JPH0622745A - Device for culture - Google Patents

Abstract

PURPOSE:To obtain a culture device having high culture efficiency capable of readily carrying out inoculation change operation of culture organism. CONSTITUTION:Organism retainers 20 to hold a culture organism are made into a basket shape and a culture solution is made to freely go in and out baskets. A culture organism is stored in the organism retainers 20 and the organism retainers are fixed to an inner wall of a culture solution tank with retaining members 12. The culture solution tank 10 is revolved around an approximately horizontal shaft. Oxygen can be efficiently fed to the culture organisms without reducing the culture solution and change of inoculation of culture organisms is readily carried out.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a culture device which has a high culture efficiency and can be easily replanted.

[0002]

2. Description of the Related Art 1970
Biotechnology that greatly advanced in the 1980s and 1980s
Now, in the 90's, the achievements are being put to practical use in many fields. In this way, it becomes important to put into practical use the technology that was carried out only in the laboratory.
Device ”.

A culture device is one of such "devices". The culture device is a device for breeding and culturing organisms while precisely controlling culture conditions. Conventionally, for example, the products shown in FIGS. 12 and 13 have been used.

FIG. 12 shows a rotary culture type culture device. In the culture device, a test tube having heat resistance and chemical resistance such as glass, a flask or the like (hereinafter, simply referred to as “bottle”) 140 is filled with a liquid medium and a culture organism, and the culture medium is opened in a cylindrical support member 142. Hole 1
Insert in 44 and fix. Then, the support member 142 is rotated by the motor 146 or the like in a state in which it is inclined. By rotating in such an inclined state, it is possible to eliminate the direction of growth that may be caused by light or gravity. Along with this, the culture solution can be stirred. Needless to say, the entire bottle is sterilized with an autoclave or the like before the culture is started. In addition, air free of miscellaneous bacteria can be injected into the bottle as needed.

As another type, there is a jafermentor type culture device shown in FIG. The culture device is
The culture solution is agitated by a so-called air lift system in which air is delivered from the bottom of the culture tank. This method has an advantage of not damaging the cultured organism unlike the method of stirring using a propeller or the like.

[0006]

However, each of the above-mentioned conventional culture devices has the following drawbacks.

In the rotary culture type shown in FIG. 12, it is difficult to increase the size of the device. That is, when the culture medium became large, the culture solution could not be sufficiently stirred.
In addition, since the inside of each bottle is sealed, the amount of medium must be reduced in order to supply sufficient oxygen to the cultured organism. However, if the amount of medium is reduced, it is necessary to reduce the amount of organisms to be cultivated, resulting in poor culturing efficiency. There is also a type in which air is forcibly sent, but this is not preferable because the device becomes more complicated and larger than necessary.

Also for the jar-famentor type shown in FIG. 13, it was necessary to reduce the amount of medium in order to supply sufficient air to the culture.

Further, in any of the above-mentioned methods, it is necessary to take out the cultured organisms one by one at the time of transplantation accompanying the growth of the cultured organisms.

Further, in the above conventional apparatus, since the cultured organism is always in the culture solution, it is always in contact with the waste products generated during the culture. Therefore, it may be unfavorable for the growth of the cultured organism.

The present invention solves the above-mentioned problems, and provides a culture device which is easy to replant a culture organism and which can supply sufficient air to the culture organism without increasing the size of the device. The purpose is to

[0012]

Means for Solving the Problems The present invention has been made to achieve the above object, and in one aspect thereof, a culture solution tank for holding a culture solution, a culture organism holding means for holding a culture organism, The culture organism held in the culture organism holding means has a moving means for alternately moving between the culture solution existing in the culture solution tank and the gas phase existing in the culture solution tank. A culture device is provided.

In this case, the moving means changes the relative positional relationship between the culture solution in the culture solution tank and the culture organism held by the culture organism holding means, and performs movement including at least a vertical component. It is preferable that it is made possible.

The moving means comprises a rotation driving means for rotating the culture solution tank, and a locking means for locking the culture organism holding means in the culture solution tank. Good.

It is preferable that the axis of rotation by the rotation driving means is substantially horizontal.

The above-mentioned culture organism holding means is a tank having a hole through which the culture liquid passes, and the size of the hole may be such that the culture organism cannot pass through. Alternatively, at least a part thereof may be composed of a mesh member.

[0017]

The culture organism is held by the culture organism holding means constituted by a net-like member such as a basket. And
The culture organism holding means is locked to the culture solution tank by the locking means. In this case, not only the culture solution but also a gas such as air exists in the culture solution tank. Then, as it is, it is rotated about the substantially horizontal axis by the rotation driving means.

[0018]

An embodiment of the present invention will be described with reference to the drawings.

As shown in FIG. 1, the culture apparatus of this embodiment is mainly composed of a culture section 1 and a pedestal 8.

The pedestal 8 has support pedestals 9 on both ends of its upper surface. Then, by the shaft provided on the support base 9,
It supports recesses 17 provided at both ends of the culture unit 1 which will be described later. The pedestal 8 is not shown in the figure,
Has a drive mechanism consisting of a printer, pulley, belt, etc.
The driving mechanism is configured to rotate the culture section 1 about the above axis.

As shown in FIGS. 2 and 3, the culture section 1 is mainly composed of a cylindrical culture solution tank 10 and a living organism holder 20 arranged in the culture solution tank 10.

The culture solution tank 10 is a tank for holding the culture solution. The shape is cylindrical as described above, and a removable lid 15 is provided on one bottom surface thereof. An O-ring or the like is arranged around the lid 15 so that the culture solution in the culture solution tank 10 does not leak outside. The above-described recess 17 is provided at the center of the lid 15 and at the center of the other bottom surface. The culture section 1 is configured to be kept substantially horizontal by supporting the recess 17 by the support base 9 and to be rotated by the force received in the recess 17.

The biological holder 2 is provided on the inner wall of the culture solution tank 10.
Locking members 12 for holding 0 are provided at various places. As shown in FIG. 4, the locking member 12 has arcuate claws having a curvature that substantially matches the outer peripheral curved surface of the biological holder 20, and the claws hold the biological holder 20 between them. This is a fixed structure (see FIGS. 2 and 3). The locking member 12 is fixed to the inner wall of the culture medium tank 10 by screws 120, washers 121, O-rings 122, etc., as shown in FIG. Therefore, the culture solution tank 10
Since it does not have a protrusion itself, the culture solution tank 10 can be easily and surely washed. However, the shape and the fixing method of the locking member 12 are not limited to this, and the biological holder 20 in the culture solution tank 10 is not limited thereto.
There is no limitation as long as it can be fixed. For example, a suction cup may be provided in the biological holder 20 so that the biological holder 20 can be adsorbed to the inner wall of the culture solution tank 10. In this case, it is preferable that the portion for sucking the suction cup is flat. Further, in this embodiment, two biological holders 20 can be attached at one time, but a larger number or only one biological holder 20 may be attached.

The shape of the culture solution tank 10 is not particularly limited as long as the culture solution does not leak outside.
Also, the material used is glass in the present embodiment, but is not particularly limited as long as it does not react with a chemical substance that may be used in a culture experiment.

The organism holder 20 is for accommodating cultured organisms. As shown in FIG. 6, the biological holder 20 is a cylindrical basket made of stainless steel, and the culture solution can easily enter and leave between the inside and the outside of the biological holder 20. The size of the cage eye is smaller than that of the cultured organism, and the cultured organism contained in the organism holder 20 does not go out of the organism holder 20. In addition, the culture organism is taken in and out by opening the lid 24 provided on the bottom surface thereof.

The specific shape of the biological holder 20 and the method for holding the cultured organisms are not limited to this. For example, if the cultured organism is relatively large, the mechanism may be such that it sandwiches and holds it.

The operation of the culture device will be described.

First, the lid 15 is opened and the culture solution is put into the culture solution tank 10. In this case, the amount of the culture solution is set so that a certain amount of space, that is, air remains in the culture solution tank 10 even when the biological holder 20 is attached later. continue,
The lid 24 is opened to store the cultured organism in the organism holder 20.
Then, after that, this time, the biological holder 20 is replaced with the culture solution tank 1.
It is put in 0 and fixed by the locking member 12. Then, the lid 15 is used for sealing. The procedure of storing the culture solution and the like up to this point is not limited to this.

Then, after that, the recess 17 of the culture solution tank 10 is supported by the support base 9 of the pedestal 8. And the culture solution tank 10
Rotate the whole. The state inside the culture solution tank 10 at this time is shown in FIG.

In the culture solution tank 10, of course,
The culture solution x is on the lower side, while the air a remaining in the culture solution tank 10 is on the upper side. Since the position and the like of the culture solution x are not particularly limited in the culture solution tank 10, this state is substantially maintained regardless of the rotation of the culture solution tank 10. On the other hand, since the biological holder 20 is fixed to the inner wall of the culture solution tank 10, its position moves with the rotation of the culture solution tank 10 (when viewed from the outside). In this case, since the culture solution tank 10 has its axis of rotation substantially horizontal, the biological holder 20 moves so as to draw a circular trajectory in a certain vertical plane. Therefore, with the movement, the biological holder 2
The cultured organism z contained in 0 has the opportunity to come into contact with both the culture solution x and the air a. in this case,
By changing the rotation speed of the culture solution tank 10, it is possible to adjust the time of contact with the culture solution x or the air a.
The nutrients and oxygen can be sufficiently supplied without reducing the amount of the culture organism z. Further, the culture medium x is sufficiently stirred by the biological holder 20.

When the culture is carried out for a long period of time, it is necessary to replace the culture solution x with a new one halfway.
In this case, another culture solution tank 1 containing a new culture solution x
By preparing 0 and transferring the cultured organism z together with the organism holder 20, the work can be completed easily and in a short time. In this case, since the cultured organism z is not exposed to the air for a long time, the influence of the work on the cultured organism z is small. Moreover, since the cultured organism z is not grasped one by one with tweezers or the like, there is no possibility of damaging the cultured organism z.

Similar to the conventional rotary culture type, the irradiation direction of light and the direction in which gravity acts can be made uniform.

Another embodiment will be described with reference to FIG. This embodiment is characterized in that the stirring effect is further enhanced by rotating the culture solution tank 10 and changing the angle of the rotating shaft thereof.

In the present embodiment, a rotary shaft 18 is provided on the culture solution tank 10 side. The rotary shaft 18 is supported by a guide base 92, a support base 94 and the like.

The support base 94 has a built-in motor and generates a driving force for rotating the culture solution tank 10. Also, the connection between the motor shaft 940 and the rotary shaft 18b is free.
The joint 96 is used so that the angle of the culture solution tank 10 can be changed.

The guide base 92 has a vertical groove 920 in the center thereof. And here, the rotary shaft 18
A bearing member 924 connected to a is arranged so as to be movable in the vertical direction.

A motor 9 is provided below the guide base 92.
A rotary plate 928 that is rotated by 6 is provided. The rotating plate 928 is connected to the bearing member 924 and the connecting arm 92.
6 are connected to form a crank. That is,
As the rotary plate 928 rotates, the bearing member 924 reciprocates in the vertical direction, and the angle of the culture solution tank 10 changes. In this case, when the bearing member 924 is at the uppermost position, the side of the rotary shaft 18a is on the rotary shaft 1 side.
8b side, and conversely, when the bearing member 924 is at the lowest position, the rotating shaft 18a side is on the rotating shaft 18a side.
It should be lower than the b side. As a result, it is possible to prevent a situation in which only the cultured organisms on either side of the rotating shaft 18a or the rotating shaft 18b are in contact with air and the cultured organisms on the other side cannot contact the air.

In this embodiment, not only the culture solution tank 10 rotates, but also its angle changes, so that stirring can be performed more effectively. This effect is particularly effective when the culture solution held in the culture solution tank 10 is increased.

Another embodiment will be described with reference to FIG. This embodiment is characterized by a driving method for rotating the culture section 1.

A drive roller 30 and a support roller 32 are provided on the pedestal 8 '. Both of these have their axis of rotation in the horizontal direction. The rotation axes of both are parallel. The driving roller 30 rotates by itself by receiving a driving force from a motor or the like not shown in this figure. On the other hand, the support roller 32 does not receive a driving force by a motor or the like, and is simply configured to be rotatable.

During the culture, the culture solution tank 10 is placed on the two drive rollers 30 and the support roller 32, and the drive roller 30 is rotated by the motor. The culture fluid tank 10 is also rotated by the force received from the drive roller 30. At this time, the support roller 32 itself does not rotate spontaneously, but since the support roller 32 is also rotated by the force received from the culture solution tank 10, the rotation of the culture solution tank 10 is not hindered. .

Although only two rollers are provided in this embodiment, if the number of rollers is further increased, one driving roller 30 can be used to provide a larger number of culture solution tanks 10.
Can be rotated. Therefore, the cost of the device can be reduced. Further, the rotation axes of the drive roller 30 and the support roller 32 do not necessarily have to be horizontal,
Similar to the embodiment shown in FIG. 8, the culture organism z in the organism holder 20 may have an inclination as long as it can be exposed to air.

Another embodiment will be described with reference to FIG.

This embodiment is characterized in that the culture medium can be exchanged without taking out the biological holder 20 from the culture medium tank 10 and without interrupting the culture. In addition,
Since the basic configurations of the culture solution tank 10 and the biological holder 20 of this embodiment are the same as those of the above embodiment, FIG. 10 is a schematic diagram in which only the points that the embodiment is different from the above embodiment are extracted. I am trying.

The culture solution tank 10 has a discharge pipe 43 which also serves as a rotating shaft at one end thereof. The discharge pipe 43 is provided with a discharge cock 40 and a discharge cock 42.

On the other hand, an injection tube 47 having a sterilizing filter 48 is provided at the other end of the culture solution tank 10. The injection tube 47 is connected to the culture solution bottle 44 via a free joint 46. Like the discharge pipe 43,
The injection tube 47 or the sterilizing filter 48 also serves as the rotating shaft of the culture solution tank 10.

By adopting such a constitution, during culturing,
The culture solution can be exchanged without stopping the rotation of the culture solution tank 10. In this case, due to the presence of the sterilization filter 48, the discharge cock 40, and the discharge cock 42, the culture solution can be injected and discharged outside the clean bench. Further, since the culture solution bottle 44 and the sterilizing filter 48 are connected by the free joint 46, the culture solution bottle 44
The connection with does not hinder the rotation of the culture solution tank 10.

Another embodiment is shown in FIG.

In the present embodiment, a tank 54 for storing a culture solution is provided separately from the culture solution tank 10, and an injection pump 50, between the tank 54 and the culture solution tank 10, is provided. The discharge pump 52 circulates the culture solution.
The injection pipe 47 and the discharge pipe 49 also serve as rotation axes, and the free joint 46a and the free joint 46b are provided so as not to hinder the rotation of the culture solution tank 10. This is similar to the embodiment shown in FIG. In addition, the same applies to the fact that sterilizing filters 48 are provided on the inflow side and the outflow side of the culture solution into the culture solution tank 10 to prevent invasion of various bacteria.

In this embodiment, since the amount of culture solution can be increased, the frequency of culture solution exchange can be reduced.

[0051]

As described above, according to the culture apparatus of the present invention, the amount of air supplied to the culture apparatus can be increased without increasing the size of the apparatus. In addition, replanting work is easy, and the possibility of contamination by various bacteria is reduced.

[Brief description of drawings]

FIG. 1 is a perspective view showing an embodiment of the present invention.

FIG. 2 is a perspective view showing details of the culture unit 1.

FIG. 3 is an explanatory diagram showing an arrangement state of biological holders 20 in the culture solution tank 10.

FIG. 4 is a perspective view showing an internal configuration of the culture solution tank 10.

FIG. 5 is an explanatory view showing a mounting structure of a locking member 12.

FIG. 6 is a perspective view showing a configuration of a biological holder 20.

FIG. 7 is an explanatory diagram showing a state in the culture solution tank 10 during culturing.

FIG. 8 is a perspective view showing another embodiment of the present invention.

FIG. 9 is an explanatory diagram showing another embodiment of the present invention.

FIG. 10 is a schematic view showing another embodiment of the present invention.

FIG. 11 is a schematic view showing another embodiment of the present invention.

FIG. 12 is a perspective view showing a conventional technique.

FIG. 13 is a perspective view showing a conventional technique.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 1 ... Culture part, 8 ... Pedestal, 9 ... Support stand, 10 ... Culture solution tank, 12 ... Locking member, 15 ... Lid, 17 ... Recessed part,
20 ... Biological cage, 24 ... Lid, 30 ... Drive roll
La, 32 ... Support roller, 40 ... Discharge cock, 42
... discharge cock, 43 ... discharge pipe, 44 ... culture solution bottle,
46 ... Free joint, 47 ... Injection tube, 48 ...
Sterilization filter, 49 ... Discharge pipe, 50 ... Infusion pump,
52 ... Discharge pump, 54 ... Tank, 120 ... Screw, 121 ... Washer, 122 ... O-ring, 12
3 ... washers, 124 ... nuts, 130 ... pumps,
132 ... Plating filter, 134 ... Culture solution tank,
136 ... Air vent hole, 140 ... Bottle, 142 ... Support member, 144 ... Hole, 146 ... Motor, x ... Culture solution, z ... Culture organism

Claims (6)

[Claims] 1. A culture solution tank for holding a culture solution, a culture organism holding means for holding a culture organism, and a culture solution for holding the culture organism retained by the culture organism holding means in the culture solution tank. And a moving means for moving between the gas phase existing in the culture solution tank and the culture solution tank. 2. The moving means changes the relative positional relationship between the culture solution in the culture solution tank and the culture organism held by the culture organism holding means, and performs movement including at least a vertical component. The culture apparatus according to claim 1, wherein the culture apparatus is a culture apparatus. 3. The moving means is configured to include a rotation driving means for rotating the culture solution tank, and a locking means for locking the culture organism holding means in the culture solution tank. The culture device according to claim 1, which is characterized in that. 4. The culture apparatus according to claim 2, wherein the axis of rotation by the rotation driving means is substantially horizontal. 5. The culture according to claim 1, wherein the culture organism holding means is a tank having a hole through which the culture solution passes, and the size of the hole is such that the culture organism cannot pass through. apparatus. 6. The culture apparatus according to claim 1, wherein at least a part of the culture organism holding means is constituted by a mesh member.