JP2002101867A - Culture container - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a culture container which is inexpensive, is easily handleable, has excellent ventilability and excellent culturing ability, and can bubble a culture product. SOLUTION: This culture container comprising a self-standing package bag obtained by inserting a bottom film between mutually facing body portion films in a mountain shape and then fusing the peripheral portion, characterized by disposing a gas-charging port comprising a fused portion and a non-fused portion in the bottom sealed portion.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a culture vessel used for culturing various organisms, and more particularly to a culture vessel for culturing while supplying gas to a culture solution.

[0002]

2. Description of the Related Art Conventionally, when culturing various organisms such as microorganisms, cells and plant tissues, cells, tissues and microorganisms are put into a culture solution to improve the contact between a gas such as oxygen and carbon dioxide and the culture solution. And go. As a container for performing this culture, a tube is connected to a hard container made of stainless steel, synthetic resin, glass, or the like, and gas is introduced and exhausted.

[0003] Such a container made of stainless steel, synthetic resin or glass is expensive, so that it cannot be disposable and there is a problem that the washing operation is troublesome. In addition, since culturing is usually performed in a space such as an incubator where temperature control is well controlled, when culturing in large quantities, hard culture vessels are heavy, bulky, easily broken, and it is difficult to install many culture vessels. There was also a problem.

[0004]

The inventors of the present invention have conducted various studies to improve the above-mentioned points, and as a result, the present inventors have found that the bubbling of a culture vessel is made of a self-contained plastic packaging bag having a gas inlet. Have been found, and the present invention has been completed.
It is an object of the present invention to provide an inexpensive and easy-to-handle bubbling culture vessel having excellent ventilating ability and culture ability.

[0005]

SUMMARY OF THE INVENTION The gist of the present invention is to fold a bottom film into a chevron between opposed body films,
What is claimed is: 1. A culture container comprising a self-standing packaging bag having a peripheral portion fused thereto, wherein a gas inlet comprising a fused portion and an unfused portion is provided in a bottom seal portion.

That is, the present invention relates to a culture container comprising a self-supporting packaging bag. By using a transparent plastic film as a material constituting the culture container, the inside of the culture container can be observed. Sometimes you can get the light you need. And, by blowing gas such as air, oxygen and carbon dioxide from the gas inlet at the bottom, it is possible to make sufficient contact between the culture solution and the gas, and to generate convection in the culture solution, thereby culturing the culture. Can be promoted.

Here, a plurality of gas inlets are formed in the culture vessel, and the plurality of gas inlets are connected by a gas passage comprising an unfused portion formed in a bottom seal portion of the packaging bag. Is preferable because the contact between the culture solution and the gas becomes better, and the stirring by the convection of the culture solution can be performed more smoothly.

[0008] Further, the gas introducing port of the culture container of the present invention is opened to the outside at the side end of the packaging bag by a gas passage formed of an unfused portion formed in the bottom seal portion of the packaging bag. In this case, a tube for blowing gas into the gas passage at the side end can be inserted and fitted.

Here, it is preferable that the culture vessel of the present invention has a connecting member capable of connecting a tube to the gas passage at the side end thereof, which is inserted and clamped in advance, so that the tube can be easily fitted. .

Further, when the culture vessel of the present invention is a culture vessel in which a cutout portion formed by cutting out a body film is provided at an upper portion of a packaging bag, and the cutout portion is covered with a filter member, the aseptic condition is improved. Ventilation can be performed without loss.

Here, the notch is formed in a slit shape over the entire width of the packaging bag, and the notch is covered with a filter member from the inside of the packaging bag. This is preferred because the production of the culture vessel of the invention is facilitated.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail. The culture container of the present invention is a self-standing packaging bag made of a film. In this specification, what is expressed as a self-contained packaging bag or simply as a packaging bag refers to the culture container of the present invention, and what is expressed as a film refers to a flexible web-like material that includes a film and a sheet. is there.

The material of the film used in the culture vessel of the present invention is not particularly limited as long as it has a heat-fusible layer as the innermost layer, and may be a single layer or a multilayer. Examples of the layer having heat fusion properties include a polyolefin-based resin layer such as polyethylene (PE) or polypropylene (PP). As the multilayer film, a multilayer film in which these resin layers are used as an inner layer, and a polyamide-based resin layer or a polyester-based resin layer having heat resistance and large physical strength is laminated on the outer layer is used. These outer layers are preferably uniaxially or biaxially stretched films because of their excellent heat resistance and physical strength. These multilayer films may be used, if necessary, in an ethylene-vinyl alcohol copolymer (EVO).
H) A layer having high gas permeability resistance, such as a resin layer or a silica vapor deposition layer, may be laminated. Further, the film used in the present invention may be laminated with aluminum foil or paper, but it is transparent from any viewpoint that promotes culture by transmitting light and allows visual observation of the state of the culture solution as contents. It is preferably a film.

The packaging bag to be the culture container of the present invention is formed by inserting a bottom film forming a bottom surface between the body films of the packaging bag in a chevron-shaped manner and heat-sealing the peripheral portion. . Each of these body film and bottom film,
The same material or different materials may be used.

[0015] The size of the culture vessel of the present invention is not particularly limited, and can be used for a plant from several ml, which is used in a laboratory, like a commonly used culture flask or culture vessel. Although it may be up to several tens of kiloliters, a small one of about 1 to 3 L is preferable in consideration of the risk of contamination.

In the culture container of the present invention, a gas inlet is provided in such a packaging bag, and various gases such as air and carbon dioxide are introduced from the gas inlet to perform bubbling from the bottom of the bag. The gas inlet is provided at the interface between the body film and the bottom film of the self-supporting packaging bag serving as a culture container. Specifically, it is composed of a portion where the body film and the bottom film are fused and a portion where the film is not fused. That is, since the unfused portion is only the film overlapping, gas can pass therethrough. Since the gap between the unfused portions is small, the gas becomes small bubbles and can be diffused into the culture solution.

By providing a plurality of gas inlets, bubbling can be made uniform. The plurality of gas inlets are connected by a gas passage. The gas passage is
As with the gas inlet, it is composed of a portion where the body film and the bottom film are fused and a portion where the film is not fused.

A gas is supplied to the gas inlet from the outside. In the culture container of the present invention, the gas supply release portion is provided at a side end of a packaging bag serving as a culture container. This release portion is also connected by a gas passage, and the gas passage is also composed of a portion where the body film and the bottom film are fused and a portion where the film is not fused. Then, a tube is inserted into the release portion to send gas, and bubbles are generated from the gas inlet. Leakage of the culture solution can be prevented by sealing the periphery of the inserted tube and the release portion with an adhesive or by heat sealing.

In the culture vessel of the present invention, it is preferable that a connecting member for connecting a tube is previously sandwiched between the release portions, since the connection of the tube is facilitated. There is no limitation on the shape or material as long as the connecting member has a sandwiching portion and a tube connecting portion, but after the sandwiching portion is sandwiched, a body film and a bottom film that can be heat-sealed, It is preferable from the viewpoint of preventing the leakage of the culture solution. The heat-sealable material is preferably made of the same resin as the innermost resin layer of the packaging bag. In the case of heat fusion, a boat-like shape in which both ends are thinly formed can be reliably fused, which is preferable.

[0020] The culture vessel of the present invention is used for introducing gas.
The upper portion may be left open to prevent the invasion of various bacteria with absorbent cotton or the like. However, disadvantages such as lack of reliability and troublesome work may occur. In such a case, cut off part of the body film,
Preferably, a gas outlet is provided. It is preferable to attach a filter member for preventing invasion of various bacteria to the gas outlet. The sticking method is not particularly limited as long as the entire surface of the gas outlet is covered with the filter member, and an adhesive may be used or heat fusion may be performed. From the viewpoint of workability and reliability, a method of performing heat fusion using a hot plate sealer, an impulse sealer, or the like is preferable. Usually, at the time of heat fusion, since the outer layer of the packaging bag is formed of a resin layer having high heat resistance, the filter member having heat fusion is thermally fused with the inner layer of the packaging bag having heat fusion. Is preferred. Examples of such a filter member include a woven fabric, a nonwoven fabric, and a thermoplastic resin film having a large number of through holes formed by a laser or the like. Among them, PP-based and PE-based nonwoven fabrics are preferably used from the viewpoint that they are inexpensive, have excellent heat-fusibility, prevent bacteria from entering, and have excellent gas permeability. As the air permeability, a value measured by the Gurley method is preferably 10 to 100 seconds.

The cutout portion may be formed with a punched hole in a circular or rectangular shape. However, if a slit is formed in the entire width direction of the packaging bag and the packaging bag is lined with a filter member, manufacturing is easy. The attachment of members is also possible, which is preferable.

The culture vessel according to the present invention will be described with reference to the drawings. FIG. 1 shows a front view of an example of a culture vessel according to the present invention in a folded state. In FIG. 1, a bottom film 2 is inserted between the body films 1 and 1 by folding in a mountain-fold shape, and a peripheral portion 3 thereof is fused to form a self-supporting packaging bag.
When performing sealing (bottom sealing) of a portion corresponding to the bottom portion, a fused portion 8 is formed except for a gas passage 6, a gas inlet 7, and an unfused portion serving as a release portion 9.

Before the packaging bag is put on the bag making machine, a continuous slit is made in the flow direction of the raw material, and a continuous filter member is stuck from the inside, or the filter member is cut on the bag making machine while making a slit. When heat sealing is performed, a slit-shaped notch 4 to which the filter member 5 is adhered is formed inside the packaging bag when the packaging bag is made. As previously mentioned,
The notch 4 may be formed in a through-hole and the filter member 5 may be adhered and placed. However, since it is necessary to control the position on a bag making machine and it becomes complicated, the notch 4 has a slit shape. Is preferred. The slit may be simply cut, but if the evacuation capacity required for the culture vessel is large, it can be adjusted by providing a gap.

In the culture vessel of the present invention, a tube is inserted into the release section 9 and the culture solution is filled. At the time of filling, a tube whose outside end is closed in advance is used, or the end of the tube is closed with a pinch cock or a clip. When inserting the tube, it is preferable to provide a hard pipe at the end of the tube. Thereafter, the packaging bag is filled with the culture solution, the release section 9 is connected to a gas supply source, and for example, sterilized air (oxygen) is pumped and introduced into the culture solution through the gas inlet 7. Bubbling occurs, the culture solution is agitated and the culture proceeds, the carbon dioxide gas generated by the cells and tissues and the blown air are exhausted through the upper slit 4, and the ventilation in the culture vessel is performed smoothly. At this time, since the filter member 6 is provided, external germs do not enter.

However, depending on the material of the packaging bag, it may be difficult to insert the tube into the release portion 9 or the culture solution may leak from the insertion portion. In such a case, it is preferable that the connecting member 10 as shown in FIG. In FIG. 3, the holding portion 11 is a portion to be held by the release portion 9 of the packaging bag. This portion may be a simple cylinder, but if high liquid tightness is required, the holding portion 11 Alternatively, it is preferable to perform heat fusion while being sandwiched between the body film and the bottom film on a bag making machine, and a boat shape as shown in FIG. 3 is preferable.

The tube connecting member 12 may be a simple pipe, but is preferably a bamboo shoot having ring-shaped irregularities for preventing the tube from falling off as shown in FIG. In addition, it is preferable that the tube connecting member 12 be extended to the opposite side of the sandwiching portion 11 as shown in FIG.

FIG. 2 shows another example of a culture vessel according to the present invention. In FIG. 2, a plurality of gas inlets 7 are formed, and they are connected by a gas passage 6. With this configuration, bubbling can be performed more smoothly.

FIG. 4 is a perspective view of an example of a culture vessel according to the present invention in use, in which a packaging bag is filled with a culture solution and a tissue to be cultured, and the upper portion is sealed by heat sealing. Have been. Then, the tube 13 is connected to a gas supply source (not shown), gas is press-fitted into the packaging bag, and bubbling is performed.

[0029]

Next, the present invention will be described more specifically with reference to examples of the present invention.

Example 1 12 μm biaxially stretched polyethylene terephthalate (PE
T) film, 15 μm biaxially stretched nylon film,
A 100 μm unstretched polypropylene film was dry-laminated using a urethane-based adhesive to prepare a raw laminate. The raw laminate was slit into a body film and a bottom film of the same material. These films and a PP nonwoven fabric having a Gurley air permeability of 50 seconds and a width of 20 mm were set in a bag making machine, and the bottom film and the nonwoven fabric were sandwiched between the two body films.
A packaging bag of 220 mm × 300 mm to which no is attached was prepared. The bottom has a fold diameter of 70 mm and is 4 mm from the fold.
A non-fused portion was formed at a position 5 mm below, which was used as a gas inlet, a gas passage, and a release portion. For the fusion of the nonwoven fabric, two seal bars with a width of 5 mm are placed 50 mm from the upper end of the packaging bag.
The culture was performed at the same time as the bottom seal by using the ones provided in parallel at an interval of mm to prepare a culture vessel.

One end having a length of 30 cm was inserted into a PP pipe, and the other end of the closed tube was inserted into a release portion, the inserted portion was heated and brought into close contact, and the release portion was sealed. . The inside of the packaging bag to which the tube was attached was sterilized with γ-rays, and 2 L of a culture solution and laver cells were filled in an aseptic environment, and the upper portion of the packaging bag was heat-sealed and sealed. Open the end of the tube outside the packaging bag, connect it to the aseptic air generator, and when aseptic air is pumped, many fine bubbles are generated inside the packaging bag and bubbling is performed efficiently Was observed.

Example 2 The connecting member shown in FIG. 3 was previously attached to the open part of the packaging bag of Example 1, and was heat-sealed to prepare the culture vessel shown in FIG. A tube of PP having a length of 30 cm and one end of which is closed is fitted to the tube connecting portion of the connecting member, 2 L of a culture solution is added, the upper portion is heat-sealed and sealed, and heated at 121 ° C. for 30 minutes. Pressure heat sterilization was performed. Under an aseptic environment, the upper fusion part of the packaging bag was cut open, the seaweed cells were put in, and resealed. Open the end of the tube outside the packaging bag, connect it to the aseptic air generator, and when aseptic air is pumped, many fine bubbles are generated inside the packaging bag and bubbling is performed efficiently Was observed.

[0033]

As described above, the culture container of the present invention is constituted by a self-standing packaging bag, and the fusion portion and the non-fusion portion form the gas inlet, the gas passage, and the release portion. Lightweight and not bulky. Moreover, since it can be folded and stored, it is extremely compact during storage. And it can be manufactured at lower cost than conventional glass or stainless steel. Further, since the culture solution is bubbled by introducing gas from the gas inlet provided in the packaging bag, stirring is sufficiently performed, and unnecessary gas is exhausted from the filter member, so that the ventilation effect and the oxygen supply effect are high, and Result The cultivation ability is high.

[Brief description of the drawings]

FIG. 1 is a front view of an example of a culture container according to the present invention in a folded state.

FIG. 2 is a front view of another example of the culture vessel according to the present invention in a folded state.

FIG. 3 is a perspective view of an example of a connection member 10 used in the present invention.

FIG. 4 is a perspective view of an example of a culture vessel according to the present invention in a used state.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 1 Body film 2 Bottom film 3 Peripheral part 4 Notch part 5 Filter member 6 Gas passage 7 Gas introduction port 8 Fusion part 9 Release part 10 Connection member 11 Nipping part 12 Tube connection part 13 Tube

Continuation of the front page (72) Inventor Yutaka Fujimura 1-4-16 Nihonbashi Bakurocho, Chuo-ku, Tokyo Inside Fujimori Industries Co., Ltd. (72) Inventor Sadako 1-4-16 Nihombashi Bakurocho, Chuo-ku, Tokyo Fujimori Kogyo Co., Ltd. F term in the company (reference) 4B029 AA02 DB16 4B065 AA83X BC05

Claims (6)

[Claims] 1. A culture container comprising a self-standing packaging bag in which a bottom film is folded in a chevron between opposed body films and a peripheral portion is fused, wherein a fused portion and an unfused portion are formed on a bottom seal portion. A culture vessel provided with a gas inlet comprising: 2. The gas inlet according to claim 1, wherein a plurality of the gas inlets are formed, and the plurality of the gas inlets are connected by a gas passage formed of an unfused portion formed in a bottom seal portion of the packaging bag. Culture vessels. 3. The packaging bag according to claim 1, wherein the gas introduction port is connected to the outside at a side end of the packaging bag by a gas passage formed of an unfused portion formed in a bottom seal portion of the packaging bag.
The culture vessel according to any one of the preceding claims. 4. The culture container according to claim 3, wherein a connection member capable of connecting a tube is sandwiched in the gas passage at a side end of the packaging bag. 5. The culture container according to claim 1, wherein a cutout portion formed by cutting out a trunk film is provided at an upper portion of the packaging bag, and the cutout portion is covered with a filter member. 6. The culture vessel according to claim 5, wherein the cutout portion is formed in a slit shape over the entire width of the packaging bag, and the cutout portion is covered with a filter member from the inside of the packaging bag. .