JPH0240315B2 - KEISHASHITARASENHANEGURUMAOMOCHIITABAIYOSOCHI - Google Patents

Abstract

PURPOSE:To obtain a culture apparatus capable of stirring a culture liquid in high efficiency and supplying oxygen, by covering a spiral blade fixed to a rotary shaft with a cylinder to obtain a stirrer and placing the stirrer slantly in an inclined culture tank. CONSTITUTION:A culture tank 21 is placed in inclined state. A stirrer is produced by fixing a spiral blade 24 to a rotary shaft 22 and covering the blade with a cylinder 25 opened at both ends. The stirrer is placed in the culture tank in inclined state. The rotary shaft 22 is slowly rotated in the direction to lift the culture liquid and the liquid is fallen from the upper end of the cylinder 25. An oxygen-containing gas (a) in the culture tank is mixed into the falling culture liquid.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to an apparatus for cultivating animal and plant cells and microorganisms such as bacteria, and the present invention relates to an apparatus for culturing microorganisms such as animal and plant cells and bacteria, and the present invention relates to an apparatus for culturing microorganisms such as animal and plant cells and bacteria. The present invention relates to a culture device that can supply oxygen to the cells and agitate the culture solution.

[Conventional technology and problems]

In order to cultivate animal and plant cells and microorganisms such as bacteria using the suspension culture method, oxygen is supplied to the cells through a culture medium, and the culture medium and cells are mixed and stirred to facilitate mass transfer of metabolic products. need to be promoted.

Conventionally, for example, a culture apparatus as shown in FIG.
It was mixed. In addition, in the figure, 3 is a culture tank (container),
Reference numeral 4 indicates a baffle plate protruding from the inner wall surface in order to enhance the stirring effect of the agitator 2, and g indicates air sent into the aeration tube 1.

However, the above-mentioned conventional apparatus has the disadvantage that cells and the like are easily damaged by the rotation of the stirrer, and the culture efficiency is low. In particular, when the device is enlarged, the speed at the tip of the stirring blade increases even if the rotation speed is the same, which increases the probability of damaging cells, etc., which is not preferable. Furthermore, 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 flow resistance and the water pressure of the culture medium and pass a large amount of air. There was a problem in that the impeller for the engine also required separate power.

In addition, as shown in FIG. 4, air g is sent from below the culture tank 3 to generate air bubbles b in the tank,
In a culture device in which the culture solution L is fluidized and stirred only by the bubbles b, the culture progresses and the culture solution L
When the viscosity of the air increases, a sufficient stirring effect cannot be obtained and the power required to pump the air becomes even more excessive.

Therefore, in order to address the above-mentioned problems, the present applicant previously applied for a culture apparatus using an inclined helical impeller in Japanese Patent Application No. 62-248075. This thing is
As shown in FIG. 5, a cylinder 15 with both ends open is installed at an angle inside the culture tank 11.
The lower open end 15b of the cylinder is immersed in the culture solution L, the upper open end 15a is positioned above the culture solution surface W, and a spiral blade 14 is provided over the entire length inside the cylinder. By slowly rotating the cylinder 14 and the cylinder 15 if necessary, the culture solution is transferred from the bottom to the top of the cylinder 15, and the culture solution is allowed to fall from the upper open end 15a of the cylinder to the liquid surface W. The oxygen-containing gas a above the culture solution is drawn into the culture solution L to supply oxygen to the culture solution, and the culture solution and cells are mixed by stirring while moving in the direction of arrow d. be. In the figure, 12 is a rotating shaft, 13 is a bearing, 1
6 is a lid, 17 is a sealing member, and l ₁ to l ₅ are
Each segment of liquid has a free surface in the area surrounded by the spiral vane 14 and the inner surface of the cylinder in the cylinder 15, and during operation, the liquid is raised (pumped) sequentially by the pump principle called Archimedes pump. It shows.

However, the above method also had the following problems. In (i), the blades 14 etc. are installed diagonally with respect to the culture tank 11, so the entire culture tank becomes large, and the total amount of culture is large compared to the flow rate circulated per unit time by the impeller, making it more efficient. It is not possible to perform good stirring. In addition, (ii) the culture tank 11 needs to be sterilized sequentially to prevent contamination with bacteria;
The culture tank is larger than necessary, making sterilization difficult. Furthermore, (iii) it is structurally difficult to adjust the optimum inclination angle of the blades.

The technical object of the present invention is to provide a culture device that supplies oxygen while efficiently agitating animal and plant cells and bacteria with a culture solution, does not damage the cells, and can miniaturize the entire culture tank. There is.

[Means for solving problems]

In order to solve the problems and technical problems of the prior art described above, the present invention has provided a spiral blade and a cylinder provided on the outer periphery of the blade with both ends open, facing the side wall of the culture tank, that is, the cylinder. The culture tank is supported along with the blades and the cylinder in a tilted manner with respect to the vertical direction, and when the culture tank is tilted, at least one of the lower ends of the blades and the cylinder is supported. The part is immersed in the culture solution, and its upper end is placed in the oxygen-containing gas above the culture solution, and in this state, at least the spiral blade and, if necessary, the cylinder are rotated to remove the culture solution. It is characterized in that the liquid is pumped up and dropped onto the surface of the culture liquid from the upper end of the cylinder.

In addition, in implementation, it is desirable to make the inclination angle of the culture tank variable.

[Effect]

Since the present invention is configured as described above, in a culture tank supported at an angle with respect to the vertical direction,
When the culture solution is poured into the spiral blade and the cylinder provided on its outer periphery until at least half of both lower ends are submerged in the culture solution surface, the culture solution will flow inside the cylinder and out of the cylinder. Approximately the same level is reached and within the cylinder the culture medium has a free surface in the region bounded by the spiral vanes and the inner surface of the cylinder.

Next, when the spiral blade is rotated in this state, each culture solution having a free surface divided by the spiral blade and the slanted cylindrical inner surface as described above is sequentially rotated while maintaining the free surface. It rises along the bottom of the inclined cylinder.

The above action is based on the principle of a so-called Archimedes pump.

At this time, in order to prevent the culture solution from flowing downward and backward through the gap between the inner surface of the cylinder and the circumferential surface of the spiral blade, it is recommended to tightly fit the spiral blade inside the cylinder and rotate them together. Most desirably, when the spiral blade is rotated within a stationary cylinder, the gap between the two must be made as narrow as possible.

The culture solution transferred from the bottom to the top of the cylinder as described above falls below the liquid surface of the culture tank from the upper open end of the cylinder, but at this time, the oxygen-containing gas present above the liquid surface of the culture tank As it falls into the culture solution, oxygen is supplied to the culture solution, and
The culture solution, cells, etc. are stirred.

The culture solution that has fallen as described above moves inside the culture tank toward the lower open end of the cylinder, and the culture solution in the entire culture tank also flows into the cylinder again while being stirred, and the above-described action is repeated.

As mentioned above, it is sufficient that at least half of both the lower end surfaces of the spiral blade and the cylinder are submerged in the culture solution surface, and the area near the drop point of the culture solution falling from the top end of the cylinder is , there must be a culture solution surface.

〔Example〕

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

FIG. 1 is a longitudinal sectional view of a culture apparatus shown in a first embodiment of the present invention, and in the figure, the same reference numerals as those shown in FIG. 5 indicate the same or similar parts.

In the figure, in a culture tank 21, there is a rotating shaft 22 supported by a bearing 23, a spiral blade 24 attached to the rotating shaft 22, and a gap on the outside (outer periphery) of the blade 24 over almost the entire length. The culture tank 21 is a cylinder 25 with both ends open, which is tightly fitted.
The spiral blades 24 and the cylinder 25 are arranged in parallel with the side wall 21a facing the cylinder 25, and the spiral blades 24 and the cylinder 25 are slowly rotated together by the rotating shaft 22. .

The culture tank 21 has a lid 26 on one side (the right side in the figure).
It is fixedly supported on a stand 29 via a grip 28 whose inclination angle can be changed with respect to the vertical direction.

Further, the rotating shaft 22 is connected to a power cable 31 driven by a motor (not shown) via a magnetic coupling 30 in order to prevent the spiral blades 24 from being contaminated by germs. ing.

The magnetic coupling 30 is connected to the inner side of a bowl-shaped drive member 30a which is connected to a power cable 31 and has a magnet _M1 attached to the inner surface thereof, and is connected to the outer periphery via a partition member 30b made of a non-magnetic material such as stainless steel. It is configured by arranging a driven member 30c to which magnet _M2 is attached, and both magnets
Due to the mutual magnetic action of M ₁ and M ₂ , both the driving and driven members 30a and 30c are magnetically coupled to transmit power (torque). In addition,
In the figure, 27 is a sealing member, and 32 is a sensor insertion position.

Next, the effect will be explained.

When the culture tank 21 is tilted by the grip 28 as shown, the internal spiral blades 24 and cylinder 25 are also tilted together.

When the rotating shaft 22 is in a stopped state, the culture solution l enters the lower open end 25b of the cylinder 25 and enters a plurality of spaces l ₁ ~ surrounded by the spiral blade 24 and the cylinder 25.
It is divided into ₃ parts, all of which maintain the same liquid level (free surface) W.

Next, the external motor connects the rotating shaft 22 via the power cable 31 and the magnetic coupling 30.
When the spiral blades 24 and the outer tube 25 slowly rotate together, each culture solution divided by the spiral blades 24 as described above is
The divided liquid _l3 , which was gradually moved upward along the bottom surface of the inclined cylinder 25 and was at the top when it stopped, is successively moved upward like _l4 , _l5 , etc. while maintaining its free surface. , and eventually reaches the upper open end 25a of the cylinder 25. From the open end 25a, the culture tank 2
1 flows out into the oxygen-containing gas a above the liquid level W and falls into the culture solution l.

As it falls, the oxygen-containing gas a is drawn into the culture solution l, supplies oxygen to the culture solution l, and
The culture solution 1, cells c, etc. are mixed by stirring. Also,
The fallen culture solution 1 moves toward the lower open end 25b of the cylinder as shown by the arrow d, and the culture solution 1 in the entire culture tank 21 is also stirred, and the culture solution 1 is transferred to the cylinder 2 again.
5.

According to this embodiment, the spiral blade 24 and the cylinder 25 are fitted without any gaps and are rotated as one, so that the liquid is pumped efficiently without backflowing downward within the cylinder 25. In addition, in principle, the blades perform their functions at extremely slow rotation speeds, so the blades do not damage soft cells, etc.
By rotating the inclined cylinder 25, the lower surface of the cylinder 25 is mixed with the culture solution l and the oxygen-containing gas a.
The oxygen supply effect is promoted because the oxygen supply layer alternately contacts the inside and outside.

In addition, since the spiral blade 24, the cylinder 25, etc. are configured to be able to maintain an arbitrary inclination angle together with the culture tank 21, the liquid level W of the culture solution l
Also, the spiral blade 24 can have an arbitrary inclination angle. Therefore, it is possible to operate at an optimal inclination angle depending on the amount of culture solution, the amount of aeration required, etc.

Furthermore, since the rotating shaft 22 and the spiral blades 24 are driven via a magnetic coupling 30 which is isolated from the outside air by the partition wall 30b, the culture tank 21 is isolated from the outside air and is driven by the magnetic coupling 30. It is possible to prevent contamination caused by various bacteria. Furthermore, since the rotating shaft 22 is driven by an external motor via a power cable 31,
It is possible to absorb relative position changes with the external motor.

FIG. 2 is a longitudinal sectional view of a culture apparatus 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.

In this embodiment, the upper open end 25a of the cylinder 25
Inside the lid 26 facing the inner cylinder 25
A guide plate 35 is attached to the side, and controls the direction of the flow flowing from the upper open end 25a of the cylinder toward the liquid surface W, and directs the flowing (falling) culture solution to a deeper liquid depth. By reaching the position, more oxygen-containing gas a is drawn into the culture solution. Also, a window (opening) is provided at the bottom of the cylinder 25 slightly below the upper open end 25a.
36 are drilled.

According to this embodiment, the guide plate 35 is connected to the cylinder 25.
Since it is provided below the upper open end 25a so as to face the upper open end 25a, the culture solution that has flowed out and fallen from the upper open end 25a of the cylinder is transferred to a position near the center of the culture tank 211 with a deeper liquid depth. By reaching this point, more oxygen-containing gas a can be drawn into the culture solution.

Further, since a window 36 is provided slightly below the upper end of the cylinder 25, a part of the culture solution is allowed to fall from the window 36 onto the liquid surface W, and the oxygen-containing gas a is drawn into the culture solution over a wider range. can be set.

Note that the guide plate 35 described above may be formed into a conical plate shape, provided on the rotating shaft 22, and rotated together with the spiral blade 24. Furthermore, by providing a flange-shaped guide plate (not shown) on the outside of the upper open end 25a of the cylinder 25, it is possible to prevent the culture solution from flowing along the outer surface of the cylinder 25 from the upper open end 25a. .

In both the first and second embodiments described above, the structure in which the cylinder 25 and the spiral blade 24 are rotated as one body has been described. You may also do so. In that case, it is necessary to make the gap between the cylinder 25 and the blade 24 as narrow as possible.

In addition, the rotating shaft 22 is connected to the magnetic coupling 3.
Although a structure has been described in which the shaft is driven through a shaft 0, it is also possible to drive the shaft through a rotary shaft that passes through a shaft sealing device as usual.

〔Effect of the invention〕

As explained above, according to the present invention, the following effects are achieved.

(i) A spiral blade and a cylinder provided on the outer periphery of the blade are installed parallel to the side wall of the culture tank, and the culture tank is tilted with respect to the vertical direction together with the blades and the cylinder. The entire culture tank can be downsized compared to the previous application of the present applicant in which spiral blades and the like are provided at an angle to the culture tank. Therefore, efficient stirring can be performed and the sterilization work inside the culture tank becomes easy. Furthermore, by changing the inclination angle of the culture tank, it becomes easy to adjust the inclination angle of the spiral blade with respect to the liquid level depending on the situation and to pump the liquid efficiently.

(ii) By tilting the culture tank, both the lower ends of the spiral blade and the cylinder are immersed in the culture solution, and the upper ends are placed in the oxygen-containing gas. It functions at an extremely slow rotational speed, so the blades do not damage soft cells. Moreover, since the surface of the lower part of the cylinder comes into contact with the culture solution and the oxygen-containing gas alternately, the supply of oxygen is promoted.

[Brief explanation of drawings]

1 and 2 are longitudinal sectional views of the culture apparatus showing the first and second embodiments of the present invention, FIGS. 3 and 4 are explanatory views showing the conventional example, and FIG. FIG. 2 is a longitudinal cross-sectional view of a culture device according to the applicant's earlier application. 21... Culture tank, 21a... Side wall, 22... Rotating shaft, 24... Spiral blade, 25... Cylinder, 2
5a... Upper open end, 25b... Lower open end, 2
6... Lid, 28... Grip, 29... Stud, 30... Magnetic coupling, 31...
Power cable, 35... Guide plate, 36...
Window, a...Oxygen-containing gas, b...Bubbles, c...Cells, l...Culture solution.

Claims (1)

[Claims] 1. A spiral blade and a cylinder provided on the outer periphery of the blade and open at both ends are provided parallel to the side wall of a culture tank, and the culture tank is tilted with respect to the vertical direction together with the blade and cylinder. with the culture tank tilted, at least a portion of both the lower ends of the blade and the cylinder are immersed in the culture solution, and the upper ends are positioned in the oxygen-containing gas above the culture solution, In this state, at least the above-mentioned spiral blade is rotated to lift the culture solution and drop it from the upper end of the cylinder onto the surface of the culture solution, thereby entraining oxygen-containing gas into the culture solution in the culture tank. 1. A culture device using an inclined helical impeller, characterized in that the culture solution is supplied with oxygen and agitated at the same time. 2. A culture device using an inclined helical impeller according to claim 1, wherein the inclination angle of the culture tank is variable. 3. Culture using the inclined helical impeller according to claim 1, wherein the culture solution is guided and controlled by a guide member in the outflow direction from the upper end of the cylinder and falls onto the surface of the culture solution. Device. 4. A culture device using an inclined helical impeller according to claim 1 or 3, wherein a window is provided at the bottom of the cylinder slightly below the upper open end. 5. A culture device using an inclined helical impeller according to any one of claims 1 to 4, wherein the cylinder is rotated together with the helical impeller.