JPS62289176A - Culture apparatus having rotary culture device and means for controlling gaseous atmosphere - 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] 3. Detailed Description of the Invention] j-1 [Invention's] Field of Use (Industrial Application Field) This invention provides a method for culturing materials such as cells (including protoplasts), group a, etc. This invention relates to a device for culturing.

(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 superior 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 are differentiated from the callus. By the way, culture until callus is formed and subsequent culture require not only changing the composition of the culture solution, but also changing the physical conditions. For example, until cell walls are formed, it is necessary to Perform in situ culture,
After cell wall formation, tilted rotation culture is 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 to culture cells, tissues, etc., but all of them can only be used under fixed, constant 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, an object of the present invention is to provide a culture device that can achieve consistent culture from the cultured material 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. An optical system capable of adjusting the amount and wavelength of the irradiated light is installed to irradiate the light necessary for the growth of the cultured material, and a means for controlling the gas atmosphere composition in the thermostatic chamber is installed. There is. 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 is
A culture tube containing a culture medium and a culture medium is irradiated 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, the culture apparatus 10 of the present invention includes a constant temperature bath 11 and a control bag M21 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. The culture tube holder 13 accommodates a plurality of culture tubes 14, for example up to 26 tubes, which accommodate the culture material to be cultured together with a culture solution.

The culture tube holder 13 supports the culture tube 14 almost vertically in a horizontal state (indicated by a broken line in the figure). Further, the culture tube holder 13 is as shown in the figure.

The inclination angle can be set arbitrarily from a horizontal state to an inclined state according to the growth process of cells, and the rotary drive motor 15
It can be rotated by This rotation speed is LOr
It can be adjusted up to pm and can be rotated in either the left or right direction.

As shown in Figure 2, the culture tube holder! 3 has a circular shape, for example, and a culture tube holder is disposed on the concentric circle thereof so that the culture tube 14 can be accommodated therein.

1 rotating incubator 3 as shown in detail in Figures 3 and 4.
0 includes a support stand 12 and a culture tube holder 13, and a fixing plate 31 is mounted on the support stand 12. The culture tube holder 13 is placed on the fixed plate 31 in the state shown in FIG. 3 (that is, the culture tube holder 13 is horizontal).
A 31-section member 32 with an inclination angle of 31 and a turntable support member 33 are installed.

The turntable 3 is mounted on the turntable support member 33.
4 is supported, and the culture tube holder 13 is removably mounted on this turntable 34. A zero rotation drive motor 15 is fixed to the turntable support member 33,
The turntable 34 and therefore the culture tube holder 13 placed thereon are rotated.

As best shown in FIG.
3 to perform tilted culture, first rotate the turntable support member 33 upward around the hinge 35, and rotate the A node member 32 around the hinge 36 to achieve this angle. The speed regulating member 32 locks and supports the turntable support member 33. That is, the one-angle member 32 is a plate-shaped body in which guide slits 7l-32a are formed in the longitudinal direction, and locking portions 32b are formed at predetermined intervals in communication with the slits 32a. By locking a locking member 37, such as a bottle or a nut, provided on the turntable support member 33 to the locking portion 32b, the inclination angle of the culture tube holder 13 is changed from 0 to 8, for example, by 5 degrees.
It is constructed so that 31 sections can be made in stages up to 5 degrees.

Note that the angle A node member 32 may be a plate-shaped body having a guide slit in the longitudinal direction, and may be formed with an index of the inclination angle (angle scale). In this case, the locking member 37
By forming a convex indicator on the surface, the inclination angle can be set continuously to match the angle.

Inclined rotational culture may be necessary when cells, callus, etc. are placed in a non-polar state to differentiate organs, but in the culture device of the present invention, the culture tube holder 13 is a rotating i8T moat and the inclination angle is adjusted. The reason for the circular part is to accommodate this.

Note that in order to control the optical system, which will be described in detail later, a constant temperature bath 11 is used.
For example, as shown in FIG. Illuminance sensors 62 can be provided around the area at predetermined intervals. As shown in FIGS. 3 and 4, this illuminance sensor 52 is an illuminance meter installed on a support member 39 that is rotatable around a support member 39 and a shaft 38a. Can be done. This installation is part 3
8, the illumination meter 62 can 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 gas etc.), 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). The connecting terminal 26 is connected to the piping accommodating portion 21b of the control device 21 via the line L1 via the container 25. Also, the nitrogen gas supply source 23 and the carbon dioxide gas supply source 24 are supplied through lines L2 and L3, respectively. It is connected to If magnetic valves 28 and 29, and these lines are connected to the piping housing portion 21b at a connecting terminal 27 via a common line L4. The solenoid valves 28 and 29 are connected to the control unit 21a of the control device 21 via a common control line L5.
is connected to.

Next, the control system for gas atmosphere composition will be explained in more detail with reference to FIG. 5. The example shown in FIG.
1-1, 112, 11-3, and 11-4) are arranged in series and are controlled by the control device 21.

The air supply source 22 is constituted by a near compressor, and the nitrogen supply source 23 and the carbon dioxide supply source 24 are constituted by gas cylinders each containing these gases. Also,
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 located within the pipe housing section 21b.
Supply air pressure regulating valve 40, solenoid valve 41 and three-way solenoid valve 46
It is connected to a common connection line L6 to each thermostat via. A pressure gauge 42 is connected between the pressure regulating valve 40 and the solenoid valve 41. In addition, a common connection line L4 between the nitrogen gas supply source 23 and the carbon dioxide gas supply source 24 is connected to the piping housing section 21.
In b, an air supply line Ll is connected between the solenoid valve 41 and the three-way solenoid valve 46 via the pressure regulating valve 43 and the solenoid valve 44.
is connected to. A pressure gauge 45 is connected between the J pressure valve 43 and the solenoid valve 44.

Each common supply line L6 has a solenoid valve 47a.

47b, 47c and 47d to constant temperature bath 11-
1, 11-2, 11-3 and 11-4.

'i[The control line L5 connecting the magnetic valves 28 and 29 is connected to the gas concentration control section 48 installed in the control section 21b.
is connected to. The solenoid valve 41 is connected to the control section 48 via the line L8, and the solenoid valve 44 is connected to the control section 4 via the line L8. Furthermore, a three-way solenoid valve 46
is connected to line L8 via line L9.

Note that the control unit 48 controls each thermostat 11-1.11-2.1.
A gas sensor is installed to detect the concentration of the sampling gas from 1-3 and 11-4, and the opening and closing of each electromagnetic valve is controlled by signals from a controller driven based on information from each gas sensor.

Furthermore, each thermostatic oven has a control unit 4 that controls the atmospheric gas in the thermostatic oven.
It has a common line L12 for sampling 8 times.

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 consists of a wood storage container (
water vapor source) 48, and in the water contained therein,
A line L10 connected to the three-way solenoid valve 46 and blowing air from the near compressor 22 is inserted. By this air blowing, air containing a predetermined amount of water vapor is passed through the drain separator 51 and the line Lll to the line L6.
supplied to A water supply tank 52 is installed at the bottom of the water storage container 43 via a pump 50, and a drain separator 5
A drain water storage container 53 is installed at the bottom of the tank 1.

Each gas supply source and water vapor source 49 can be switched as appropriate depending on the type of gas whose composition needs to be changed in the thermostatic chamber 11. Cultivation can be carried out under normal atmospheric conditions, but for example, the oxygen concentration,
It is preferable to appropriately adjust the carbon dioxide concentration, humidity, etc. by introducing nitrogen gas, carbon dioxide gas, and water vapor into the constant temperature bath 11 according to each stage of culture.

For example, when lowering the oxygen concentration in the thermostatic chamber 11, a nitrogen source is used to dilute the oxygen concentration.

That is, the nitrogen gas from the nitrogen gas pump 23 is reduced in pressure by the pressure regulating valve 43, and the solenoid valves 28, 44, 46, 47 and 54 (54a) are opened by the control unit 48.

54b, 54c, 5Ld)
Flows from the inside to the outside. At this time, the /lN elementary 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 the 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, use near compressor 2.
2 is purified by an air purifier 25, and is supplied into each thermostatic chamber 11 via 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 pump 24 is passed through the line 3, the solenoid valve 29, the pressure regulating valve 43, the solenoid valve 44, and the three-way fi solenoid valve 6 to the line L6.
carbon dioxide gas is supplied from the magnetic valve 47 into each constant temperature bath 11 through '7M. Note that as another method for increasing the carbon dioxide concentration, since it is sufficient for the carbon dioxide concentration to be 5000 ppm or less, after setting the oxygen gas concentration to a predetermined concentration by the method described in -F, adapters 55a, 55 installed in each thermostatic chamber are used.
A fixed amount of carbon dioxide gas may be injected from b, 55c and 55d using a gas syringe.

Furthermore, in order to increase the humidity in the thermostatic chamber, the three-way solenoid valve 4B is switched to the direction of the main air source 49, and after removing the water droplets entrained with the humidified air by the drain separator 51, the line Lll and each thermostatic chamber l via line L6.
Supply within l.

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, in the thermostatic chamber IJ, an optical system 16 is provided north of the culture tube holder 13 for irradiating the culture tube with light necessary for cell growth. is removably installed. This optical system 16 includes a light source consisting of, for example, a plurality of lamps 17, and these lamps 17 are configured to be able to adjust the amount of irradiation light individually or for each group. Thereby, it is possible to equalize the illuminance over the entire surface of the culture tube holder 13 according to each stage of culture, to change the illuminance depending on the position, or to make light and dark states of day and night appear. Furthermore, when the culture tube holder 13 is tilted, a difference in illuminance of about 10,000 lux can be provided 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 culture, 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 2121. 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 0 group unit control sliders 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 installed to cover the entire surface of the transparent window member 11a of the thermostatic oven 11 and to block light from outside. By covering the window member 11a with the shutter 81, the lamp 17 can be turned off and the inside of the thermostatic chamber 11 can be brought into a night state, or external light can be blocked, so that culture can be performed using only the light from the lamp 17. You can also do that. This shutter 81 faces the constant temperature bath 11 (back side).
is made of the above-mentioned highly reflective material. Constant temperature bath 11
If the inner wall of the tube 81 and the back surface of the sheath 81 are made of a highly reflective material, the light from the lamp 17 will be reflected and the reflected light will be irradiated onto the culture tube 14 again, thereby reducing the output of the lamp 17. Can be done. Note that the shutter 81 may have a double structure for the window member 11a and be installed in the space between them.Furthermore, the shutter 81 may be set at a facing angle so as to face the culture tube holder 13 when it is tilted. The spherical light condensing plate 82 may be freely provided in the thermostatic chamber 11, thereby reducing the gJf! of the irradiated light! I can measure 5.

Further, a blind 83 is provided below the wavelength selection filter 61.
The lamp 17 is opened and closed by opening and closing the blind 83.
It is also possible to control the irradiation angle of the light from the culture tube holder 13 to the culture tube holder 13.

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. Further, the flash can also be generated in synchronization with the rotation of the culture tube holder 13. Furthermore, the distance between the light source 17 and the culture tube holder 13 can be set to be 2111rr1 relatively. 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 illumination intensity or the dregs concentration fluctuates from a predetermined set value.

These controls are controlled by a control W? attached to the thermostatic chamber 11. c21
21.

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, herbaceous plants such as habropuffgus, tobacco, petunia, rice, corn, carrot, potato, wheat, barley, sugar cane, and diver, and woody plants such as pine, eucalyptus, poplar, coffee, paragum, grape, elm, and apple. Examples include cells and tissues. In addition, the cells (including protoplasts), tissues, and organs to be cultured include mesophyll cells, cells such as cultured cells derived from various tissues, tissues such as cup disks, embryos, mesophyll, cortex, and pith, shoot tips, meristems such as plate edges,
In addition, organs such as stems, leaves, rot, roots, and flowers can be exemplified.

For example, in order to culture plant cells using the culturing apparatus of the present invention, the temperature is 0 to 30°C, the illuminance is 0 to 20,000 lux, the oxygen concentration is 2 to 20%, and the carbon dioxide gas is 0 to 50%. Plant cells and tissues were grown in Gamborg B5 basal medium (Gamborg
et al., 1968), Murashige-Skoog MS
Basic cultivation i1! (Murashige *Skoog 1
It is also possible to suspend in a liquid medium such as 962) and perform static culture or rotational culture, or to perform static culture or rotational culture by placing the above medium in a medium solidified with agar or the like. . 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), and indole-3-propionic acid ( I
PA), indole-3-acetic acid (IBA).

Phenyl acid 8 (FAA), benzofuran-3-acetic acid (
BFA), nonaxins such as PBA), and cytokinins such as KT-30 (manufactured by Kyowa Hakko Co., Ltd.), 6-benzylaminopurine (BA), zeatin (Z), and kinetin. Can 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. For example, as shown in FIG.
A culture tube holder containing a culture solution is placed on a culture solution exchanger 80 equipped with a rotor pump 92 having a rotor pump 1 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. 13 to Jt 27. One end of the drain tube 34 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. Further, one end of the supply tube 93 is introduced into a supply container S5 containing a new culture solution,
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 feeding table discharge can also be automated.

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

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 +02 fixed therein. The filter member 102 allows the culture solution to pass through, but does not allow the culture solution (
(Including cultured materials and those proliferated and grown by culture)
It is made of a porous material that does not allow water to pass through. 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 vibrator 104 passes through the filter member 102
is provided. This pipe 104 is fixed to the ceramic filter 102 by welding, and its lower end reaches near the bottom 101b of the main body 101 so that the culture solution 106 inside the main body 101 can be sufficiently discharged. Further, the upper end of the vibrator 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 the opening of the main body 101 is closed.

Culture using the above-mentioned culture tube will be described below, taking the culture of protoplasts as an example.

First, in order to culture protoplasts, the culture tube 14 is set up vertically, and the protoplasts 105 are placed in the filter member 10.
2, and the culture solution is placed on the filter member 102 about 1 to 2 times.
The culture solution 1 is poured into the main body 101 to the extent that it reaches a position of about 1.0 mm.
Add 5. If culture is performed in this state, culture can be performed in the same way as when using a conventional petri dish. Moreover,
In that case, since the culture solution can freely move up and down the filter member +02, metabolic substances such as polyphenols produced as the culture progresses will also be transferred to the filter member +02 along with the culture solution.
02 and is replaced with fresh culture solution 1013 from the lower part of the filter member 102, the time period for replacing the culture solution can be significantly extended.

Furthermore, when the culture progresses and the culture solution 106 needs to be replaced, the culture solution can be easily supplied and drained by connecting the supply and discharge pipe of the culture solution exchange device to the upper end of the pipe 104. can. in this case.

Even if the culture solution 10B is discharged, the culture remains in the filter member 10.
2, so if a new culture solution is injected into the main body 101, the culture can be continued.

Once the protoplasts have progressed to callus formation,
A required amount of culture solution can be injected into the upper part of the filter member 102, and the culture solution can be directly subjected to tilted/rotated 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 may have a culture solution reservoir 101c at the bottom that is approximately spherical as shown in FIG. 10, rectangular parallelepiped as shown in FIG. 11, or has any other suitable shape. It's okay. With this bulging-shaped reservoir 101c, when the culture tube 14 is subjected to slant culture, the culture solution enters the reservoir lot c, which further reduces the risk of the culture solution 106 spilling out from the main body 101. be done.

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

Experimental example 1 Annual plant Haplopagus (HaplopappuSg)
After sterilization, the growing points of S. racilis were cut out and added to Ganpaug B5 basal medium with 2 mg/L of 6-benzylaminopurine (BA) and 30.00 mg/L as a carbon source.
Modified medium (pH 5, 6) with 0 mg/7 sucrose added
and cultured by rotation using the apparatus of the present invention (rotating incubator tilt angle 65°). Culture conditions are temperature 28℃
The illuminance was 2000 to 12.0°00 lux, the oxygen concentration in the culture atmosphere was 15%, and the rotation speed was 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 (Pinu Sdensiflora), the embryos were removed and placed in 2 m of Ganpaug B5 basic medium.
g/f naphthaleneacetic acid (NAA), 4 mg/p-
A medium containing NoBA and 30.000 mg/℃ sucrose as a carbon source and solidified with agar (agar concentration 0.8
%, pH 5, 6) and cultured stationary using the culture apparatus of the present invention (tilt angle 0°). Culture condition is temperature 2
The temperature was 8°C, the illuminance was 3000 lux, and the oxygen concentration in the culture atmosphere was 5%. Under these conditions, the calli formed from the embryos were cultured for one month, suspended in a liquid medium of the same composition (no agar added), and continuously tilted with the same device (tilt angle 60°).
) By rotating culture, we succeeded in producing a large number of pale green cultured cells that were uniform and showed vigorous growth.

For comparison, we cultured the cells statically for one month under the same conditions, subcultured them to the same medium, and continued to culture them statically. In this plot, the proliferation rate was about 50% of that in the plot where we cut to rotary culture and suppressed the growth. there were.

[Effects of the Invention] As described above, in the culture apparatus of the present invention, a rotary incubator and an optical system capable of adjusting the amount of irradiation light are installed in a constant temperature chamber, and the gas atmosphere composition within the constant temperature chamber is adjusted. In addition, 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 composition of the culture gas atmosphere can be adjusted at each stage of culture. You can adjust it to suit your needs. Therefore, the culture conditions during the entire growth process of cells etc. can be consistently controlled within the same device.

[Brief explanation of the drawing]

FIG. 1 is a front view showing an embodiment of the culture device of the present invention, FIG. 2 is a plan view of a culture tube holder incorporated into the culture vessel 2t of the present invention, and FIG. FIG. 4 is a side view showing the rotating incubator of the apparatus of the present invention in a horizontally arranged state; FIG. , a circuit diagram of the gas control system of the culture device of the present invention, FIGS. 6 and 7 are front views showing other examples of the culture device of the present invention, and FIG. 8 is a schematic diagram of the culture medium exchange device. , FIGS. 9 to 11 are cross-sectional views showing culture tubes suitable for use in the culture apparatus of the present invention. 0... Constant temperature bath, 12-φφ support stand. 13...Culture container holder, 14...Culture tube, 15-
・War drive motor, 1611...Optical system. 17φ...Lamp, 21...Fist control device, 22, 23.
24.49...φ gas supply source, 32.Fist/angle adjustment member, 48...φ gas concentration control unit, 62---Illuminance sensor, 61...Wavelength selection filter, 71.72---Light source control Controller, 81Φ・Fushi shutter, 82・
...Reflector, 83...Contest blind, 90...Culture solution exchanger Figure 6 section C) faction

Claims (14)

[Claims] (1) A constant temperature bath that can set and maintain the internal temperature at a desired temperature suitable for the growth process of the cultured material, and a culture tube installed in the constant temperature bath and containing the cultured material to be cultured together with the culture solution. A rotatable culture vessel for storing a culture tube, the culture vessel comprising a culture tube holder for holding a plurality of culture tubes, a rotation drive unit for rotationally driving the culture tube holder, and a cover for the culture tube holder. a rotating incubator equipped with an inclination angle adjusting member for arbitrarily setting the inclination angle from a horizontal state to an inclined state according to the growth process of the culture, and a support body that removably supports the culture tube holder; A gas atmosphere control means for controlling the gas atmosphere composition in a thermostatic chamber according to the growth process of a cultured object, the gas atmosphere control means including a supply source of oxygen gas, nitrogen gas, and carbon dioxide gas to be supplied to the thermostatic chamber. a gas atmosphere control means having a gas 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 a 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. (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 illuminometer for measuring the amount of irradiated light. (3) A culture apparatus comprising a rotating incubator 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 according to claim 1, wherein the culture tube holder has a circular shape, and a accommodating section for accommodating a plurality of culture tubes is arranged concentrically therein. and a culture device having gas atmosphere control means. (5) The inclination angle adjusting member is made of a plate-shaped body with a slit for guiding a locking member provided on the culture tube holder, and the locking position of the locking member determines the position of the culture tube holder. 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) Any one of claims 1 to 6, wherein the control mechanism includes 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 apparatus comprising the rotary culture vessel and gas atmosphere control means according to 1. (8) The optical system includes a plurality of light sources removably provided above the thermostatic chamber, and a wavelength selection filter provided below the light sources that adjusts the wavelength of light irradiated to the culture tube. Claims 1 to 7 include a light source control unit that controls light irradiation from a light source.
A culture apparatus comprising the rotary culture vessel according to any one of Items 1 and 2 and gas atmosphere control means. (9) Cultivation comprising a rotating culture vessel 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 culture vessel. Device. (10) The rotary incubator and gas atmosphere control according to 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 device having means. (11) Claims 1 to 10 further include 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 onto the culture tube. A culture apparatus comprising the rotary culture vessel according to any one of the above and gas atmosphere control means. (12) A culture apparatus comprising a rotary incubator 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 apparatus comprising the rotary culture vessel according to item 1 and gas atmosphere control means.