JPH10201467A - Judgement of growth of material to be cultured and culture apparatus therefor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To judge the growth of bulbs of golden-banded lily, trumpet lily, etc., as a material to be cultured from the state of reduction in electrical conductance by measuring the amount of an electrical conductive substance existing in a liquid medium by the electrical conductance and to produce systematically lilies. SOLUTION: For example, a bioreactor 1 is charged with a liquid medium A and bulbs B of golden-banded lily as materials to be cultured and the materials B to be cultured are cultured while being properly controlled. The temperature of the bioreactor 1 is controlled by a jacket 2. An electrical conductance controller 3 is connected to the bioreactor 1 and measures the electrical conductance of the liquid medium A and the controller 3 is equipped with a sensor 3a. The sensor 3a is immersed in the liquid medium A, measures the electrical conductance of the liquid medium A and is connected to a control part C and a medium supplying means 4. The bioreactor 1 is further equipped with a heating medium supplying means 5, a pH detecting means 6, an acid supplying means 7, an alkali supplying means 8, a level sensor 9, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and a culture apparatus for judging the growth of an object to be cultured, and is a technique suitable for use in the planned production of bulbs such as lily of the valley and lily of the valley.

[0002]

2. Description of the Related Art Lily plants (Lilium) include yamali lily (Lilium auratum) and tepow lily (Lilium eleganc).
Many species are known, including e), and there are many valuable varieties for appreciation, and some of the bulbs are used as high quality foods.

[0003] As a technique related to the culture of lily plants,
Technical Example 1: Japanese Patent Application Laid-Open No. 55-15734, "Large Propagation Method of Lily Seedlings", Technical Example 2: Japanese Patent Application Laid-Open No. Sho 61-285928, "Method of Proliferating Lily Seedlings", Technical Example 3: Japanese Patent Application Laid-Open No. 2-4
No. 6239, "Lily Plant Growth Method" and the like.

In Technical Example 1, a stem, leaf, petal, anther, ovary, inflorescence, bud, scaly, root or other tissue of a plant of the genus Lily is cut into small pieces and subjected to a sterilization treatment to obtain a tissue piece. Used as a culture. The tissue piece is cultured in a solid medium, and when a sphere is formed, the scale is separated. The isolated scales are liquid-cultured to enlarge and grow into scale masses, and then the scales are separated from the scale masses and cultured in a solid or liquid medium to form small bulbs. . Thereafter, these small bulbs are grown as seeds and seedlings by soil cultivation and are allowed to flower.

[0005] In Technical Example 2, a growing point mass having a shoot apex structure is formed from a tissue piece or a cultured cell, and is transplanted to a seedling medium to produce a seedling. Further, in the technical example 3, when a liquid culture medium in which an oxygen-containing gas is aerated is used for culturing tissue pieces or cultured cells, for example, a tube having many holes placed in the liquid culture medium is used. The oxygen-containing gas is pumped through a culture layer that performs a gyro-like swirling motion or a technique for pumping the oxygen-containing gas from the air.

[0006]

However, when examining the growth and the like of the bulb, it is common to measure the wet weight of the bulb taken out from the culture medium.
The relationship between the growth of the bulb and the condition of the culture medium has not been sufficiently established, and for example, it has been difficult to determine whether or not the culture medium has sufficient capacity during culture. For this reason, there was a place to be improved in optimizing the culture medium conditions and systematically producing bulbs of lily plants.

[0007] As a method of measuring the biomass in the culture tank, Technical Example 4: Japanese Patent Application Laid-Open No. 2-57954, entitled "Method of Measuring Biomass" has been proposed, in which one or more electrodes are provided in the culture tank. However, the electric conductivity during this period is measured at two different frequencies. However, based on the detected results, the technology has not yet been developed to maintain the optimal conditions for the culture medium and grow the culture. .

[0008] The present invention has been made in view of such problems, and has the following objects. To easily grasp the growth status of bulbs. To provide and maintain an effective culture environment for bulb growth. Judgment of bulb growth status is not limited by the type of culture medium. To enable the planned production of bulbs. To allow adaptation of other plants to culture medium.

[0009]

In order to culture the seedling bulbs of the genus Lily and monitor the growth of the bulbs, the amount of the electrically conductive substance present in the liquid medium is measured by the electric conductivity, and the electric conductivity is reduced. The presence or absence of the effectiveness of the medium is determined depending on the situation described above. That is, the rate of decrease in the electrical conductivity is detected, and when the change in the rate of decrease is small, it is determined that the liquid medium is not effective. When it is determined that the liquid medium is not effective, a technique for supplying the concentrated medium according to the decrease in the electric conductivity in the liquid medium and determining the degree of recovery of the electric conductivity is employed. Further, as a culture device for cultivating seedling bulbs of a lily genus plant, a bioreactor containing a liquid medium for culturing an object to be cultured, and an electric conductor connected to the bioreactor for measuring the electric conductivity of the liquid medium A technique having a degree sensor and a medium supply means connected to the electric conductivity sensor and supplying a medium into the bioreactor when the electric conductivity decreases is applied. A jacket for adjusting the temperature of the bioreactor by circulating a heating medium (temperature-regulated water: temperature-regulated water) is provided outside the bioreactor. A technique is applied in which a heat medium supply means for storing the medium is connected, the heat medium is supplied into the jacket through a supply port, and the heat medium is returned from the discharge port to the heat medium supply means after heat exchange. Also,
A control unit that keeps the temperature of the heat medium stored in the heat medium supply unit constant is added. The liquid medium stored in the bioreactor is operated by operating a medium supply means having a concentrated medium supply system for supplying a concentrated medium and a pure water supply system for supplying pure water. The bioreactor is provided with a pH detecting means for detecting the pH inside the liquid, and the pH detecting means is provided with an acid supply system and an alkali supply system for supplying an acid and an alkali. The liquid level management of the liquid culture medium stored in the bioreactor is performed by driving the culture medium supply means based on the detection signal of the level sensor that detects the liquid level, and when the liquid culture medium is excessively supplied, The liquid level is kept constant by overflowing the liquid medium with a liquid level setting means (overflow section) arranged on the side of the bioreactor. Oxygen is supplied to the liquid medium by supplying air from the air supply means to the sparger disposed at the bottom of the bioreactor to perform aeration.At this time, a sterilizing filter is provided between the air supply means and the sparger. By disposing in the intervening state, the supplied air is sterilized. The detection of the amount of dissolved oxygen dissolved in the liquid medium is performed by arranging the dissolved oxygen detecting means in the bioreactor, and the dissolved oxygen detecting means is connected to a mass flow controller for setting the air supply amount, whereby the dissolved oxygen is detected. The adjustment is performed by increasing the ventilation speed when the measured value falls below the set value, and conversely, by decreasing the ventilation speed when the measured value exceeds the set value. The bioreactor is provided with a control unit for setting the conditions of the culture medium based on internal information such as dissolved oxygen, electric conductivity, acidity, liquid level, and air volume. Further, in a culture apparatus for cultivating seedling bulbs of a lily plant, a bioreactor having a pair of culture tanks may be employed. A culture medium transfer means for sucking the liquid culture medium stored in the culture tank and transferring the liquid culture medium to another culture tank is provided, and a suction and exhaust means for sucking and discharging air is provided above the pair of culture tanks. Technology is applied. Similarly, when a bioreactor having a pair of culture tanks is used as a bioreactor having a pair of culture tanks, for example, the lower part of the upper culture tank is connected via a siphon tube, and A sparger for sending out air is provided at the upper part, and a technology is applied to this sparger in which air supplied from the air supply means is sent through a lower culture tank.

[0010]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a first embodiment of a method for judging the growth of a body to be cultured and a culture apparatus according to the present invention will be described with reference to FIG. In FIG. 1, reference symbol A denotes a liquid medium, B denotes a bulb (Bulb), C denotes a control unit, F denotes a sterilization filter, RL denotes a liquid level, S denotes a sparger, 1 denotes a bioreactor, 2 denotes a jacket, and 3 denotes electrical conductivity. Controller, 4
Is a medium supply means, 5 is a heating medium supply means, 6 is a pH controller, 7 is an acid supply system, 8 is an alkali supply system, 9 is a level sensor, 10 is a liquid level setting means, 11 is an air supply means,
2 is dissolved oxygen detection means, 13 is a mass flow controller,
Reference numeral 14 denotes an exhaust unit.

The bioreactor 1 is a container for accommodating a culture medium together with a culture medium and culturing the culture medium under appropriate management. In this case, a bulb (Bulb) B of a lily of the valley is used as the culture medium. A liquid medium A is used as a medium for culturing the bulb B.

The jacket 2 is disposed outside the bioreactor 1 and adjusts the temperature of the bioreactor 1 using a heat medium or the like. In this case, the heat medium may be, for example, temperature-controlled water (temperature control). Water is circulated inside.

The electric conductivity controller 3 is connected to the bioreactor 1 and measures the electric conductivity of the liquid medium A. The electric conductivity controller 3 includes an electric conductivity sensor 3a provided with electrodes. The electric conductivity sensor 3a is immersed in the liquid medium A, and the electric conductivity (ion concentration) in the liquid medium A is determined. However, since the inside of a living cell is filled with electrolyte, the electric conductivity increases as compared to an inanimate medium, It focuses on the fact that the ion concentration changes with the growth of the bulb (Bulb) B, and is connected to the control unit C and the medium supply means 4.

The medium supply means 4 has a concentrated medium supply system 4A for supplying a concentrated medium and a pure water supply system 4B for supplying pure water, which are arranged in a connected state to the bioreactor 1. , The respective solutions are supplied to the bioreactor 1 by the operation of the pump P.

The heat medium supply means 5 stores a heat medium maintained at an appropriate temperature or a high temperature in advance, supplies the heat medium to the inside of the jacket 2 through the supply port 2a, and penetrates the inside of the jacket 2. Recirculation for recovering from the outlet 2b is performed later. Further, the heating medium supply means 5 includes a temperature sensor 5a, which detects the temperature by immersing the temperature sensor 5a in the liquid medium A, and based on the temperature detection data, the control unit C controls the heating medium supply means. A control signal for adjusting the temperature of the heat medium to an appropriate value is output to the control unit 5.

The pH detecting means 6 is provided with a pH sensor 6a, which detects the pH of the culture medium by immersing it in a liquid culture medium A. The system 7 and the alkali supply system 8 are operated to supply an acid or an alkali as needed.

The acid supply system 7 is connected to the bioreactor 1, and operates the pump P to supply the acid to the liquid medium A when the acid is insufficient based on a command from the control unit C.

The alkali supply system 8 is connected to the bioreactor 1 and operates the pump P to supply the alkali to the liquid medium A when the alkali is insufficient based on a command from the control unit C.

The level sensor 9 is immersed in the liquid medium A to detect the liquid level RL of the bioreactor 1 and is connected to the pure water supply system 4B so that the liquid level RL of the bioreactor 1 falls below a set value. At this time, the pure water supply system 4B is operated to supply pure water to the bioreactor 1, thereby keeping the liquid level RL constant.

The liquid level setting means 10 is provided on the side of the bioreactor 1, and sets the liquid level RL of the bioreactor 1 to a constant level by overflowing the liquid medium A.

The air supply means 11 is for feeding fresh air to the bioreactor 1 and is connected to a sparger S at the bottom of the bioreactor 1 and blows air (aeration) from the sparger S to the liquid medium A. Is supplied. The sparger S is provided with a sterilizing filter F for sterilizing supplied air.

The dissolved oxygen detecting means 12 comprises a liquid medium A
The measurement data is transmitted to the mass flow controller 13 and is also connected to the control unit C.

The mass flow controller 13 is interposed between the air supply means 11 and the sparger S,
This is for setting the amount of air supply to the bioreactor 1. On the basis of the dissolved oxygen amount data from the dissolved oxygen detection means 12, the control unit C increases the ventilation speed when it determines that the airflow rate falls below the set value, and increases the airflow rate when it determines that the airflow rate exceeds the set value. Is adjusted to reduce.
The air supplied excessively inside the bioreactor 1 and the carbon dioxide generated due to the growth of the bulbs are
It is discharged from the exhaust means 14.

Next, a second embodiment of the apparatus for culturing a body to be cultured according to the present invention will be described with reference to FIG.

The bioreactor 1 according to the second embodiment
Inside, the heat exchanging means 21 is arranged in the exhaust pipe 14a of the exhaust means 14 (or in the middle thereof). The heat exchange means 21 has a refrigerant supply port 21a for sending refrigerant, and a refrigerant discharge port 21b for discharging refrigerant after heat exchange,
By cooling the exhaust pipe 14a, the bioreactor 1
The steam generated along the inside is condensed by the heat exchanging means 21 to form water droplets and return to the bioreactor 1 along the inner wall of the exhaust pipe 14a. With this technique, the water consumption of the bioreactor 1 is reduced.

FIG. 3 shows a third embodiment of the apparatus for culturing a cultured object according to the present invention.

The bioreactor 1 according to the third embodiment
, A pair of culture tanks 22 are arranged, a pair of culture tanks 22 are connected via a pair of pumps P in a parallel state, and a timer T is arranged for each pump P. . Then, intermittent communication for periodically sucking the liquid culture medium A stored in one culture tank 22 and transferring it to the other culture tank 22 is performed. That is, the culture medium transfer means 23 is constituted by the pump P and the timer T, and the suction and exhaust means 24 for sucking and discharging air is arranged above each culture tank 22 so that the liquid culture medium A is alternately transferred. The bulb (Bulb) B stored in each culture tank 22 repeats a liquid phase atmosphere and a gas phase atmosphere,
The nutrients and oxygen required for growth are effectively supplied.

FIG. 4 shows a fourth embodiment of the apparatus for culturing a cultured object according to the present invention.

The bioreactor 1 according to the fourth embodiment
Is a pair of culture tanks (25A, 25A,
25B), the lower part of the upper culture tank 25A is connected to the lower culture tank 25B via the siphon pipe 26, and the liquid culture medium A is stored in the upper culture tank 25A, and the bulb is stored. (Bulb) B.

When air is supplied to the upper culture tank 25A, air is supplied from the air supply means 11 to the lower culture tank 25B via the siphon tube 26 and ejected from the sparger S. Will be

The liquid culture medium A in the upper culture tank 25A can flow down to the lower culture tank 25B via the siphon tube 26 by introducing air from the vent 27. At this time, by introducing air from the ventilation port 27 via the sterilizing filter F, contamination of bacteria and the like is prevented. On the other hand, when the liquid culture medium A in the lower culture tank 25B is returned to the upper culture tank 25A, the air is supplied from the air supply means 11 and pushed up via the siphon tube 26. Note that a sterilizing filter F is also provided in the air supply means 11. The movement of the liquid medium A is performed intermittently as necessary.

FIG. 5 shows a fifth embodiment of the apparatus for culturing a body to be cultured according to the present invention.

The bioreactor 1 according to the fifth embodiment
Is rotated up and down by a motor 28 as shown by an arrow so as to uniformly supply oxygen and nutrients to the liquid medium A. A sparger S connected to the air supply means 11 is disposed inside the bioreactor 1 to perform aeration. The air in the bioreactor 1 is discharged by the exhaust means 14 together with carbon dioxide and the like generated as the bulb (Bulb) B grows.

【Example】

[Culture conditions] The spheres and seedling bulbs of Yamayuri were used as the culture target. As a medium, a liquid medium and a solid medium are used.
%, 0.1 ppm NAA (α-naphthaleneacetic acid) was added to the MS medium, and a solid medium was used by adding 0.2% Gelrite to a liquid medium and solidifying it.

FIG. 6 shows the difference in the growth rate (g / l) of the pupa of the lily of the valley when cultured in the liquid medium and the solid medium, between the bulb fresh weight (g) and the bulb fresh weight concentration (g / l). This was compared by examining the amount of increase. In the figure, the horizontal axis indicates the number of days of culture, and the vertical axis indicates the fresh bulb weight (g / bulb) and the fresh bulb weight concentration (g / l). ○, △, 6 in the figure
● is as follows. [Sample 1 (circle)] Medium; liquid medium Volume of culture medium; 50 ml spheres inoculated; 5 pcs [Sample 2 (marked with triangles)] Medium; solid medium Medium volume; 50 ml spheres inoculated; 5 pcs [sample 3 ( ● mark)] Medium; liquid medium Amount of medium: 150ml Inoculated spheres; 10

Examining the fresh bulb weight (g / bulb), the weight of each of Sample 1, Sample 2 and Sample 3 rapidly increased from the time when the culturing days passed 50 days. Further, when the weight increase is at a speed (gradient), sample 3 is the fastest, followed by sample 1 and sample 2.

Examining the bulb fresh weight concentration (g / l), the growth rate (slope) of the culture days from 30 days to 80 days shows that Sample 1 and Sample 3 showed almost the same growth (ratio). (Growth rate) tendency, always exceeding the value of sample 2. The specific growth rate at this time was 0.032 day-1 in the liquid medium and 0.024 day-1 in the solid medium.

That is, it is shown that the use of a liquid medium rather than a solid medium is more suitable for efficient cultivation of spheres in both weight and growth rates than in a solid medium.

FIG. 7 shows the effect of the difference in the concentration of sucrose employed as a main nutrient component on the growth of seedlings when cultivating the seeds of yamali lily in a liquid medium. Raw weight (g / bul
b) shows the measurement results of the bulb fresh weight concentration (g / l) and the conductivity of the medium (electric conductivity; ms / cm). In addition,
For sucrose, sample A is 3%, sample B is 6%
%, Sample C is 9%, and
Regarding the seedling concentration, those in which 10 seedlings were inoculated in 150 ml of a liquid medium were used. The bulb fresh weight concentration (g / l) is indicated by a logarithm as in FIG.

Examining the bulb fresh weight (g / bulb), each sample showed a similar tendency to increase in weight from the start of cultivation, and the growth rate became slightly slow after 60 days. In any case, no significant difference is observed between the samples. At this time, the growth rate was 0.07 day-1.

When the conductivity (ms / cm) of the medium was examined, each sample showed 5 ms / cm to 7 ms / cm at the start of cultivation, but gradually decreased as the number of days of cultivation increased. When the number of days of culture is 100 days, all of them show 3 ms / cm or less. In other words, it is clear that the conductivity (ms / cm) of the medium tends to decrease as the fresh bulb weight (g / bulb) and the fresh bulb concentration (g / l) increase.

FIG. 8 shows the change in conductivity (change amount: ms / cm) and the change in fresh weight concentration (change amount: g / l) when the seedlings were cultured in a liquid medium. Degree change (m
s / cm), and the vertical axis indicates the change in fresh weight concentration (g / l). In addition, the same sample as that shown in FIG. 7 was used.

In any of the samples, it was observed that the change in fresh weight concentration (g / l) increased in direct proportion to the change in the conductivity (ms / cm) of the medium. . Also, in this case, since there is almost no difference in the slope of each sample, it can be said that the effect of the amount of sucrose added to the medium on the relationship between the change in conductivity and the change in fresh weight concentration is extremely small. The following empirical formula was determined from the relationship shown in FIG. Raw weight concentration change = 32.6 × conductivity change (Equation 1) However, the unit is as shown in FIG.

FIG. 9 shows the results of a culture test performed in various bioreactors in order to identify a bioreactor capable of growing bulbs in large quantities and effectively. In this case, a seedling or a bulb is adopted as the bulb,
The maximum bulb inoculation concentration was 67 cells / l. The maximum number of culture days was 63 days.

The highest growth rate among the bioreactors was obtained when the cells were cultured in an air-lift type bioreactor. Next, a gas-liquid exchange type bioreactor and a rotary drum type bioreactor were used. In the case of a bulb, the specific growth rate when seedlings are used is 0.034 day-1 to 0.071 day-1, whereas when a bulb is used, 0.005 day-1 to 0.
It was 028 day-1.

Although not shown in FIG. 9, in the bulb culture employing the bioreactor, the aeration rate is also involved in the growth of the bulb, and the growth of the bulb is hindered when excessive oxygen is supplied. Was admitted. As described above, the reactor type most suitable for bulb culture is an air-lift type, and in this case, it is desirable to supply dissolved oxygen sufficiently, and to adjust the aeration to such an extent that the bulb hardly flows. Is obtained.

According to the method for judging the growth of a cultured object and the culture apparatus according to the present invention, the following effects can be obtained. (1) By culturing a bulb (a body to be cultured) while detecting the electric conductivity in the medium, the growth state of the bulb can be easily grasped. (2) The control unit sets the conditions of the culture medium based on the internal information such as dissolved oxygen, electric conductivity, acidity, liquid level, air volume, etc. during culturing, and the An effective culture environment for growth can be provided and maintained. (3) By measuring the electric conductive substance contained in the medium, the growth status of the bulb can be determined without being limited by the type of the medium. (4) By performing the culture while grasping the growth state of the bulb, the planned production of the bulb can be enabled. (5) By providing an appropriate culture environment while measuring the electric conductive substance contained in the culture medium, adaptation to culture of culture of other plants or animals can be made possible.

[Brief description of the drawings]

FIG. 1 is a front sectional view showing a first embodiment of a method for judging the growth of a cultured object and a culture apparatus according to a first embodiment of the present invention.

FIG. 2 is a front cross-sectional view of a second embodiment of a method for judging the growth of a body to be cultured and a culture apparatus according to the present invention, with a part thereof being omitted.

FIG. 3 is a front cross-sectional view of a third embodiment of a method for judging the growth of a body to be cultured and a culture apparatus according to the present invention, with a part thereof omitted;

FIG. 4 is a front cross-sectional view of a fourth embodiment of a method for judging the growth of a body to be cultured and a culture apparatus according to the present invention, with a part thereof omitted;

FIG. 5 is a front view, partially omitted, showing a fifth embodiment of the method for judging the growth of a cultured object and the culture apparatus according to the present invention.

FIG. 6 is a growth comparison diagram of lily bulbs (balls) cultured in a liquid medium and a solid medium.

FIG. 7 is a diagram showing the relationship between the growth of lily bulbs (seedlings) and the change in the conductivity of the medium when cultured at different sucrose concentrations.

FIG. 8 is a graph showing the relationship between a change in fresh weight concentration of lily bulbs (seedlings) and a change in conductivity of a medium.

FIG. 9 is a relationship diagram showing the results of a culture test of lily bulbs when cultured in different bioreactors.

[Explanation of symbols]

 Reference Signs List A liquid medium B bulb C control unit F sterilization filter P pump RL liquid level S sparger T timer 1 bioreactor 2 jacket 2a supply port 2b outlet 3 electric conductivity controller 3a electric conductivity sensor 4 medium supply means 4A concentration Medium supply system 4B Pure water supply system 5 Heat medium supply means 5a Temperature sensor 6 pH controller 6a pH detection means 7 Acid supply system 8 Alkali supply system 9 Level sensor 10 Liquid level setting means 11 Air supply means 12 Dissolved oxygen detection means 13 Mass flow Controller 14 Exhaust means 14a Exhaust pipe 21 Heat exchange means 21a Refrigerant supply port 21b Refrigerant discharge port 22 Culture tank 23 Medium transfer means 24 Intake / exhaust means 25A Culture tank 25B Culture tank 26 Siphon tube 27 Vent 28 Motor

Claims (17)

[Claims] 1. A method for monitoring the growth of a bulb (B) during culturing, wherein the amount of an electrically conductive substance present in a liquid medium (A) is measured by the electric conductivity, and the state of the decrease in the electric conductivity is measured. And determining whether the liquid medium is effective. 2. The method according to claim 1, wherein the rate of decrease in the electrical conductivity is detected, and when the change in the rate of decrease is small, it is determined that the liquid medium (A) is not effective. A method for judging the growth of a body to be cultured. 3. The method according to claim 1, wherein a concentrated medium is supplied in accordance with a decrease in the electric conductivity in the liquid medium (A), and a degree of recovery of the electric conductivity is determined. A method for determining the growth of a culture. 4. A bioreactor (1) containing a liquid culture medium (A) for culturing a bulb (B), and an electric conductivity sensor connected to the bioreactor and measuring the electric conductivity of the liquid culture medium (1). 3a) and a medium supply means (4) connected to the electric conductivity sensor and supplying a concentrated medium into the bioreactor when the electric conductivity decreases. 5. The cultivation of the cultured body according to claim 4, wherein a jacket (2) for circulating a heat medium to adjust the temperature of the bioreactor is provided outside the bioreactor (1). apparatus. 6. A heating medium supply means (5) for storing a heating medium is connected to the jacket (2), and the heating medium is supplied to the supply port (2).
6. The apparatus for culturing a body to be cultured according to claim 4, wherein the apparatus is supplied into the jacket via a), and is returned from the outlet (2b) after heat exchange. 7. The culture medium according to claim 6, wherein a control unit (C) for keeping the temperature of the stored heat medium constant is added to the heat medium supply means (5). Culture device. 8. The medium supply means (4) has a concentrated medium supply system (4A) for supplying a concentrated medium and a pure water supply system (4B) for supplying pure water. ,
8. The apparatus for culturing a body to be cultured according to 5, 6 or 7. 9. The bioreactor (1) is provided with a pH detecting means (6a) for detecting the pH of a culture medium in the bioreactor, and an acid supply system (7) for supplying an acid and an alkali in a state connected to the pH detecting means. ) And an alkali supply system (8), wherein the apparatus for culturing a body to be cultured according to claim 4, 5, 6, 7 or 8. 10. A level sensor (9) for detecting a liquid level (RL) of a stored liquid medium (A) in a bioreactor (1), and a medium supply means (4) is provided by a detection signal of the level sensor. ) Is driven.
The apparatus for culturing a body to be cultured according to 5, 6, 7, 8 or 9. 11. A liquid level setting means (10) for maintaining a liquid level (RL) constant by overflowing a liquid medium (A) is provided on a side of the bioreactor (1). The apparatus for culturing a body to be cultured according to claim 4, 5, 6, 7, 8, 9, or 10. 12. A sparger (S) for supplying oxygen to the liquid medium (A) by being connected to an air supply means (11) and aerating the liquid medium (A) is provided at the bottom of the bioreactor (1). Claims 4, 5, 6, 7, 8,
12. The apparatus for culturing a body to be cultured according to 9, 10 or 11. 13. The sterilizing filter (F) for sterilizing supplied air is interposed between the air supply means (11) and the sparger (S). An apparatus for culturing a body to be cultured. 14. The bioreactor (1) is provided with a dissolved oxygen detecting means (12) for detecting an amount of dissolved oxygen dissolved in the liquid medium (A), and the dissolved oxygen detecting means sets an air supply amount. It is connected to the controller (13) and performs an adjustment to increase the ventilation rate when the measured value of the dissolved oxygen falls below the set value, and to decrease the ventilation rate when the measured value of the dissolved oxygen exceeds the set value. Claims 4, 5,
The apparatus for culturing a body to be cultured according to 6, 7, 8, 9, 10, 11, 12, or 13. 15. The bioreactor (1) has dissolved oxygen,
A control unit (C) for setting conditions of a culture medium based on each internal information such as electric conductivity, acidity, liquid level, air volume, etc., is provided. 6, 7, 8,
The apparatus for culturing a body to be cultured according to 9, 10, 11, 12, 13, or 14. 16. The bioreactor (1) has a pair of culture tanks (22), between which the liquid medium (A) contained in one of the culture tanks is sucked and the other is sucked. The medium transfer means (23) for transferring to a culture tank is provided, and a suction and exhaust means (24) for sucking and discharging air is provided above the pair of culture tanks.
The apparatus for culturing a body to be cultured according to 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15. 17. The bioreactor (1) has a pair of culture tanks (25A, 25B) vertically separated from each other, and the lower part of the upper culture tank is connected via a siphon pipe (26). A sparger (S) for sending air to the upper part, and an air supply means (11) for sending air to the lower culture tank is provided. 10, 11, 12, 13, 1
17. The apparatus for culturing a body to be cultured according to 4, 15, and 16.