【０００１】[0001]
【発明の属する技術分野】本発明は振盪培養用の容器で
あって、さらに詳しくは、容器の外内部または培養液と
容器内気体とのガス交換能が優れ培養効率のよい振盪培
養容器に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a container for shaking culture, and more particularly to a shaking culture container having excellent gas exchange ability between the inside and outside of the container or between a culture solution and gas in the container and having high culture efficiency.
【０００２】[0002]
【従来技術】従来より微生物、植物組織の振盪培養を行
うに際しては、培養液中に細胞（組織）を入れ振盪させ
て気体と培養液との接触を良くして行っている。そし
て、この培養を行う容器としては、三角フラスコ、試験
管、ビーカー等のガラス容器や硬質プラスチック製容器
が使用されている。しかし、ガラス容器は破損の恐れが
あり、また、これらの容器を反復使用する場合、菌の混
ざり合いを防止するため十分な洗滌或は殺菌を行う必要
があったが、この洗滌操作は手間がかかり、殊に試験管
や三角フラスコのような開口部の狭い容器の場合は洗浄
ばかりではなく、内容物の出し入れが不便であった。そ
こで、本発明者らは先にこの点を改良して作業性の良好
な植物培養用容器を提案した（実開平３−１２６８５０
号公報参照）。この容器は内面に熱接着性合成樹脂層を
有する２枚のシート状の表裏面材及びこれらの表裏面材
の間にあって、それぞれの下縁部に接続した内方につつ
折り状にした底面材とからなり、底面材を開くと図４に
示したような形状をし、使用に際しては上方の開口部を
封口治具によって封じていた。この容器により先に述べ
た作業性は大いに改善されたが、この容器は、未使用時
には底面材がつつ折り状になって折り畳んであるため使
用時には底面を開いて容器を形成しなければならず、ま
た、容器の上方になるにしたがって表面材と裏面材との
間隔がせばまり、開口部は封口治具によって封じられて
いるので容器内の空間が狭く、培養に必要な換気は単に
面材に設けた透孔によって行われるだけであるため培養
細胞が縊死したり、或は培養代謝ガスの発生のため容器
が膨張し破袋を生ずるおそれがあった。2. Description of the Related Art Conventionally, when shaking culture of a microorganism or plant tissue is performed, cells (tissue) are placed in a culture solution and shaken to improve the contact between gas and the culture solution. As a container for performing this culture, a glass container such as an Erlenmeyer flask, a test tube, or a beaker or a hard plastic container is used. However, glass containers may be damaged, and when these containers are used repeatedly, it is necessary to perform sufficient washing or sterilization to prevent mixing of bacteria, but this washing operation is troublesome. In particular, in the case of a container having a narrow opening such as a test tube or an Erlenmeyer flask, not only washing but also taking out and in of the contents is inconvenient. Therefore, the present inventors have previously improved this point and proposed a container for plant culture having good workability (Japanese Utility Model Application Laid-Open No. 3-126850).
Reference). This container has two sheet-like front and back materials having a heat-adhesive synthetic resin layer on the inner surface, and a bottom material folded between the front and back materials and inwardly connected to the lower edges thereof. When the bottom material was opened, it had a shape as shown in FIG. 4, and when used, the upper opening was sealed with a sealing jig. Although the above-described workability has been greatly improved by this container, the bottom of the container is folded and folded when not in use, so the bottom must be opened to form the container when in use. In addition, the distance between the front and back materials is narrowed toward the top of the container, and the opening is closed by a sealing jig, so the space in the container is narrow, and the ventilation required for culture is merely a surface. There is a risk that cultured cells may be hung or crushed due to the generation of metabolic gas for cultivation, and that the container may expand and break the bag, since it is performed only through the holes provided in the material.
【０００３】[0003]
【発明が解決しようとする課題】そこで、本発明者は上
記の欠点を改良し、容易に容器を形成することができる
と共に、容器内の容積を広くし、容器内の気体と培養液
内の気体とのガス交換が十分行えるような容器について
種々検討した結果、本発明を完成したもので、本発明の
目的は、容易に培養液を攪拌することが出来、培養液と
容器内気体とのガスとのガス交換能にすぐれた培養効率
のよい振盪培養容器を提供することである。Therefore, the present inventor has improved the above-mentioned drawbacks, can easily form a container, and can increase the volume in the container so that the gas in the container and the culture medium in the culture solution can be easily formed. As a result of various studies on a container capable of sufficiently performing gas exchange with gas, the present invention has been completed, and an object of the present invention is to enable easy stirring of the culture solution, and to allow the culture solution and the gas in the container to be mixed. An object of the present invention is to provide a shaking culture vessel having excellent culture efficiency and excellent gas exchange ability with gas.
【０００４】[0004]
【課題を解決するための手段】本発明の要旨は、内層に
熱接着性合成樹脂層を有し、底面がほぼ正四方形である
振盪培養用容器であって、該容器の上面に底面の断面積
より小さい断面積を有する注出口を前記容器の中央部に
突設させ、且つ、該容器の前面又は後面若しくはその両
面の上方の位置に透孔を穿設し、該透孔部分をフィルタ
ーで接着閉塞することを特徴とする振盪培養用容器であ
る。即ち、本発明においては、容器の底面を正四方形に
するため底部中央から側縁部に対して４５度の角度とな
るように熱接着してあり、従って、単に培養液を注入す
るのみで開底して底面正方形の容器となり、また、底面
から容器上方の注出口の肩部にいたるまでの各側壁面
（前面、後面及びヒダ部をいう）は同形であるため容器
内の空間は広がり、且つ底面が正四方形であるので培養
液を攪拌して円流を生じることができ、その結果培養液
と容器内気体との接触が十分に行えることができる。SUMMARY OF THE INVENTION The gist of the present invention is to provide a container for shaking culture having a thermoadhesive synthetic resin layer as an inner layer and a substantially square bottom surface. A spout having a cross-sectional area smaller than the area is projected at the center of the container, and a through hole is formed at a position above the front or rear surface or both surfaces of the container. This is a container for shaking culture characterized by adhesively closing. That is, in the present invention, in order to make the bottom of the container into a square, the container is heat-bonded at an angle of 45 degrees from the center of the bottom to the side edge, and thus is opened only by injecting the culture solution. The bottom is a square bottom container, and the side walls from the bottom to the shoulder of the spout above the container (referred to as front, back and folds) are the same shape, so the space in the container expands, In addition, since the bottom surface is a square, the culture solution can be stirred to generate a circular flow, and as a result, the culture solution and the gas in the container can be sufficiently contacted.
【０００５】[0005]
【発明の実施の態様】本発明について更に詳細に説明す
る。本発明において使用する内層に熱接着性合成樹脂層
を有する包装材とは一方の面に熱融着性合成樹脂層を有
する耐熱性及び耐水性のある合成樹脂シートであって、
例えばポリエチレンまたはポリプロピレンなどのポリオ
レフィン樹脂、ポリエステル樹脂、ポリアミド樹脂など
の耐熱性及び耐水性を有する合成樹脂シートの一方の面
にポリエチレンなどの熱接着性樹脂層を積層したもの
で、容器を形成する際に熱接着性樹脂層を内面とする。BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in more detail. The packaging material having a heat-adhesive synthetic resin layer in the inner layer used in the present invention is a heat- and water-resistant synthetic resin sheet having a heat-fusible synthetic resin layer on one surface,
For example, when a container is formed by laminating a heat-adhesive resin layer such as polyethylene on one surface of a heat- and water-resistant synthetic resin sheet such as a polyolefin resin such as polyethylene or polypropylene, a polyester resin, or a polyamide resin. The heat-adhesive resin layer is used as the inner surface.
【０００６】本発明では上記の包装材をもって底面がほ
ぼ正四方形で、上面に底面の断面積より小さい断面積を
有する注出口を中央部に突設させた容器を形成するので
あるが、その形成手段は特に限定されるものではない。
例えば、中央部を突出させ、底辺の中央よりそれぞれ両
側縁に対し４５度に切断した包装材の４枚をもってそれ
ぞれの側縁部、底辺部及び肩辺部の必要個所を熱接着し
たり、或いは、中央部に相当する部分及び底面を形成す
る部分を有する形状をした２枚の包装材より必要個所を
熱接着したり、或いは、ほぼ同形の包装材でもって形成
したガゼット型容器を折り畳んだ状態で、底部の中央か
ら両側縁に対し４５度に切断、融着したりする等の手段
によって形成する。得られた容器の注出口は底面に比し
て狭く、この注出口にはスポンジ等の多孔体で通気性を
保持しながら外部からのごみ、雑菌等の侵入を防止す
る。また容器を形成する包装材の少なくとも１枚には上
半分の位置に透孔を穿設し、この透孔の内面又は外面か
ら通気性を有するが菌の混入を防止するフィルターを貼
着する。透孔の大きさとしては、容器の大きさによって
適宜に決定すれば良いが、通常タテ約３０ｍｍ程度、ヨ
コ約６０ｍｍ程度であり、また、フィルターとしてはポ
リエステル系繊維、ポリエチレン系繊維、ポリプロピレ
ン系繊維等からなる不織布を使用する。フィルターの貼
着手段としては熱融着や接着剤等を使用する。本発明に
おいては底面はその中央から側縁部に対して４５度の角
度をもって熱融着してあるので培養液を容器内に注入す
ると容易に開底し、形成された容器は安定に直立する。
そして、底面は正方形を形成するので培養液を攪拌する
と、円流となり攪拌しやすく、培養液と容器内の気体と
の交換能は優れている。According to the present invention, a container having the above-described packaging material and having a spout having a substantially square bottom surface and a cross-sectional area smaller than the cross-sectional area of the bottom surface protruding from the center at the upper surface is formed. Means are not particularly limited.
For example, the center portion is protruded, and the necessary portions of the side edge portion, the bottom side portion and the shoulder side portion are thermally bonded with four pieces of the packaging material cut at 45 degrees to both side edges from the center of the bottom side, or A state in which a required portion is heat-bonded from two pieces of packaging material having a portion corresponding to a central portion and a portion forming a bottom surface, or a gusset-type container formed by using packaging materials having substantially the same shape is folded. Then, it is formed by means such as cutting and fusing at 45 degrees from the center of the bottom to both side edges. The spout of the obtained container is narrower than the bottom surface, and the spout prevents the invasion of dust, germs and the like from the outside while maintaining air permeability with a porous material such as sponge. In addition, at least one of the packaging materials forming the container is provided with a through hole in the upper half position, and a filter is attached from the inner or outer surface of the through hole that has air permeability but prevents bacteria from entering. The size of the through-hole may be appropriately determined depending on the size of the container, and is usually about 30 mm in length and about 60 mm in width, and the filter is made of polyester fiber, polyethylene fiber, polypropylene fiber. Use a nonwoven fabric consisting of As a means for attaching the filter, heat fusion, an adhesive or the like is used. In the present invention, the bottom is heat-sealed at an angle of 45 degrees from the center to the side edge, so that when the culture solution is injected into the container, it easily opens and the formed container stands upright. .
Since the bottom surface forms a square, when the culture solution is stirred, it becomes a circular flow and is easily stirred, and the exchange ability between the culture solution and the gas in the container is excellent.
【０００７】次に図をもって本発明を説明する。図１は
本発明にかかる振盪培養用容器の斜視図であり、図２は
その容器を折り畳たんだ状態の平面図であり、図３は先
に提案した培養容器である。図において、容器１を構成
する包装材２は、その上部中央部に突出した注出口３を
有し、下部の底部中央４から側縁部５に対して４５度の
角度をもって融着されている。容器１は側縁部５、底部
６、および肩部７において融着されている。また、側壁
面の少なくとも１つには透孔８が穿設されており、この
透孔８にはフィルターが貼着されている。このような容
器において、注出口から培養液を注入すると、容器は直
ちに開底して図１の状態になる。Next, the present invention will be described with reference to the drawings. FIG. 1 is a perspective view of a container for shaking culture according to the present invention, FIG. 2 is a plan view of the container in a state where the container is folded, and FIG. 3 is a culture container previously proposed. In the figure, a packaging material 2 constituting a container 1 has a spout 3 protruding in the upper central portion thereof, and is fused from a lower central portion 4 of the lower portion to a side edge portion 5 at an angle of 45 degrees. . The container 1 is fused at a side edge 5, a bottom 6, and a shoulder 7. Further, at least one of the side wall surfaces is provided with a through hole 8, and a filter is attached to the through hole 8. In such a container, when a culture solution is injected from the spout, the container is immediately opened to the state shown in FIG.
【０００８】[0008]
実施例１ 幅１１０ｍｍ、奥行き１１０ｍｍの底面で高さが１６０
ｍｍの直方体の前面及び後面の２か所に横６０ｍｍ，縦
３０ｍｍの透孔を設け、これにポアサイズ０．２〜０．
７μｍを有するポリプロピレン不織布よりなるフィルタ
ーを貼着した本発明にかかる培養用容器を用いてパン酵
母（Ｓ．ｃｅｒｅｖｉｓｉａｅ）を振盪培養した。即
ち、市販のペースト状パン酵母より分離したパン酵母
（Ｓ．ｃｅｒｅｖｉｓｉａｅ）を使用し、Ｙ−Ｍ寒天斜
面培地上で、３０℃、２４時間培養した後に、２白金の
菌体を酵母増殖用培地２００ｍｌに接種し、本発明の培
養容器を用いて３０℃、１８時間、２２０ｒｐｍの回転
で培養した。酵母増殖用培地には、０．２％ＭｇＳＯ4
・７Ｈ2Ｏ、０．４％（ＮＨ4）2ＳＯ4、０．８％ＫＨ2
ＳＯ4、０．８％ｙｅａｓｔ ｅｘｔｒａｃｔ、３％ｇ
ｌｕｃｏｓｅからなる液体培養池（ＰＨ ６）を用い
た。本培養は酵母増殖用培地に対して５％（Ｖ／Ｖ）の
前培養液を加えた後、培養温度３０℃で行なった。培養
の結果を図３に示した。なお、比較例として５００ｍｌ
容の三角フラスコを使用して同様に行なった。その結果
を図３に示した。図３における分析方法は次の方法で測
定した。Ｕ．Ｏ．Ｄ．（ｕｎｉｔ ｏｐｔｉｃａｌ ｄ
ｅｎｎｓｉｔｙ）単位菌体濃度は、波長５８０ｎｍを用
いてで分光分析計（島津ボシュロム分光分析計スペクロ
ニック２０Ａ）で定量した。エタノール（ｇ／ｌ）定量
は、Ｃｈｒｏｍｏｓｏｒｂ １０５（６０−８０メッシ
ュ）を充填した内径５ｍｍ、長さ２ｍガラスカラムを使
用した。Ｇｌｕｃｏｓｅ（ｇ／ｌ）は、ムスロターゼ・
ＧＯＧ（和光純薬グルコース−ＣII−テストワコー）を
用いて定量した。Example 1 A bottom surface having a width of 110 mm and a depth of 110 mm and a height of 160 mm
A rectangular hole having a width of 60 mm and a length of 30 mm is provided at two places on the front and rear sides of a rectangular parallelepiped having a diameter of 0.2 mm to 0.2 mm.
Baker's yeast (S. cerevisiae) was shake-cultured using the culture vessel according to the present invention to which a filter made of a polypropylene nonwoven fabric having a thickness of 7 μm was adhered. That is, after using a baker's yeast (S. cerevisiae) isolated from a commercially available pasty baker's yeast and culturing it at 30 ° C. for 24 hours on a Y-M agar slant medium, the two platinum cells were transformed into a yeast growth medium. 200 ml was inoculated and cultured in the culture vessel of the present invention at 30 ° C. for 18 hours at 220 rpm. The yeast growth medium contains 0.2% MgSO4
7H2 O, 0.4% (NH4 )2 SO4 , 0.8% KH2
SO4 , 0.8% yeast extract, 3% g
A liquid culture pond (PH6) made of lucose was used. The main culture was performed at a culture temperature of 30 ° C. after adding a 5% (V / V) preculture liquid to the yeast growth medium. The results of the culture are shown in FIG. In addition, 500 ml as a comparative example
The same procedure was performed using a conical Erlenmeyer flask. The result is shown in FIG. The analysis method in FIG. 3 was measured by the following method. U. O. D. (Unit optical d
The unit cell concentration was quantified using a spectrophotometer (Shimadzu Bausch & Lomb spectrometer 20A) using a wavelength of 580 nm. For ethanol (g / l) quantification, a glass column with an inner diameter of 5 mm and a length of 2 m packed with Chromosorb 105 (60-80 mesh) was used. Glucose (g / l) is
Quantification was performed using GOG (Wako Pure Chemical Industries Glucose-CII-Test Wako).
【０００９】[0009]
【発明の効果】以上述べたように、本発明においては、
底部中央から側縁部に対して４５度の角度となるように
熱接着したことによって正方形の安定した底面を形成
し、容器の上部に至るまで側壁面が狭まらず容器内の容
積が大きく、従って、容器内の気体と培養液殻発する気
体の交換能は優れている。As described above, in the present invention,
Formed a stable bottom surface of a square by heat bonding from the bottom center to the side edge at an angle of 45 degrees to the side edge, the side wall surface does not narrow down to the top of the container and the volume in the container is large Therefore, the exchange ability between the gas in the container and the gas emitted from the culture solution shell is excellent.
【図１】本発明にかかる培養用容器の斜視図FIG. 1 is a perspective view of a culture vessel according to the present invention.
【図２】折り畳んだ状態の本発明にかかる培養法容器の
平面図FIG. 2 is a plan view of the culture method container according to the present invention in a folded state.
【図３】実施例（Ａ）及び比較例（Ｂ）の培養結果を示
した図FIG. 3 is a diagram showing the culture results of Example (A) and Comparative Example (B).
【図４】先に提案された容器の斜視図FIG. 4 is a perspective view of a previously proposed container.
１ 容器 ２ 包装材 ３ 注出口
４ 底部中央 ５ 側縁融着部 ６ 底部融着部 ７ 肩部融着
部 ８ 透孔 ９ 封口治具1 Container 2 Packaging material 3 Spout
4 bottom center 5 side edge fusion part 6 bottom fusion part 7 shoulder fusion part 8 through hole 9 sealing jig
───────────────────────────────────────────────────── フロントページの続き (72)発明者 藤村 豊 東京都中央区日本橋馬喰町１丁目４番16号 藤森工業株式会社内 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Yutaka Fujimura 1-4-16-1 Nihombashi Bakurocho, Chuo-ku, Tokyo Inside Fujimori Kogyo Co., Ltd.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP25844296AJP4042167B2 (en) | 1996-09-30 | 1996-09-30 | Shake culture container |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP25844296AJP4042167B2 (en) | 1996-09-30 | 1996-09-30 | Shake culture container |
| Publication Number | Publication Date |
|---|---|
| JPH1099071Atrue JPH1099071A (en) | 1998-04-21 |
| JP4042167B2 JP4042167B2 (en) | 2008-02-06 |
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP25844296AExpired - Fee RelatedJP4042167B2 (en) | 1996-09-30 | 1996-09-30 | Shake culture container |
| Country | Link |
|---|---|
| JP (1) | JP4042167B2 (en) |
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| JP2002330750A (en)* | 2001-05-11 | 2002-11-19 | Hideo Tanaka | Flask for shaking culture and device for shaking culture |
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| US9376655B2 (en) | 2011-09-29 | 2016-06-28 | Life Technologies Corporation | Filter systems for separating microcarriers from cell culture solutions |
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| JP2002330750A (en)* | 2001-05-11 | 2002-11-19 | Hideo Tanaka | Flask for shaking culture and device for shaking culture |
| US9005971B2 (en) | 2005-04-22 | 2015-04-14 | Life Technologies Corporation | Gas spargers and related container systems |
| US9259692B2 (en) | 2005-04-22 | 2016-02-16 | Life Technologies Corporation | Gas spargers and related container systems |
| US9475012B2 (en) | 2005-04-22 | 2016-10-25 | Life Technologies Corporation | Gas spargers and related container systems |
| US9682353B2 (en) | 2005-04-22 | 2017-06-20 | Life Technologies Corporation | Gas spargers and related container systems |
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