JP2024032920A - Closed 3D fractal culture system and its culture - Google Patents

Abstract

[Problem] Cultivating microorganisms and cells under any gas atmosphere or under any pressure, culturing adherent microorganisms, biofilm-forming microorganisms, complex aerobic and anaerobic microorganism groups, and cells that require contact. To provide a device that can be organized in large quantities and used for any purpose. [Solution] The device is capable of supplying any gas or providing any pressure, and is equipped with a buffer region and a support that can separate microorganisms or cells within the device, so that any microorganisms, cells, Alternatively, it is equipped with a line that efficiently collects those metabolites. [Selection diagram] Figure 1

Description

The present invention is a novel technology that can be applied to the cultivation of aerobic microorganisms, extremophilic microorganisms, complex microorganisms consisting of effective microbial groups forming intestinal flora, complex microorganisms for environmental purification, and the efficient cultivation of special cells. Regarding the culture system.

A wide variety of microorganisms exist in the natural world, and it is said that more than one million types of microorganisms, with billions of microorganisms living in each gram of soil, are present. % or more) are known to be difficult-to-cultivate microorganisms that are difficult to cultivate. In addition, various extremophile microorganisms that have adapted to the environmental conditions inhabit the deep sea, where the temperature and pressure are high, and the cold regions. Under these circumstances, there is a need for technology to search for various microbial groups that can contribute to the preservation of the global environment and life activities.

On the other hand, microorganisms that are relatively easy to culture, such as natto, yeast, and lactic acid bacteria, are being used in various industries, mainly in the food industry (see Non-Patent Document 1). It can be said that the technology is almost established (for example, see Patent Documents 1 to 3).

However, in an animal weighing 60 kg, there are about 1-2 kg of microorganisms, and it is said that the number of microorganisms exceeds the number of somatic cells. For example, in humans, it is said that more than 1,000 types of intestinal bacteria, totaling more than 40 trillion, live in the intestinal tract, which exceeds the 37 trillion human body cells. In order to control more intestinal flora than the cells of a living body, it is necessary to understand the microorganisms in the intestine, but conventional technology focuses on culturing a single microorganism. Therefore, it is difficult to analyze the behavior of complex microorganisms that mimic the intestine, and furthermore, there is no established technology to culture the mimicked intestinal flora itself.

As shown in Non-Patent Documents 2-6, the importance of intestinal flora has been recognized in recent years, and when the balance of the ecosystem is disrupted, so-called dysbiosis can cause various diseases such as inflammatory bowel disease, cancer, and allergies. It is known to lead to the onset of the disease. As for epidemiological findings, for example, the microbial structure in human feces is divided into groups based on the area of residence and the content of the diet, and other factors include diseases, nutritional conditions, indicator molecules of inflammation, and metabolism in the feces. Knowledge such as correlation with analysis data of products etc. has been reported. It does not take into account the control of intestinal flora by type, taking into account the genetic background of the host. In recent years, research data has been reported worldwide showing that both diet and gender influence the composition of the intestinal microbiota, and the diversity of the intestinal microbiota is important for controlling the human metabolome. It has been pointed out that For these reasons, fecal transplantation has been clinically applied as a therapeutic method of transplanting feces from healthy humans.

Furthermore, bacteriophages are known to be involved in killing pathogenic bacteria, and can also contribute to the maintenance of intestinal flora homeostasis in the intestines (see Non-Patent Document 7). Therefore, there are expectations for the application of phage medicine, which utilizes bacteriophages that parasitize specific pathogenic bacteria, but the technology for cultivating and maintaining large quantities of phages has not been established.

As biology progresses, the scheme of cell tissue formation is becoming clearer. Functional differentiation progresses from a single fertilized egg through the stages of differentiation into undifferentiated stem cells, endoderm, mesoderm, and ectoderm. Furthermore, as regenerative medicine progresses, it is necessary to utilize these mechanisms to understand various life activities.

In particular, in the process of functional differentiation of cells, there are environmental factors such as intercellular adhesion and oxygen concentration gradients that determine the structure of cell populations, and compared to conventional two-dimensional culture and single-phase culture, three-dimensional culture The idea that this is more physiological is gaining acceptance. The importance of multicellular aggregate structures called spheroids and the importance of model experiments using organoids formed by different cell groups are being understood, but there is no way to obtain them in large quantities semi-automatically, and how to use them themselves. There is no established method to use it. Currently, it is difficult to efficiently culture and maintain target cells and tissues in large quantities.

Japanese Patent Application Publication No. 2001-120252 Japanese Patent Application Publication No. 2002-85051 Japanese Unexamined Patent Publication No. 10-191965

Keishi Horiuchi, "Development of yogurt deoxygenation fermentation technology", Journal of the Society of Bioengineering, Japan Society of Bioengineering, 2010, Vol. 88, No. 11, p. 594-600 Hirokuni Miyamoto et al., “Production and academic elucidation of highly functional fermented products from unused biomass resources using thermophilic microorganisms,” Journal of the Society of Bioengineering, Japan Society of Bioengineering, 2018, Volume 96, No. 2 , p.56-63 Shinji Fukuda, "Understanding and controlling the intestinal ecosystem through metabologenomics", Biochemistry, Japanese Biochemical Society, 2016, Vol. 88, No. 1, p.61-70 Marcus J. Claesson, et al., “Gut microbiota composition correlates with diet and health in the elderly”, Nature, 2012, Vol.488, No.7410, p.178-184 Daniel I. Bolnick, et al., “Individual diet has sex-dependent effects on vertebrate gut microbiota”, Nature Communications, 2014, Vol.5, No.4500 Emmanuelle Le Chatelier, et al., “Richness of human gut microbiome correlates with metabolic markers”, Nature, 2013, Vol.500, No.7464, p.541-549 Jeremy J. Barr, et al., “Bacteriophage adhering to mucus provide a non-host-derived immunity”, Proceedings of the National Academy of Sciences of the United States of America, 2013, Vol.110, No.26, 10771- 10776

As mentioned above, when culturing microorganisms in large quantities, it has been carried out using liquid media. Therefore, there is a possibility that microorganisms that can be cultured only on solid media cannot be cultured efficiently. In addition, since there is no support in the liquid medium to which adhesion molecules on the cell surface of microorganisms can adhere, it is difficult to cultivate cells that take into account the characteristics of adhesion molecules derived from microorganisms. Furthermore, it is not necessarily suitable for culturing microorganisms that can contribute to the formation of intestinal flora, where both anaerobic and aerobic microorganisms may exist.

The same can be said for cell culture. In other words, it is necessary to consider efficient culture conditions when culturing stem cells that require feeder cells, undifferentiated cells in the process of tissue formation, single cells, or organoids. There was no mechanical way to do that.

Furthermore, there has been no attempt to use the culture system itself to purify the environment or apply it to medical equipment itself.

The present invention has been made in view of the above circumstances, and aims to cultivate microorganisms and cells in any gas atmosphere or under any pressure, and to cultivate sessile microorganisms, biofilm-forming microorganisms, aerobic and anaerobic microorganisms. The purpose of the present invention is to provide a device that organizes a complex group of microorganisms and cells that require contact, and can be used in large quantities for any purpose.

In order to solve the above problems, the present invention is a device capable of supplying any gas or providing any pressure, and also includes a buffer region and a support that can separate microorganisms or cells in the device. The system is characterized by being equipped with a line for efficiently collecting any microorganisms, cells, or their metabolites.

Preferably, the liquid medium is characterized by containing a porous carrier or support.

Preferably, the porous carrier is characterized by containing thermoplasticity, an organic material having the property of being slowly decomposed in nature, or a biodegradable plastic. Preferably, agar, agar, gelatin, cellulose mixed ester, etc. are used as the support.

Related microorganisms include useful microorganisms that form intestinal flora, environmental purification microorganisms that do not function alone, complex microorganisms/biofilms that can contribute to wastewater treatment, and difficult microorganisms that can only grow in special environments. Cultivable microorganisms and extremophiles include special groups of microorganisms that live in the deep sea, such as thermophiles, psychrophiles, halophiles, alkaliphiles, acidophiles, hyperbaric bacteria, and organic solvent-resistant bacteria. It will be done.

Furthermore, as the cells and tissues, preferably single cells, organoids, and a mixed culture system of feeder cells and undifferentiated cells can be selected. In addition, culture vessels or supports can be selected in order to repeat stimulation multiple times at the differentiation stages of not only mesenchymal stem cells, which are somatic stem cells of the mesodermal lineage, but also endodermal and ectodermal lineages. do.

These allow data collection as a model system. Furthermore, it is expected that it will be able to accumulate itself as a database, compare it with reaction information in existing living organisms, and utilize it as a medical device that has the potential to serve as a model artificial organ.

The culture system according to the present invention allows each target organism after culture to adapt to the environment in which each organism originally inhabits, and to be physiologically self-similar, that is, by setting culture conditions. The purpose is to be able to culture in a fractal manner. For this purpose, for example, it is possible to culture living organisms coexisting with a porous carrier under any gas atmosphere, and it is also possible to culture them in a form that is adhered to the support or on a solid state. In addition, by enabling the exchange of physiologically active substances such as hormones, efficient mass culture is possible. In addition, by connecting culture tanks that can set multiple types of culture conditions such as temperature, pH, pressure, and gas atmosphere, it is possible to culture different microorganisms or cells. A feature of this method is that it is possible to separate different microorganisms or cells by adjusting properties such as pore size of the support. Furthermore, cell tissues and organoids are expected to be used not only as model systems for physiological reactions, but also as medical devices such as artificial organs.

A conceptual diagram regarding the present invention. A conceptual diagram showing a continuous petri dish culture device. A conceptual diagram showing a continuous culture device utilizing a porous carrier. A conceptual diagram showing a continuous culture device utilizing a membrane module. Conceptual diagram showing the role of the support in the culture system. FIG. 2 is a conceptual diagram showing the connection mode of the culture system. The conceptual diagram which shows the connection style of the culture system outside the culture tank. FIG. 2 is a conceptual diagram showing the information processing hierarchical structure of the control system. Conceptual diagram showing usage for soil improvement. Conceptual diagram showing the uses of wastewater treatment. A conceptual diagram showing the uses within the wastewater treatment facility. Conceptual diagram showing the use of intestinal flora cocktail for transplantation. Conceptual diagram showing the uses of medical bacteriophage. Conceptual diagram of general cell differentiation and tissue formation. Conceptual diagram of gastrointestinal and lung tissue formation by common endoderm-derived progenitor cells.

An embodiment of the present invention will be described below with reference to the drawings. However, the following is merely an illustrative example of an embodiment of the present invention, and the scope of the present invention is not limited to the following embodiment, and may be modified as appropriate without departing from the spirit of the present invention. It is.

The culture system according to this embodiment efficiently cultivates the target organism under any gas atmosphere, under any pressure, or under any temperature, depending on the characteristics of the support, and Can be maintained stably. In the culture system according to this embodiment, by using a highly pressure-resistant material such as pressure-resistant glass for the culture device, it is possible to supply any gas or provide any pressure. The pressure can be varied from 30 MPa to 60 MPa, equivalent to the pressure at depths of 3000 m to 6000 m, up to approximately 100 MPa at the maximum. In addition, the culture system according to the present embodiment utilizes hollow fiber membranes, artificial cell membranes, carbon nanotubes, etc. in the device, and also dissolves gas components in the hydrosphere and creates a concentration difference to eliminate microorganisms or cells. It includes a separable buffer area as well as a support. The culture system according to this embodiment is further equipped with a line for efficiently collecting any microorganisms, cells, or their metabolites, and in a region on the collection side separated by the support, a culture medium solution, a collection Use sterile water or sterile saline, flow them into the device through a line, and collect them. Therefore, as shown in FIG. 1, the culture device according to this embodiment can be used for probiotics, prebiotics, and transplants, which have been difficult to culture and maintain with conventional devices. , producing or maintaining a complex cocktail of effective microorganisms in the intestine, bacteriophages that are involved in killing pathogenic bacteria, biofilms that can contribute to environmental purification, and extremophiles that can live under high temperature, high pressure, or low temperature. can be done. Further, the device itself can be expected to be utilized as an environmental purification system. In cell culture, unlike traditional culture devices, gas exchange and support properties can be adapted to mimic the physiological conditions appropriate for individual cells and tissues, making it possible to culture tissues such as organoids. It is expected that they will be implemented stably and in large quantities, and that they will be used to construct an evaluation system as a model system, or to produce transplanted tissues based on the data. Furthermore, the device itself can be expected to be used as a medical device that imitates some of the biological reactions.

As the material for the culture device according to this embodiment, any one of heat-resistant/pressure-resistant glass, stainless steel, and heat-resistant polycarbonate is suitable.

Next, the present invention will be further explained by showing design examples, but the present invention is not limited thereto.

(Example 1) FIG. 2 shows a culture apparatus 10 used in this example. The culture device 10 includes a culture tank 11 made of stainless steel, a support 12 , a recovery solution feeding pipe 13 for recovering the culture, and a stirring blade 14 for stirring the inside of the culture tank 11 . A microbial fermentation solution containing coffee grounds, which is a porous material sterilized in an autoclave, is sealed in a culture tank 11 as a support 12, and a spore-forming bacillus, Bacillus coagulans, is prepared at a concentration of 10 ⁶ per ml. was added, cultured in an oxygen atmosphere for 2 days in a liquid culture, maintained at a heating temperature of 60°C using a planar heater, and adhered to a porous body of 10 ⁸ or more per ml using sterilized yeast extract as a raw material. It became possible to collect a mixed bacterial culture of Bacillus coagulans. The sterilized microbial fermentation solution used was deposit number PTA-1773, which was internationally deposited with the ATCC. Furthermore, as Bacillus coagulans, the deposit number BP-02066 at the National Institute of Technology and Evaluation deposited by Miyamoto, one of the inventors of the present invention, was used on June 17, 2015. Using a sterilized solution containing dead bacteria called PTA-1773, it is possible to culture the target complex microorganism as a complex microorganism while controlling the culture of other microorganisms by utilizing the quorum sensing function. did it.

(Example 2) FIG. 3 shows a culture apparatus 20 used in this example. The culture device 20 includes a culture tank 21, a stainless steel plate 22 for a solid medium, a liquid supply pipe 23 for a recovery solution for recovering the culture, and an agitation blade 24 for stirring the medium solution and the like. Using a tank that has only a stainless steel dish 22, only one stirring blade 24 at the bottom, and no liquid feed pipe 23, an agar solution that has been heat sterilized in an autoclave is placed on the surface of the stainless steel dish 22 as a support. I put it in. On the surface of the stainless steel plate 22, agar as a support is solidified at room temperature, and then Bacillus bacteria, which is a spore bacteria adjusted to 10 ⁶ per ml, is mixed with cold water, and then the solidified agar is mixed with cold water. 10 ⁸ or more of the bacteria per ml can be obtained by adding the bacteria to the water, collecting the cold water containing the bacteria, setting the heating temperature to 50°C or higher, and then introducing air through a sterile filter for aerobic culture. It became possible to culture in 24 hours. In order to collect the bacterial cells, it is possible to obtain concentrated bacterial cells by pouring cold water and centrifuging the cold water containing the bacterial cells. In addition, in order to remove agar, it is possible to recover it by circulating sterilized water at 70° C. or higher. In addition, in order to equalize the conditions on the stainless steel dishes 22, stirring blades 24 are installed on each stainless steel dish 22, and in order to adjust and equalize the amount of bacterial cells input, a structure having a liquid feeding pipe 23 is provided. It is also possible to have

The support according to this embodiment may be a dish having a large number of depressions smaller than the size of the microorganism and having a diameter and depth between 0.22 μm and 10 μm, or a temperature-dependent support as shown in Example 2. Agar, agar, and gelatin, which have variable solidification and liquefaction properties, are suitable as thermoplastic materials that can change their shape in the following applications: That is, it is convenient because the target organism can be attached to the surface under solidified conditions and cultured in any gas atmosphere, and the culture medium can be mixed. In addition, when culturing using both sides of the support, as shown in Figure 4, materials that can be selected by pore size include hollow fiber membranes, carbon nanotubes, and cellulose mixed esters with filter pores. etc. are appropriate. This makes it possible to exchange useful microorganisms and metabolites with different pore sizes between culture system X and culture system Y. Further, if a biological membrane, an artificial biological membrane, or a material having such characteristics is included in the support, arbitrary gas exchange is possible by osmotic pressure, gas partial pressure, etc.

(Example 3) By applying the above-mentioned concept and constructing the culture apparatus 30 shown in FIG. It is also possible to culture complex microorganisms whose cells can only be cultured in liquid. The culture device 30 includes a culture tank 31 and a tube 32 as a support. Specifically, the pH is adjusted, or the tip 36 of the tube 32 is sealed, and a small number of pores are provided in the tube 32 with a fine pore size (0.22 μm or less in diameter) and an interval of 1 mm or more between the pores. The inside of the tube 32 is filled with bacterial cells that can be cultured under arbitrary gas conditions, the gas is forcibly introduced into the tube 32, the outside 37 of the tube 32 is filled with a highly viscous liquid medium, and a trace amount of the gas component is However, it is expected that it will be possible to cultivate bacterial cells that are compatible with the conditions in which they are mixed into a highly viscous liquid medium, and to simultaneously maintain and manage multiple types of bacterial cells. Using the culture device 30, it was possible to culture a culture of NP-1 strain, which is a closely related species of Bacillus subtilis, and Bacillus coagulans BP-02066, with the former strain being preferential. The results are shown in Table 1.

These microorganism groups include useful microorganisms that form intestinal flora, environmental purification microorganisms that do not function as a single group, complex microorganisms/biofilms that can contribute to wastewater treatment, and difficult-to-cultivate microorganisms that can only grow in special environments. Thermophilic bacteria, psychrophilic bacteria, halophilic bacteria, alkaliphilic bacteria, acidophilic bacteria, hyperbaric bacteria, and organic solvent-resistant bacteria are expected to be active microorganisms and extremophilic microorganisms. In addition, when biodegradable plastic is used as a porous material, by culturing microorganisms that can decompose L-type lactic acid or succinic acid, which are the raw materials, biodegradable plastic or temperature-dependent shape change can be achieved. It is possible to efficiently produce co-cultures of microorganisms with agar, agar, gelatin, etc., which are capable of oxidation, and is expected to create materials that can be used in many fields such as agriculture, livestock, and environmental purification.

In addition, by adjusting the pore size of the tube 32 and setting it to a pore size (0.22 μm or less in diameter) that does not allow bacteria to pass through, it is possible to prevent bacteria from entering the tube 32 on one side and cells or cells on the other side. By culturing organoids, bacteria or cells/organoids can be cultured in a symbiotic system of bacteria, cells, and organoids, with material circulation through the pores of the tube 32, and in a system similar to reactions in living organisms. This culture system can also be used as a model system, and is effective in understanding the formation of bacteria or tissues/organoids that can only be cultured in vivo. It is also expected that once this data is accumulated, it will be possible to apply it to artificial organs.

Furthermore, for the purpose of isolating bacteriophages that can infect only a given pathogen, in order to propagate them, the pathogen is introduced into one area, and a medium in which they can proliferate is introduced, and while culturing, the pore size is Due to the sieving effect, it is possible to efficiently separate and collect only bacteriophages without collecting pathogenic bacteria.

If the application is to culture and maintain arbitrary cells/tissues, feeder cells and stem cells may be co-cultivated in a physically separated form within the tube 32 or in the external area 37 to provide an efficient feeder. By utilizing cell culture and their metabolites, it is possible to culture stem cells, organize stem cells, or culture organoids of target tissues.

These so-called separated co-culture systems of feeder cells and stem cells, as well as co-culture systems of feeder cells and organoids, etc., can be used as experimental models in place of animal experiments. Furthermore, by maintaining the culture, the metabolites can be obtained efficiently, and by accumulating experimental data, it is expected that it will be used as a model system for replacing artificial organs or as part of medical equipment.

In addition, as shown in Figure 6, by completely dividing the culture tank and semi-separating it using a filter or support, various conditions such as temperature, gas, and pressure of the culture tank can be dramatically changed. It is also possible to construct a co-culture system and collect each culture while avoiding contamination.

In order to construct these culture systems as a system, equipment outside the culture tank shown in FIG. 7 is required, and as a control system, it is preferable to construct the control system shown in FIG. 8.

7 and 8 illustrate a control system used in the culture system according to this embodiment. As shown in FIG. 7, in the first control 110, a liquid culture medium can be supplied. By adding agar to the culture medium, it is supplied in liquid form at a temperature of 60°C or higher, and then solidified after lowering the temperature to 60°C or lower in a tank. It is possible to supply living organisms such as necessary microorganisms to the plant. It is also possible to use the line in conjunction with gas sharing. Regarding gas exchange, mainly by utilizing the second control 120, it is possible to supply or discharge any gas other than air, such as oxygen, carbon dioxide, nitrogen, hydrogen sulfide, etc., by changing the concentration gradient. The third control 130 is a recovery line and allows input and discharge of recovery liquid. Regarding the recovery liquid, when dissolving the agar, add sterilized water or sterile physiological saline at a temperature of 60°C or higher, and when recovering without dissolving the agar, add 30°C or lower, preferably 4-10°C. Add sterile water or sterile physiological saline at a temperature range of In the culture device connection line, the flow rate of the liquid supply line is controlled by considering the relationship with the microorganisms or cells to be cultured in a complex manner. The fourth control 140 also controls the temperature of the target culture tank or the stirring conditions in the culture tank in the fifth control 150, and also controls the first control 110, second control 120, and third control. 130 and fourth control 140.

The control system as shown above is summarized as an information processing base 100, as shown in FIG. The information processing base 100 can comprehensively control the control system factors described from the first control 110 to the fifth control 150 based on the accumulated culture condition big data. In particular, it is preferable that the big data processing of the information processing base 100 be performed under conditions that can be optimized by machine learning and artificial intelligence. Therefore, the information processing base 100 is equipped with an external terminal input such as a LAN cable terminal, Wi-Fi, or USB terminal that allows communication over the Internet, and is provided with a function that allows input of the latest culture conditions for microorganisms, cells, etc. It is preferable that more efficient culture is possible by this method.

Regarding the control flow within the information processing base 100, as shown in FIG. A signal is issued, and the control is also linked with the third control 130 (S13). Next, in the second control 120, a gas supply switch is received (S21), the gas supply concentration and frequency are checked (S22), and adjustment is made possible to receive a stop signal (S23). The third control 130 is set so that OD or pH can be used as an index to check the degree of cultivation, and upon completion of the cultivation (S31), the collected liquid can be introduced (S32). In addition, the speed of collection is adjusted in consideration of the fragility of the culture (S33), and the collection can be stopped when the collection is completed (S34). The fourth control 140 is interlocked with the second control 120 (S41), with the third control 130 (S42), with another connected culture device (S43), and with the flow rate and flow rate. Adjust. Further, the fifth control 150 is linked with the first control 110 to the fourth control 140 (S51-S54), and enables smooth cultivation and recovery in the culture device (S55).

As shown in Figure 9, when a mixed culture of porous biodegradable plastic and microorganisms is applied to farmland, when lactic acid or succinic acid is used as the composition of the biodegradable plastic, microorganisms The decomposition of minerals can slowly release acids in the soil, which can be beneficial to crops whose mineral uptake is enabled by acidification of the local pH of the soil.

As shown in Figure 10, when a mixed culture of porous biodegradable plastic or other degradable carriers and wastewater purification biofilm is applied upstream of wastewater, microorganisms that contribute to wastewater purification are As the specific surface area increases, it is expected that the decomposition efficiency will increase and downstream purification will progress. Similarly, as shown in FIG. 11, if used in an aeration tank in a wastewater treatment facility, it is expected that the decomposition efficiency will similarly increase and the volume of sludge will be reduced.

As shown in Figure 12, if a complex microorganism formed by effective microorganisms of intestinal flora is used as a cocktail, it can be used as an alternative technology such as the current fecal transplantation, as a medical material for humans, or as a material for animal intestinal flora. It can be used as an intestinal flora cocktail to be transplanted into the immature intestine shortly after birth to improve the condition.

As shown in FIG. 13, by culturing bacteriophages that selectively kill pathogenic bacteria in the intestines, they can be used as a safe alternative to antibiotics. Reducing the use of artificial antibiotics is an urgent global issue, so it can be expected to be a safe prescription.

With the progress of regenerative medicine, many technological developments are progressing, but there is still room for improvement in the construction of systems that automate cell organization. FIG. 14 shows general cell differentiation, but as shown in FIG. 15, the form of final differentiation changes depending on the environmental stimulation factors given to each cell. By utilizing this device, we will modify these environmental stimuli, construct target tissues and organoids, and proceed with the production of organoid model systems, transplanted tissues, and use as medical devices.

As described above, by utilizing the culture system according to the present invention, it can be used in a wide range of fields such as microbial culture, soil improvement, crop production, animal production, environmental purification, and medical fields.

10, 20, 30 Culture device 11, 21, 31 Culture tank 12 Support (porous body)
13 Liquid feeding tube 14 Stirring blade 22 Stainless steel plate 23 Liquid feeding tube 24 Stirring blade 32 Tube (support body)
36 Tip (of the tube) 37 Outside (of the tube) 100 Information processing base 110 First control (control related to the culture medium supply line)
120 Second control (control related to gas exchange line)
130 Third control (control related to collection line)
140 4th control (control related to culture device connection line)
150 Fifth control (control related to culture tank control line)

Claims (5)

A system for efficiently cultivating organisms in large quantities through the surface of a support under any gas atmosphere, or under any pressure or temperature, as well as recovery of cultures derived from the organisms concerned. A culture system including a device that enables
two culture tanks for culturing the organisms;
a stirring blade that stirs the inside of the culture tank;
a medium supply line connected to the culture tank and supplying the medium into the culture tank;
a culture device connection line that connects the two culture tanks and controls the flow rate and flow rate of the liquid sent between the two culture tanks;
the support provided in the culture device connection line;
A recovery solution recovery line and a recovery solution input line connected to the culture tank and for recovering the culture;
an air supply line and an exhaust line that are connected to the culture tank and perform gas exchange within the culture tank;
and an information processing base that comprehensively controls control system factors based on accumulated culture condition big data,
The information processing base includes a first control unit that controls supply of the medium in the culture tank;
a second control for controlling gas exchange in the culture tank;
a third control for controlling input and discharge of the recovery solution in the culture tank;
a fourth control that performs interlocking of the second control and the third control and control of the culture device connection line;
A fifth control that performs interlocking of the first control, the second control, the third control, and the fourth control, and controls the stirring conditions of the culture tank and the temperature of the culture tank. A culture system featuring: 2. The culture system according to claim 1, further comprising an apparatus for continuously culturing anaerobic organisms in a liquid medium in order to culture and maintain a complex group of aerobic organisms and anaerobic organisms. 3. The culture system according to claim 1, wherein a living organism that promotes the growth of the organism to be cultured is used as the support. 4. The culture system according to claim 1, wherein a porous carrier with a large surface area or a tube-shaped filtration membrane is used as the support. 4. The culture system according to claim 1, wherein the support or its side surfaces are in a highly viscous liquid state, so that an air-liquid interface can be used for culturing organisms.

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