JP2558458Y2 - Medium container - Google Patents

Description

[Detailed description of the invention]

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a medium container used for preserving a medium for providing nutrients to cells in culturing the cells.

[0002]

2. Description of the Related Art Japanese Patent Application Laid-Open No.
As described in Japanese Patent Publication No. 7, a solvent injection tube and a culture medium outlet in which a sterilization filter is disposed in the middle of the upper end of a bag-shaped main body made of a gas-impermeable flexible synthetic resin sheet, It is known that a freeze-dried medium is enclosed inside a main body.

When using such a medium container, the protector is broken to expose the medium outlet, to which a connecting needle attached to one end of a connecting tube is connected, and to the other end of the connecting tube. The connected needle is connected to the culture medium inlet of the culture bag (or concentrated culture bag), and the culture medium is cultured through the connection tube (or concentrated culture bag).
Was derived.

[0004]

Therefore, in order to continuously dispense the culture medium into a plurality of cell culture vessels, the connecting needle attached to the other end of the connecting tube must be carefully so as not to be contaminated by touching the floor or the like. Had to pay attention.

[0005] The present invention has been made in view of the above circumstances, and has as its object to solve the above-mentioned problems and to provide a medium container which allows continuous and easy dispensing of a medium. And

[0006]

The bag used in the present invention is made of a gas-impermeable plastic film or sheet.
m is ² · 24hr · atm, ² · carbon dioxide 200～1000Ml / m
24 hr · atm is preferred. The raw material of the plastic film or sheet is not particularly limited, and ethylene-vinyl alcohol; polyvinylidene chloride; polyamide, polyester; a graft polymer of butadiene-acrylonitrile rubber with acrylonitrile and methyl acrylate; polyvinyl chloride, vinyl chloride-ethylene Copolymer,
A non-plastic soft vinyl chloride composition comprising a vinyl chloride resin such as an ethylene-vinyl acetate-vinyl chloride copolymer and an ethylene-carbon monoxide-vinyl acetate copolymer is exemplified. Further, two or more kinds of the plastics may be laminated and used.

[0007] The bag-like body has at least one side so that the discoloration of the buffer from orange to crimson when the pH of the culture medium is inclined to an alkalinity unsuitable for culturing bacteria or turbidity when microorganisms are mixed can be visually recognized. It is preferable to have a transparent portion.

[0008] In order to connect one end of the culture medium outlet tube to the opening of the bag-like body and to form a culture medium inlet, for example,
A tube (for example, an inner diameter of 3 to 5 mm) made of the same material as the bag-shaped body can be provided on the bag-shaped body by high-frequency fusion, heat fusion, adhesion, or the like.
It is preferable to integrate the tube and the bag-like body by high-frequency fusion or heat fusion because complete sealing is maintained.

In the culture medium container of the present invention, the stopper for closing the connection between the bag-shaped body and the medium lead-out tube has a blind hole opened to the medium lead-out tube side. A groove in the circumferential direction is engraved on the outer peripheral wall located closer to the tube side to form a small-diameter portion, and the bag-shaped body and the culture medium lead-out tube can be communicated by breaking at the small-diameter portion. Have been like.

In the culture medium container of the present invention, at least a part of the culture medium inlet is provided so that serum or chemicals rich in nutrients such as proteins can be added to the culture medium later using a syringe. Is preferably screwed with a co-injected rubber stopper made of rubber.

In the medium container of the present invention, the protective cover is
It is composed of a main body and a lid, so that even when the remaining medium is taken out of a clean bench and re-stored in a refrigerator, it can be fitted airtight so that the tip of the medium lead-out tube does not come into contact with the outside air and become contaminated. ing. The material for forming the protective cover main body is not particularly limited, but includes polycarbonate, polystyrene, polymethyl methacrylate, acrylonitrile-styrene copolymer, hard polyvinyl chloride,
Transparent synthetic resins such as polypropylene, polymethylpentene, and polyethylene terephthalate are preferred.
The shape is not particularly limited, and may be any of a bell shape, a western bell shape, a bowl shape, a cup shape, and the like.
Preferably, it is set to 〜80 mm. Further, it is preferable that the tip of the culture medium lead-out tube is positioned 0.5 to 50 mm inward from the opening edge of the protective cover main body. The material for forming the protective cover lid is not particularly limited, but a thermoplastic resin such as polyethylene, polypropylene, polyvinyl chloride, and polystyrene is preferable. In addition, the lid is also sterilized at the end of the medium outlet tube during sterilization work in the closed lid state,
A filter made of a filter paper, a nonwoven fabric, or the like, which has gas permeability but does not allow microorganisms to pass through, is fixed to the hole by means such as adhesion and fusion.

In the medium container of the present invention, a grip portion having a diameter larger than that of the medium lead-out tube is provided between the protective cover main body and the clamp. This is preferable because the clamp can be operated with the forefinger. The material forming the grip portion is not particularly limited, but is preferably flexible. The cross-sectional shape is not particularly limited, and may be any of a cylinder, an elliptic cylinder, and a polygonal cylindrical body. The thickness is preferably about 8 to 25 mm, and the length is preferably about 50 to 150 mm. In order to form the gripping part, even if the cylindrical body is inserted into the medium guiding tube, the cylindrical body having an inner diameter substantially equal to the outer diameter of the medium guiding tube is integrally connected by induction heating, dielectric heating, adhesion, or the like. Or any of them.

Before use, the medium container of the present invention is subjected to radiation sterilization, high-pressure steam sterilization, and ethylene oxide gas (EOG).
It is used after being sterilized by sterilization, electron beam sterilization or the like.

The medium container of the present invention is used, for example, as follows. After EOG sterilization, the medium is injected into the bag from the medium injection port in a clean bench, and the medium injection port is sealed with a rubber injection stopper. In taking out the culture medium, first, the tube is temporarily sealed by a clamp inserted into the culture medium lead-out tube, and then the stopper that closes the connection between the bag-shaped body and the medium lead-out tube is closed with a small diameter part. The rupture is made to connect the bag-shaped body and the medium outlet tube. Next, the bag is lifted with the medium outlet tube side down, the clamp is loosened, and the medium is supplied to the cell culture container.

[0015]

In the medium container of the present invention, as described above, the connection between the bag-shaped body and the medium leading-out tube can be connected to the medium leading-out tube by breaking at the small diameter portion. The medium is not contaminated by microorganisms and the pH does not change because it is closed by the stopper.
In addition, since a protective cover consisting of a main body and a lid is provided at the other end of the medium outlet tube, even if the medium outlet tube is left as it is on the floor of the clean bench after dispensing, the tip of the tube can be removed. Is not contaminated by contact with clean bench floors and equipment.

[0016]

Next, the medium container of the present invention will be further described with reference to the embodiments shown in the drawings. FIG. 1 is a side view of the culture medium container of the present invention, FIG. 2 is a partially enlarged view of a connection portion between the bag-like body and the culture medium lead-out tube, and FIG. It is a partial enlarged view which shows the state which connected the bag-shaped body and the culture medium lead-out tube. 1 is a bag-shaped body, 2 is a medium lead-out tube connected by fusing one end to an opening of the bag-shaped body 1, 3
Is a medium injection port formed in the opening of the bag-like body 1.

The bag 1 is made by fusing a non-plastic soft vinyl chloride resin sheet having a thickness of 0.15 mm. The soft vinyl chloride resin sheet is composed of 100 parts by weight of a vinyl chloride-ethylene copolymer (average degree of polymerization: 1300, ethylene content: 4% by weight) and ethylene-carbon monoxide-
It is formed by extrusion molding a resin composition containing 100 parts by weight of a vinyl acetate copolymer (carbon monoxide content: 10% by weight, vinyl acetate content: 24% by weight). The oxygen gas permeability of this sheet is 170 ml / m ² · 24 hr · atm, and the carbon dioxide gas permeability is 890 ml / m ² · 24 hr · atm.

As shown in FIG. 2, a blind hole 41 is formed in the connection between the bag-shaped body 1 and the medium outlet tube 2 so as to open toward the medium outlet tube 2. The V-shaped groove in the circumferential direction is formed in the outer peripheral wall located on the tube side, and is closed by the hard polyvinyl chloride stopper 4 having the small-diameter portion 42 formed therein. As shown in FIG. By breaking at the portion 42, the bag-like body 1 and the culture medium outlet tube 2 can be communicated.

At the other end of the culture medium outlet tube 2 having an outer diameter of 6 mm, a substantially bowl-shaped polycarbonate protective cover main body 5 is mounted. 62 made of "Tyvek")
Is hermetically fitted with a lid body 6 to which is thermally bonded. A male luer type connector 21 is attached to the tip of the culture medium outlet tube 2, and the tip of the connector 21 is located 5 mm inward from the opening end of the protective cover main body 5. In the vicinity of the protective cover main body 5, a clamp 7 which can sandwich the tube 2 from both sides and temporarily seal the outflow of the culture medium is attached, and an inner diameter of 6.5 between the protective cover main body 5 and the clamp 7 is provided. A holding portion 8 is provided by externally fitting a cylinder having a diameter of 13 mm, an outer diameter of 13 mm, and a length of 90 mm. Reference numeral 11 denotes a hole for hanging the bag-like body 1.

In use, the medium container is sterilized by EOG, and after sufficient EOG removal, the powdered MEM medium (F
4.8 g of “AUTOPAW” manufactured by Low Corporation) is dissolved in 500 ml of purified water, sterilized by high-pressure steam at 121 ° C. for 20 minutes, and cooled.
In a clean bench, a solution obtained by mixing 14 ml of a 7.5% sodium bicarbonate solution and 5 ml of a 200 mM glutamine solution was entirely injected from the medium inlet 3 using a sterilized funnel, and the mixed rubber stopper 31 was screwed on. Next, fetal bovine serum was injected from the co-injected rubber stopper 31 using a syringe so as to be 10% V / V, and a medium was prepared and stored in a refrigerator. The pH at the time of preparation of the medium was 7.61, and the pH after refrigerated storage for 2 months was 7.63, which was almost unchanged, and no microbial contamination was observed. Further, storage at -80 ° C was also possible.

The stopper 4 of the medium container taken out of the refrigerator is broken off from the small-diameter portion 42, the hanging hole 11 of the bag-shaped body 1 is hung by a hook (not shown), and the grip 8 is moved to the middle finger or lower of the right hand. While grasping with three fingers and opening and closing the clamp 7 with the thumb and forefinger, about 5 ml was continuously dispensed into 10 Petri dishes having a diameter of 60 mm. Next, the tip of the medium outlet tube 2 temporarily sealed by the clamp 7 is left on the floor of the clean bench, and is replaced with another 10 Petri dishes. Dispensed. After the work was completed, the lid 6 was fitted to the protective cover main body 5 and stored again in the refrigerator. After the lapse of 5 days, no microbial contamination was observed in any of the medium container and the replaced petri dish.

[0022]

[Effect of the Invention] The medium container of the present invention is configured as described above, so that the sterility of the medium can be maintained and the pH during storage is maintained.
Since there is no change and there is no need to bring the entire container into the clean bench and only the lead-out tube needs to be retracted, it is possible to effectively use the inside of the clean bench. Since it can be left on the surface, the dispensing operation can be performed continuously, easily and quickly.

[Brief description of the drawings]

FIG. 1 is a side view of the culture medium container of the present invention.

FIG. 2 is a partially enlarged view of a connection portion between a bag-shaped body and a medium lead-out tube in FIG.

FIG. 3 is a partially enlarged view showing a state in which the plug of FIG. 2 is broken at a small diameter portion and a bag-shaped body and a medium leading-out tube are communicated with each other.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Bag-shaped body 2 Culture medium lead-out tube 3 Medium injection port 4 Plug 5 Protective cover main body 6 Lid 7 Clamp 8 Grip part 41 Small diameter part

Claims (1)

(57) [Scope of request for utility model registration] 1. An end of a medium outlet tube is connected to an opening of a bag made of a gas-impermeable plastic film or sheet, and a medium inlet is formed, and the connection between the bag and the medium outlet tube is formed. The part is closed by a stopper that can communicate the bag-shaped body and the medium lead-out tube by breaking at the small diameter part, and a protective cover consisting of a main body and a lid is provided at the other end of the medium lead-out tube. A culture medium container, wherein a clamp is attached near a protective cover.