JP2013048567A - Gas barrier closed container - Google Patents
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Abstract
Description
本発明は、雰囲気調整剤、及び雰囲気調整剤と内容物とを隔離する機構を備えたガスバリア性密閉容器、前記容器を用いたガス濃度の調整方法、並びに前記容器を用いた細胞培養方法に関する。 The present invention relates to an atmosphere adjusting agent, a gas barrier closed container provided with a mechanism for isolating the atmosphere adjusting agent and contents, a gas concentration adjusting method using the container, and a cell culture method using the container.
容器内のガス濃度を調整する方法の１つに、雰囲気調整剤を用いる方法がある。例えば、食品、医薬品、電子材料部品等の品質保持のためには、脱酸素剤や乾燥剤が用いられている。また、植物の生育制御を目的としてエチレン等の植物ホルモン等の濃度を調整する雰囲気調整剤が、抗菌目的では、ヒノキチオールやアリルイソチオシアネート等の抗菌効果を有する揮発性ガスを発生する雰囲気調整剤が、それぞれ用いられている。さらに、細胞等の培養の際は、酸素濃度と二酸化炭素濃度と、場合によってはアルデヒド濃度とを調整する雰囲気調整剤が用いられている。特に、近年では、多くの研究分野で細胞等の低酸素培養が注目されており、特許文献１にはこれらの例が多数報告されている。例えば、体外受精胚は酸素濃度５容量％下、癌細胞は酸素濃度１〜２容量％下、虚血実験や虚血再灌流実験は酸素濃度０．１容量％下で行われることが多い。 One method for adjusting the gas concentration in the container is to use an atmosphere adjusting agent. For example, oxygen scavengers and desiccants are used to maintain the quality of foods, pharmaceuticals, electronic material parts, and the like. In addition, an atmosphere conditioner that adjusts the concentration of plant hormones such as ethylene for the purpose of plant growth control, and for antibacterial purposes, an atmosphere conditioner that generates volatile gases having antibacterial effects such as hinokitiol and allyl isothiocyanate Are used respectively. Furthermore, when culturing cells or the like, an atmosphere regulator that adjusts the oxygen concentration, the carbon dioxide concentration, and, in some cases, the aldehyde concentration is used. In particular, in recent years, low-oxygen culture of cells and the like has attracted attention in many research fields, and Patent Document 1 reports many examples of these. For example, in vitro fertilized embryos are often performed at an oxygen concentration of 5% by volume, cancer cells at an oxygen concentration of 1 to 2% by volume, and ischemia experiments and ischemia reperfusion experiments at an oxygen concentration of 0.1% by volume.
しかし、この方法の場合、ガス濃度は雰囲気調整剤の種類と量、及びガスバリア性密閉容器の容積との組み合わせによってガス濃度が決まるため、ガス濃度の微調整が困難であり、任意のガス濃度に調整することが難しい。また、真空ポンプ等の脱気装置と、ガスボンベ等のガス供給装置を備えたガスバリア性密閉容器によりガス濃度を調整する方法もあるが、装置が大掛かりになり、手間とコストがかかる課題があった。 However, in this method, since the gas concentration is determined by the combination of the type and amount of the atmosphere adjusting agent and the volume of the gas barrier closed container, it is difficult to finely adjust the gas concentration. Difficult to adjust. There is also a method of adjusting the gas concentration by using a gas barrier closed container equipped with a degassing device such as a vacuum pump and a gas supply device such as a gas cylinder, but there is a problem that the device becomes large and labor and cost are high. .
本発明では、簡便かつ安価に、目的のガス濃度に調整する容器、該容器を用いたガス濃度調整方法、及び該容器を用いた細胞等の培養方法を提供することを目的とする。 It is an object of the present invention to provide a container that can be easily and inexpensively adjusted to a target gas concentration, a gas concentration adjusting method using the container, and a method for culturing cells and the like using the container.
本発明者らは、雰囲気調整剤、及び雰囲気調整剤と内容物とを隔離する機構を備えているガスバリア性密閉容器を用い、目的のガス濃度に到達した時点で雰囲気調整剤と内容物とを隔離することにより、内容物の存在する空間のガス濃度を簡便に調整できる事を見出し、本発明を完成した。 The inventors use an atmosphere adjusting agent and a gas barrier closed container equipped with a mechanism for separating the atmosphere adjusting agent and the contents, and when the target gas concentration is reached, the atmosphere adjusting agent and the contents are removed. By isolating, it was found that the gas concentration in the space where the contents exist can be easily adjusted, and the present invention has been completed.
本発明により、任意のガス濃度に簡便かつ安価に調整することが可能となる。 According to the present invention, it is possible to easily and inexpensively adjust an arbitrary gas concentration.
本発明のガスバリア性密閉容器は、少なくとも、雰囲気調整剤、雰囲気調整剤と内容物とを隔離する機構、及びガスバリア性密閉容器本体から構成される。 The gas barrier closed container of the present invention includes at least an atmosphere adjusting agent, a mechanism for isolating the atmosphere adjusting agent and the contents, and a gas barrier closed container main body.
［ガスバリア性密閉容器本体］
本発明のガスバリア性密閉容器本体は、その内外の気体の流通を妨げ、容器内の雰囲気調整剤により調整されたガス濃度を長期間維持する事ができるものであれば特に制限はなく、材質、形状、容積等いずれも適した任意のものを採用することができる。
[Gas barrier sealed container body]
The gas barrier closed container body of the present invention is not particularly limited as long as it prevents the gas flow inside and outside and can maintain the gas concentration adjusted by the atmosphere adjusting agent in the container for a long period of time. Any suitable shape, volume, etc. can be employed.
容器本体の材質は、例えば、硬質の容器であれば、ガラス、金属、ポリカーボネート等のプラスチック、等を用いることができる。また、軟質の容器であれば、ガスバリア性を付与する層及びシーラント層を有するガスバリア性多層フィルム等を用いることができる。 As the material of the container body, for example, glass, metal, plastic such as polycarbonate, etc. can be used if it is a hard container. Moreover, if it is a soft container, the gas-barrier multilayer film etc. which have the layer which provides gas-barrier property, and a sealant layer can be used.
ガスバリア性を付与する層には、例えば、(1)一軸ないし二軸延伸された、ポリエチレンテレフタレート等のポリエステルフィルム、ＭＸＤ６等のポリアミドフィルム、又はビニルアルコール共重合体フィルム、(2)アルミニウム箔、(3)一軸ないし二軸延伸されたポリエチレンテレフタレートフィルム、ポリアミドフィルム、ポリオレフィン系フィルムなどの延伸フィルム上にアルミニウム等の金属薄膜を蒸着した金属蒸着プラスチックフィルム、(4)一軸ないし二軸延伸されたポリエチレンテレフタレートフィルム、ポリアミドフィルム、ポリオレフィン系フィルムなどの延伸フィルム上に、酸化アルミニウムや酸化ケイ素などの無機化合物の薄膜を設けた無機化合物蒸着プラスックフィルム、(5)上記のプラスチックフィルムに塩化ビニリデン樹脂、無機層状化合物とポリビニルアルコール等の水溶性高分子の混合物等を適量塗工したバリア層コーティングプラスチックフィルム、等を用いることができる。更に、ガスバリア性を付与する層を構成する材料は、単層として、または組み合わせて多層として用いることもできる。 Examples of the layer imparting gas barrier properties include (1) uniaxially or biaxially stretched polyester film such as polyethylene terephthalate, polyamide film such as MXD6, or vinyl alcohol copolymer film, (2) aluminum foil, ( 3) Metal-deposited plastic film obtained by depositing a metal thin film such as aluminum on a stretched film such as a uniaxially or biaxially stretched polyethylene terephthalate film, polyamide film or polyolefin film, and (4) uniaxially or biaxially stretched polyethylene terephthalate. An inorganic compound vapor-deposited plastic film in which a thin film of an inorganic compound such as aluminum oxide or silicon oxide is provided on a stretched film such as a film, a polyamide film, or a polyolefin-based film. , Inorganic layered compound and the polyvinyl barrier layer coating a plastic film a mixture of a water-soluble polymer and an appropriate amount coated of such alcohols, and the like can be used. Furthermore, the material which comprises the layer which provides gas-barrier property can also be used as a multilayer as a single layer or in combination.
シーラント層には熱可塑性樹脂を用いることができ、例えば、ポリエチレン、ポリプロピレン、ポリアミド、ポリエステル等を用いることができる。調整対象のガスが酸素である場合は、該容器の酸素透過度は１０ｍＬ／（ｍ２・ｄａｙ・ａｔｍ）以下であることが好ましい。また、該容器内で細胞等を培養する際には、該容器は透明な材質である方が好ましい。これは、容器外部から内容物の視認を可能とするためである。 A thermoplastic resin can be used for the sealant layer, and for example, polyethylene, polypropylene, polyamide, polyester, or the like can be used. When the gas to be adjusted is oxygen, the oxygen permeability of the container is preferably 10 mL / (m 2 · day · atm) or less. Further, when culturing cells or the like in the container, the container is preferably made of a transparent material. This is because the contents can be visually recognized from the outside of the container.
［雰囲気調整剤を隔離する機構］
雰囲気調整剤と内容物とを隔離する機構は何ら限定されず公知の機構を用いることが出来る。例えば硬質容器の場合、容器内を２以上の区域に分け、境界部にバルブ等の繰り返し開閉可能な機構を取り付け、開放していた前記機構を閉めることで雰囲気調整剤と内容物とを隔離する機構を用いることができる。また、軟質容器の場合は、雰囲気調整剤を含む箇所をヒートシールする事で雰囲気調整剤と内容物とを隔離することが出来る。また、クリップ、若しくはチャック等、あるいはこれらを組み合わせて隔離する機構は、繰り返し開閉できるため好ましく用いられる。凹部と凸部の組み合わせで構成されるクリップ等は密封した際の密閉性が高く好ましい。また、チャックは、ダブルチャックとするとより高い密閉性を保って剤を隔離することができるため好ましい。
[Mechanism for isolating the atmosphere modifier]
There is no limitation on the mechanism for separating the atmosphere adjusting agent from the contents, and a known mechanism can be used. For example, in the case of a hard container, the inside of the container is divided into two or more areas, a mechanism that can be repeatedly opened and closed, such as a valve, is attached to the boundary part, and the atmosphere adjusting agent and contents are isolated by closing the opened mechanism. A mechanism can be used. Moreover, in the case of a soft container, an atmosphere regulator and a content can be isolated by heat-sealing the location containing an atmosphere regulator. Further, a clip, a chuck, or the like, or a mechanism for separating them in combination is preferably used because it can be repeatedly opened and closed. A clip or the like composed of a combination of a concave portion and a convex portion is preferable because of its high sealing performance when sealed. The chuck is preferably a double chuck because the agent can be isolated while maintaining higher sealing performance.
本発明の容器には、撹拌羽根等のガスの拡散効率を上げるための機器やガス濃度を測定するための機器を入れてもよい。また、容器内のガスの採取や外部から容器内にガスを注入するための機構を設けてもよい。この機構を設けた場合、外部からガスを注入することで、雰囲気調整剤により調整されたガス濃度をさらに調整することができるため、例えば、内容物によるガスの発生や吸収によるガス濃度の変動を調整することが可能となり、好ましい。 The container of the present invention may be equipped with a device such as a stirring blade for increasing the gas diffusion efficiency and a device for measuring the gas concentration. Further, a mechanism for collecting gas in the container and injecting gas into the container from the outside may be provided. When this mechanism is provided, the gas concentration adjusted by the atmosphere adjusting agent can be further adjusted by injecting gas from the outside. It is possible to adjust, which is preferable.
［雰囲気調整剤、調整対象ガス］
本発明に用いる雰囲気調整剤及び調整対象ガスには特に制限はなく、調整対象ガスに適した雰囲気調整剤を使用できる。例えば、酸素濃度の低減には脱酸素剤を、二酸化炭素濃度の低減にはアルカリ成分を含有した雰囲気調整剤を、水蒸気濃度調整には乾燥剤や調湿剤を用いることが出来る。また、アルコール類、アルデヒド類、アミン類、エステル類、エーテル類、ケトン類、ラクトン類、テルペン類、チオール類等の有機物低減には活性炭などの吸着剤や化学反応を利用した各種除去剤を、それぞれ使用することができる。更に、ガス濃度を高める場合、各種のガス発生剤を用いることが出来る。ガス発生剤としては、目的のガスを吸着させたものや、化学反応によりガスを発生させるものを用いることが出来る。特に、細胞培養の際には、酸素濃度と二酸化炭素濃度とを調整することができるアスコルビン酸等の有機物を主剤とする雰囲気調整剤が好ましく、特許文献１にはこれらの雰囲気調整剤の例が多数報告されている。本発明のガスバリア性密閉容器により、酸素濃度を０〜５容量％に、かつ二酸化炭素濃度２〜１０容量％に、±０．２容量％の精度で調整する事が可能となり好ましい。
[Atmosphere conditioner, gas to be adjusted]
There is no restriction | limiting in particular in the atmosphere adjusting agent and adjustment object gas used for this invention, The atmosphere adjustment agent suitable for adjustment object gas can be used. For example, an oxygen scavenger can be used to reduce the oxygen concentration, an atmosphere conditioner containing an alkali component can be used to reduce the carbon dioxide concentration, and a desiccant or a humidity control agent can be used to adjust the water vapor concentration. In addition, to reduce organic substances such as alcohols, aldehydes, amines, esters, ethers, ketones, lactones, terpenes, thiols, etc., various adsorbents such as activated carbon and various removal agents using chemical reactions are used. Each can be used. Furthermore, when increasing the gas concentration, various gas generating agents can be used. As the gas generating agent, one that adsorbs a target gas or one that generates gas by a chemical reaction can be used. In particular, in the case of cell culture, an atmospheric conditioner mainly composed of an organic substance such as ascorbic acid capable of adjusting the oxygen concentration and the carbon dioxide concentration is preferred. Patent Document 1 discloses examples of these atmosphere conditioners. Many reports have been made. The gas barrier closed container of the present invention is preferable because the oxygen concentration can be adjusted to 0 to 5% by volume and the carbon dioxide concentration can be adjusted to 2 to 10% by volume with an accuracy of ± 0.2% by volume.
［ガス濃度分析方法］
容器内のガス濃度分析方法に特に制限はなく、容器内に蛍光式のセンサー部を取り付け、密閉した容器の外部からその蛍光の消光量を測定することでガス濃度が分析できる非破壊式の分析機器を用いる方法や、容器に取り付けたガス採取機構からガスを採取し、ガスクロマトグラフィー等の分析機器で分析する方法、あるいは、小型のガス濃度分析機器を容器内に入れる方法や、検知剤等を用いてもよい。小型のガス濃度分析機器や検知剤等を用いた場合は、ガス濃度変化を連続的に観察できるため、雰囲気調整剤を隔離するタイミングを把握しやすく好ましい。
[Gas concentration analysis method]
There is no particular restriction on the gas concentration analysis method in the container. A non-destructive analysis that can analyze the gas concentration by attaching a fluorescence sensor inside the container and measuring the extinction amount of the fluorescence from the outside of the sealed container. A method of using an instrument, a method of collecting gas from a gas sampling mechanism attached to a container and analyzing it with an analytical instrument such as gas chromatography, a method of putting a small gas concentration analyzer in a container, a detecting agent, etc. May be used. When a small gas concentration analyzer or a detection agent is used, the change in gas concentration can be continuously observed.
［ガス濃度調整方法、細胞培養方法］
本発明のガス濃度調整方法を用いると、既存の雰囲気調整剤を用いたガス濃度調整方法や、炭酸ガスインキュベーターやマルチガスインキュベーターでは実現が困難であった、酸素濃度０〜１容量％の範囲においても酸素濃度を調整することが可能となる。したがって、簡便かつ安価に上記酸素濃度下における細胞培養実験を実施することができる。
[Gas concentration adjustment method, cell culture method]
When the gas concentration adjusting method of the present invention is used, it is difficult to realize the gas concentration adjusting method using an existing atmosphere adjusting agent, a carbon dioxide incubator, or a multi-gas incubator. It is also possible to adjust the oxygen concentration. Therefore, the cell culture experiment under the oxygen concentration can be carried out simply and inexpensively.
［細胞］
本発明の細胞培養方法で用いられる細胞には特に制限はなく、微生物細胞、動物細胞、植物細胞、昆虫細胞等、任意の細胞を用いることができる。
[cell]
The cells used in the cell culture method of the present invention are not particularly limited, and any cell such as a microbial cell, an animal cell, a plant cell, or an insect cell can be used.
［培養容器］
本発明の細胞培養方法で用いられる培養容器は、容器外との通気性が確保されていれば特に制限はなく、容積、形状、材質等いずれも培養に適した任意のものを採用することができる。また、蓋部を有する培養容器が好ましく用いられるが、この際は、培養容器外との通気性を確保する必要がある。
[Culture container]
The culture vessel used in the cell culturing method of the present invention is not particularly limited as long as air permeability to the outside of the vessel is ensured, and any volume, shape, material, etc. suitable for culture can be adopted. it can. In addition, a culture container having a lid is preferably used, but in this case, it is necessary to ensure air permeability outside the culture container.
以下に実施例をあげて本発明を詳細に説明するが、本発明はこれによって限定されるものではない。 Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited thereto.
（酸素及び二酸化炭素濃度測定方法）
袋内のガスを注射器で吸引し、酸素濃度はジルコニア式酸素濃度計（東レエンジニアリング株式会社製）を用いて、二酸化炭素濃度は赤外線式二酸化炭素濃度計（光明理化学工業株式会社製）を用いて測定した。
(Oxygen and carbon dioxide concentration measurement method)
The gas in the bag is sucked with a syringe, the oxygen concentration is measured using a zirconia oxygen analyzer (manufactured by Toray Engineering Co., Ltd.), and the carbon dioxide concentration is measured using an infrared carbon dioxide concentration meter (manufactured by Komyo Chemical Co., Ltd.). It was measured.
（実施例１）
酸素バリア層が二軸延伸ナイロン（ＯＮＹ）とエチレンビニルアルコール共重合体系フィルム（ＥＶＯＨ）からなり、熱可塑性樹脂として低密度ポリエチレン（ＬＬＤＰＥ）を用いた酸素透過度が１０ｍＬ／（ｍ２・ｄａｙ・ａｔｍ）の多層フィルムからなる三方がシールされた袋に、２５００ｍLの空気を封入し、雰囲気調整剤（商品名「アネロパック・ケンキ５％」、三菱ガス化学株式会社製）1つを入れて袋の口をシールし、密閉した。これを３７℃の恒温層に入れ、酸素濃度が約３容量％に到達した時点（１５分後）で雰囲気調整剤を袋の隅に寄せ、ヒートシールしてこれを袋内で隔離した。その後、３日間３７℃で保管し、雰囲気調整剤を含まない区域内の酸素濃度と二酸化炭素濃度とを測定した。結果を表１に示した。
Example 1
The oxygen barrier layer is made of biaxially stretched nylon (ONY) and an ethylene vinyl alcohol copolymer film (EVOH), and the oxygen permeability using low density polyethylene (LLDPE) as a thermoplastic resin is 10 mL / (m 2 · day · Atm) A three-sided sealed bag made of multilayer film is filled with 2500 mL of air, and one atmosphere control agent (trade name “Aneropack Kenki 5%”, manufactured by Mitsubishi Gas Chemical Co., Ltd.) is placed in the bag. The mouth was sealed and sealed. This was placed in a constant temperature layer at 37 ° C., and when the oxygen concentration reached about 3% by volume (after 15 minutes), the atmosphere modifier was brought to the corner of the bag and heat sealed to isolate it in the bag. Then, it stored at 37 degreeC for 3 days, and measured the oxygen concentration and carbon dioxide concentration in the area which does not contain an atmospheric conditioner. The results are shown in Table 1.
（実施例２）
酸素濃度が約１容量％に到達した時点（４３分後）で雰囲気調整剤を袋内で隔離した以外は実施例１と同様にして、酸素濃度と二酸化炭素濃度とを測定した。結果を表１に示した。
(Example 2)
The oxygen concentration and carbon dioxide concentration were measured in the same manner as in Example 1 except that the atmosphere adjusting agent was isolated in the bag when the oxygen concentration reached about 1% by volume (after 43 minutes). The results are shown in Table 1.
（実施例３）
酸素濃度が約０．５容量％に到達した時点（５１分後）で雰囲気調整剤を袋内で隔離した以外は実施例１と同様にして、酸素濃度と二酸化炭素濃度とを測定した。結果を表１に示した。
(Example 3)
The oxygen concentration and carbon dioxide concentration were measured in the same manner as in Example 1 except that the atmosphere adjusting agent was isolated in the bag when the oxygen concentration reached about 0.5% by volume (after 51 minutes). The results are shown in Table 1.
（比較例１）
雰囲気調整剤を袋内で隔離しないこと以外は実施例１と同様にして、１５分後と３日後の袋内の酸素濃度と二酸化炭素濃度とを測定した。結果を表１に示した。
(Comparative Example 1)
The oxygen concentration and carbon dioxide concentration in the bag after 15 minutes and 3 days were measured in the same manner as in Example 1 except that the atmosphere modifier was not isolated in the bag. The results are shown in Table 1.
実施例１〜３から明らかなように、本発明のガスバリア性密閉容器を用いると、複雑で高価な装置を用いることなく、酸素及び二酸化炭素濃度を±０．２容量％という非常に高い精度で調整することが可能であった。また、炭酸ガスインキュベーター等でも調整が困難であった酸素濃度１容量％以下のガス濃度も調整することが可能であった。 As is apparent from Examples 1 to 3, when the gas barrier closed container of the present invention is used, the oxygen and carbon dioxide concentrations are as high as ± 0.2% by volume without using a complicated and expensive apparatus. It was possible to adjust. In addition, it was possible to adjust the gas concentration with an oxygen concentration of 1% by volume or less, which was difficult to adjust with a carbon dioxide incubator or the like.
一方、比較例１では、雰囲気調整剤を隔離していないため、酸素濃度、二酸化炭素濃度ともに低下し続け、特定のガス雰囲気を安定的に維持することができなかった。 On the other hand, in Comparative Example 1, since the atmosphere adjusting agent was not isolated, both the oxygen concentration and the carbon dioxide concentration continued to decrease, and the specific gas atmosphere could not be stably maintained.
（実施例４）
ガスバリア性密閉容器として、アネロ専用パウチ袋（三菱ガス化学株式会社製）を用い、これにＭＥＭ培地（製品名；インビトロジェン社製 ＧＩＢＣＯ １１０９５-０８０ Ｍｉｎｉｍｕｍ Ｅｓｓｅｎｔｉａｌ Ｍｅｄｉｕｍ， ｌｉｑｕｉｄ Ｅａｒｌｅ塩含有）を１０ｍＬ入れた９０ｍｍ径ディッシュ２枚（品名；ＡＧＣテクノグラス社製 ＩＷＡＫＩ ３０２０−１００ ９０ｍｍ／Ｔｉｓｓｕｅ Ｃｕｌｔｕｒｅ Ｄｉｓｈ）、雰囲気調整剤（商品名「アネロパック・ケンキ５％」、三菱ガス化学株式会社製、及び商品名「アネロパック・微好気」、三菱ガス化学株式会社製）各１個を封入し、凹部と凸部とを組み合わせた着脱自在な密閉性の高いクリップでパウチ袋を密閉した。しばしばディッシュのふたを持ち上げてディッシュ内の空気とパウチ袋内の空気とが均一になるように空気の拡散を促し、酸素濃度が約０．７容量％に到達した時点（１０分後）で、クリップを用いてディッシュと雰囲気調整剤とを隔離した。その後、３日間３７℃で保管し、ディッシュが封入されている区域内の酸素濃度と二酸化炭素濃度とを測定した。結果を表２に示した。
Example 4
90 mm containing 10 mL of MEM medium (product name: GIBCO 11095-080 Minimum Essential Medium, liquid Earle salt, manufactured by Invitrogen Corporation) was used as an anero barrier container for the air barrier (Mitsubishi Gas Chemical Co., Ltd.). 2 diameter dishes (product name: IWAKI 3020-100 90 mm / Tissue Culture Dish manufactured by AGC Techno Glass), atmosphere conditioner (trade name “Aneropack Kenki 5%”, manufactured by Mitsubishi Gas Chemical Co., Ltd., and trade name “Aneropack “Microaerobic” (Mitsubishi Gas Chemical Co., Ltd.) each was enclosed, and the pouch bag was sealed with a detachable and high-tightness clip combining concave and convex portions. Often when the lid of the dish is lifted to promote air diffusion so that the air in the dish and the air in the pouch bag are uniform, the oxygen concentration reaches about 0.7% by volume (after 10 minutes), A dish was used to isolate the dish from the atmosphere modifier. Then, it stored at 37 degreeC for 3 days, and measured the oxygen concentration and the carbon dioxide concentration in the area where the dish was enclosed. The results are shown in Table 2.
（比較例２）
ディッシュと雰囲気調整剤とをパウチ袋内で隔離しないこと以外は実施例４と同様にして、１０分後と３日後の袋内の酸素濃度と二酸化炭素濃度とを測定した。結果を表２に示した。
(Comparative Example 2)
The oxygen concentration and carbon dioxide concentration in the bag after 10 minutes and 3 days were measured in the same manner as in Example 4 except that the dish and the atmosphere modifier were not separated in the pouch bag. The results are shown in Table 2.
細胞培養用の培地を含むディッシュを同封した実施例４でも、雰囲気調整剤隔離後３日間は±０．２容量％の範囲で目的の酸素濃度と二酸化炭素濃度とを維持していた。 Even in Example 4 in which a dish containing a cell culture medium was enclosed, the target oxygen concentration and carbon dioxide concentration were maintained within a range of ± 0.2% by volume for 3 days after isolation of the atmosphere control agent.
一方、比較例２では、雰囲気調整剤を隔離していないため、酸素濃度、二酸化炭素濃度ともに低下し続け、特定のガス濃度を安定的に維持することができなかった。 On the other hand, in Comparative Example 2, since the atmosphere adjusting agent was not isolated, both the oxygen concentration and the carbon dioxide concentration continued to decrease, and the specific gas concentration could not be stably maintained.
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| JP6026707B1 (en) * | 2016-02-08 | 2016-11-16 | 花王株式会社 | Method for producing mature sebaceous cells |
| WO2018003073A1 (en) * | 2016-06-30 | 2018-01-04 | 株式会社サンプラテック | Set for transporting culture container and unit for transporting cell or biological tissue |
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| JPS62265979A (en) * | 1986-05-13 | 1987-11-18 | I S O:Kk | Method for cultivating anaerobic mold and tool therefor |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP6026707B1 (en) * | 2016-02-08 | 2016-11-16 | 花王株式会社 | Method for producing mature sebaceous cells |
| WO2017138078A1 (en) * | 2016-02-08 | 2017-08-17 | 花王株式会社 | Method for producing mature sebaceous gland cells |
| WO2018003073A1 (en) * | 2016-06-30 | 2018-01-04 | 株式会社サンプラテック | Set for transporting culture container and unit for transporting cell or biological tissue |
| JPWO2018003073A1 (en) * | 2016-06-30 | 2019-04-18 | 株式会社サンプラテック | Culture container transport set and cell / body tissue transport unit |
| US11312933B2 (en) | 2016-06-30 | 2022-04-26 | Sanplatec Corporation Ltd. | Set for transporting culture container and unit for transporting cell or biological tissue |
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