JPS63251077A - Culture device using axial-flow impeller - Google Patents

Abstract

PURPOSE:To efficiently cultivate cells, etc., by reduced power, by setting an axial-flow impeller in a cylinder in a culture tank, rotating the impeller, making a culture solution, cells, etc., flow from the top of the cylinder on liquid surface of the culture solution outside the cylinder. CONSTITUTION:A cylinder 13 is set in a culture tank 11 in such a way that the top of the cylinder is protruded from liquid surface W of a culture solution and an axial-flow impeller 15 is laid in the cylinder 13. The impeller 15 is rotated, the culture solution (l), cells (c), etc., are made to flow from the top of the cylinder 13 on the liquid surface W of culture solution outside the cylinder 13. Consequently, an oxygen-containing gas in the upper part of the culture tank 11 is entangled in the culture solution (l) to feed oxygen to the culture solution (l) and to stir the culture solution (l), the cells (c), etc. Consequently, since stirring in the culture tank and feed of oxygen can be simultaneously carried out only by rotating the axial-flow impeller, the cells, etc., can be cultivated efficiently and simply by less power.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to an apparatus for culturing microorganisms such as animal and plant cells and bacteria, and a device for culturing microorganisms such as animal and plant cells and bacteria.
The present invention relates to a culture device capable of supplying oxygen to cells, etc. with a small amount of power, and stirring a single culture solution to promote mass transfer of metabolic products.

[Conventional technology and problems]

Generally, in order to culture animal and plant cells and microorganisms such as bacteria, oxygen is supplied to the cells etc. through a culture solution, and
It is necessary to mix and stir the culture solution and cells to promote mass transfer of metabolic products.

Conventionally, for example, a culture apparatus as shown in FIG. 7 was provided, and bubbles were discharged into the culture solution from an aeration pipe 1, and the culture solution was stirred and mixed by a stirrer 2. In addition,
In the figure, 3 is a culture tank (container), 4 is a baffle plate protruding from the inner wall surface to enhance the stirring effect of the agitator 2, and g is a diffuser pipe 1.
This is the air that is sent to the

However, in the conventional apparatus described above, since external air is forced into the culture tank 3, it is essential to use a sterilization filter to prevent bacteria from the external air from entering the culture tank. However, since the sterilization filter is a porous material with extremely fine gaps, a large amount of power is required to overcome the excessive flow resistance and water pressure of the culture solution and pass a large amount of air. There was a problem in that the impeller for stirring also required separate power.

In addition, as shown in FIG. 8, air g is added from below the culture tank 3.
In a culture device in which air bubbles are generated in the tank by sending the air bubbles into the tank, and the culture solution T is fluidized and stirred only by the bubbles, as the culture progresses and the viscosity of the culture at increases, a sufficient stirring effect cannot be obtained. However, there was a problem in that the power required to pump the air was even more excessive.

The technical objective of the present invention is to provide a culture device that can supply R element to animal and plant cells and bacteria while stirring them with less power without contaminating them with germs.

[Means for solving problems]

In order to solve the problems and technical problems of the prior art described above, the present invention provides a cylinder inside a culture tank with its upper part protruding from the culture solution surface, and an axial flow impeller inside the cylinder. By providing a wheel and rotating the axial flow impeller,
The culture solution and cells, etc. are allowed to flow out from the top of the cylinder and fall onto the surface of the culture solution outside the cylinder, thereby entraining the oxygen-containing gas at the top of the culture tank into the culture solution to supply culture solution pressure oxygen. , it is characterized by stirring the culture solution, cells, etc.

Note that the cylinder may be fixed or may be rotated together with the axial flow impeller.

[Effect]

Since the present invention is configured as described above, when the culture solution is poured into the culture tank to a predetermined level, the culture solution is absorbed into the culture tank through the lower end opening of the cylinder provided with the upper part protruding above the culture solution surface. The inside and outside of the cylinder reach approximately the same level. Next, when the axial flow impeller is rotated in this state, the culture solution cells, etc. rise inside the cylinder due to the liquid lifting action of the axial flow impeller, overflow from the opening at the top of the cylinder to the outside of the cylinder, and the culture solution fall onto the surface.

As mentioned above, when culture solution cells, etc. fall from the opening at the top of the cylinder onto the culture solution surface, the oxygen-containing gas from the upper part of the culture tank is drawn into the culture fX solution, and oxygen is supplied to the culture solution. . The dropped liquid is mixed with the culture liquid in the tank, sucked into the cylinder from the lower end opening by the action of the axial impeller, rises again inside the cylinder, and flows out from the upper opening.

In this way, a circulating flow is formed inside and outside the cylinder, and in the process, oxygen is supplied, and the culture solution, cells, etc. are agitated and mixed by the circulating flow.

〔Example〕

Next, embodiments of the present invention will be described with reference to the drawings.

FIG. 1 is a longitudinal sectional view of a cell culturing apparatus showing a first embodiment of the present invention.

In the figure, a cylinder 13 integrated with a lid 12 is fixedly attached to culture #J11 near the center of a culture tank (container) 11, and a rotating shaft 1 is attached inside the fixed cylinder 13.
An axial flow impeller 15 mounted at 4 is provided.

The axial impeller 15 is configured by, for example, having flat blades installed at an angle with respect to the rotation direction (direction perpendicular to the axis). Note that the axial flow impeller 15 is a spiral blade 5 as shown in FIG.
It is also possible to configure it with 5.

A plurality of outflow holes 16 are bored in the upper part of the fixed cylinder 13 at positions above the liquid level W in the tank,
As the axial flow impeller 15 rotates, as shown by the double arrow ■,
The pumped culture solution t, cells C, etc. flow out of the cylinder 13 from the outflow hole 16 (wet field) and fall onto the liquid surface W in the culture tank 11 as shown by the double arrow @. Along with this fall,
Air a above the liquid level W of the culture tank 11 becomes bubbles and is drawn into the culture liquid reservoir, and oxygen is supplied to the culture liquid t.

The culture solution t mixed with air bubbles in this way follows the circulation flow by the axial impeller 15 and passes through the lower end of the cylinder 13 and the curved upper surface of the bottom plate 12a, as shown by the double arrow ■. It flows into the inside of the cylinder 13.

On the other hand, the entire inside of the culture tank 11 is sterilized, and the air hood above the culture tank is also sterile. Therefore, cells C and the like in the culture solution are not contaminated with germs.

In addition, the water level difference between the outflow hole 16 at the top of the cylinder and the liquid level W of the culture solution is small, and the rotation speed of the axial flow impeller 15 for flowing out the culture solution etc. from the outflow hole 16 may be small. There is little damage to the cells C etc. due to the rotation of the impeller 15.

According to this embodiment, it is possible to simultaneously stir the inside of the culture tank 11 and supply oxygen by simply rotating the axial flow impeller 15, so cells, etc. can be cultured efficiently and easily with less power. .

In addition, if the amount of oxygen supplied is insufficient only by entraining air a by dropping the culture solution from the outflow hole 16 at the top of the cylinder 13 onto the liquid surface W, axial flow? Oxygen-containing air g, such as sterilized air, may be supplementarily mixed in through the air diffuser hole 17 provided below the pump 15.

FIG. 2 is a longitudinal cross-sectional view of a cell culture device showing a second embodiment of the present invention, and in the figure, the same reference numerals as those shown in FIG. 1 indicate the same or similar parts. do.

In this embodiment, a cylinder 23 with an outflow hole 26 facing downward at its upper end is attached to a rotating shaft 14 via a support disk 23a, and an axially long cylinder 23 located at the center of the culture tank 11 It rotates integrally with the outer periphery of the impeller 25. In the figure, 16 is a ventilation hole provided in the upper lid 12, and 17 is a sterilization filter provided in the middle of the ventilation hole 16.

According to this embodiment, the cylinder 23 rotates together with the axial flow impeller 25, so as shown in the first embodiment (FIG. 1)
As shown in FIG.
There is no chance of it getting caught in the gap between the inner surface of the body and destroying it.

In addition, since the axial flow impeller 25 is longer in the axial direction than in the first embodiment, the purpose can be achieved at a lower rotation speed.
Therefore, the degree of damage to cells etc. is further reduced.

FIG. 3 is a vertical cross-sectional view of a main part showing the v13 embodiment of the present invention, and in the figure, the same reference numerals as those shown in FIG. 2 indicate the same or similar parts.

In the figure, the point that the cylinder 23 rotates integrally with the outer circumference of an axially long axial flow impeller 25 located approximately at the center of the culture tank 11 is the same as in the second embodiment (FIG. 2). In this embodiment, stirring blades 35 are particularly provided on the outside of the rotating cylinder 23.

According to this embodiment, when the axial flow impeller 25 and the cylinder 23 rotate together as a result of the rotation of the rotating shaft 14, the liquid in the culture tank 11 flows into the cylinder 23 as shown by the double arrow ■○. Due to the stirring blades 35 provided on the outside of the outer cylinder 23 where a circulating flow flows from the outside to the inside, a pressing flow as shown by the double arrow O and a swirling flow as shown by the double arrow ■ are generated. Cells C, etc. that tend to stay near the bottom of the culture tank 11 can be caused to flow by the flow, and can be efficiently sucked into the axial flow impeller 25.

FIG. 4 is a longitudinal sectional view showing a fourth embodiment of the present invention, and in the figure, the same reference numerals as those shown in FIG. 2 indicate the same or similar parts.

In the figure, the point that the cylinder 23 rotates integrally with the outer circumference of an axially long axial flow impeller 25 located approximately at the center of the culture tank 11 is the same as in the second embodiment (FIG. 2). In this embodiment, the rotating shaft 44 that keeps the axial flow impeller 25 stationary is used.
is driven via a magnet cassolin 4P (electromagnetic joint).

That is, a thin plate (partition plate) 42 a made of stainless steel or other non-magnetic material is formed at the center of the bottom plate 42 of the culture tank 11 , and a thin plate 42 a is formed in the center of the bottom plate 42 of the culture tank 11 .
1, a rotating disk 41 directly connected to a rotating shaft 44 maintains a gap between its outer circumferential surface and the inner circumferential surface of a thick recess 42b of the bottom plate 42 formed on the outer circumferential portion of the thin plate 42a. It is inserted so that it can be inserted.

On the other hand, on the lower side of the thin plate 42a, that is, on the outside of the culture tank 11, a driving disk 43 in which a driving magnet 43a is embedded with a narrow gap is provided to face the driving disk 43, and the driving disk 43 has a driving shaft 43b. The drive source is connected to the drive source via the drive source.

A rotating disk 4 in which each driven or driving magnet is embedded, which is arranged opposite to each other with a gap between the thin plates 42a mentioned above.
1 and the drive disk 43 form a magnetic coupling. In the magnetic coupling,
In order to rotate the rotary disk 41 while maintaining a narrow gap with the thin plate 42L, the rotary disk 41 is supported by a fixed shaft 42cK implanted in the thin plate 42a.

In addition, as mentioned above, the rotating disk 41 arranged in the culture tank
In order to rotate the rotary disc 41 while maintaining a narrow gap with the thin plate 42a, a large number of radial grooves are formed on the surface of the rotating disk 41 facing the thin plate 42a, which are engraved in a direction that causes rotation in the direction of rotation. During operation, as the magnetically coupled rotating disk 41 rotates in the same direction as the W-shaped disk 43 rotates, dynamic pressure is generated by the fluid interposed between the spiral grooved surface and the thin plate 42a. It is also possible to cause the rotating disk 41 to float and support thrust by a pressure fluid film that generates the dynamic pressure. In the figure, 45 is the rotation circle u
This is a stirring blade provided on the top surface of i41.

According to this embodiment, the axial flow impeller 25 is driven via a magnetic coupling, and the rotating shaft 44 is connected to the culture tank 11.
Since the shaft does not penetrate through the bottom plate 42 of the shaft, a shaft seal is unnecessary.

Therefore, there is no risk of bacteria entering into the culture tank 11 from the shaft sealing part and contaminating the cells C, etc. By the rotation of the stirring blade 45 provided on the upper surface of the rotating disk 41, A flow as shown is generated, and there is no fear that the cells C and the like will stay at the bottom of the culture tank 11.

FIG. 6 is a longitudinal sectional view showing an example of the use of the culture device of the present invention, and in the figure, the same reference numerals as those shown in FIG. 2 indicate the same or similar parts.

In this usage example, an axial flow impeller 25 that rotates together with a plurality of cylinders 23 (three in the figure) is arranged in a large culture tank 61 so as to be driven by the rotating shaft 14.

According to this example of use, the action of the plurality of axial flow impellers effectively supplies oxygen to the culture solution in a large culture tank and agitates the culture solution, cells, etc.

In the above usage example, an axial flow impeller and a cylinder that rotate together as shown in the second implementation row (Fig. 2) are used, but other implementations described above are also used. Of course, it is also possible to use a plurality of culture devices (units) of the type described in the example in parallel.

〔Effect of the invention〕

As explained above, according to the present invention, a cylinder is provided inside the culture tank, and an axial flow impeller is provided inside the cylinder, and by rotating the impeller, the culture solution, cells, etc. are caused to flow out from the top of the cylinder. By allowing the liquid to fall onto the surface of the culture liquid outside the cylinder, it is possible to stir the culture tank and supply oxygen at the same time just by rotating the axial flow impeller, which is efficient and informative with less power. Cells, etc. can be cultured in stool.

[Brief explanation of the drawing]

1 to 4 are longitudinal cross-sectional views of a culture device showing the first to fourth embodiments of the present invention, FIG. 5 is a cross-sectional view of main parts, and FIG. 6 is a longitudinal cross-sectional view showing an example of use of the present invention. Front view, Figures 7 and 8
The figure is an explanatory diagram showing a conventional example. 11...Culture tank, 13.23...Cylinder, 1
4゜44... Rotating shaft, 15.25.55... Axial flow impeller, 16.26... Outflow hole, 35.4
5... Stirring blade - a... Air, b... Bubbles,
C...Cells, g...Air, t...Culture solution. Figure 1 Figure 5 Figure 6

Claims (1)

[Claims] 1. A cylinder is provided inside the culture tank with its upper part protruding from the surface of the culture solution, an axial flow impeller is provided inside the cylinder, and the impeller is rotated. The culture solution and cells, etc. are allowed to flow out from the top of the cylinder and fall onto the surface of the culture solution outside the cylinder, thereby drawing the oxygen-containing gas from the top of the culture tank into the culture solution and supplying oxygen to the culture solution. A culture device using an axial flow impeller, characterized in that the culture solution, cells, etc. are agitated at the same time. 2. A culture device using an axial flow impeller according to claim 1, wherein the cylinder is fixed. 3. A culture device using an axial flow impeller according to claim 1, wherein the cylinder is integrated with the outer periphery of the axial flow impeller and rotated together with the impeller. 4. A culture apparatus using an axial flow impeller according to claim 3, wherein stirring means is provided on the outside of the cylinder that rotates together with the axial flow impeller. 5. The axial flow impeller described in any one of claims 1 to 3, wherein the axial flow impeller is driven by a magnetic coupling provided below the axial flow impeller. A culture device using an axial flow impeller. 6. A culture device using an axial flow impeller according to claim 5, wherein stirring means is provided on a rotating disk that covers the driven magnet of the magnetic coupling.