and environment. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 is a diagram showing a schematic configuration of a culture apparatus of the present invention. [Diagram 2] 10 is a graph showing the result of temperature control when the auxiliary device is driven in the present embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Hereinafter, the present invention will be described based on an embodiment. Fig. 1 is a diagram showing a schematic configuration of a culture device of the present invention. As shown in Fig. 1, a culture device 10 of the present invention has a culture device main body 11, and a door 12 attached so as to be rotatable about a rotation axis provided at the end of the front opening of the culture device main body 11 as one faces the left side. The culture device main body 11 constitutes a culture chamber S, and the door 12 constitutes a partition wall for separating the culture chamber S from the outside world. The door 12 may be arranged so as to be rotatable about a rotation axis provided at the right side of the front opening of the culture device main body 11 as one faces the right side.

[0014] In addition, in the culture device main body 11, i.e., in the culture chamber S, an auxiliary device 13 as an observation device or the like and an auxiliary device control unit 14 disposed below the auxiliary device 13 are disposed. A culture vessel T containing at least cells and a culture medium is placed on the auxiliary device control unit 14. The power source of the auxiliary device control unit 14 is configured to be taken in from the outside through a connector receptacle disposed on the inner wall surface of the culture device main body 11 and a connector 15. In addition, a heater for heating the culture chamber S is embedded in the wall surface of the culture device main body 11, and further a temperature control device for controlling the temperature of the heater is embedded. In addition, a temperature sensor 17 is attached to the inner wall surface of the culture device main body 11.

[0015] The auxiliary device 13 is configured so as to be detached from the auxiliary device control unit 14 and freely inserted into and removed from the culture device main body 11, i.e., the culture chamber S. The auxiliary device control unit 14 controls the auxiliary device 13 and can further display the control process by the auxiliary device 13. The display results can be appropriately observed on a monitor 12A provided at the upper center of the door 12.

[0016] In the culture device 10, the auxiliary device 13 may be driven and operated while cells are being cultured in the culture chamber S of the culture device 10. When this happens, the auxiliary device 13 generates heat, and the temperature in the culture chamber S may rise above a predetermined set temperature due to the heat generation. In addition, gases such as oxygen and carbon dioxide may be released from the cells and culture medium in the culture container T due to this temperature rise, and further gases may be generated from impurities attached to the culture device main body 11, the door 12, and the auxiliary device 13.

[0017] In this way, when the auxiliary device 13 is operated within the culture chamber S, the temperature and environment within the culture chamber S change, causing the culture conditions to deviate from the initially set conditions, and it may become impossible to culture the cells within the culture container T as planned.

[0018] However, in the present invention, the temperature in the culture chamber S is set to a temperature somewhat lower than the set temperature in advance, in anticipation of the above-mentioned temperature rise, by a temperature control device disposed within the wall surface of the culture device main body 11. Therefore, the difference between this set temperature and the set temperature after control corrects and offsets the temperature rise in the culture chamber S accompanying the operation of the auxiliary device 13, so that even when the auxiliary device 13 is operated within the culture chamber S, the temperature and environment can be maintained uniform, and the cells can be cultured in the culture container T as planned.

[0019] For example, when the temperature inside the culture chamber S is to be maintained at 37° C., the temperature inside the culture chamber S is lowered by about 1 to 2° C. below the culture set temperature when the auxiliary device 13 is driven, that is, when the auxiliary device 13 is turned on.

[0020] On the other hand, when the operation of the auxiliary device 13 is turned off in the above-mentioned state, the temperature in the culture chamber S is set to a low temperature so as to compensate for the temperature rise caused when the operation of the auxiliary device 13 is turned on. Therefore, if the state is left as it is, the cell temperature and the associated environmental culture conditions will again deviate from the initial set conditions. For this reason, when the auxiliary device 13 is turned off, in order to compensate for the temperature drop in the culture chamber S caused by the turning off of the auxiliary device 13, the temperature of the culture chamber S is set in advance, prior to turning off the operation of the auxiliary device 13, from the state in which the temperature has been reduced as described above to a temperature slightly higher than the initial culture set temperature by a temperature control device (not shown). Then, after a predetermined time has elapsed, the culture set temperature is set.

[0021] By performing such operations, it is now possible to correct the excessive drop in temperature caused by the turning off of the auxiliary device 13 and the accompanying environmental changes in the environmental gases, etc., and it becomes possible to culture cells in the culture container T as scheduled.

[0022] For example, when the temperature inside the culture chamber S is to be maintained at 37° C., the temperature inside the culture chamber S is raised by about 1 to 2° C. above the culture set temperature when the auxiliary device 13 is driven, that is, when the auxiliary device 13 is turned off.

[0023] As described above, according to the present invention, the temperature rise and fall that accompanies the on / off operation of the auxiliary device 13 disposed in the culture chamber S is corrected in advance by the temperature control device. Therefore, the temperature in the culture chamber S can always be kept uniform, and further, the environment in the culture chamber S can be kept uniform by maintaining the uniform temperature. In other words, it is possible to provide a novel culture device 10 and a culture method that can accurately culture cells while eliminating the effects of disturbances in temperature and environment.

[0024] Strictly speaking, the above-mentioned temperature decrease and temperature increase depend on the set temperature in the culture chamber S and the type and operation time of the auxiliary device 13, so it is preferable to obtain a correlation between the set temperature in the culture chamber S and the type and operation time of the auxiliary device 13 for each culture device 10, and to determine in advance the degree of temperature increase and decrease in the culture chamber S based on this correlation.

[0025] Fig. 2 is a graph showing the results of temperature control when a camera is driven as the auxiliary device 13. As shown in the upper part of Fig. 2, when the camera is kept on, the temperature in the culture chamber S rises over time from 37°C to 37.8°C, but by controlling the camera to be on and off, it is possible to maintain the temperature at 37°C over time.

[0026] After this correlation is obtained, the temperature increase and decrease of the auxiliary device 13 can be easily realized by PID control in the temperature control device.

[0027] In addition, when the auxiliary device 13 is unknown and the correlation as described above cannot be obtained, the temperature is appropriately measured using a temperature sensor 17 arranged in the culture device main body 11, and the temperature is linked to the temperature control device, and the temperature in the culture chamber S is appropriately monitored using the temperature sensor 17 while the temperature control device decreases or increases the temperature in the culture chamber S as described above.

[0028] Furthermore, in the present invention, as shown in Fig. 1, the connector 15 of the auxiliary device control unit 14 is covered with a protective member 18. In general, connectors are difficult to clean and prone to contamination. However, as described above, such contamination can be prevented by covering the connector 15 with the protective member 18. Considering use under humid conditions or dry heat sterilization, examples of the protective member 18 include corrosion-resistant metals such as aluminum and stainless steel, or resin materials such as silicon and PPS that have low water absorption and are resistant to hydrolysis.

[0029] Specific examples of the auxiliary device 13 include a camera as an observation device, a pH detection element, a shaker, a temperature sensor for monitoring, a CO2 concentration sensor, an O2 concentration sensor, a germicidal lamp, a sterilization device, and the like.

[0030] In view of the above embodiments, the present application also includes the following inventions. [1] A culture device in which a culture vessel containing at least cells and a culture medium is placed in a culture chamber and cultured, An auxiliary device that can be freely inserted into and removed from the culture chamber to assist in culturing the cells; an auxiliary device control unit for controlling the auxiliary device; A temperature control device for controlling the temperature inside the culture chamber, A culture apparatus characterized in that the temperature inside the culture chamber is kept constant by on / off control of the auxiliary device. [2] A culture device in which a culture vessel containing at least cells and a culture medium is placed in a culture chamber and cultured, An auxiliary device that can be freely inserted into and removed from the culture chamber to assist in culturing the cells; an auxiliary device control unit for controlling the auxiliary device; A temperature control device for controlling the temperature inside the culture chamber, A culture device, characterized in that the temperature in the culture chamber is kept constant by the temperature control device. [3] The culture device described in [1] or [2], characterized in that the temperature control device is configured to pre-compensate for temperature changes caused by turning the auxiliary device on and off. [4] The culture device described in any one of [1] to [3], comprising a temperature sensor for measuring temperature changes within the culture chamber. [5] The culture device according to any one of [1] to [4], wherein a protective cap is provided to cover the connector of the auxiliary device control unit to protect the connector. [6] The culture apparatus according to any one of [1] to [5], wherein the auxiliary device is an image capturing device that captures images of cells. [7] The culture apparatus according to any one of [1] to [5], wherein the auxiliary device is a detection element for measuring the pH of the culture medium. [8] A culture method in which a culture vessel containing at least cells and a medium is placed in a culture chamber and cultured, Heating the culture chamber to a predetermined temperature by a temperature control device to culture cells; A step of assisting the culture of the cells with an auxiliary device that can be freely inserted into and removed from the culture chamber; and maintaining a constant temperature in the culture chamber by on / off control of the auxiliary device. [9] A culture method in which a culture vessel containing at least cells and a medium is placed in a culture chamber and cultured, Heating the culture chamber to a predetermined temperature by a temperature control device to culture cells; A step of assisting the culture of the cells with an auxiliary device that can be freely inserted into and removed from the culture chamber; and maintaining the temperature in the culture chamber constant using the temperature control device.

[10] The culture method described in [8] or [9], wherein the temperature control device compensates in advance for temperature changes caused by turning on and off the operation of the auxiliary device.

[11] The culture method described in [8] or [9], wherein the temperature within the culture chamber is measured by a temperature sensor disposed within the culture chamber, and the temperature within the culture chamber is corrected while monitoring the temperature within the culture chamber with the temperature sensor.

[0031] Although several embodiments of the present invention have been described above, these embodiments are presented as examples and are not intended to limit the scope of the invention. These novel embodiments can be implemented in various other forms, and various omissions, substitutions, and modifications can be made without departing from the spirit of the invention. These embodiments and their modifications are included in the scope and spirit of the invention, and are included in the scope of the invention and its equivalents described in the claims.

[0032] For example, in the above embodiment, the temperature control device is disposed inside the culture device main body 11, but as long as heat resistance is ensured, it may be disposed outside the culture device main body 11. Also, although the auxiliary device control unit 14 is disposed inside the culture device main body 11, it may be disposed outside the culture device main body 11. [Explanation of symbols]

[0033] 10 Culture device 11 Culture device main body 12 Doors 12A Monitor 13 Auxiliary equipment 14 Assistive Device Control Unit 15 Connectors 17 Temperature Sensor 18 Protective materials

Claims

1. A culture device in which a culture vessel containing at least cells and a culture medium is placed in a culture chamber and cultured, An auxiliary device that can be freely inserted into and removed from the culture chamber to assist in culturing the cells; an auxiliary device control unit for controlling the auxiliary device; A temperature control device for controlling the temperature inside the culture chamber, A culture apparatus characterized in that the temperature inside the culture chamber is kept constant by on / off control of the auxiliary device.

2. A culture device in which a culture vessel containing at least cells and a culture medium is placed in a culture chamber and cultured, An auxiliary device that can be freely inserted into and removed from the culture chamber to assist in culturing the cells; an auxiliary device control unit for controlling the auxiliary device; A temperature control device for controlling the temperature inside the culture chamber, A culture device, characterized in that the temperature in the culture chamber is kept constant by the temperature control device.

3. The incubation device according to claim 1 or 2, wherein the temperature control device is configured to compensate in advance for a temperature change caused by turning on and off the operation of the auxiliary device.

4. The culture apparatus according to claim 1 , further comprising a temperature sensor for measuring a temperature change in the culture chamber.

5. The culture device according to claim 1 or 2, wherein a protective cap for protecting a connector of the auxiliary device control section covers the connector.

6. The culture apparatus according to claim 1 or 2, wherein the auxiliary device is an image capturing device that captures an image of the cells.

7. The culture apparatus according to claim 1 or 2, wherein the auxiliary device is provided with a detection element for measuring the pH of the culture medium.

8. A culture method in which a culture vessel containing at least cells and a medium is placed in a culture chamber and cultured, Heating the culture chamber to a predetermined temperature by a temperature control device to culture cells; A step of assisting the culture of the cells with an auxiliary device that can be freely inserted into and removed from the culture chamber; and maintaining a constant temperature in the culture chamber by on / off control of the auxiliary device.

9. A culture method in which a culture vessel containing at least cells and a medium is placed in a culture chamber and cultured, Heating the culture chamber to a predetermined temperature by a temperature control device to culture cells; A step of assisting the culture of the cells with an auxiliary device that can be freely inserted into and removed from the culture chamber; and maintaining the temperature in the culture chamber constant using the temperature control device.

10. The culture method according to claim 8 or 9, wherein the temperature control device compensates in advance for a temperature change caused by turning on and off the operation of the auxiliary device.

11. 10. The culture method according to claim 8, further comprising: measuring the temperature inside the culture chamber by a temperature sensor disposed inside the culture chamber; and correcting the temperature inside the culture chamber while monitoring the temperature inside the culture chamber with the temperature sensor.

Images