WO2017013974A1

WO2017013974A1 - Stirring device

Info

Publication number: WO2017013974A1
Application number: PCT/JP2016/067939
Authority: WO
Inventors: 惟杉田; 勝難波; 良一芳賀; 拓司安藤
Original assignee: 株式会社日立製作所
Priority date: 2015-07-22
Filing date: 2016-06-16
Publication date: 2017-01-26
Also published as: JP6764221B2; JP2017023053A

Abstract

Provided is a stirring device that is capable of efficiently generating a vertical circulating flow, even at a low stirring speed, in a liquid enclosed in a flexible bag. The stirring device (100) comprises a flexible bag (1)which has flexibility and in which a liquid (7) can be enclosed, a stirrer (2) which is disposed within the flexible bag (1) and generates a swirling flow in the liquid (7), and a flow-blocking part (5) which is suspended from the top of the flexible bag (1) in such a manner as to block the swirling flow of the liquid (7), wherein the flow-blocking part (5) is disposed in such a manner that the bottom end thereof is positioned below the surface of the liquid (7) and above the bottom part of the flexible bag (1).

Description

Stirrer

The present invention relates to a stirring device provided with a flexible bag for culture.

In fields such as the biochemical industry and the pharmaceutical industry, an agitation tank is often used for culturing floating cells and producing useful substances using the cells while agitating the culture solution. The stirring tank is usually provided with a stirring bar and filled with a culture solution together with various cells. By rotating the stirrer while aerating air or oxygen to the culture solution, a horizontal swirling flow is generated in the culture solution, so that nutrients, oxygen and the like are efficiently supplied to the cells.

The cell growth efficiency and the productivity of useful substances tend to be greatly influenced by the supply efficiency of nutrients and oxygen to the cells, that is, the stirring state of the culture solution. Therefore, a baffle (baffle plate) is generally provided inside the stirring tank. The baffle is usually suspended in the stirring tank so as to block a part of the swirling flow generated by the rotating motion of the stirring bar. A part of the swirl flow is blocked by the baffle to be converted into a vertical circulation flow, and a vertical flow is generated in the culture solution, thereby increasing the stirring efficiency of the culture solution.

By the way, in recent years, a single-use flexible bag has been used as an agitation tank in order to reduce the man-hours and costs associated with washing and sterilization of the agitation tank, and to avoid contamination of the culture medium due to contamination with foreign substances. It has come to be. Since the flexible bag is formed of a resin material having flexibility, at the time of use, the flexible bag is supported by a rigid support member to hold the container shape. Since the flexible bag has poor rigidity and does not include a mechanism for sterilizing the inside, the installation method of the stirrer is restricted.

As a stirrer that stirs a culture solution in a flexible bag, a configuration in which a magnetically driven stirrer (magnet stirrer) is installed at the bottom of the flexible bag or a motor shaft-coupled stirrer is flexible A form to be inserted into a bag is known. Unlike the motor shaft connection type, the magnetic force drive type has the advantage that it is not necessary to provide a high-cost shaft seal mechanism at the connecting portion between the flexible bag and the shaft. On the other hand, according to the motor shaft connection type, unlike the magnetic drive type, the position of the stirrer is not limited to the bottom of the flexible bag, so it is suitable even when the culture medium is deep. Used.

In a stirrer that stirs a culture solution in a flexible bag, it is desired to install a baffle from the viewpoint of increasing the stirring efficiency of the culture solution. For example, U.S. Patent No. 6,057,051 discloses a stirred tank reactor system comprising a flexible bag, a shaft, an impeller and a bearing. Patent Document 1 describes that a housing is provided outside the flexible bag, and the flexible bag is folded around a plurality of baffles provided in the housing (see paragraph 0022 and the like).

Special table 2007-534335 gazette

However, with the stirring device described in Patent Document 1, it is difficult to efficiently circulate the culture solution in the vertical direction inside the flexible bag. Even if the flexible bag is shaped by the baffle provided outside the flexible bag, the flow velocity of the swirling flow due to the rotational movement of the stirrer is not sufficiently maintained in the vicinity of the inner surface of the wall of the flexible bag. is there. That is, in the stirring apparatus described in Patent Document 1, although the baffle is provided, the vertical circulation flow of the culture solution cannot be efficiently generated, and a good stirring efficiency is achieved. Not suitable for.

In addition, since the flexible bag does not have sufficient rigidity, in a stirrer that stirs the culture solution in the flexible bag, attention must be paid to the installation position and installation method of the baffle. In addition, in recent years, there has been an increasing demand for a stirring device that supports the cultivation of animal cells and the like. When culturing large cells such as animal cells, it is necessary to slow the stirring speed from the viewpoint of preventing the cells from being damaged by stirring. Therefore, in such a case, it is required that good stirring efficiency is realized at a low stirring speed. Moreover, in a stirrer that stirs a culture solution in a flexible bag, a magnetically driven stirrer is often used. In the magnetic drive type configuration, since the installation position of the stirrer is restricted by the bottom of the flexible bag, it is desired to generate the vertical circulation flow more efficiently.

Therefore, an object of the present invention is to provide a stirring device capable of efficiently generating a vertical circulation flow in a liquid sealed in a flexible bag even at a low stirring speed.

In order to solve the above problems, a stirrer according to the present invention has flexibility, a flexible bag that can enclose a liquid therein, and is disposed inside the flexible bag and swirls around the liquid. A stirrer that generates a flow and a flow blocking unit that is suspended from the top of the flexible bag and that blocks the swirling flow of the liquid, and the lower end of the flow blocking unit is the liquid liquid It is provided so that it may be located below the surface and above the bottom of the flexible bag.

According to the present invention, it is possible to provide a stirring device capable of efficiently generating a vertical circulation flow in a liquid sealed in a flexible bag even at a low stirring speed. Specifically, since the stirring efficiency can be improved even at a low stirring speed of about 100 rpm, it is possible to prevent the cells from being damaged by the stirring. In addition, it is possible to realize good stirring efficiency without being strongly influenced by the position of the stirring bar installed in the stirring tank.

It is sectional drawing which shows the whole structure of the stirring apparatus which concerns on 1st Embodiment of this invention. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 1st Embodiment of this invention. It is the II sectional view taken on the line in FIG. 2A. It is sectional drawing which shows the fixing method of a baffle. It is a perspective view which shows the effect | action of a baffle. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 2nd Embodiment of this invention. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 3rd Embodiment of this invention. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 4th Embodiment of this invention. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 5th Embodiment of this invention. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 6th Embodiment of this invention. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 7th Embodiment of this invention. It is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 8th Embodiment of this invention. It is a perspective view of the baffle with which the stirring apparatus which concerns on 8th Embodiment of this invention is equipped. It is a figure which shows the result of having analyzed the relationship between the distance from the wall part inner surface of a flexible bag to a baffle, and the flow volume integral value of an up-and-down circulating flow. It is a figure which shows the relationship between the height from the bottom inner surface of a flexible bag to the lower end of a baffle, and the flow volume integrated value of an up-and-down circulating flow. It is a figure which shows the effect of the flow volume integrated value of the up-and-down circulation flow in the stirring apparatus which concerns on the Example of this invention. It is a figure which shows the relationship between the installation number of a baffle, and the flow-rate volume integral value of an up-and-down circulation flow. It is sectional drawing which shows the principal part structure of the stirring apparatus which installed the rectangular flexible bag. It is the II-II sectional view taken on the line in FIG. 15A. It is a figure which shows the relationship between the height from the bottom part inner surface of a flexible bag in a rectangular type flexible bag to the lower end of a baffle, and the flow volume integrated value of an up-and-down circulating flow.

[First Embodiment]
First, the stirring device according to the first embodiment of the present invention will be described. In addition, about the structure which is common in each following figure, the same code | symbol is attached | subjected and the overlapping description is abbreviate | omitted.

FIG. 1 is a cross-sectional view showing an overall configuration of a stirring device according to a first embodiment of the present invention.
As shown in FIG. 1, the stirring device 100 according to the first embodiment includes a flexible bag 1, a stirring blade (stirring bar) 2, a measuring device 3, a control device 4, and a baffle (flow blocking unit). 5, top plate 6, port 8, shaft 9, stirring motor 10, gas control device 11, sparger 12, temperature control heater 13, support member 14, pedestal 15, and submerged aeration Gas supply pipe 16, gas-phase aeration gas supply pipe 17, culture medium discharge pipe 18, exhaust pipe 19, exhaust filter 20, gas control valve 21, sealing member 22, sensor 23, baffle A support (internal support) 24 and a support member 25 are provided.

The flexible bag 1 is made of a synthetic resin and is a substantially transparent container having flexibility. The flexible bag 1 is folded and flattened before use. On the other hand, as shown in FIG. 1, a predetermined container shape is obtained by enclosing a culture solution (liquid) 7 or gas inside during use. Specifically, the cross-sectional shape of the flexible bag 1 according to the present embodiment is a substantially circular shape, and is a container shape having a substantially cylindrical body portion and a bottom portion.

The circumference and bottom of the flexible bag 1 are supported by a rigid support member 14. The support member 14 is provided in a box shape having a bottom portion and a wall portion, and is fixed to the gantry 15. The flexible bag 1 is placed on the bottom of the support member 14, and stands upright while maintaining its container shape by being supported by the wall portion of the support member 14.

The flexible bag 1 is provided with a plurality of pipe portions that communicate between the inside and the outside of the flexible bag 1, and various pipes are connected to these pipe portions. For example, as shown in FIG. 1, the submerged gas supply pipe 16 and the gas-phase gas supply pipe 17 are connected to a pipe portion provided in the flexible bag 1. A sparger 12 for generating fine bubbles is attached to the tip of the gas supply pipe 16 for in-liquid ventilation, and the sparger 12 is arranged in the liquid phase on the bottom side in the flexible bag 1. The tip of the gas supply pipe 17 for gas-phase ventilation is arranged in the gas phase on the top side in the flexible bag 1. On the other hand, a gas regulator 11 is connected to the proximal end side of the gas supply pipe 16 for submerged gas supply and the gas supply pipe 17 for gas-phase ventilation.

The submerged aeration gas supply pipe 16 and the gas phase aeration gas supply pipe 17 are each a culture solution (in the figure, in which various gases such as air, oxygen, nitrogen, carbon dioxide, etc. are sealed in the flexible bag 1. And may be used for aseptically supplying the gas phase on the culture medium. The gas control device 11 is connected to the control device 4 through a signal line as shown by a broken line in FIG. 1, and the gas control device 11 supplies gas through the liquid supply gas supply pipe 16 and the gas supply gas supply pipe 17 for gas phase ventilation. The supply amount is adjusted by the gas adjusting device 11 under the control of the control device 4.

The flexible bag 1 has an exhaust pipe 19 at the top and a culture medium discharge pipe 18 at the bottom. The tip of the exhaust pipe 19 is disposed in the gas phase in the flexible bag 1. On the other hand, the base end of the exhaust pipe 19 is connected to the gas control valve 21 via the exhaust filter 20. As shown by a broken line in FIG. 1, the gas regulating valve 21 is connected to the control device 4 through a signal line, and the opening and closing of the gas regulating valve 21 is controlled by the control device 4. The gas from which the splashes have been removed by the exhaust filter 20 is exhausted through the gas control valve 21 so that the internal pressure of the flexible bag 1 is maintained. Moreover, the culture solution discharge pipe 18 is connected to the flexible bag 1 through a valve so that the culture solution can be discharged to the outside of the flexible bag 1.

The flexible bag 1 is provided with a pipe for supplying a culture solution, a pipe for seeding cells, and a culture in the same manner as the gas supply pipe 16 for in-liquid aeration and the gas supply pipe 17 for a gas-phase aeration. Pipes (not shown) such as a pipe for sampling the liquid, a pipe for supplying the pH adjusting agent, and a pipe for exchanging the culture solution are connected.

Various pipes such as the gas supply pipe 16 for submerged aeration gas and the gas supply pipe 17 for gas-phase aeration, and the pipe portion provided in the flexible bag 1 are made of silicone rubber or other synthetic resin. And has flexibility. These pipes are disposable parts that are used after being sterilized in advance and are discarded after being used once. Moreover, the pipe part provided in the flexible bag 1 has the pipe end sealed before use, and various pipes and pipe parts are aseptically connected during use.

The temperature adjusting heater 13 is provided to heat the culture solution sealed in the flexible bag 1 and is installed around the lower portion of the flexible bag 1. The temperature adjusting heater 13 is connected to the control device 4 through a signal line as indicated by a broken line in FIG. 1 so that the heating of the culture solution by the temperature adjusting heater 13 is controlled by the control device 4. It has become.

The sensor 23 is inserted into the port 8 that communicates the flexible bag 1 and the support member 14. Although one sensor 23 is installed in FIG. 1, a plurality of sensors such as a pH meter, a thermometer, a dissolved oxygen meter, and a carbon dioxide concentration meter are usually installed. The sensor 23 is generally an electrode type sensor that uses an electrochemical principle for various measurements, an optical sensor that uses an optical principle, or the like. The port 8 is a member that maintains sterility and liquid tightness inside the flexible bag 1 when the sensor 23 is inserted from the outside of the support member 14.

The sensor 23 is connected to the measuring device 3 via a signal line as shown by a broken line in FIG. 1, and the measuring device 3 is connected to the control device 4 via a signal line. When the measurement device 3 receives the measurement signal from the sensor 23, the measurement device 3 transmits the measured information to the control device 4. And the control apparatus 4 controls the gas control apparatus 11, the heater 13 for temperature control, the gas control valve 21 etc. based on those information, the temperature of the culture solution enclosed with the flexible bag 1, Manage oxygen concentration and other conditions within an appropriate range.

The stirring blade (stirring bar) 2 stirs the culture solution and generates a horizontal swirling flow (rotating flow) in the culture solution. A top plate 6 having rigidity is joined to the top of the flexible bag 1, and the top plate 6 is provided with a through hole. The shaft 9 is inserted into the through hole of the top plate 6 through a sealing member 22 that seals the flexible bag 1 aseptically. The stirring blade 2 is provided inside the flexible bag 1 in a state of being fixed to the distal end side of the shaft 9. On the other hand, a stirring motor 10 is connected to the proximal end side of the shaft 9. A rotational flow of the stirring blade 2 is driven by the stirring motor 10 so that a swirling flow can be generated in the culture solution.

In FIG. 1, two stirring blades 2 are fixed to the shaft 9. However, one stirring blade 2 may be fixed, or three or more stirring blades 2 may be fixed. The blade shape and the number of the stirring blades 2 can be set appropriately. Furthermore, the stirring blade 2 is inserted into the bottom side of the flexible bag 1 instead of being driven by a stirring motor 10 installed above the top of the flexible bag 1 as shown in FIG. It is good also as a form driven with the stirring motor connected with a shaft and installed in the downward direction of the bottom part of the flexible bag 1. FIG. Further, the stirring blade 2 may be in a magnetic drive type without being fixed to a shaft or the like.

FIG. 2A is a cross-sectional view showing a configuration of a main part of the stirring apparatus according to the first embodiment of the present invention, and FIG. 3A is a cross-sectional view showing a baffle fixing method, and FIG. 3B is a perspective view showing the operation of the baffle.
In FIG. 2A and FIG. 2B, the principal parts, such as the flexible bag 1, the stirring blade 2, the baffle 5, the baffle support 24, etc. with which the stirring apparatus 100 is equipped, are shown, and the other structure is abbreviate | omitted. 2A and 2B, the dimension H is the height from the bottom inner surface of the flexible bag 1 to the lower end of the baffle 5, and the dimension L is the height from the bottom inner surface of the flexible bag 1 to the culture liquid level. The dimension d is the distance between the baffle 5 and the inner surface of the wall of the flexible bag 1, and the dimension D is the inner diameter of the flexible bag 1.

As shown in FIGS. 2A and 2B, the baffle (flow blocking unit) 5 hangs down from the top of the flexible bag 1 toward the inside, and the swirling flow of the culture solution (indicated by an arrow in FIG. 2B). It is provided to block a part. Specifically, the lower end of the baffle 5 is provided so as to be positioned below the liquid level of the culture solution sealed in the flexible bag 1. Moreover, the baffle 5 is provided in the cross-sectional shape which opposes the flow of the swirl flow of a culture solution, and exerts resistance. In addition, although the cross-sectional shape of the baffle 5 is a rectangular shape in FIG. 2B, it can be provided in an appropriate shape such as a square shape, a circular shape, or an elliptical shape. The dimensions of the width and thickness of the baffle 5 are not particularly limited. For example, the width of the baffle 5 is about 0.1 times the width of the culture tank (inner diameter of the flexible bag 1), as in a general baffle. It can be provided as a width or a thickness of about 1 mm or more.

The lower end of the baffle 5 is provided so that it may be located above the bottom part of the flexible bag 1, as shown to FIG. 2A. That is, the baffle 5 is provided separately from the inner surface of the bottom of the flexible bag 1, and the swirling flow of the culture solution is provided in the height section from the inner surface of the bottom of the flexible bag 1 to the lower end of the baffle 5. It is in a state where resistance is hardly exerted against. Therefore, as shown in FIG. 3A, below the lower end of the baffle 5, a horizontal swirling flow (indicated by an arrow in FIG. 3A) generated by the rotational motion of the stirring blade 2 flows while maintaining the flow velocity. Become. Then, above the lower end of the baffle 5, a part of the swirl flow is blocked by the baffle 5, thereby being converted into a vertical circulation flow (indicated by white arrows in FIG. 3A). Since the flow velocity of the swirling flow is maintained at a high speed below the lower end of the baffle 5, a strong vertical circulation flow is generated by being blocked by the baffle 5, and the stirring efficiency of the culture solution is increased. .

The baffle 5 is a ratio of the height (H) from the bottom inner surface of the flexible bag 1 to the lower end of the baffle 5 and the height (L) from the bottom inner surface of the flexible bag 1 to the culture liquid level. The value (H / L) is preferably provided so as to be greater than 0 and 0.8 or less, and more preferably 0.4 or more and 0.7 or less. If the ratio value (H / L) is 0.8 or less, the effect of the baffle 5 that converts the swirl flow into the up-and-down circulation flow is effectively exhibited, so that the stirring efficiency of the culture solution can be improved satisfactorily. . When the ratio value (H / L) is in the range of 0.4 to 0.7, the amount of generated vertical circulating flow increases, so that the stirring efficiency of the culture solution becomes better.

The baffle 5 is preferably provided separately from the inner surface of the wall portion of the flexible bag 1. By providing a gap between the outer end of the baffle 5 in the radial direction of the flexible bag 1 and the inner surface of the wall portion of the flexible bag 1, the flexible bag 1 can be located above the lower end of the baffle 5. A swirling flow of the culture solution can be generated along the inner wall surface. Therefore, it becomes possible to keep the flow velocity of the swirl flow higher, and a strong vertical circulation flow is generated by blocking the faster swirl flow by the baffle 5.

The baffle 5 has a ratio value (d / D) between the distance (d) from the inner wall surface of the flexible bag 1 to the outer end of the baffle 5 and the inner diameter (D) of the flexible bag 1 is 0. It is preferable to be provided so that it may exceed 0.08 or less, and more preferably 0.01 to 0.03. If the ratio value (d / D) is 0.08 or less, the effect of the baffle 5 that converts the swirl flow into the up-and-down circulation flow is effectively exhibited, so that the stirring efficiency of the culture solution can be improved satisfactorily. . When the ratio value (d / D) is in the range of 0.01 or more and 0.03 or less, the amount of vertical circulation flow increases, so that the stirring efficiency of the culture solution becomes better.

As shown in FIGS. 1, 2 A, and 2 B, the stirring device 100 according to this embodiment includes a baffle 5 that is provided integrally with the flexible bag 1. The baffle 5 has flexibility and forms a convex shape having a hollow structure opened toward the outside of the flexible bag 1. A rigid support member 24 is inserted into the flexible bag 1.

The baffle 5 is formed of a synthetic resin as with the flexible bag 1. Specifically, the baffle 5 may be formed of the same type of synthetic resin as that of the flexible bag 1, or a synthetic resin that is foldable and has higher rigidity than the flexible bag 1. May be formed. Moreover, you may form with thickness rather than the flexible bag 1. FIG. In FIG. 2, the baffle 5 is formed seamlessly on the top of the flexible bag 1, but can be integrally provided by joining by welding or the like.

The baffle support (internal support) 24 is made of a material having higher rigidity than the baffle 5 and the flexible bag 1. Further, the baffle support 24 is provided so as to substantially match the shape of the hollow structure that is recessed at the top of the baffle 5. In FIG. 1, FIG. 2A and FIG. 2B, the baffle support 24 is made of synthetic resin, but may be made of metal such as aluminum or stainless steel, glass or the like. When the baffle support 24 is loosely inserted into the baffle 5, the shape of the baffle 5 formed of a synthetic resin having flexibility is maintained, and the baffle support 24 can resist a swirling flow caused by stirring. The baffle support 24 is fixed to the gantry 15 via a support member 25 attached to the upper end side (see FIG. 1). The support member 25 may be provided integrally with the baffle support 24 in advance, or may be attached when the baffle support 24 is fixed.

The flexible bag 1 is installed in the stirring device 100 according to the following procedure, for example. Before the flexible bag 1 is installed in the stirrer 1, the liquid or gas is usually not sealed and the inside thereof is in a substantially sterile state and is folded and stored. When installing in the stirring apparatus 100, first, the flexible bag 1 is placed on the bottom of the support member 14 fixed to the gantry 15. At this time, other support members may be used together with the support member 14 in order to assist the support of the flexible bag 1. For example, it is possible to install a hoist, a crane, or the like on the gantry 15 and suspend the flexible bag 1 and place it.

The flexible bag 1 placed on the bottom of the support member 14 is connected to a gas supply pipe 16 for in-liquid ventilation and a gas supply pipe 17 for gas-phase ventilation. That is, the submerged gas supply pipe 16 and the gas-phase gas supply pipe 17 are connected to a pipe part that is pre-assembled in the flexible bag 1 and sealed aseptically. As a method for aseptically connecting the pipes, various methods such as a method using an aseptic connection type connector and a method using welding connection can be used. In general, a sterile connection type connector has a structure in which a tube end is sealed with a filter or a seal. In the case of an aseptic connection type connector, connectors sealed with a filter or a seal are connected in an open space, and the pipes are connected aseptically by removing the filter and the seal after the connection. Also, in the case of welding connection, piping is connected aseptically using a dedicated device capable of welding the pipe ends together in a closed space.

Subsequently, air is aseptically injected into the flexible bag 1 through the gas supply pipe 16 for in-liquid ventilation and the gas supply pipe 17 for gas-phase ventilation. The amount of air injected may be an amount that allows the flexible bag 1 to stand on its own. Then, the side surface of the flexible bag 1 that is infused with air to some extent is fixed to the wall portion of the support member 14. At this time, the various sensors 23 and the port 8 are aligned.

Subsequently, a stirring motor 10 is attached to the flexible bag 1. The stirring motor 10 may be supported by the sealing member 22 or may be supported by the gantry 15 having a relatively high load resistance. After the agitation motor 10 is attached to the flexible bag 1, air is aseptically injected again. By injecting air again, the flexible bag 1 has a container shape substantially equivalent to that in use. The flexible bag 1 is connected to other pipes including a pipe for supplying the culture solution. Other pipes are connected by using various methods for aseptically connecting the pipes in the same manner as the liquid supply gas supply pipe 16 and the gas supply gas supply pipe 17.

In the stirring apparatus 100 according to the present embodiment, the baffle 5 is fixed after the flexible bag 1 is fixed to the support member 14. As shown in FIG. 3B, the base end side of the baffle 5 is connected to the top of the flexible bag 1, and the inside of the hollow structure of the baffle 5 has a structure opened toward the outside of the flexible bag 1. ing. When the baffle 5 is fixed, the baffle support 24 is inserted into the baffle 5 from this opening. Then, the inserted baffle support 24 is supported by the support member 25 on the base end side protruding outside the flexible bag 1 and is fixed to the gantry 15 (see FIG. 1).

Subsequently, the sensor 23 is inserted into the port 8 of the flexible bag 1. In general, an electrode-type sensor that is used in direct contact with a culture solution is sterilized in advance before being inserted into the port 8. As the sterilization treatment, an appropriate treatment method such as steam sterilization or heat sterilization can be used. However, it is also possible to use a single-use sensor that has been sterilized in advance and packaged aseptically, instead of a normal sensor.

When using the stirring device 1, the culture solution is aseptically injected into the flexible bag 1 through a pipe (not shown) connected to the flexible bag 1. Next, the agitation motor 10 is actuated by the control device 4, and the agitation of the culture medium is started by driving the rotational motion of the agitation blade 2. The temperature of the culture solution is adjusted to a predetermined temperature range by the temperature adjusting heater 13, and the dissolved oxygen concentration is deoxygenated by supplying air, oxygen or the like through the submerged gas supply pipe 16 or by supplying nitrogen. To adjust to a predetermined concentration range. In addition, the pH of the culture solution is supplied by supplying carbon dioxide through the gas supply pipe 17 for gas-phase aeration or supplying an alkaline pH adjuster through a pipe (not shown) connected to the flexible bag 1. Do.

The desired cells to be cultured are seeded in a culture solution that is sealed in the flexible bag 1 and whose temperature, dissolved oxygen concentration, pH, and the like are adjusted to predetermined conditions. For cell seeding, cells that have been pre-cultured in advance in another cell culture vessel or the like are generally used. For example, the pre-cultured cell suspension or culture solution is injected into the culture solution through a pipe (not shown) connected to the flexible bag 1.

The cell culture is performed for a predetermined period while maintaining the temperature, dissolved oxygen concentration, pH and the like in a predetermined range and stirring the culture solution by operating the stirring blade 2. The temperature, dissolved oxygen concentration, and the like of the culture solution are monitored by the measuring device 3 connected to the sensor 23 and adjusted and controlled by the control device 4. Further, the control device 4 can also control the supply of carbon dioxide and the pH adjusting agent. And after completion | finish of culture | cultivation, a culture solution is discharged | emitted from the flexible bag 1 through the culture solution discharge pipe 18, and the cultured cell, the useful substance produced in the culture solution, etc. are collect | recovered.

The stirrer 100 can be used for applications such as cell culture and production of useful substances. Examples of cells to be cultured include Chinese hamster ovary cells (CHO cells), baby hamster kidney cells, mouse myeloma cells and other animal cells, Escherichia coli and yeast microbial cells, plant cells, insect cells, etc. And various types of cells. Examples of useful substances include various physiologically active substances, particularly antibody drugs, and other functional substances having industrial utility. Examples of the physiologically active substance include tissue-type plasminogen activator, erythropoietin, interferon and the like that have applications as thrombolytic agents. Antibody drugs include, for example, monoclonal antibodies, polyclonal antibodies, humanized antibodies, human antibodies, other immunoglobulins, and the like.

According to the stirring device 100 according to this embodiment, the lower end of the baffle (flow blocking unit) 5 is below the liquid level of the culture solution (liquid) 7 and above the bottom of the flexible bag 1. Therefore, the baffle does not greatly disturb the flow velocity of the swirling flow generated by the operation of the stirring blade (stirring member) 2 and the flow velocity of the swirling flow is constant even at a low stirring speed. It can be kept above. Therefore, the high-speed swirling flow is converted into the vertical circulation flow by the baffle 5, and the vertical circulation flow can be efficiently generated inside the flexible bag 1.

When culturing animal cells or the like in the stirrer 100, it is preferable to set the rotational speed of the stirring blade 2 to be slow from the viewpoint of preventing the cells from being damaged by the shearing force due to stirring. Specifically, it is preferably 500 rpm or less, more preferably 400 rpm or less, and it is not hindered to be about 100 rpm. In the stirring device 100 according to the present embodiment, even when the rotation speed of the stirring blade 2 is set to about 100 rpm, for example, the baffle installation position is appropriately set as described above to efficiently circulate vertically. Since the flow can be generated, it is advantageous in that the situation where the animal cell or the like is damaged or killed is prevented.

Further, in the stirring device 100 according to the present embodiment, the baffle 5 is provided integrally with the flexible bag 1 so as to have flexibility. Therefore, the flexible bag 1 can be handled in a folded state before use, and is excellent in convenience. In addition, if the flexible bag 1 that is sterilized in advance is used, it is suitable for ensuring the sealing performance and sterility of the flexible bag 1, and it is necessary to perform sterilization separately after the baffle 5 is fixed. There is an advantage that there is no.

Moreover, in the stirring device 100 according to the present embodiment, the baffle 5 having flexibility is supported by the baffle support 24 having rigidity. Therefore, when the flexible bag 1 is installed in the stirring device 100, the baffle support 24 is fixed to the mount 15 or the like, so that the flexible baffle 5 is placed on the support member 14. This is advantageous in that the flexible bag 1 can be securely fixed.

[Second Embodiment]
Next, a stirring device according to a second embodiment of the present invention will be described.

FIG. 4 is a cross-sectional view showing a main configuration of a stirring device according to the second embodiment of the present invention.
As shown in FIG. 4, the stirring device 100 according to the second embodiment includes a baffle (flow blocking unit) 5 A having rigidity separate from the flexible bag 1. Other configurations in the stirring device 100 are the same as those in the first embodiment.

The baffle 5A is made of a material having higher rigidity than the flexible bag 1. In FIG. 4, the baffle 5A is formed of a synthetic resin. However, the baffle 5A is a resin that is foldable and has higher rigidity than the flexible bag 1, a hard resin, or a metal. Alternatively, it may be formed of glass or the like. The baffle 5 A is provided so as to penetrate the top of the flexible bag 1 so that the baffle 5 A can be supported from the outside of the flexible bag 1. The baffle 5 A may be inserted into a port provided at the top of the flexible bag 1 or may be previously welded to the top of the flexible bag 1. From the viewpoint of simplifying the sterilization process separately performed after the baffle 5A is fixed, a form of welding in advance is preferable.

As shown in FIG. 4, the baffle 5 A is supported by a support member 25 on the base end side protruding to the outside of the flexible bag 1. And the support member 25 is fixed to the mount frame 15 (refer FIG. 1). The baffle 5A is fixed in this manner, so that the baffle 5A is installed at a predetermined position inside the flexible bag 1, and functions to convert the swirling flow caused by the stirring into a vertical circulation flow. The height (H) from the bottom inner surface of the flexible bag 1 in the baffle 5A, the distance (d) from the inner wall surface of the flexible bag 1, the width and the thickness are the same as those in the baffle 5. It can be set similarly to the dimensions.

According to the stirrer 100 according to this embodiment, it is possible to efficiently generate a vertical circulation flow inside the flexible bag 1 by the baffle 5A. Further, the baffle 5A has high rigidity and has resistance to the swirling flow, which is advantageous in that it can reliably convert a high-speed swirling flow into a vertical circulation flow.

[Third Embodiment]
Next, a stirring device according to a third embodiment of the present invention will be described.

FIG. 5 is a cross-sectional view showing a main configuration of a stirring device according to the third embodiment of the present invention.
As shown in FIG. 5, the stirring device 100 according to the third embodiment includes a baffle (flow blocking unit) 5 B having rigidity separate from the flexible bag 1. Moreover, it replaces with the said top plate 6 provided independently of a baffle, and the top plate 6B which can fix the baffle 5B is provided. Other configurations in the stirring device 100 are the same as those in the first embodiment.

The baffle 5B is formed of a material having higher rigidity than the flexible bag 1, like the baffle 5A. In FIG. 5, the baffle 5B is made of a synthetic resin. However, the baffle 5B is a resin that is foldable and has higher rigidity than the flexible bag 1, a hard resin, or a metal. Alternatively, it may be formed of glass or the like. The baffle 5B is stored in advance in the flexible bag 1, and is fixed to the inner surface side of the top plate 6B. The height (H) from the bottom inner surface of the flexible bag 1 in the baffle 5B, the distance (d) from the wall inner surface of the flexible bag 1, the width and the thickness are the same as those in the baffle 5. It can be set similarly to the dimensions.

The top plate 6B is formed of a material having rigidity. The top plate 6B may have a circular cross-sectional shape like the top of the flexible bag 1 or be flexible so that the baffle 5B installed symmetrically with respect to the central axis can be fixed. You may provide only on the diameter line of the top part of the bag 1. FIG. Moreover, you may fix the top plate 6B to the mount 15 through the support member 25 (refer FIG. 1). According to the form in which the top plate 6B is fixed, the weight of the agitating motor 10 can be supported together with the baffle 5B, so that the structure of the agitating device 100 becomes more stable.

According to the stirrer 100 according to this embodiment as described above, it is possible to efficiently generate a vertical circulation flow inside the flexible bag 1 by the baffle 5B. In addition, the baffle 5B is rigid and has resistance to the swirling flow, which is advantageous in that a high-speed swirling flow can be reliably converted into a vertical circulation flow. Further, since the baffle 5B is held inside the flexible bag 1 before use, the baffle 5B is suitable for ensuring the sealing property and sterility of the flexible bag 1 and is sterilized in advance. If the processed flexible bag 1 is used, there is an advantage that it is not necessary to separately sterilize after the baffle 5B is fixed.

[Fourth Embodiment]
Next, a stirring device according to a fourth embodiment of the present invention will be described.

FIG. 6 is a cross-sectional view showing a main configuration of a stirrer according to the fourth embodiment of the present invention.
As shown in FIG. 6, the stirring device 100 according to the fourth embodiment includes a baffle (flow blocking unit) 5C having rigidity separate from the flexible bag 1, and magnetic coupling members 24C1 and 24C2. ing. The magnetic coupling members 24C1 and 24C2 include a first magnetic member 24C1 provided inside the flexible bag 1 and a second magnetic member 24C2 provided outside the flexible bag 1. Other configurations in the stirring device 100 are the same as those in the first embodiment.

The baffle 5C is formed of a material having higher rigidity than the flexible bag 1, similarly to the baffle 5A. In FIG. 6, the baffle 5C is formed of a synthetic resin. However, the baffle 5C is a resin that is foldable and has higher rigidity than the flexible bag 1, a hard resin, or a metal. Alternatively, it may be formed of glass or the like. The first magnetic member 24C1 is attached to the upper end of the baffle 5C in advance before being stored in the flexible bag 1. A second magnetic member 24C2 is disposed outside the top of the flexible bag 1. The first magnetic member 24C1 and the second magnetic member 24C2 are each formed of a magnetic material, and can attract each other by magnetic force.

The baffle 5 C is fixed with the flexible bag 1 sandwiched between the attached first magnetic member 24 C 1 and the second magnetic member 24 C 2 by magnetic force. The second magnetic member 24C2 may be fixed to the gantry 15 via the support member 25 (see FIG. 1). In addition, a rigid top plate is disposed at the installation position of the baffle 5C at the top of the flexible bag 1, and a concave portion or a convex portion that engages with the second magnetic member 24C2 is provided on the top plate to provide the baffle 5C. It is good also as a form which prevents this position shift. The height (H) from the bottom inner surface of the flexible bag 1 in the baffle 5C, the distance (d) from the wall inner surface of the flexible bag 1, the width and the thickness are the same as those in the baffle 5. It can be set similarly to the dimensions.

According to the stirrer 100 according to this embodiment as described above, it is possible to efficiently generate a vertical circulation flow inside the flexible bag 1 by the baffle 5C. Further, the baffle 5C is rigid and has resistance to the swirling flow, and is advantageous in that it can reliably convert a high-speed swirling flow into a vertical circulation flow. Further, since the baffle 5C is held inside the flexible bag 1 before use, the baffle 5C is suitable for ensuring the sealing property and sterility of the flexible bag 1 and is sterilized in advance. If the processed flexible bag 1 is used, there is an advantage that it is not necessary to sterilize separately after fixing the baffle 5C. In addition, according to the fixation using magnetic force, the baffle 5C can be placed at an arbitrary position inside the flexible bag 1 before use, so that the flexible bag 1 becomes difficult to be bulky, and the baffle 5C It is also possible to make the installation position of the adjustable.

[Fifth Embodiment]
Next, a stirring device according to a fifth embodiment of the present invention will be described.

FIG. 7: is sectional drawing which shows the principal part structure of the stirring apparatus which concerns on 5th Embodiment of this invention.
As illustrated in FIG. 7, the stirring device 100 according to the fifth embodiment includes a flexible baffle (flow blocking unit) 5D integrated with the flexible bag 1, a reinforcing member 26, and a sealing member 27. I have. Other configurations in the stirring device 100 are the same as those in the first embodiment.

The baffle 5D has a hollow convex shape that opens toward the outside of the flexible bag 1. The baffle 5 D is formed of a synthetic resin with flexibility like the flexible bag 1. Specifically, the baffle 5 may be formed of the same type of synthetic resin as that of the flexible bag 1, or a synthetic resin that is foldable and has higher rigidity than the flexible bag 1. May be formed. Moreover, you may form with thickness rather than the flexible bag 1. FIG. The inside of the baffle 5 D is isolated from the inside of the flexible bag 1, but has an open structure toward the outside of the flexible bag 1.

In the baffle 5D, air is sealed in the internal space, and the opening is sealed by the sealing member 27. The baffle 5D is in a flat state before use, and is baffled by air pressure when air is sealed when the baffle 5D is fixed. The height (H) from the bottom inner surface of the flexible bag 1 in the baffle 5D, the distance (d) from the inner wall surface of the flexible bag 1, the width and the thickness are the same as those in the baffle 5. It can be set similarly to the dimensions.

The reinforcing member 26 has higher rigidity than the flexible bag 1 and reinforces the strength of the joint portion between the baffle 5 D and the top of the flexible bag 1. The reinforcing member 26 may be formed of the same synthetic resin as that of the flexible bag 1 with a thickness, or may be formed of a synthetic resin having higher rigidity than the flexible bag 1. Further, it may be provided in the form of a port with a hard resin or the like. By providing the reinforcing member 26, the baffle 5D receives pressure due to the agitation of the culture solution, and the baffle 5D breaks when stress concentrates on the joint between the baffle 5D and the top of the flexible bag 1. Is to be prevented.

According to the stirrer 100 according to this embodiment, it is possible to efficiently generate a vertical circulation flow inside the flexible bag 1 by the baffle 5D. Further, since the baffle 5D has flexibility, the baffle 5D can be handled in a folded state before use in which air is not sealed inside, and is excellent in convenience. Moreover, it is suitable for ensuring the sealing property and sterility of the inside of the flexible bag 1, and if the flexible bag 1 sterilized in advance is used, it is necessary to sterilize after fixing the baffle 5D. There is an advantage that there is no.

[Sixth Embodiment]
Next, a stirring device according to a sixth embodiment of the present invention will be described.

FIG. 8 is a cross-sectional view showing a main configuration of a stirrer according to the sixth embodiment of the present invention.
As shown in FIG. 8, the stirring device 100 according to the sixth embodiment includes a baffle 5 F that is suspended from the top of the flexible bag 1 and fixed to the bottom of the flexible bag 1. Specifically, the baffle 5F includes a plate-like part (flow blocking part) 5F1 provided on the upper side and a perforated part (connecting part) 5F2 provided on the lower side. Other configurations in the stirring device 100 are the same as those in the first embodiment.

The baffle 5 F is formed of a synthetic resin with flexibility similar to the flexible bag 1. Specifically, the baffle 5F may be formed of the same type of synthetic resin as that of the flexible bag 1, or a synthetic resin having a foldable flexibility and higher rigidity than the flexible bag 1. May be formed. Moreover, you may form with thickness rather than the flexible bag 1. FIG. The plate-like portion 5F1 (flow blocking portion) of the baffle 5F is provided so as to hang from the top of the flexible bag 1 and block the flow of the swirling flow of the culture solution. On the other hand, the perforated portion (connecting portion) 5F2 is connected to the lower end of the plate-like portion 5F1 so that the swirl flow of the culture medium can flow through the liquid passage hole 28. In addition, the shape of the liquid passage hole 28 can be provided in an appropriate shape such as a rectangular shape, a square shape, a circular shape, or an elliptical shape. In addition, an appropriate number of liquid passage holes 28 can be provided.

The upper end (the upper end of the plate-like portion 5F1) and the lower end (the lower end of the perforated portion 5F2) of the baffle 5F may be joined and integrated with the flexible bag 1 by welding or the like, or a magnetic coupling member ( 24C1, 24C2) may be used. That is, the plate-like portion 5F1 is fixed to the bottom portion together with the top portion of the flexible bag 1 through the perforated portion 5F2. By fixing the plate-like portion 5F1 also on the lower end side, the baffle 5F is pushed by the flow of the swirling flow and is easily bent, and a situation in which the efficiency of generating the vertical circulation flow is reduced is prevented. . At this time, since stress is concentrated by the flow of the swirl flow at the joint between the baffle 5F and the flexible bag 1, a reinforcing member (26) having higher rigidity than the flexible bag 1 may be installed. . The height (H) from the bottom inner surface of the flexible bag 1 in the baffle 5F, the distance (d) from the inner wall surface of the flexible bag 1, the width and the thickness are the same as those in the baffle 5 described above. It can be set similarly to the dimensions. However, the height (H) from the bottom inner surface of the flexible bag 1 is indicated by a broken line in FIG. 8 from the bottom inner surface of the flexible bag 1 to the lower end of the plate-like portion 5F1, that is, the uppermost end of the liquid passage hole 28. )) Is defined as the height.

According to the stirrer 100 according to this embodiment as described above, it is possible to efficiently generate a vertical circulation flow inside the flexible bag 1 by the baffle 5F. Further, since the baffle 5F has flexibility, it can be handled in a folded state before use, and is excellent in convenience. Moreover, if the flexible bag 1 which is suitable for ensuring the sealing property and sterility of the inside of the flexible bag 1 and sterilized in advance is used, it is necessary to sterilize separately after fixing the baffle 5F. There is an advantage that there is no. Further, since the baffle 5F is fixed to the bottom of the flexible bag 1, the baffle 5F has resistance to the swirling flow while having flexibility, and the high-speed swirling flow is surely provided in the vertical circulation flow. This is advantageous in that it can be converted into

[Seventh Embodiment]
Next, a stirring device according to a seventh embodiment of the present invention will be described.

FIG. 9 is a cross-sectional view showing a main configuration of a stirrer according to the seventh embodiment of the present invention.
As shown in FIG. 9, the stirring device 100 according to the seventh embodiment includes a baffle (flow blocking unit) 5G that hangs down from the top of the flexible bag 1 and the baffle 5G with respect to the bottom of the flexible bag 1. A fixing string (connecting portion) 29 to be fixed is provided. Other configurations in the stirring device 100 are the same as those in the first embodiment.

The baffle 5G is formed with flexibility by a synthetic resin like the flexible bag 1. Specifically, the baffle 5G may be formed of the same type of synthetic resin as that of the flexible bag 1, or a synthetic resin that is foldable and has higher rigidity than the flexible bag 1. May be formed. Moreover, you may form with thickness rather than the flexible bag 1. FIG. The upper end of the baffle 5G may be joined and integrated with the flexible bag 1 by welding or the like, or may be fixed using a magnetic coupling member (24C1, 24C2). The height (H) from the bottom inner surface of the flexible bag 1 in the baffle 5G, the distance (d) from the inner wall surface of the flexible bag 1, the width and the thickness are the same as those in the baffle 5. It can be set similarly to the dimensions.

The fixed string 29 is connected to the lower end of the baffle 5G so that the swirl flow of the culture solution can flow. One end of the fixed string 29 is welded to the lower end of the baffle 5G and the other end is welded to the bottom of the flexible bag 1, and the lower end side of the baffle 5G is fixed with tension. The fixed string 29 can be formed of an appropriate material, but is preferably made of a synthetic resin. Although the fixed string 29 can be provided with an appropriate diameter and number, it is preferable to provide at least two or more baffles 5G per baffle 5G so that the baffle 5G is difficult to bend with respect to the swirling flow.

According to the stirrer 100 according to this embodiment as described above, it is possible to efficiently generate a vertical circulation flow inside the flexible bag 1 by the baffle 5G. Moreover, since the baffle 5G and the fixed string 29 have flexibility, they can be handled in a folded state before use, and are excellent in convenience. In addition, it is suitable for ensuring the sealing performance and sterility of the inside of the flexible bag 1, and if the flexible bag 1 sterilized in advance is used, it is necessary to sterilize it separately after fixing the baffle 5G. There is an advantage that there is no. Further, since the baffle 5G is fixed to the bottom of the flexible bag 1 by the fixing string 29, the baffle 5G has resistance against swirling flow while having flexibility, and the lower side of the baffle 5G. Is advantageous in that the swirling flow is difficult to be blocked, and a high-speed swirling flow can be efficiently converted into a vertical circulation flow.

[Eighth Embodiment]
Next, a stirrer according to the eighth embodiment of the present invention will be described.

FIG. 10A is a cross-sectional view showing a main configuration of a stirring device according to an eighth embodiment of the present invention. FIG. 10B is a perspective view of a baffle provided in the stirring device according to the eighth embodiment of the present invention.
As illustrated in FIG. 10A, the stirring device 100 according to the eighth embodiment includes a baffle (flow blocking unit) 5H that hangs down from the top of the flexible bag 1 and the baffle 5H with respect to the bottom of the flexible bag 1. And a fixing plate (connecting portion) 30 to be fixed. Other configurations in the stirring device 100 are the same as those in the first embodiment.

The baffle 5H is formed of a synthetic resin so as to be flexible like the flexible bag 1. Specifically, the baffle 5H may be formed of the same type of synthetic resin as that of the flexible bag 1, or a synthetic resin that is foldable and has higher rigidity than the flexible bag 1. May be formed. Moreover, you may form with thickness rather than the flexible bag 1. FIG. The upper end of the baffle 5H may be joined and integrated with the flexible bag 1 by welding or the like, or may be fixed using a magnetic coupling member (24C1, 24C2). The height (H) from the bottom inner surface of the flexible bag 1 in the baffle 5H, the distance (d) from the inner wall surface of the flexible bag 1, the width and the thickness are the same as those in the baffle 5. It can be set similarly to the dimensions.

Similarly to the baffle 5H, the fixing plate 30 may be formed of the same type of synthetic resin as that of the flexible bag 1, and has higher rigidity than the flexible bag 1 while having foldable flexibility. You may form with a synthetic resin. As shown in FIG. 10B, the fixing plate 30 is connected to the lower end of the baffle 5H so as to allow a swirling flow of the culture medium to flow through by being installed perpendicular to the baffle 5H. Since the baffle 5H is arranged perpendicular to the flow direction of the swirl flow of the culture solution, the baffle 5H blocks the swirl flow of the culture solution and converts the swirl flow into a vertical circulation flow. On the other hand, since the fixed plate 30 is provided in parallel with the flow direction of the swirling flow, the fixed plate 30 does not greatly disturb the flow velocity of the swirling flow.

The upper end of the baffle 5H and the lower end of the fixing plate 30 may be integrated by welding or the like to the flexible bag 1, or may be fixed using magnetic coupling members (24C1, 24C2). . Further, the lower end of the baffle 5H and the upper end of the fixing plate 30 may be joined by welding or the like, or may be integrally formed in advance. That is, the baffle 5H is fixed to the bottom of the flexible bag 1 through the fixing plate 30.

According to the stirrer 100 according to this embodiment as described above, it is possible to efficiently generate a vertical circulation flow inside the flexible bag 1 by the baffle 5H. Moreover, since the baffle 5H has flexibility, it can be handled in a folded state at the joint between the baffle 5H and the fixed plate 30 before use, and is excellent in convenience. It will be. In addition, it is suitable for ensuring the sealing property and sterility of the inside of the flexible bag 1, and if the flexible bag 1 sterilized in advance is used, it is necessary to sterilize separately after fixing the baffle 5H. There is an advantage that there is no. Further, since the baffle 5H is fixed to the bottom of the flexible bag 1 by the fixing plate 30, the baffle 5H has resistance against swirling flow while having flexibility, and below the baffle 5H. Is advantageous in that the swirling flow is not easily obstructed and a high-speed swirling flow can be reliably converted into a vertical circulation flow. In addition, outside the fixed plate 30 in the radial direction of the flexible bag 1, it is possible to increase the flow velocity of the swirling flow by adjusting the installation position of the fixed plate 30.

As mentioned above, although the example of the stirring apparatus which concerns on embodiment of this invention was demonstrated, this invention is not limited to these specific forms. For example, the stirrer can be configured by combining the configurations in each of the embodiments described above. If the baffle is provided so as to block the swirling flow of the liquid and the lower side is provided not to block the flow of the swirling flow of the liquid, the baffle is relatively swirling at a predetermined stirring speed. It is possible to generate a strong vertical circulation flow.

Further, the container shape of the flexible bag 1 is not limited to a cylindrical shape, and the stirring efficiency can be improved even in a rectangular shape, a polygonal shape or the like. In addition, the internal diameter (D) in the square-shaped flexible bag 1 means the width dimension which passes the installation position of a baffle. The number of baffles installed can be an appropriate number of 2 or more. In particular, the configuration in which the two baffles are arranged symmetrically with respect to the center of the flexible bag 1 can suppress the stress concentration sites that are likely to occur around the joint portion of the baffle, so that the agitation efficiency and the flexible bag are good. It is suitable for coexistence with the structural stability of 1. The flexible bag 1 can be applied by being provided on an appropriate scale in the range of several L to several thousand L.

Hereinafter, the present invention will be specifically described with reference to examples, but the technical scope of the present invention is not limited thereto.

[Example 1]
As Example 1, in the stirring device in which the cylindrical flexible bag was installed, the influence of the baffle installation position on the liquid stirring efficiency was evaluated.

The stirring efficiency of the liquid was analyzed by the finite volume method using the general-purpose fluid analysis program “FLUENT” for the internal circulation efficiency of the liquid sealed in the flexible bag. In this finite volume method, the space inside the flexible bag was divided into minute control volumes, and the flow velocity, viscosity, turbulent kinetic energy, etc. were determined for each control volume. The stirring efficiency of the liquid was evaluated using as an index the volume integral value of the flow rate of the upward flow and the downward flow in the flexible bag. The vertical circulation flow is a flow that circulates in the vertical direction inside the flexible bag by converting the horizontal swirling flow by the rotational movement of the stirring bar by the baffle.

The stirring device was in the form shown in FIG. The flexible bag 1 was a cylindrical container with a capacity of 4L. The inner diameter (D) of the flexible bag 1 is 150 mm. The height (L) from the bottom inner surface of the flexible bag 1 to the liquid level of the culture solution is 230 mm. The baffle 5 was a flat plate having a thickness of 2 mm and a width of 12 mm, and two pieces were installed symmetrically with respect to the central axis of the flexible bag 1. Further, only one flat paddle type was installed as the stirring blade 2. The installation position of the stirring blade 2 is the center of the flexible bag 1, and the installation height is 25 mm from the bottom inner surface of the flexible bag 1. The rotation speed of the stirring blade in stirring the culture solution was set to 100 rpm.

First, the distance (d) from the wall portion inner surface of the flexible bag 1 to the outer end of the baffle 5 and the ratio value (d / D) of the inner diameter (D) of the flexible bag 1 are changed to circulate up and down. The flow velocity volume integral value of the flow was analyzed. At this time, the value (H) of the height (H) from the bottom inner surface of the flexible bag to the lower end of the baffle and the height (L) from the bottom inner surface of the flexible bag to the liquid level of the culture solution / L) was 0 (a state where the lower end of the baffle was in contact with the bottom inner surface of the flexible bag).

FIG. 11 is a diagram showing a result of analyzing the relationship between the distance from the wall portion inner surface of the flexible bag to the baffle and the flow velocity volume integral value of the vertical circulation flow.
In FIG. 11, the relative value with respect to the flow volume integrated value in d / D = 0 is shown. As shown in FIG. 11, as the distance between the baffle and the inner surface of the wall portion of the flexible bag increases, the flow velocity volume integral value of the up-and-down circulating flow increases and becomes maximum near d / D = 0.02. ing. As the distance between the baffle and the inner surface of the wall of the flexible bag further increases, the flow volume integrated value of the vertical circulation flow gradually decreases, and the flow velocity at d / D = 0 when d / D = 0.08. It is equivalent to the volume integral value.

Accordingly, the ratio value (d / D) between the distance (d) from the inner surface of the wall portion of the flexible bag 1 to the outer end of the baffle 5 and the inner diameter (D) of the flexible bag exceeds 0 and is 0. It can be said that it is preferable to provide a value of 0.08 or less. In particular, when the ratio value (d / D) is in the range of 0.01 or more and 0.03 or less, a high flow rate volume integral value is realized, which can be said to be suitable for increasing the stirring efficiency.

Next, the ratio value (H) of the height (H) from the bottom inner surface of the flexible bag to the lower end of the baffle and the height (L) from the bottom inner surface of the flexible bag to the liquid level of the culture solution / L) was changed, and the flow velocity volume integral value of the vertical circulation flow was analyzed. At this time, the value (d / D) of the distance (d) from the wall portion inner surface of the flexible bag 1 to the outer end of the baffle 5 and the inner diameter (D) of the flexible bag 1 is 0. .04.

FIG. 12 is a diagram showing the relationship between the height from the bottom inner surface of the flexible bag to the lower end of the baffle and the flow velocity volume integral value of the vertical circulation flow.
In FIG. 12, the relative value with respect to the flow velocity volume integral value in H / L = 1 is shown. As shown in FIG. 12, when the height from the bottom inner surface of the flexible bag to the lower end of the baffle increases, the flow volume integral value of the vertical circulation flow increases and reaches a maximum near H / L = 0.5. It has become. When the height from the bottom inner surface of the flexible bag to the lower end of the baffle is further increased, the flow volume integrated value of the vertical circulation flow is gradually decreased. It can be confirmed that this tendency is the same even when the installation height of the stirring blade is set to 10 mm from the bottom inner surface of the flexible bag. That is, even when the installation height of the stirring blade is set to 10 mm from the inner surface of the bottom of the flexible bag, it is confirmed that the flow velocity volume integral value of the vertical circulation flow is maximum near H / L = 0.5. Is done. In particular, in the vicinity of H / L = 0.8 or less, the flow velocity volume integral value exceeds the flow velocity volume integral value at H / L = 0 (the state where the lower end of the baffle is in contact with the bottom inner surface of the flexible bag). It has become.

Therefore, the value (H / H) of the height (H) from the bottom inner surface of the flexible bag to the lower end of the baffle and the height (L) from the bottom inner surface of the flexible bag to the culture liquid level. It can be said that L) is preferably provided so as to be more than 0 and 0.8 or less. In particular, when the ratio value (H / L) is in the range of 0.4 to 0.7, a high flow velocity volume integral value is realized, which can be said to be suitable for increasing the stirring efficiency.

FIG. 13 is a diagram showing the effect of the flow volume integrated value of the vertical circulation flow in the stirring device according to the example of the present invention.
In FIG. 13, the effect of the flow volume integrated value of the vertical circulation flow in the stirring apparatus according to the example is shown as a relative value with respect to the comparative example. Comparative Example 1 is a stirrer in which no baffle is installed, that is, a stirrer in which the distance ratio value (d / D) is set to 0 and the height ratio value (H / L) is set to 1.0. is there. Comparative Example 2 is a stirrer in which the baffle is placed in close contact with the bottom inner surface and the wall inner surface of the flexible bag, that is, the distance ratio value (d / D) is 0, and the height ratio value. This is a stirring device in which (H / L) is set to 0. On the other hand, the example is an agitation apparatus in which the distance ratio value (d / D) is set to 0.04 and the height ratio value (H / L) is set to 0.5.

As shown in FIG. 13, the stirring device (Comparative Example 2) in which the baffle is placed in close contact with the bottom inner surface and the wall inner surface of the flexible bag is compared with the stirring device (Comparative Example 1) in which the baffle is not installed. Thus, the flow velocity volume integral value of the vertical circulation flow shows an increase of 16%. On the other hand, the stirrer (Example) in which the baffle is installed separately from the bottom inner surface and the wall inner surface of the flexible bag shows an increase of 41% in the flow volume integral value of the vertical circulation flow. Therefore, it can be said that it is preferable to separate the baffle installation position from the bottom inner surface and the wall inner surface of the flexible bag from the viewpoint of increasing the stirring efficiency by generating a strong vertical circulation flow.

Next, the integrated volume value of the velocity of the vertical circulation flow was analyzed by changing the number of baffles installed. The number of baffles installed was set to 0, 2, 4, and 8, and a plurality of baffles were equally installed along the circumferential direction of the flexible bag. At this time, the value (d / D) of the distance (d) from the wall portion inner surface of the flexible bag to the outer end of the baffle and the inner diameter (D) of the flexible bag is 0.04. did. Further, the ratio value (H /) of the height (H) from the bottom inner surface of the flexible bag to the lower end of the baffle and the height (L) from the bottom inner surface of the flexible bag to the liquid level of the culture solution. L) was set to 0.5.

FIG. 14 is a diagram showing the relationship between the number of baffles installed and the flow velocity volume integral value of the vertical circulation flow.
As shown in FIG. 14, the stirring device with two baffles installed shows an increase in the integrated volume of flow velocity of the vertical circulation flow of about 40% compared with the stirring device with zero baffles installed. ing. On the other hand, it is shown that even if the number of baffles installed is 4 or more, a large increase in the flow volume integrated value of the vertical circulation flow is not recognized. Therefore, it can be said that two baffles are sufficient from the viewpoint of increasing the stirring efficiency by generating a strong vertical circulation flow. In particular, since the installation of the baffle is restricted because the rigidity of the flexible bag is poor, it can be said that the number of baffles installed is preferably two.

[Example 2]
As Example 2, in the stirring device in which a rectangular flexible bag was installed, the influence of the baffle installation position on the liquid stirring efficiency was evaluated. As in Example 1, the liquid stirring efficiency was analyzed by the finite volume method using the general-purpose fluid analysis program “FLUENT” for the internal circulation efficiency of the liquid sealed in the flexible bag.

FIG. 15A is a cross-sectional view illustrating a configuration of a main part of a stirring device provided with a rectangular flexible bag. FIG. 15B is a cross-sectional view taken along the line II-II in FIG. 15A.
The flexible bag 1A shown in FIGS. 15A and 15B is a rectangular container having a capacity of 4L. The flexible bag 1A has a square shape with a width (X) of 140 mm and a depth (Y) of 140 mm. Moreover, the height (L) from the bottom inner surface of the flexible bag 1A to the liquid level of the culture solution is 200 mm. Moreover, the baffle 5 was made into a flat plate shape having a thickness of 2 mm and a width of 12 mm, and two pieces were installed symmetrically with respect to the central axis on the bisector of the side of the flexible bag 1A. Further, the stirring blade 2 was a flat paddle type as in Example 1. The installation position of the stirring blade 2 is the center of the flexible bag 1A, and the installation height is 25 mm from the inner surface of the bottom of the flexible bag 1A. The rotation speed of the stirring blade in stirring the culture solution was set to 100 rpm.

Value of the ratio (H / L) between the height (H) from the bottom inner surface of the flexible bag to the lower end of the baffle and the height (L) from the bottom inner surface of the flexible bag to the culture liquid level The volume integral value of the up-and-down circulation flow was analyzed by changing. At this time, the value (d / X) of the distance (d) from the wall portion inner surface of the flexible bag 1 to the outer end of the baffle 5 and the width (X) of the flexible bag 1 is 0. (The baffle is in contact with the inner surface of the wall of the flexible bag).

FIG. 16 is a diagram showing the relationship between the height from the bottom inner surface of the flexible bag to the lower end of the baffle and the flow velocity volume integral value of the vertical circulation flow in the rectangular flexible bag.
In FIG. 16, the relative value with respect to the flow volume integrated value in H / L = 1 is shown. As shown in FIG. 16, when the height from the bottom inner surface of the flexible bag to the lower end of the baffle increases, the flow volume integral value of the vertical circulation flow increases and reaches a maximum near H / L = 0.7. It has become. Further, when the height from the bottom inner surface of the flexible bag to the lower end of the baffle increases, the flow velocity volume integral value of the vertical circulation flow decreases. Particularly, when H / L = 0.9 to below 0.8, the flow velocity volume integral value is H / L = 0 (the flow velocity volume integral when the lower end of the baffle is in contact with the bottom inner surface of the flexible bag). It exceeds the value.

Therefore, even in the rectangular flexible bag, the height (H) from the bottom inner surface of the flexible bag to the lower end of the baffle and the height from the bottom inner surface of the flexible bag to the liquid level of the culture solution ( It can be said that the ratio (H / L) to L) is preferably provided so as to be greater than 0 and not greater than 0.8. In particular, when the ratio value (H / L) is in the range of 0.4 to 0.7, a high flow velocity volume integral value is realized, which can be said to be suitable for increasing the stirring efficiency.

Further, in the rectangular flexible bag, the ratio value (d / X) between the distance (d) from the wall inner surface of the flexible bag to the outer end of the baffle and the width (X) of the flexible bag. ) And the volume integral value of the vertical circulation flow was analyzed. As the distance (d) from the inner wall surface of the flexible bag to the outer end of the baffle increased, the flow volume integral value of the vertical circulation flow increased. , D / X = 0.03 was found to be the maximum. In the present specification, the width (X) of the flexible bag has the same significance as the inner diameter (D) of the flexible bag with respect to the installation position of the baffle.

Therefore, even in a rectangular flexible bag, the ratio value (d) between the distance (d) from the wall portion inner surface of the flexible bag 1 to the outer end of the baffle 5 and the width (X) of the flexible bag ( It can be said that d / X) is preferably provided so as to be more than 0 and 0.08 or less. In particular, when the ratio value (d / X) is in the range of 0.01 or more and 0.03 or less, a high flow rate volume integral value is realized, which can be said to be suitable for increasing the stirring efficiency.

1 Flexible bag 2 Stirring blade (stirring bar)
3 Measuring device 4 Control device 5 Baffle (flow blocking part)
6 Top plate 7 Culture solution 8 Port 9 Shaft 10 Stirring motor 11 Gas control device 12 Sparger 13 Temperature control heater 14 Support member 15 Base 16 Submerged gas supply tube 17 Gas phase supply gas supply tube 18 Culture solution discharge tube 19 Exhaust pipe 20 Exhaust filter 21 Gas control valve 22 Sealing member 23 Sensor 24 Baffle support (internal support)
25 Support member 26 Reinforcement member 27 Sealing member 28 Fluid passage hole 29 Fixed string (connecting portion)
30 Fixing plate (connection part)
100 Stirrer

Claims

A flexible bag having flexibility and capable of enclosing a liquid therein;
A stirrer disposed inside the flexible bag for creating a swirling flow in the liquid;
A flow blocking unit provided to hang from the top of the flexible bag and block the swirling flow of the liquid;
The stirring device according to claim 1, wherein a lower end of the flow blocking unit is provided below the liquid level of the liquid and above the bottom of the flexible bag.
The value (d / D) of the distance (d) from the inner surface of the wall portion of the flexible bag to the flow blocking portion and the inner diameter (D) of the flexible bag exceeds 0 and is 0.08. The stirring device according to claim 1, wherein:
The ratio value (d / D) between the distance (d) from the inner wall surface of the flexible bag to the flow blocking portion and the inner diameter (D) of the flexible bag is 0.01 or more and 0.00. The stirring device according to claim 2, wherein the stirring device is 03 or less.
The value of the ratio between the height (H) from the bottom inner surface of the flexible bag to the lower end of the flow blocking portion and the height (L) from the bottom inner surface of the flexible bag to the liquid level of the liquid The stirring device according to claim 1, wherein (H / L) is more than 0 and 0.8 or less.
The value of the ratio between the height (H) from the bottom inner surface of the flexible bag to the lower end of the flow blocking portion and the height (L) from the bottom inner surface of the flexible bag to the liquid level of the liquid The stirring apparatus according to claim 4, wherein (H / L) is 0.4 or more and 0.7 or less.
The stirrer according to claim 1, wherein the flow blocking part is formed of a flexible material.
The stirrer according to claim 1, wherein the flow blocking portion is provided integrally with the flexible bag and has a hollow structure opened toward the outside of the flexible bag.
The stirrer according to claim 7, wherein a rigid internal support is inserted into the flow blocking part.
The stirring device according to claim 8, wherein an upper end of the internal support is fixed by a support member having rigidity.
The stirrer according to claim 7, wherein air is sealed inside the flow blocking part.
2. The flow blocking part is formed of a material having higher rigidity than the flexible bag, and an upper end of the flow blocking part is fixed by a support member having rigidity. Stirring device.
A first magnetic member is attached to an upper end of the flow blocking portion, and the flow blocking portion is flexed by an attractive force between the first magnetic member and a second magnetic member disposed outside the flexible bag. The stirring device according to claim 1, wherein the stirring device is fixed with a pouch interposed therebetween.
The flow blocking part is fixed to a bottom part of the flexible bag through a connecting part that is connected to a lower end of the flow blocking part so that a swirl flow of the liquid can flow. The stirring apparatus according to 1.