JPH0427835B2 - - Google Patents

Abstract

PURPOSE:To enable integrated culture work from the preparation of a material to be cultured to the differentiation of tissue, by using a thermostatic chamber, a rotatable incubator having arbitrarily adjustable inclination angle, an optical system for irradiating the material with light having adjustable wavelength and intensity and a means for controlling the composition of gaseous atmosphere in the thermostatic chamber. CONSTITUTION:The culture apparatus is composed of a thermostatic chamber 11 having arbitrarily adjustable inner temperature and a rotatable incubator placed in the thermostatic chamber 11 and holding culture tubes 14 each containing a material to be cultured. Said rotatable incubator 30 is furnished with a culture tube holding member 13 to hold a plurality of culture tubes 14, a driving means 15 for rotating the holding member, an inclination angle adjusting means 32 to arbitrarily adjust the inclination angle of the culture tube holding member 13 from horizontal state to inclined state according to the growth of the material to be cultured and a supporting member 34 to detachably support the culture tube holding member 13. The apparatus is further provided with a gaseous atmosphere controlling means for controlling the composition of gaseous atmosphere in the thermostatic chamber 11 and having gas sources 22, 23, 24 of O2, N2 and CO2 and means 21 for controlling the feed of the gases and provided with an optical system 16 capable of adjusting the intensity of light radiated to the culture tube 14.

Description

DETAILED DESCRIPTION OF THE INVENTION [Object of the Invention] (Industrial Application Field) The present invention relates to an apparatus for culturing cultured materials such as cells (including protoplasts) and tissues.

(Prior Art) Cultivation and propagation of cells, tissues, etc. are not only important for basic research in biotechnology, but also are known to be useful for mass propagation of high-quality plants. Taking plant cells as an example, when protoplasts are immersed in a culture solution and allowed to proliferate, a callus (amorphous cell mass) is formed, and by changing the composition of the culture solution, various organs can be differentiated from the callus. By the way, culture until callus formation and subsequent culture require not only changing the composition of the culture solution but also changing the physical conditions. For example, static culture may be performed until cell walls are formed, and after cell wall formation, tilted rotation culture may be performed. Furthermore, it may be necessary to change the culture atmosphere gas composition during the growth process of the cultured material.

Conventionally, various devices such as incubators have been used for culturing cells, tissues, etc., but all of them can only be used under certain fixed culture conditions.

(Problems to be Solved by the Invention) Conventional culture equipment can only be used under fixed culture conditions, so it is not possible to set various culture conditions that occur during the growth process of the cultured material using these equipment alone. Can not. Therefore, in order to cope with this problem, it is necessary to increase the number of devices used and furthermore to complicate the control of culture conditions at each stage of culture. That is, since the culture conditions vary considerably during the growth process of the cultured object, when conventional culturing equipment is used alone, it is not possible to perform consistent culture according to the growth process of the cultured object.

Therefore, it is an object of the present invention to provide a culture device that can achieve consistent culture from culture to tissue differentiation.

[Structure of the Invention] (Means for Solving the Problems) The culture apparatus of the present invention includes a rotatable incubator for storing a culture tube in a constant temperature bath whose internal temperature can be arbitrarily set. In addition to installing an optical system that can adjust the amount and wavelength of irradiation light to irradiate the light necessary for the growth of the cultured object, a means for controlling the composition of the gas atmosphere within the thermostatic chamber is installed. . In addition, the inclination angle of the incubator can be arbitrarily set from a horizontal state to an inclined state.

(Function) The inside of the constant temperature bath is set at a desired temperature suitable for each stage of culture, and the gas atmosphere control means provides a gas composition suitable for each stage of culture.
In addition, the incubator is tilted in the culture stage where tilted culture is required. The optical system irradiates the culture tube containing the cultured material together with the culture solution with light at an amount corresponding to each stage of culture.

(Example) Hereinafter, an example of the present invention will be described in detail with reference to the drawings.

As shown in FIG. 1, a culture device 10 of the present invention
is equipped with a constant temperature bath 11 and a control device 21 attached thereto. The thermostatic chamber 11 can arbitrarily set and maintain its internal temperature according to each stage of culturing the cultured material.

Inside the constant temperature bath 11, a rotary incubator 30 including a support stand 12 and a culture tube holder 13 supported on the support stand 12 is installed. This culture tube holder 1
3 accommodates a plurality of culture tubes 14, for example, up to 26 culture tubes 14, which accommodate the culture material to be cultured together with a culture solution. The culture tube holder 13 is in a horizontal state (in the figure,
(indicated by a broken line), the culture tube 14 is supported almost vertically. Further, as shown in the figure, the inclination angle of the culture tube holder 13 can be arbitrarily set from a horizontal state to an inclined state according to the growth process of cells, and it can be rotated by a rotary drive motor 15. This rotation speed can be adjusted up to 10 rpm,
Furthermore, the rotation direction can be either left or right.

As shown in FIG. 2, the culture tube holder 13 has, for example, a circular shape, and a culture storage section is arranged on the concentric circle thereof so that the culture tube 14 can be accommodated therein.

As shown in detail in FIGS. 3 and 4, the rotating incubator 30 includes a support stand 12 and a culture tube holder 1.
3, and a fixed plate body 31 is placed on the support stand 12. The inclination angle adjustment member 32 and turntable support member 33 of the culture tube holder 13 are installed on the fixed plate 31 in the state shown in FIG. 3 (that is, the state in which the culture tube holder 13 is horizontal). ing. A turntable 34 is supported on the turntable support member 33, and the culture tube holder 13 is removably placed on the turntable 34. Rotary drive motor 15
is fixed to the turntable support member 33 and rotates the turntable 34 and therefore the culture tube holder 13 placed thereon.

As best shown in FIG. 4, when performing tilted culture by tilting the culture tube holder 13, first rotate the turntable support member 33 upward about the hinge 35, and
The angle adjustment member 32 is rotated about the hinge 36 to lock and support the turntable support member 33 on the angle adjustment member 32. That is, the angle adjusting member 32 is a plate-shaped body in which a guide slit 32a is formed in the longitudinal direction, and locking portions 32b are formed at predetermined intervals in communication with the slit 32a. A locking member 37, such as a pin or a nut, provided on the turntable support member 33 is connected to the locking portion 32.
By locking with b, the inclination angle θ of the culture tube holder 13 can be adjusted stepwise, for example, in 5° increments up to 85°.

Note that the angle adjustment member 32 may be a plate-like member having a guide slit in the longitudinal direction, and may be formed with an index (angle scale) of the inclination angle. In this case, if a convex indicating portion is formed on the locking member 37, the inclination angle can be continuously set in accordance with the angle scale.

Inclined rotational culture may be necessary when cells, callus, etc. are placed in a nonpolar state to differentiate organs, but in the culture device of the present invention, the culture tube holder 13 can be rotated and the inclination angle can be adjusted. This is to accommodate this.

In order to control the optical system, which will be described in detail below, an illuminance sensor for detecting the illuminance on the culture tube holder 13 can be installed in the constant temperature bath 111, and this sensor can be linked with the optical system. For example, as shown in FIG. 2, illuminance sensors 62 can be provided around the culture tube holder 13 at predetermined intervals. This illuminance sensor 62 is shown in FIGS.
As shown in the figure, the illumination meter mounting part 38 can be installed on the support member 39 and the support member 39 which is rotatable about the shaft 39a.
This attachment portion 38 allows the illumination meter 62 to be maintained in a horizontal state regardless of the inclination angle of the culture tube holder.

Now, the culture gas atmosphere in the constant temperature chamber 11 is based on the atmospheric atmosphere, and is normally under the atmospheric atmosphere, but the composition of this gas atmosphere (oxygen concentration, carbon dioxide concentration, etc.) may vary depending on the growth process of the cultured material. ), an air supply source 22, a nitrogen gas supply source 23, and a carbon dioxide gas supply source 24 are installed outside the thermostatic chamber 11 (FIG. 1). Air supply source 2
2 is a connection terminal 2 connected to the piping housing portion 21b of the control device 21 via the air purifier 25 via the line L1.
Connected at 6. Further, the nitrogen gas supply source 23 and the carbon dioxide gas supply source 24 are connected to electromagnetic valves 28 and 29 through lines L2 and L3, respectively, and these lines are connected to the piping accommodation portion 21b at the connection terminal 27 via a common line L4. ing. The solenoid valves 28 and 29 are connected to the control section 21a of the control device 21 via a common control line L5.

Next, the control system for the gas atmosphere composition will be explained in more detail with reference to FIG. In the illustrated example, four thermostats 11-1, 11-2, 11-3 and 11
- shows a mechanism in which four are installed in series and controlled by the control device 21.

The air supply source 22 is constituted by an air compressor, and the nitrogen supply source 23 and carbon dioxide supply source 24 are constituted by gas cylinders each containing these gases. Further, the air cleaner 25 includes an air filter 25a, a mist separator 25b, and a micro mist separator 25c.

The air supply line L1 is connected to a common connection line L6 to each thermostatic chamber through a supply air pressure regulating valve 40, a solenoid valve 41, and a three-way solenoid valve 46 in the piping housing portion 21b. Pressure regulating valve 40 and solenoid valve 4
A pressure gauge 42 is connected between the pressure gauge 1 and the pressure gauge 42 . In addition, a nitrogen gas supply source 23 and a carbon dioxide gas supply source 2
4 common connection line L4 is connected within the piping housing part 21b via a pressure regulating valve 43 and a solenoid valve 44,
The solenoid valve 41 and the three-way solenoid valve 46 are connected to the air supply line L1. Pressure regulating valve 43 and solenoid valve 44
A pressure gauge 45 is connected between the two.

The common supply line L6 each has a solenoid valve 74.
It is connected to constant temperature baths 11-1, 11-2, 11-3 and 11-4 via a, 47b, 47c and 47d.

Control line L connecting solenoid valves 28 and 29
5 is connected to a gas concentration control section 48 installed in the control section 21b. Solenoid valve 41 is connected to line L7
The electromagnetic valve 44 is connected to the control section 4 through a line L8. Furthermore, the three-way solenoid valve 46 is connected to L8 via line L9.
is connected to.

Note that the control unit 48 controls each thermostat 11-1, 11
A gas sensor is installed to detect the concentration of sampling gas from -2, 11-3, and 11-4, and the opening and closing of each solenoid valve is controlled by a signal from a controller driven based on information from each gas sensor. .

Further, each thermostatic oven has a common line L12 for sampling the atmospheric gas in the thermostatic oven to the control unit 48.

In addition to these, a humidity adjustment mechanism may be installed to adjust the humidity of the atmosphere within the thermostatic chamber. As shown in FIG. 5, this humidity control mechanism has a water storage container (water vapor source) 49, which is connected to a three-way solenoid valve 46, and supplies air from the air compressor 22 into the water contained therein. A blowing line L10 is inserted. As a result of this air blowing, air containing a predetermined amount of water vapor is supplied to the line L6 via the drain separator 51 and the line L11. A water supply tank 52 is installed at the bottom of the water storage container 49 via a pump 50, and a drain water storage container 53 is installed at the bottom of the drain separator 51.

Each gas supply source and water vapor source 49 are connected to the constant temperature bath 1
1, the composition can be changed as appropriate depending on the type of gas that needs to be changed.
Cultivation can be carried out under normal atmospheric conditions, but for example, depending on the oxygen concentration, carbon dioxide concentration, humidity, etc. at each stage of cultivation, nitrogen gas, carbon dioxide gas,
It is preferable to introduce water vapor into the constant temperature bath 11 and adjust it appropriately.

For example, when lowering the oxygen concentration in the constant temperature bath 11, the oxygen concentration is diluted using a nitrogen source.
That is, the nitrogen gas from the nitrogen gas cylinder 23 is reduced in pressure by the pressure regulating valve 43, and is kept at a constant temperature via the solenoid valves 28, 44, 46, 47 and 54, 54a, 54b, 54c, and 54d, which are opened by the control unit 48. It flows from inside the tank 11 to the outside. At this time, the oxygen gas concentration in the thermostatic chamber 11 is detected by an oxygen gas sensor in the control section 48, and the electromagnetic valve 44 is controlled by a signal from a controller in the control section 48 in response to the detection. When the oxygen gas concentration approaches the set value, the opening degree of the electromagnetic valve 44 is automatically reduced, and the electromagnetic valves 47 and 54 are also closed to prevent excessive supply of nitrogen gas.

When increasing the oxygen gas concentration, the air from the air compressor 22 is purified by the air purifier 25, and is supplied into each thermostat 11 via the line L1 in the same manner as the supply of nitrogen gas.

When increasing the carbon dioxide concentration, the carbon dioxide gas from the carbon dioxide cylinder 24 is passed through the line L3 to the solenoid valve 29, the pressure regulating valve 43, the solenoid valve 44, and the three-way solenoid valve 4.
Carbon dioxide gas is supplied from the electromagnetic valve 47 into each constant temperature bath 11 through line L6 via line L6. In addition, as an alternative method to increase the carbon dioxide concentration, the carbon dioxide concentration can be
Since 5000 ppm or less is sufficient, after setting the oxygen gas concentration to a predetermined concentration using the above method, the adapters 55a, 55b, 55c installed in each thermostatic chamber
From 55d onwards, a certain amount of carbon dioxide gas may be injected using a gas syringe.

Furthermore, in order to increase the humidity in the thermostatic chamber, the three-way solenoid valve 46 is switched to the direction of the water vapor source 49,
After the water droplets entrained by the humidified air are removed by the drain separator 51, the humidified air is supplied into each thermostatic chamber 11 via the line L11 and the line L6.

As described above, the gas atmosphere composition within the thermostatic chamber is controlled according to the atmosphere composition required for the growth process of the cultured object.

Returning to FIG. 1 to explain the culture apparatus of the present invention, an optical system 16 is provided in the constant temperature bath 11 above the culture tube holder 13 for irradiating the culture tube with light necessary for cell growth. is removably installed. This optical system 16 includes, for example, a plurality of lamps 1.
These lamps 17 have a structure in which the amount of irradiation light can be adjusted individually or in units of several groups. As a result, the culture tube holder 1 can be adjusted according to each stage of culture.
3. It is possible to equalize the illuminance on the entire surface, change the illuminance depending on the position, or make light and dark states of day and night appear. In addition, the culture tube holder 13
When tilted, it is possible to provide an illuminance difference of about 10,000 lux between the upper and lower parts.

The optical system and its control mechanism will be explained below with reference to FIGS. 6 and 7, but the gas atmosphere control means are not shown in these figures because they are the same as described above.

Referring to FIG. 6, a wavelength selection filter 61 is replaceably installed below the light source 17, and when obtaining the wavelength necessary for culturing, the filter that selectively transmits only that wavelength is replaced as appropriate. You can deal with it by doing this. Further, an illuminance sensor 62 is installed at an appropriate position around the culture tube holder 13, and this illuminance sensor 62 is connected to an illuminance meter 71 of the control device 21. While observing the illuminance detected by the illuminance sensor 62 with the illuminance meter 71, the lamps 17 are controlled as a whole, individually, or in groups. This control can be performed by a controller 72 consisting of all control sliders of the control device 21 and a controller 73 consisting of group unit control sliders. The group control slider can also control the lamps 17 individually.

Referring to FIG. 7, the inner wall of the thermostatic chamber 11 is made of a highly reflective material such as aluminum vapor deposition film, stainless steel, or chrome plating film. Further, a shutter 81 is provided to cover the entire surface of the transparent window member 11a of the thermostatic chamber 11 to block light from the outside.
is installed so that it can be stored freely. By covering the window member 11a with the shutter 81, the lamp 1
The lamp 7 can be turned off to make the inside of the thermostatic chamber 11 night-like, or the culture can be carried out using only the light from the lamp 17 since external light can be blocked. This shutter 81 is
The surface (back surface) facing 1 is made of the above-mentioned highly reflective material. Inner wall of constant temperature chamber 11, shutter 8
If the back surface of the tube 1 is made of a highly reflective material, the light from the lamp 17 will be reflected and the culture tube 14 will be irradiated with the reflected light again, so that the output of the lamp 17 can be reduced. In addition, shutter 81
Alternatively, the window member 11a may have a double structure and be installed in the space between them. Furthermore, when the culture tube holder 13 is tilted, a spherical light condensing plate 82 may be provided in the constant temperature bath 11 so as to face the culture tube holder 13 at a freely facing angle. This makes it possible to adjust the irradiation light.

Furthermore, it is also possible to install a blind 83 below the wavelength selection filter 61 and to control the irradiation angle of the light from the lamp 17 to the culture tube holder 13 by opening and closing the blind 83.

Furthermore, in any of the culture apparatuses shown in FIGS. 6 and 7, an ultraviolet lamp can be installed in the light source 17, and the irradiation can be controlled to induce mutations. In addition, culture tube holder 1
It is also possible to generate a flash in synchronization with the rotation of No. 3. Furthermore, a light source 17 and a culture tube holder 1
It is also possible to relatively adjust the distance to 3. Alternatively, the light source 17 may generate pulsed light to irradiate the cultured object with the light while periodically changing the amount of irradiation light.

In addition, in the culture apparatus of this invention, each indicator may be provided so as to issue an alarm when the illuminance or gas concentration varies from a predetermined set value. These controls can be performed by a control device 21 attached to the constant temperature bath 11.

Examples of objects to be cultured using the culturing apparatus of the present invention include cells and tissues of animals and plants, and microorganisms. Preferably, Haplopatpus, tobacco, petiunia, rice, corn, carrot, potato, wheat, barley,
Herbaceous plants such as sugarcane and soybean, and pine,
Examples include cells and tissues of woody plants such as eucalyptus, poplar, coffee, paragum, grape, elm, and apple. In addition, cells (including protoplasts), tissues, and organs to be cultured include:
Cells such as mesophyll cells, cultured cells derived from various tissues, tissues such as scutellum, embryo, mesophyll, cortex, and pith;
Meristem tissues such as the shoot apex and root tip, as well as stems, leaves, anthers,
Examples include organs such as roots and flowers.

For example, in order to culture plant cells using the culturing apparatus of the present invention, the temperature of 0 to 30°C and the temperature of 0 to 30° C.
Illuminance of 20000 lux, oxygen concentration of 2-20%, 0
~50% carbon dioxide concentration, rotating incubator tilt angle 0
Plant cells and tissues are suspended in a liquid medium such as Gamborg B5's basic medium (Gamborg et al. 1968) or Murashige-Skoog's MS basic medium (Murashige-Skoog 1962) at ~85°C, and cultured statically and rotated. It can be cultured, or it can be placed in a medium solidified with agar or the like to perform stationary culture or rotational culture. The plant growth hormones added to the medium used include naphthalene acetic acid (NAA),
2,4-dichlorophenoxyacetic acid (2,4-D),
Indole-3-acetic acid (IAA), indole-3
-Propionic acid (IPA), indole-3-butyric acid (IBA), phenyl acetic acid (PAA), benzofuran-
3-Auxins such as acetic acid (BFA) and phenylbutyric acid (PBA) and KT-30 (manufactured by Kyowa Hakko Co., Ltd.), 6
- Cytokinins such as benzylaminopurine (BA), zeatin (Z), kinetin, etc. may be used.

During or after the culture described above, the culture solution in the culture tube 14 may be replaced. In this case, remove the culture tube holder 13 for rotary culture from the turntable 34, attach it to the culture solution exchanger, and culture. Fluid can be exchanged. For example, the 8th
As shown in the figure, a culture solution exchanger 90 is provided with a rotor pump 92 having a plurality of rotors 91, and a supply tube 93 and a discharge tube 94 that are squeezed by the rotor pump 92 to supply and discharge the culture solution, respectively. The culture tube holder 13 containing the culture solution is placed on the tube. One end of the drain tube 94 is introduced into the drain container 96, and the other end is introduced into the culture tube 14 from which the culture solution is to be drained. Also,
One end of the supply tube 93 is introduced into the supply container 95 containing fresh culture solution, and the other end is introduced into the culture tube 14 from which the culture solution has been drained. By driving the rotor pump 92, the culture solution can be continuously supplied and discharged. Note that this supply/discharge can also be automated.

Note that it is preferable to use the culture tubes 14 shown in FIGS. 9 to 11. Referring to FIG. 9, the culture tube 14 has a bottomed cylindrical body 101 made of, for example, a test tube. This main body 101
is made of a material that transmits the light necessary for culturing, such as glass.

A filter member 102 is provided inside the main body 101 so as to traverse the inside of the main body 101 via an annular seal member 103 installed on the inner wall thereof. The seal member 103 is made of Teflon, for example, and is fitted and locked into the main body 101 with the filter member 102 fixed therein. The filter member 102 is made of a porous material that allows the culture solution to pass through, but does not allow the culture (including the cultured material and those that have proliferated and grown by culture) to pass therethrough. The pores of the porous material are smaller than the size of the cultured material. Preferably, filter member 102 comprises a ceramic filter.

A culture solution supply/discharge pipe 104 is provided to penetrate the filter member 102 . This pipe 104
is fixed to the ceramic quilter 102 by welding, and its lower end is connected to the culture solution 106 in the main body 101.
It reaches near the bottom 101b of the main body 101 so that it can be sufficiently discharged when discharging. Further, the upper end of the pipe 104 is set below the opening end of the main body 101 so as not to interfere with the installation of a closing member (not shown) when closing the opening 101a of the main body 101.

The culture using the above-mentioned culture tube will be explained below using the culture of protoplasts as an example.

First, in order to culture protoplasts, the culture tube 14 is set up vertically, the protoplasts 105 are placed on the filter member 102, and the main body is heated to the extent that the culture solution reaches a position of about 1 to 2 mm above the filter member 102. Add culture solution 15 into 101. If culture is performed in this state, the culture can be performed in the same way as when using a conventional shear dish. Moreover, in that case, since the culture solution can freely move up and down the filter member 102, metabolites such as polyphenols produced as the culture progresses also move to the lower part of the filter member 102 together with the culture solution. Since the culture solution 106 is replaced with fresh culture solution 106 from the lower part of the culture solution 102, the time period for replacing the culture solution can be extended considerably.

Furthermore, when the culture progresses and the culture solution 106 needs to be replaced, the culture solution can be easily supplied and discharged by connecting the supply and discharge pipe of the above-mentioned culture solution supply and discharge device to the upper end of the pipe 104. I can do it. In this case, even if the culture solution 106 is discharged, the culture is not removed from the filter member 102.
Next, if a new culture solution is injected into the main body 101,
Culture can be continued as is.

When the protoplasts have progressed to callus formation, a necessary amount of the culture solution is injected into the upper part of the filter member 102, and the culture solution can be directly subjected to tilt/rotation culture.

Note that the main body of the culture tube 14 is not limited to that shown in FIG. 9. For example, the main body 101
The culture solution reservoir 101c may have a substantially spherical shape as shown in FIG. 0, a rectangular parallelepiped shape as shown in FIG. 11, or any other appropriate shape at the bottom. With this bulging-shaped reservoir 101c, when the culture tube 14 is subjected to slant culture, the culture solution enters the reservoir 101c.
06 spilling out of the main body 101 is further reduced.

Examples of culture experiments using the culture apparatus of the present invention will be described below.

Experimental example 1 The growing point of the annual plant Haploppus gracilis was cut off after sterilization, and 2 mg/6 was added to the basic medium of Gamborg B5.
- Suspended in a modified medium (PH5.6) containing benzylaminopurine (BA) and 30,000 mg of sucrose as a carbon source, and rotary cultured using the apparatus of this invention (rotary incubator tilt angle 65°) did. The culture conditions were a temperature of 28° C., an illuminance of 2,000 to 12,000 lux, an oxygen concentration of 15% in the culture atmosphere, and a rotation speed of 3 rpm.

As a result, 5 weeks after the start of culture, pale green shoot primordium aggregates, which could not be obtained conventionally with an inclination angle of 0°, were obtained.

Experimental example 2 After sterilizing the seeds of Japanese red pine (Pinus densiflora), the embryos were extracted and added to Gamborg B5 basic medium at 2 mg/kg.
of naphthalene acetic acid (NAA), 4 mg/BA of
and a medium containing 30,000 mg of sucrose as a carbon source and solidified with agar (agar concentration 0.8%; PH
5.6) The cells were placed on a bed and cultured stationary using the culture apparatus of the present invention (tilt angle of 0°). Culture conditions are temperature 28
℃, illuminance was 3000 lux, and oxygen concentration in the culture atmosphere was 5%. Under these conditions, the callus formed from the embryos is suspended in a liquid medium of the same composition (no agar added) and continuously cultured in the same apparatus with rotation (tilt angle of 60°). According to
We succeeded in producing a large number of pale green cultured cells that were uniform and proliferated vigorously.

For comparison, we performed static culture for one month under the same conditions, subcultured them to the same medium, and continued static culture, but the proliferation rate in this area was about 50% of that in the area where we switched to rotary culture. Ta.

[Effects of the Invention] As described above, in the culture apparatus of the present invention, a rotatable incubator and an optical system capable of adjusting the amount of irradiation light are installed in a thermostatic chamber, and the gas atmosphere composition within the thermostatic chamber is adjusted. In addition to being equipped with a control means, the inclination angle of the culture tube holder of the rotary incubator can be set arbitrarily, so the inclination angle of the incubator, the amount of light irradiation, and the culture gas atmosphere composition can be adjusted according to each stage of culture. You can adjust it to the best condition. Therefore, the culture conditions during the entire growth process of cells etc. can be consistently controlled within the same device.

[Brief explanation of drawings]

FIG. 1 is a front view showing one embodiment of the culture device of the present invention, FIG. 2 is a plan view of a culture tube holder incorporated into the culture device of the present invention, and FIG. 3 is a culture device of the present invention. FIG. 4 is a side view showing a state in which the rotary incubator of the culture apparatus of the present invention is arranged horizontally. FIG. A circuit diagram of the gas control system of the culture device, FIGS. 6 and 7 are front views showing other examples of the culture device of the present invention, FIG. 8 is a schematic diagram of the culture solution exchange device, and FIG. 9 11 to 11 are cross-sectional views showing culture tubes suitable for use in the culture apparatus of the present invention. 10... Constant temperature bath, 12... Support stand, 13... Culture container holder, 14... Culture tube, 15... Drive motor, 16
...Optical system, 17...Lamp, 21...Control device, 2
2, 23, 24, 49...Gas supply source, 32...Angle adjustment member, 48...Gas concentration control unit, 62...Illuminance sensor, 61...Wavelength selection filter, 71, 72...
Light source control controller, 81...Shutter, 8
2...Reflector, 83...Blind, 90...Culture solution exchanger.

Claims (1)

[Scope of Claims] 1. A thermostatic chamber capable of setting and maintaining an internal temperature at a desired temperature suitable for the growth process of a cultured material, and a cultured material placed in the thermostatic chamber and to be cultured together with a culture solution. A rotatable incubator for accommodating culture tubes, comprising a culture tube holder for holding a plurality of culture tubes, a rotation drive unit for rotationally driving the culture tube holder, and the culture tube. A rotary culture system comprising an inclination angle adjusting member for arbitrarily setting the inclination angle of the holder from a horizontal state to an inclined state according to the growth process of the cultured object, and a support body that removably supports the culture tube holder. a gas atmosphere control means for controlling the gas atmosphere composition in the constant temperature bath according to the growth process of the cultured material, the gas atmosphere control means for controlling the gas atmosphere composition in the constant temperature bath, the gas atmosphere control means for controlling the gas atmosphere composition in the constant temperature bath, the gas atmosphere control means for controlling the gas atmosphere composition in the constant temperature bath, which a gas atmosphere control means having a gas supply source including a supply source and a control mechanism for controlling gas supply from the gas supply source to the thermostatic chamber; 1. A culture apparatus having a rotating culture vessel and gas atmosphere control means, characterized in that the culture tube is equipped with an optical system capable of adjusting the amount of irradiation light for irradiating the culture tube with light necessary for growth of the culture tube. 2. A culture apparatus comprising a rotary incubator and gas atmosphere control means according to claim 1, wherein the culture tube holder has an attachment part for an illuminance meter for measuring the amount of irradiated light. 3. A culture apparatus comprising a rotating culture vessel and gas atmosphere control means according to claim 2, wherein the illuminance meter mounting portion maintains a horizontal state regardless of the inclination angle of the culture tube holder. 4. The rotary incubator and gas according to claim 1, wherein the culture tube holder has a circular shape, and housing portions for accommodating a plurality of culture tubes are arranged concentrically therein. A culture device having atmosphere control means. 5. The inclination angle adjusting member is made of a plate-like body having a slit for guiding a locking member provided on the culture tube holder, and the position of the culture tube holder is adjusted depending on the locking position of the locking member. A culture apparatus comprising a rotary culture vessel according to any one of claims 1 to 4, which adjusts the inclination angle, and gas atmosphere control means. 6. A culture apparatus comprising a rotary incubator and gas atmosphere control means according to any one of claims 1 to 5, wherein the gas supply source includes a water vapor supply source. 7. According to any one of claims 1 to 6, wherein the control mechanism has a gas sensor that detects the gas concentration in the thermostatic chamber, and controls gas supply based on a signal from the gas sensor. A culture device comprising the rotary incubator and gas atmosphere control means described above. 8. The optical system includes a plurality of light sources that are removably provided above the thermostatic chamber, a wavelength selection filter that adjusts the wavelength of light irradiated to the culture tube that is provided below the light source, and the light source. A light source control section for controlling light irradiation.
A culture apparatus comprising the rotary culture vessel and gas atmosphere control means according to 1. 9. A culture apparatus comprising a rotating incubator and gas atmosphere control means according to claim 8, wherein the light source control unit controls the irradiation amount based on a signal from an illuminance sensor installed around the rotating incubator. 10. Claim 8 or 9, wherein a blind is installed below the wavelength selection filter to adjust the angle of light irradiation to the culture tube.
A culture apparatus comprising the rotary culture vessel and gas atmosphere control means as described in 1. 11. Any one of claims 1 to 10, further comprising a reflecting plate that is arranged to face the rotating culture vessel when it is in an inclined state and reflects light from the optical system to the culture tube. A culture apparatus comprising the rotary culture vessel according to item 1 and gas atmosphere control means. 12. A culture apparatus comprising a rotating culture vessel and gas atmosphere control means according to any one of claims 8 to 11, wherein the light source control unit independently controls the light source. 13. A culture apparatus comprising a rotating culture vessel and gas atmosphere control means according to any one of claims 8 to 12, wherein the light source includes an ultraviolet lamp for inducing mutations. 14. Any one of claims 1 to 13, wherein the thermostatic chamber has a transparent window member, and a light shutter is installed to cover the window member and block light from the outside. A culture device comprising the rotary culture vessel and gas atmosphere control means described in .