JP5214714B2 - Incubator - Google Patents

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JP5214714B2
JP5214714B2 JP2010274954A JP2010274954A JP5214714B2 JP 5214714 B2 JP5214714 B2 JP 5214714B2 JP 2010274954 A JP2010274954 A JP 2010274954A JP 2010274954 A JP2010274954 A JP 2010274954A JP 5214714 B2 JP5214714 B2 JP 5214714B2
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culture
flat bag
support device
container
bag
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JP2012120495A (en
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博行 松田
裕司 川畠
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Fujimori Kogyo Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/48—Holding appliances; Racks; Supports
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/14—Bags
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/26—Constructional details, e.g. recesses, hinges flexible

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Description

本発明は、微生物や細胞など（本明細書では、これらを総称して「微生物等」という場合がある。）の培養を簡便かつ能率よく行うことのできる培養装置に関する。   The present invention relates to a culture apparatus capable of simply and efficiently culturing microorganisms, cells, and the like (in the present specification, these may be collectively referred to as “microorganisms”).

微生物や細胞などの培養は、医薬品や食品などとして有用な各種物質の生産に広く用いられている。また、従来のガラス製やステンレス製の培養器に比べて安価に構成可能な培養容器として、袋状の柔軟な容器（バッグ、パウチ等）を用いた培養装置が提案されている（例えば特許文献１，２参照）。   Culture of microorganisms and cells is widely used for production of various substances useful as pharmaceuticals and foods. Further, as a culture vessel that can be constructed at a lower cost than conventional glass or stainless steel incubators, a culture device using a bag-like flexible container (bag, pouch, etc.) has been proposed (for example, Patent Documents). 1 and 2).

特開２００８−２１２０４９号公報JP 2008-212049 A 特開２００９−１５９８９０号公報JP 2009-159890 A

微生物等の大量培養には、その微生物等の生育に必要な酸素（Ｏ２）や二酸化炭素（ＣＯ２）等のガスを培養液中に効率よく供給する必要がある。培養液への通気を促進する方法として、培養容器の内部で培養液を通気攪拌する方法と、培養容器を外部から振盪させる方法（振盪培養）が知られている。振盪培養の場合、培養容器の内部に攪拌装置を収容する必要がないので、より簡便となる。しかしながら、袋状容器を振盪培養に用いる場合、ガラス製やステンレス製等の剛直な容器を振盪する場合に比べて、容器の耐久性が低く、特に振盪が長期にわたる場合、培養中に予期しないピンホールが発生し、フィルター等で浄化されていない外気等が容器内部に侵入し、あるいは、培養液が漏れ出し、さらには、漏れ出した培養液が容器内に戻るなどして、容器の内部が汚染されたり、容器の周囲が汚れたりすることがある。 For mass culture of microorganisms or the like, it is necessary to efficiently supply gas (such as oxygen (O 2 ) or carbon dioxide (CO 2 )) necessary for the growth of the microorganisms or the like into the culture solution. As a method for promoting aeration to the culture solution, a method in which the culture solution is aerated and stirred inside the culture vessel and a method in which the culture vessel is shaken from the outside (shaking culture) are known. In the case of shaking culture, since it is not necessary to house a stirring device inside the culture vessel, it becomes more convenient. However, when using a bag-like container for shaking culture, the durability of the container is lower than when shaking a rigid container made of glass or stainless steel, especially when shaking is prolonged, and an unexpected pin is used during culture. The inside of the container is caused by the occurrence of holes and the outside air that has not been purified by the filter or the like entering the inside of the container, or the culture medium leaks and the leaked culture liquid returns to the container. It may become contaminated and the area around the container may become dirty.

本発明は、上記事情に鑑みてなされたものであり、袋状容器を振盪培養に用いたときのピンホールの発生を抑制することが可能な培養装置を提供することを課題とする。   This invention is made | formed in view of the said situation, and makes it a subject to provide the culture apparatus which can suppress generation | occurrence | production of a pinhole when a bag-like container is used for shaking culture.

前記課題を解決するため、本発明は、対向する基材の内面の周囲をシールしたシール部を有する平袋からなる培養容器と、前記培養容器を支持する支持装置を備える振盪培養用の培養装置であって、前記平袋の片面を下にして前記支持装置上に支持された前記培養容器の少なくとも互いに対向する２辺を前記支持装置に固定する固定手段を備え、前記支持装置は底壁部と側壁部とを備えるトレイ状であり、前記固定手段は、前記平袋の周縁部を前記底壁部から持ち上がらせて前記支持装置の側壁部に固定していることを特徴とする培養装置を提供する。
前記培養容器は、前記平袋の周縁部に前記固定手段に係止される複数の孔を有することも可能である。
前記固定手段は、前記平袋の周縁部をつかんで固定する、つかみ具を備えることも可能である。 In order to solve the above problems, the present invention provides a culture apparatus for shaking culture, comprising a culture container comprising a flat bag having a seal portion sealed around the inner surface of an opposing base material, and a support device for supporting the culture container. A fixing means for fixing at least two opposite sides of the culture vessel supported on the support device with one side of the flat bag facing down to the support device, the support device having a bottom wall portion And a side wall portion, and the fixing means lifts the peripheral edge portion of the flat bag from the bottom wall portion and fixes the culture device to the side wall portion of the support device. provide.
The culture container may have a plurality of holes that are locked to the fixing means at the peripheral edge of the flat bag.
The fixing means may include a gripping tool that holds and fixes the peripheral edge of the flat bag.

本発明によれば、固定装置により平袋状の培養容器にテンションを加えて固定することができるので、ピンホールの要因となるシワの発生を防止でき、これにより、ピンホールの発生を抑制することができる。   According to the present invention, the flat bag-shaped culture container can be tensioned and fixed by the fixing device, so that the generation of wrinkles that cause pinholes can be prevented, thereby suppressing the generation of pinholes. be able to.

本発明の第１実施形態における（ａ）培養容器の平面図、（ｂ）培養装置の断面である。BRIEF DESCRIPTION OF THE DRAWINGS (a) Top view of culture container in 1st Embodiment of this invention, (b) It is a cross section of a culture apparatus. 本発明の第２実施形態における（ａ）培養容器の平面図、（ｂ）培養装置の断面である。(A) The top view of a culture container in 2nd Embodiment of this invention, (b) It is a cross section of a culture apparatus.

以下、好適な実施の形態に基づき、図面を参照して本発明を説明する。
図１に、本発明の培養装置２０の一形態例を示す。この培養装置２０は、培養容器１０を支持する支持装置２１と、支持装置２１を振盪する振盪手段（図示せず）を備える振盪培養用の培養装置であって、培養容器１０が図１（ａ）に示すように平袋からなり、かつ図１（ｂ）に示すように、この平袋の片面を下にして支持装置２１上に支持された培養容器１０の少なくとも互いに対向する２辺を支持装置２１に固定する固定手段２４を備えている。 The present invention will be described below based on preferred embodiments with reference to the drawings.
In FIG. 1, the example of 1 form of the culture apparatus 20 of this invention is shown. The culture apparatus 20 is a culture apparatus for shaking culture that includes a support apparatus 21 that supports the culture container 10 and a shaking means (not shown) that shakes the support apparatus 21, and the culture container 10 is shown in FIG. 1), and as shown in FIG. 1 (b), supports at least two opposite sides of the culture vessel 10 supported on the support device 21 with one side of the flat bag facing down. Fixing means 24 for fixing to the device 21 is provided.

培養容器１０となる平袋の基材１１としては、可撓性および不透水性で強度が高いことから合成樹脂フィルムやシート（本明細書では、これらを総称して「フィルム」という。）が好ましい。そして、培養容器を製造する観点からは、ヒートシール性を有するものが好ましく、ヒートシール可能な単層フィルムや、ヒートシール性樹脂層を有する積層フィルム等が挙げられる。また、培養の作業性からは、容器内部が透視可能なものが好ましい。そのようなフィルムを構成する合成樹脂としては、例えば、ポリエチレン（ＰＥ）やポリプロピレン（ＰＰ）などのポリオレフィン、軟質塩化ビニルなどが挙げられる。
特に高い強度が要求される場合は、二軸延伸ナイロンフィルムや二軸延伸ポリエチレンテレフタレート（ＰＥＴ）フィルムを補強層として積層しても良い。酸素などのガスバリア性が必要な場合は、シリカやアルミナの蒸着層や、エチレン−ビニルアルコール共重合体（ＥＶＯＨ）等のガスバリア性樹脂層が積層されたフィルムを使用しても良い。これらのフィルムが複数層積層されたり、重畳されていたりしても良い。
培養容器１０は、内容物の漏れをより厳重に防ぐため、２重袋や３重袋とすることもできる。２重袋や３重袋の内層においては、両面でヒートシール可能な単層フィルムや積層フィルムを用い、シール部１２において内層と外層とをシールして平袋を形成することができる。また、ガスの通路とするため、袋の一部にフィルターを設けることもできる。この場合、フィルターは、ＰＥやＰＰなどのポリオレフィン系不織布であると、基材１１への熱溶着が可能となり好ましい。 As the base material 11 of the flat bag used as the culture container 10, since it is flexible and water-impermeable and has high strength, a synthetic resin film or sheet (in the present specification, these are collectively referred to as “film”). preferable. And from a viewpoint of manufacturing a culture container, what has heat-sealing property is preferable, and the laminated | multilayer film etc. which have a heat-sealable single layer film and a heat-sealable resin layer are mentioned. From the viewpoint of culturing workability, it is preferable that the inside of the container can be seen through. Examples of the synthetic resin constituting such a film include polyolefins such as polyethylene (PE) and polypropylene (PP), and soft vinyl chloride.
When particularly high strength is required, a biaxially stretched nylon film or a biaxially stretched polyethylene terephthalate (PET) film may be laminated as a reinforcing layer. When a gas barrier property such as oxygen is required, a film in which a vapor-deposited layer of silica or alumina or a gas barrier resin layer such as ethylene-vinyl alcohol copolymer (EVOH) is laminated may be used. A plurality of these films may be laminated or superposed.
The culture container 10 can be a double bag or a triple bag in order to prevent leakage of contents more strictly. In the inner layer of the double bag or triple bag, a flat bag can be formed by sealing the inner layer and the outer layer at the seal portion 12 using a single-layer film or a laminated film that can be heat-sealed on both sides. In addition, a filter can be provided in a part of the bag to provide a gas passage. In this case, it is preferable that the filter is a polyolefin-based non-woven fabric such as PE or PP because it can be thermally welded to the substrate 11.

培養容器１０を構成する基材１１の厚みは特に限定されず、基材１１の材質や培養容器１０の容量に応じて適宜選択することができる。支持装置２１に支持させる際の作業性や培養時の保形性の観点からある程度の腰があることが好ましいので、通常は、５０μｍ〜１０００μｍ程度であることが好ましい。培養容器１０の容量は特に制限されないが、例えば、０．１〜１０００Ｌの容量とすることができる。
培養容器１０の内部は、必要に応じて、無菌状態に保持されることが好ましい。無菌状態とする方法としては、例えば、清浄な環境下で製造したり、製造後に滅菌したりして内部を無菌状態することが挙げられる。滅菌方法は特に制限されないが、例えば、γ線滅菌などの放射線滅菌、エチレンオキサイドガス（ＥＯＧ）滅菌、オートクレーブ（ＡＣ）滅菌などが挙げられる。 The thickness of the base material 11 constituting the culture vessel 10 is not particularly limited, and can be appropriately selected according to the material of the base material 11 and the capacity of the culture vessel 10. Since it is preferable that there is a certain amount of waist from the viewpoint of workability at the time of supporting the support device 21 and shape retention during culture, it is usually preferably about 50 μm to 1000 μm. Although the capacity | capacitance in particular of the culture container 10 is not restrict | limited, For example, it can be set as the capacity | capacitance of 0.1-1000L.
The inside of the culture vessel 10 is preferably maintained in a sterile state as necessary. As a method of making it aseptic, for example, it can be manufactured in a clean environment, or sterilized after production to sterilize the inside. The sterilization method is not particularly limited, and examples thereof include radiation sterilization such as γ-ray sterilization, ethylene oxide gas (EOG) sterilization, and autoclave (AC) sterilization.

支持装置２１は、底壁部２２と側壁部２３とを備えるトレイ状であり、培養容器１０は、平袋の片面を下にして載置される。培養容器１０の上面となる平袋の別の片面には、給気、排気、内容物のサンプリングなどに使用可能なポート１６を有するポート部材１５が設けられている。ポート部材１５は、例えば合成樹脂の成形品からなり、袋の内面に鍔状のフランジ（図示せず）を内側から接着剤や熱融着等で固着し、基材１１に形成した開口を通して外方に突出させる。ポート１６は、それぞれキャップ、蓋、栓、コック等で開閉可能にし、また、必要に応じてチューブ１７を取り付け可能にしてもよい。また、注射器（シリンジ）等を用いて培養液の採取や注入等を行うようにしてもよい。給気や排気のように継続して開放する場合には、雑菌の混入を防ぐため、フィルターを設けることが好ましい。
給気は、目的の培養を行うために、空気（あるいはＯ２含有ガス）により好気的条件としたり、ＣＯ２により培地のｐＨを一定に保ったり、窒素ガスやアルゴン等の不活性ガスにより嫌気的条件としたりする等、任意の気体を１種または２種以上供給することが可能である。 The support device 21 has a tray shape including a bottom wall portion 22 and a side wall portion 23, and the culture vessel 10 is placed with one side of a flat bag facing down. A port member 15 having a port 16 that can be used for air supply, exhaust, sampling of contents, and the like is provided on another side of the flat bag that is the upper surface of the culture vessel 10. The port member 15 is made of, for example, a synthetic resin molded article, and a flange-like flange (not shown) is fixed to the inner surface of the bag from the inside by an adhesive, heat fusion, or the like. Protruding toward. Each port 16 can be opened and closed by a cap, a lid, a stopper, a cock, etc., and a tube 17 can be attached if necessary. Further, the culture solution may be collected or injected using a syringe (syringe) or the like. In the case of continuously opening like air supply or exhaust, it is preferable to provide a filter in order to prevent contamination of germs.
In order to perform the desired culture, air supply is performed under aerobic conditions with air (or O 2 -containing gas), the pH of the medium is kept constant with CO 2 , or with an inert gas such as nitrogen gas or argon. It is possible to supply one kind or two or more kinds of arbitrary gases such as anaerobic conditions.

本発明においては、培養容器１０の周縁部１３に張力をかけた状態を維持するため、培養容器１０の少なくとも互いに対向する２辺を支持装置２１に固定する固定手段が設けられる。固定手段としては、バネ、ネジ、クリップ、リベット、係合、嵌合、磁石、接着剤など、種々の手段を利用することができる。各辺は、辺の長手方向に沿って連続した範囲を固定してもよく、また、固定箇所を複数に分散してもよい。   In the present invention, in order to maintain a state where tension is applied to the peripheral portion 13 of the culture vessel 10, a fixing means for fixing at least two sides of the culture vessel 10 facing each other to the support device 21 is provided. As the fixing means, various means such as a spring, a screw, a clip, a rivet, an engagement, a fitting, a magnet, and an adhesive can be used. For each side, a continuous range along the longitudinal direction of the side may be fixed, or the fixing points may be dispersed in a plurality.

図１に示す培養装置２０は、培養容器１０である平袋の周縁部１３に複数の孔１４を有する。孔１４は、図１（ａ）に示すように、固定手段となる係止具２４に固定されるそれぞれの辺ごとに、適宜の間隔で複数形成されることが好ましい。図１（ｂ）に示すように、係止具２４を孔１４に通すことにより平袋の周縁部１３を支持装置２１の側壁部２３にしっかりと固定することができる。固定の際には、左右に引っ張りながら孔１４の位置を係止具２４の取り付け位置に合わせることにより、平袋にテンションを加えてシワの発生しにくい状態とすることができる。そのため、対向する２辺の孔１４の距離は、支持装置２１の寸法に合わせて予め適切な位置をとるように設定することが好ましい。   The culture apparatus 20 shown in FIG. 1 has a plurality of holes 14 in a peripheral edge portion 13 of a flat bag that is a culture container 10. As shown in FIG. 1A, it is preferable that a plurality of holes 14 are formed at appropriate intervals for each side fixed to the locking tool 24 serving as a fixing means. As shown in FIG. 1B, the peripheral edge 13 of the flat bag can be firmly fixed to the side wall 23 of the support device 21 by passing the locking tool 24 through the hole 14. At the time of fixing, by adjusting the position of the hole 14 to the mounting position of the locking tool 24 while pulling left and right, it is possible to apply tension to the flat bag so that wrinkles are not easily generated. Therefore, it is preferable to set the distance between the two opposing holes 14 in advance so as to take an appropriate position in accordance with the dimensions of the support device 21.

また、図２に示す培養装置２０Ａは、培養容器１０Ａである平袋の周縁部１３が幅広とされ、つかみ具２４Ａで固定するために十分なつかみ代を確保している。これにより、図２（ｂ）に示すように、固定の際には、培養容器１０Ａを左右に引っ張りながらつかみ具２４Ａに挟持（把持）させることにより、平袋にテンションを加えてシワの発生しにくい状態とすることができる。
つかみ具２４Ａを用いると、周縁部１３に特別な加工をしなくても培養容器１０Ａを固定することができる。つかみ具は、周縁部１３の保持力を高めるため、挟持面に突起や凹凸を設けてもよい。また、十分なテンションが加わる寸法に合わせてつかみ具２４Ａでつかむ位置の目安を指示するため、周縁部１３に印刷等でマークを設けてもよい。 Further, in the culture apparatus 20A shown in FIG. 2, the peripheral edge portion 13 of the flat bag that is the culture vessel 10A is wide, and a sufficient grip margin is secured for fixing with the gripping tool 24A. Thus, as shown in FIG. 2B, when fixing, the culture container 10A is clamped (gripped) by the gripping tool 24A while pulling it left and right, thereby applying tension to the flat bag and generating wrinkles. It can be difficult.
When the gripping tool 24A is used, the culture vessel 10A can be fixed without special processing on the peripheral edge portion 13. In order to increase the holding force of the peripheral edge portion 13, the gripper may be provided with protrusions or irregularities on the clamping surface. In addition, a mark may be provided on the peripheral portion 13 by printing or the like in order to indicate a guide for a position to be grasped by the grasping tool 24A in accordance with a dimension to which sufficient tension is applied.

培養装置は、支持装置を揺動可能な揺動装置（図示せず）を備えることが好ましい。揺動方法としては、円運動や往復運動等の水平面内の運動、シーソー運動やロッキング等の上下運動が挙げられる。   The culture apparatus preferably includes a rocking device (not shown) that can rock the support device. Examples of the swinging method include movement in a horizontal plane such as circular movement and reciprocating movement, and vertical movement such as seesaw movement and rocking.

上記の培養装置２０，２０Ａは、固定手段２４，２４Ａにより平袋状の培養容器１０，１０Ａにテンションを加えて固定することができるので、ピンホールの要因となるシワの発生を防止することができる。これにより、振盪が長期にわたる場合でも、培養中のピンホールの発生を抑制し、培養の歩留まりを向上することができる。
振盪培養においては、培地として流動性のある液体培地を用い、これに微生物等の培養体を供給した培養液１の液面上に相当量の気体２を存在させ、培養液１に気体２を効率よく吸収させるため、培養液１を振盪により、よく波立たせる必要がある。従来は、この波打ちによる衝撃がシール部１２付近に断続的に加わるとき、シール部１２付近の基材１１が同期して振動することにより、シワやピンホールの原因となっていたものと考えられる。
これに対して、本形態例の培養装置２０，２０Ａによれば、培養容器１０，１０Ａの周縁部１３にテンションをかけて固定し、シワを防止しているので、培養液１の波打ちによる衝撃がシール部１２付近に断続的に加わっても、シール部１２付近の基材１１の振動が抑制される等の作用により、基材１１の損傷を効果的に抑制することができる。
また、培養容器１０の下面が底壁部２２上に接地しているので、内容物を含む培養容器１０の荷重が底壁部２２に加わり、固定手段２４，２４Ａへの負荷を軽減することができる。このため、固定手段２４，２４Ａは簡便な構成でもよい。 The culture apparatuses 20 and 20A can fix the flat bag-shaped culture containers 10 and 10A by applying tension to the flat bag-like culture containers 10 and 10A, thereby preventing wrinkles that cause pinholes. it can. Thereby, even when shaking is prolonged, generation | occurrence | production of the pinhole during culture | cultivation can be suppressed and the yield of culture | cultivation can be improved.
In shaking culture, a fluid liquid medium is used as a medium, and a considerable amount of gas 2 is present on the surface of the culture medium 1 to which a culture body such as a microorganism is supplied. In order to absorb efficiently, it is necessary to shake the culture solution 1 well by shaking. Conventionally, when the impact due to this undulation is intermittently applied in the vicinity of the seal portion 12, the base material 11 in the vicinity of the seal portion 12 vibrates synchronously, which is considered to cause wrinkles and pinholes. .
On the other hand, according to the culture apparatuses 20 and 20A of the present embodiment, since the wrinkles are prevented by fixing the peripheral portions 13 of the culture containers 10 and 10A with tension, the impact caused by the undulation of the culture medium 1 Even if intermittently applied in the vicinity of the seal portion 12, damage to the base material 11 can be effectively suppressed by an action such as suppression of vibration of the base material 11 in the vicinity of the seal portion 12.
In addition, since the lower surface of the culture vessel 10 is grounded on the bottom wall portion 22, the load of the culture vessel 10 including the contents is applied to the bottom wall portion 22, and the load on the fixing means 24 and 24A can be reduced. it can. For this reason, the fixing means 24, 24A may have a simple configuration.

以上、本発明を好適な実施の形態に基づいて説明してきたが、本発明は上述の形態例に限定されるものではなく、本発明の要旨を逸脱しない範囲で種々の改変が可能である。
培養装置には、培養容器内の培養液や培養体の温度をモニターして制御するために、温度センサーを設置してもよい。温度センサーとしては、熱電対やサーミスタなど接触式のもの、放射温度計などの非接触式のものを用いることができる。温度センサーは培養容器に直接貼り付けてもよいが、支持装置に取り付けても良い。さらには、排液管に温度センサーを接続し、例えば、培養中の培地の一部を排液管を通じて吸引し、その温度を検知できるようにしてもよい。
また、必要に応じ、温度センサーに代えて、あるいは温度センサーと併用して、培養液のｐＨや酸素濃度等をセンサーによってモニタリングしてもよい。 As mentioned above, although this invention has been demonstrated based on suitable embodiment, this invention is not limited to the above-mentioned example, Various modifications are possible in the range which does not deviate from the summary of this invention.
The culture apparatus may be provided with a temperature sensor in order to monitor and control the temperature of the culture solution and the culture in the culture vessel. As the temperature sensor, a contact type sensor such as a thermocouple or a thermistor and a non-contact type sensor such as a radiation thermometer can be used. The temperature sensor may be directly attached to the culture vessel, but may be attached to the support device. Further, a temperature sensor may be connected to the drainage pipe so that, for example, a part of the culture medium being cultured is sucked through the drainage pipe and the temperature can be detected.
Further, if necessary, instead of the temperature sensor or in combination with the temperature sensor, the pH and oxygen concentration of the culture solution may be monitored by the sensor.

培養装置には、培養容器の内部を加温して培養を促進するために加温装置を備えることが好ましい。加温装置は、培養容器の周囲や基材の内部に設置してもよい。また、培養容器が２重袋や３重袋など多重の包装袋である場合、外袋の基材の底面をくり抜いて別装置のヒーターに内袋を接触させてもよい。加温装置は培養容器に固定してもよいが、例えば、加温装置を支持装置の表面に設けるなど、培養容器と接触させるだけでもよい。加温装置としては、ニクロム線、シーズ線、カーボンやアルミ箔の回路を合成樹脂層に挟み込んだ面状発熱体などの発熱体を用いることができる。この加温装置は、培養装置に取り付けられた温度センサーの情報に基づいて制御可能とされていることが好ましい。
培養装置の加温は、設置場所の雰囲気温度を恒温にすることで制御してもよい。 The culture device is preferably provided with a heating device in order to warm the inside of the culture vessel and promote culture. The heating device may be installed around the culture container or inside the substrate. In addition, when the culture container is a multiple packaging bag such as a double bag or triple bag, the bottom of the base material of the outer bag may be cut out and the inner bag may be brought into contact with the heater of another device. The heating device may be fixed to the culture vessel, but it may be merely brought into contact with the culture vessel, for example, the heating device is provided on the surface of the support device. As the heating device, a heating element such as a sheet heating element in which a circuit of nichrome wire, sheathed wire, carbon or aluminum foil is sandwiched between synthetic resin layers can be used. This heating device is preferably controllable based on information from a temperature sensor attached to the culture device.
You may control the heating of a culture apparatus by making the atmospheric temperature of an installation place constant.

培養容器１０として、内層に厚さ１００μｍのＰＥからなる単層フィルムを用い、外層にＰＥ５０μｍ／ＥＶＯＨ２５μｍ／ＰＥ５０μｍの共押出による積層フィルムを２組用意し、対向する内面の周囲をシールして容量が５〜２００Ｌ、外寸が３８０ｍｍ×３６０ｍｍ〜１０００ｍｍ×１４００ｍｍの平袋を作製した。この平袋に、液体培地を容量の２〜５割程度入れ、ポートから容量比５〜８％程度のＣＯ２を含む空気を流通させるためのチューブを接続した。また、平袋は、テンションをかけて対向する２辺を支持装置に固定した。円運動、往復運動、シーソー運動等により振盪培養を行ったところ、２０日間以上の長時間振盪させても容器の周縁部にシワがよることがなく、ピンホールは発生しなかった。 As the culture vessel 10, a single-layer film made of PE having a thickness of 100 μm is used for the inner layer, and two sets of laminated films obtained by co-extrusion of PE 50 μm / EVOH 25 μm / PE 50 μm are prepared for the outer layer, and the capacity is sealed by sealing the periphery of the opposing inner surfaces A flat bag having a size of 5 to 200 L and an outer size of 380 mm × 360 mm to 1000 mm × 1400 mm was produced. About 20 to 50% of the volume of the liquid medium was put into this flat bag, and a tube for circulating air containing CO 2 having a volume ratio of about 5 to 8% was connected from the port. Further, the flat bag was fixed to the supporting device on the two opposite sides by applying tension. When shaking culture was performed by circular motion, reciprocating motion, seesaw motion or the like, no wrinkles were formed on the peripheral edge of the container even when shaken for a long period of 20 days or longer, and no pinholes were generated.

１０，１０Ａ…培養容器、１３…周縁部、１４…係止孔、２０，２０Ａ…培養装置、２１…支持装置、２４…係止具（固定手段）、２４Ａ…つかみ具（固定手段）。 DESCRIPTION OF SYMBOLS 10,10A ... Culture container, 13 ... Peripheral part, 14 ... Locking hole, 20, 20A ... Culture apparatus, 21 ... Supporting device, 24 ... Locking tool (fixing means), 24A ... Grazing tool (fixing means).

Claims (3)

対向する基材の内面の周囲をシールしたシール部を有する平袋からなる培養容器と、前記培養容器を支持する支持装置を備える振盪培養用の培養装置であって、前記平袋の片面を下にして前記支持装置上に支持された前記培養容器の少なくとも互いに対向する２辺を前記支持装置に固定する固定手段を備え、前記支持装置は底壁部と側壁部とを備えるトレイ状であり、前記固定手段は、前記平袋の周縁部を前記底壁部から持ち上がらせて前記支持装置の側壁部に固定していることを特徴とする培養装置。 A culture apparatus for shaking culture, comprising a culture container comprising a flat bag having a seal portion sealed around the inner surface of the opposing base material, and a support device for supporting the culture container, wherein one side of the flat bag is A fixing means for fixing at least two opposite sides of the culture vessel supported on the support device to the support device, and the support device has a tray shape including a bottom wall portion and a side wall portion, The culture device characterized in that the fixing means lifts the peripheral edge of the flat bag from the bottom wall and fixes it to the side wall of the support device. 前記培養容器は、前記平袋の周縁部に前記固定手段に係止される複数の孔を有することを特徴とする請求項１に記載の培養装置。   The culture apparatus according to claim 1, wherein the culture container has a plurality of holes that are locked to the fixing means at a peripheral portion of the flat bag. 前記固定手段は、前記平袋の周縁部をつかんで固定する、つかみ具を備えることを特徴とする請求項１に記載の培養装置。   The culture apparatus according to claim 1, wherein the fixing means includes a gripping tool that holds and fixes a peripheral edge of the flat bag.
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