JP2012115232A - Cell culturing apparatus - Google Patents

Cell culturing apparatus Download PDF

Info

Publication number
JP2012115232A
JP2012115232A JP2010270236A JP2010270236A JP2012115232A JP 2012115232 A JP2012115232 A JP 2012115232A JP 2010270236 A JP2010270236 A JP 2010270236A JP 2010270236 A JP2010270236 A JP 2010270236A JP 2012115232 A JP2012115232 A JP 2012115232A
Authority
JP
Japan
Prior art keywords
culture
gas
tube
tubes
blowing means
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
JP2010270236A
Other languages
Japanese (ja)
Other versions
JP5768365B2 (en
Inventor
Mie Mori
美栄 森
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
IHI Corp
Original Assignee
IHI Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by IHI Corp filed Critical IHI Corp
Priority to JP2010270236A priority Critical patent/JP5768365B2/en
Publication of JP2012115232A publication Critical patent/JP2012115232A/en
Application granted granted Critical
Publication of JP5768365B2 publication Critical patent/JP5768365B2/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • 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
    • C12M23/06—Tubular
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/06—Nozzles; Sprayers; Spargers; Diffusers

Landscapes

  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

PROBLEM TO BE SOLVED: To provide a cell culturing apparatus capable of efficiently stirring a culture solution with ventilation air etc. regardless of the size of a culture tank.SOLUTION: In the cell culturing apparatus, the culture solution 11 is stored in the culture tank 10 and cells are cultured by feeding a gas into the culture tank 10. A pair of vertical culture cylinders 12a and 12b are connected to form a reaction tank 10. Gas blowing means 14a and 14b are respectively connected to the bottoms of the culture cylinders 12a and 12b, and exhaust means 15a and 15b are respectively connected to the tops of the culture cylinders 12a and 12b. The gas is blown into the bottom of one culture cylinder 12a from one gas blowing means 14a, and the gas is discharged from the top of the other culture cylinder 12b via the exhaust means 15b. These steps are alternately repeated to alternately move and stir the culture solution 11 in the culture cylinders 12a and 12b.

Description

本発明は、細胞を播種した培養液を収容し、その培養液にエアーを通気しながら撹拌して細胞培養するための細胞培養装置に関するものである。   The present invention relates to a cell culture apparatus for storing a culture solution seeded with cells and culturing the cells by agitating the culture solution with aeration of air.

従来、細胞を播種した培養液を収容し、その培養液にエアーを通気しながら撹拌して細胞培養するための細胞培養装置は、内部に撹拌羽根を有するジャーファーメンターおよびタンクの構造が主流である（特許文献１）。   Conventionally, a cell culture apparatus for containing a culture medium in which cells are seeded and agitating the cell while aerating air to the culture medium and culturing the cell has a main structure of a jar fermenter and a tank having a stirring blade inside. Yes (Patent Document 1).

しかし、撹拌羽根による培養方法では、撹拌によるせん断力で細胞にダメージを与えることが懸念されるため、エアーにて撹拌することが提案されている（特許文献２，３）。   However, in the culture method using a stirring blade, it is feared that cells are damaged by a shearing force by stirring, and therefore stirring with air has been proposed (Patent Documents 2 and 3).

特許文献２では、培養容器内に、下端が開口した管を設け、その管の上部の圧力を変動させることで、培養容器内の培養液を管内に、或いは管から培養容器へ交互に繰り返し出し入れすることで撹拌するようにしている。   In Patent Document 2, a tube having a lower end is provided in a culture vessel, and the culture solution in the culture vessel is repeatedly put into and out of the culture vessel by changing the pressure at the top of the tube. To make it stir.

また、特許文献３では、培養容器内の下部から気泡を断続的に供給し、その気泡を培養容器内で成長させて容器の内径に近い大泡とし、その大泡が培養容器を上昇することで、培養液を撹拌するようにしている。   Moreover, in patent document 3, bubbles are intermittently supplied from the lower part in the culture vessel, the bubbles are grown in the culture vessel to form large bubbles close to the inner diameter of the vessel, and the large bubbles rise in the culture vessel. The culture solution is stirred.

特開平０９−１３５６８０号公報JP 09-135680 A 特開２００６−２８８２４４号公報JP 2006-288244 A 特表２００７−５１１２３０号公報Special table 2007-511230 gazette

しかしながら、特許文献２では、培養液に通気するための通気装置の他に圧力を変動させる手段が必要となり、装置が複雑になる問題があると共に、圧力を変動させるための管内の圧力変動室に気泡が混入した場合には、圧力変動室内の気体を除去することが必要となる問題もある。   However, in Patent Document 2, in addition to the aeration apparatus for aeration of the culture solution, means for varying the pressure is required, and there is a problem that the apparatus becomes complicated, and the pressure variation chamber in the tube for varying the pressure is used. When bubbles are mixed, there is a problem that it is necessary to remove the gas in the pressure fluctuation chamber.

これに対して、特許文献３では、大泡を培養容器内で上昇させるだけで、培養液を撹拌できる利点があるものの、大泡の上昇力だけで撹拌を行うため、培養容器の径が１ｃｍ以下と小さく、培養容器を大容量にすることができない問題がある。   On the other hand, in Patent Document 3, although there is an advantage that the culture solution can be stirred only by raising the large bubbles in the culture vessel, the culture vessel has a diameter of 1 cm because stirring is performed only by the rising force of the large bubbles. There is a problem that the culture container cannot be made large in volume as follows.

そこで、本発明の目的は、上記課題を解決し、培養槽の大きさにかかわらず、培養液を通気エアーなどで効率よく撹拌できる細胞培養装置を提供することにある。   Accordingly, an object of the present invention is to provide a cell culture apparatus that can solve the above-mentioned problems and can efficiently stir the culture solution with aeration air regardless of the size of the culture tank.

上記目的を達成するために請求項１の発明は、培養槽内に培養液を収容し、その培養槽にガスを通気して培養する細胞培養装置において、鉛直な一対の培養筒の下部を連結して反応槽を形成し、その両培養筒の下部にガス吹込手段をそれぞれ接続すると共に両培養筒の上部に排気手段をそれぞれ接続し、いずれか一方のガス吹込手段からその一方の培養筒の下部にガスを吹き込むと共に、他方の培養筒の上部から排気手段を介してガスを排出し、これを交互に繰り返して、両培養筒の培養液を交互に移動して撹拌することを特徴とする細胞培養装置である。   In order to achieve the above object, the invention according to claim 1 is a cell culture apparatus in which a culture solution is accommodated in a culture tank and gas is supplied to the culture tank for culturing, and the lower portions of a pair of vertical culture tubes are connected. The reaction tank is formed, and gas blowing means are connected to the lower portions of the two culture tubes and exhaust means are connected to the upper portions of the two culture tubes, respectively. The gas is blown into the lower part, and the gas is discharged from the upper part of the other culture tube through the exhaust means, and this is alternately repeated, and the culture solution in both culture tubes is alternately moved and stirred. A cell culture device.

請求項２の発明は、鉛直な一対の培養筒の下部を屈曲筒で接続してＵ字状の反応槽を形成し、ガス吹込手段と排気手段の切り換えで、屈曲筒を通して培養液を一方から他方、又はこの逆に培養液を移動させて撹拌する請求項１記載の細胞培養装置である。   In the invention of claim 2, the lower part of a pair of vertical culture tubes is connected by a bent tube to form a U-shaped reaction tank, and the culture solution is passed through the bent tube from one side by switching between the gas blowing means and the exhaust means. The cell culture device according to claim 1, wherein the culture medium is moved and stirred on the other hand or vice versa.

請求項３の発明は、ガス吹込手段には、培養筒の下部の内周壁に沿って気泡を吹き出すエアレーションノズルが設けられる請求項１又は２記載の細胞培養装置である。   A third aspect of the present invention is the cell culture apparatus according to the first or second aspect, wherein the gas blowing means is provided with an aeration nozzle that blows out bubbles along the inner peripheral wall of the lower part of the culture tube.

請求項４の発明は、培養筒の上部には、その培養筒の内周側に沿って上昇した培養液を中心側に流下させる対流促進部材が設けられる請求項３記載の細胞培養装置である。   The invention according to claim 4 is the cell culture device according to claim 3, wherein a convection promoting member is provided at the upper part of the culture tube to cause the culture solution rising along the inner periphery of the culture tube to flow down to the center side. .

本発明によれば、下部を連結した一対の培養筒のいずれか一方にガスを流し、他方の培養筒からガスを排出し、これを交互に繰り返すことで、通気するガスで培養液を培養筒間で移動させながら混合撹拌することができるという優れた効果を発揮するものである。   According to the present invention, gas is allowed to flow through one of a pair of culture tubes connected at the lower portion, gas is discharged from the other culture tube, and this is alternately repeated, so that the culture solution is transferred to the culture tube with the gas that is vented. It exhibits an excellent effect of being able to mix and stir while moving between them.

本発明の一実施の形態を示す図である。It is a figure which shows one embodiment of this invention. 本発明の他の実施の形態を示す図である。It is a figure which shows other embodiment of this invention.

以下、本発明の好適な一実施の形態を添付図面に基づいて詳述する。   A preferred embodiment of the present invention will be described below in detail with reference to the accompanying drawings.

図１（ａ）、図１（ｂ）は、本発明の細胞培養装置の一実施の形態を示したものである。   FIG. 1 (a) and FIG. 1 (b) show an embodiment of the cell culture device of the present invention.

図１において、１０は、培養液１１を収容する培養槽で、鉛直に立ち上げられた左右一対の培養筒１２ａ、１２ｂの下部を屈曲筒１３で連結してＵ字状に形成される。左右の培養筒１２ａ、１２ｂは、ＳＵＳやガラス管などで形成され、例えば１０Ｌの培養液１１を培養する場合、内径は、例えば２５ｃｍ、長さが１００ｃｍに形成されると共に屈曲筒１３は、これら左右の培養筒１２ａ、１２ｂの内径と同じ径で、左右の培養筒１２ａ、１２ｂの下端を接続するよう１８０度屈曲させた形状に形成される。   In FIG. 1, reference numeral 10 denotes a culture tank for storing a culture solution 11, which is formed in a U shape by connecting the lower portions of a pair of left and right culture tubes 12 a and 12 b that are vertically set up with a bent tube 13. The left and right culture tubes 12a and 12b are formed of SUS, glass tubes, or the like. For example, when culturing 10L of the culture solution 11, the inner diameter is, for example, 25cm, and the length is 100cm. It has the same diameter as the inner diameter of the left and right culture tubes 12a, 12b, and is formed in a shape bent 180 degrees so as to connect the lower ends of the left and right culture tubes 12a, 12b.

屈曲筒１３の下部には、左右の培養筒１２ａ、１２ｂの下部にエアーなどのガスを交互に吹き込むガス吹込手段１４ａ、１４ｂが接続され、左右の培養筒１２ａ、１２ｂの上部には、培養筒１２ａ、１２ｂ内に供給されたエアーなどのガスを交互に排出する排気手段１５ａ、１５ｂが接続される。   Gas blowing means 14a, 14b for alternately blowing gas such as air is connected to the lower part of the left and right culture tubes 12a, 12b at the lower part of the bent tube 13, and the culture tubes are provided above the left and right culture tubes 12a, 12b. Exhaust means 15a and 15b for alternately discharging gas such as air supplied into 12a and 12b are connected.

ガス吹込手段１４ａ、１４ｂは、屈曲筒１３から培養槽１０内に入り、その上端が左右の培養筒１２ａ、１２ｂの下部に臨むように設けられた気泡吹込管１６ａ、１６ｂと、屈曲筒１３の外側の気泡吹込管１６ａ、１６ｂに順次接続された供給側開閉バルブ１７ａ、１７ｂと供給側フィルタ１８ａ、１８ｂとで構成される。また供給側開閉バルブ１７ａ、１７ｂの上流側は図示していないがエアーなどのガス供給源に接続され、供給側開閉バルブ１７ａ、１７ｂを開くことで、気泡吹込管１６ａ、１６ｂの上端から左右の培養筒１２ａ、１２ｂ内の培養液１１にエアーなどのガスを吹き込めるようになっている。   The gas blowing means 14a and 14b enter the culture tank 10 from the bent tube 13, and the bubble blowing tubes 16a and 16b provided so that the upper ends thereof face the lower portions of the left and right culture tubes 12a and 12b. It comprises supply-side on-off valves 17a, 17b and supply-side filters 18a, 18b that are sequentially connected to the outer bubble blowing pipes 16a, 16b. Further, the upstream side of the supply side opening / closing valves 17a, 17b is connected to a gas supply source such as air (not shown), and the supply side opening / closing valves 17a, 17b are opened to open the left and right sides of the bubble blowing pipes 16a, 16b. A gas such as air can be blown into the culture solution 11 in the culture tubes 12a and 12b.

排気手段１５ａ、１５ｂは、左右の培養筒１２ａ、１２ｂの上面に接続された排気管１９ａ、１９ｂと、その排気管１９ａ、１９ｂに順次接続された排出側フィルタ２０ａ、２０ｂと、排出側開閉バルブ２１ａ、２１ｂとで構成される。   Exhaust means 15a, 15b includes exhaust pipes 19a, 19b connected to the upper surfaces of the left and right culture tubes 12a, 12b, exhaust side filters 20a, 20b sequentially connected to the exhaust pipes 19a, 19b, and exhaust side opening / closing valves. 21a and 21b.

次に本実施の形態の作用を説明する。   Next, the operation of this embodiment will be described.

先ず、培養槽１０内に動物細胞等を播種した培養液１１を収容し、左右いずれかのガス吹込手段１４ａ（又は１４ｂ）からエアーなどのガスを気泡Ｂとして培養液１１に吹き込み、吹き込んだ培養筒１２ａ（又は１２ｂ）とは反対側の排気手段１５ｂ（又は１５ａ）を開放しておき、これを交互に繰り返すことで、培養液１１が、一方の培養筒１２ａから屈曲筒１３を通して他方の培養筒１２ｂに移動し、これを交互に繰り返すことで、培養液１１が撹拌される。   First, a culture solution 11 in which animal cells or the like are seeded is accommodated in a culture tank 10, and a gas such as air is blown into the culture solution 11 as bubbles B from either the right or left gas blowing means 14a (or 14b). The exhaust means 15b (or 15a) on the opposite side of the cylinder 12a (or 12b) is opened, and this is alternately repeated so that the culture solution 11 passes from the one culture cylinder 12a to the other culture through the bent cylinder 13. The culture solution 11 is stirred by moving to the cylinder 12b and repeating this alternately.

これを更に説明すると、図１（ａ）に示すように、左側のガス吹込手段１４ａの開閉バルブ１７ａを開いた状態とし、右側のガス吹込手段１４ｂの開閉バルブ１７ｂを閉じた状態とし、排気側は、左側の排気手段１５ａの開閉バルブ２１ａを閉じ、右側の排気手段１５ｂの開閉バルブ２１ｂを開いた状態とする。これにより、左側のガス吹込手段１４ａから培養筒１２ａに吹き込まれた気泡Ｂはそのまま培養筒１２ａ内を上昇してその上部の気相部Ｇａに溜まり、その気相部Ｇａの圧力で、水面Ｗａが下がり、培養液１１は、屈曲筒１３を通して右側の培養筒１２ｂに流れ、培養筒１２ｂの気相部Ｇｂ内のガスが排気手段１５ｂの開閉バルブ２１ｂを通って排気される。   Further explaining this, as shown in FIG. 1A, the open / close valve 17a of the left gas blowing means 14a is opened, the open / close valve 17b of the right gas blowing means 14b is closed, and the exhaust side Is a state in which the open / close valve 21a of the left exhaust means 15a is closed and the open / close valve 21b of the right exhaust means 15b is opened. As a result, the bubbles B blown into the culture tube 12a from the gas blowing means 14a on the left side rise as they are in the culture tube 12a and accumulate in the upper gas phase portion Ga. The water surface Wa is caused by the pressure of the gas phase portion Ga. The culture solution 11 flows to the right culture tube 12b through the bent tube 13, and the gas in the gas phase portion Gb of the culture tube 12b is exhausted through the opening / closing valve 21b of the exhaust means 15b.

また培養筒１２ｂ内の培養液１１の液面Ｗｂが上昇したならば、図１（ｂ）に示すように、右側のガス吹込手段１４ｂの開閉バルブ１７ｂを開き、左側のガス吹込手段１４ａの開閉バルブ１７ａを閉じ、排気側は、右側の排気手段１５ｂの開閉バルブ２１ｂを閉じ、左側の排気手段１５ａの開閉バルブ２１ａを開いた状態に切り換える。これにより、右側のガス吹込手段１４ｂから培養筒１２ｂに気泡Ｂが吹き込まれ、培養筒１２ｂの気相部Ｇｂにガスが溜まり、その気相部Ｇｂの圧力で、水面Ｗｂが下がり、培養液１１は、屈曲筒１３を通して左側の培養筒１２ａに流れ、培養筒１２ａの気相部Ｇａ内のガスが排気手段１５ａの開閉バルブ２１ａを通って排気される。   When the liquid level Wb of the culture solution 11 in the culture tube 12b rises, as shown in FIG. 1 (b), the open / close valve 17b of the right gas blowing means 14b is opened, and the left gas blowing means 14a is opened / closed. The valve 17a is closed, and the exhaust side closes the open / close valve 21b of the right exhaust means 15b and switches the open / close valve 21a of the left exhaust means 15a to the open state. Thereby, bubbles B are blown into the culture tube 12b from the gas blowing means 14b on the right side, gas is accumulated in the gas phase portion Gb of the culture tube 12b, the water surface Wb is lowered by the pressure of the gas phase portion Gb, and the culture solution 11 Flows through the bent cylinder 13 to the left culture cylinder 12a, and the gas in the gas phase portion Ga of the culture cylinder 12a is exhausted through the open / close valve 21a of the exhaust means 15a.

このように、左右のガス吹込手段１４ａ、１４ｂと排気手段１５ａ、１５ｂを交互に切り換えることで、通気するガスだけで、培養槽１０内の培養液１１にせん断力を与えることなく撹拌することが可能となる。この切り換えは、培養液１１全体の約３割程度が、左側の培養筒１２ａと右側の培養筒１２ｂを交互に移動するように切り換えることで、効率的に撹拌することができる。また気泡吹込管１６ａ、１６ｂから培養液１１に吹き込む気泡Ｂは、気泡径が約１ｍｍ程度となるように、気泡吹込管１６ａ、１６ｂの吹出口に多孔質板などを設けて吹き込むようにする。   In this way, by alternately switching the left and right gas blowing means 14a and 14b and the exhaust means 15a and 15b, the culture medium 11 in the culture tank 10 can be agitated without applying shearing force only with the aerated gas. It becomes possible. This switching can be efficiently agitated by switching so that about 30% of the whole culture medium 11 moves alternately between the left culture cylinder 12a and the right culture cylinder 12b. The bubbles B blown into the culture solution 11 from the bubble blowing tubes 16a and 16b are blown by providing a porous plate or the like at the outlet of the bubble blowing tubes 16a and 16b so that the bubble diameter is about 1 mm.

次に、図２により本発明の他の実施の形態を説明する。   Next, another embodiment of the present invention will be described with reference to FIG.

図２の実施の形態は、基本的な構成は図１の細胞培養装置と同じであるが、ガス吹込手段１４ａ、１４ｂで、培養筒１２ａ、１２ｂに気泡Ｂを吹き込む際に、その吹き込みで培養筒１２ａ、１２ｂ内の培養液１１をさらに撹拌できるように構成したものである。   The basic configuration of the embodiment of FIG. 2 is the same as that of the cell culture apparatus of FIG. 1, but when the bubbles B are blown into the culture tubes 12a and 12b by the gas blowing means 14a and 14b, the culture is performed by blowing them. The culture solution 11 in the cylinders 12a and 12b is configured to be further stirred.

図２（ａ）に示すように、ガス吹込手段１４ａ、１４ｂの気泡吹込管１６ａ、１６ｂの先端には、エアレーションノズル２３ａ、２３ｂが設けられ、培養筒１２ａ、１２ｂの上部には対流促進部材２４ａ、２４ｂが設けられる。   As shown in FIG. 2A, aeration nozzles 23a and 23b are provided at the tips of the bubble blowing pipes 16a and 16b of the gas blowing means 14a and 14b, and a convection promoting member 24a is provided above the culture tubes 12a and 12b. , 24b are provided.

エアレーションノズル２３ａ、２３ｂは、図２（ｄ）、図２（ｅ）に示すように気泡吹込管１６ａ、１６ｂの先端に取り付けられたリング状ノズル２５の円周方向上面に、多数のノズル孔２６が穿設して形成される。リング状ノズル２５の外径は、培養筒１２ａ、１２ｂの内径と略同じ径に形成される。   As shown in FIGS. 2D and 2E, the aeration nozzles 23a and 23b have a large number of nozzle holes 26 formed on the circumferential upper surface of the ring-shaped nozzle 25 attached to the tips of the bubble blowing tubes 16a and 16b. Is formed by drilling. The outer diameter of the ring-shaped nozzle 25 is formed to be substantially the same as the inner diameter of the culture tubes 12a and 12b.

対流促進部材２４ａ、２４ｂは、図２（ｂ）、図２（ｃ）に示すように下端が開口した断面逆Ｕ字状の円環リング２７の内周にロート状のノズル２８を設け、円環リング２７の頂部にガス放出穴２９がその円周方向に沿って多数穿設して形成される。円環リング２７の外径は、培養筒１２ａ、１２ｂの内径と略同じ径に形成される。   As shown in FIGS. 2B and 2C, the convection promoting members 24a and 24b are provided with a funnel-shaped nozzle 28 on the inner periphery of an annular ring 27 having an inverted U-shaped cross-section whose lower end is opened. A large number of gas discharge holes 29 are formed at the top of the ring ring 27 along the circumferential direction thereof. The outer diameter of the annular ring 27 is formed to be approximately the same as the inner diameter of the culture tubes 12a and 12b.

この実施の形態においては、ガス吹込手段１４ａ（又は１４ｂ）からエアレーションノズル２３ａ（又は２３ｂ）にて気泡Ｂが、培養筒１２ａ（又は１２ｂ）の内周に沿って吹き出され、その気泡Ｂが培養筒１２ａ（又は１２ｂ）の内周壁に沿って上昇し、上部の対流促進部材２４ａ（又は２４ｂ）の円環リング２７内に入り、ガス放出穴２９から吹き出され、気相部Ｇａ（又はＧｂ）に放出される。これにより、培養筒１２ａ（又は１２ｂ）の培養液１１は他方の培養筒１２ｂ（又は１２ａ）に移動するようになるが、この気相部Ｇａの圧力に加えて、内周側では気泡Ｂが多く、中心側では気泡Ｂが少ないため、内周側と中心側では密度差が生じ、中心側では下降流が生じ、対流促進部材２４ａ（又は２４ｂ）の円環リング２７に気泡Ｂと共に上昇した培養液１１の一部は、ロート状のノズル２８の外周で反転して下降流となる。また気泡Ｂと共に円環リング２７内に入り、対流促進部材２４ａ（又は２４ｂ）上に放出された培養液１１は図示の矢印のように反転してロート状のノズル２８内を通って、培養筒１２ａ（又は１２ｂ）の中心を流下することとなる。   In this embodiment, bubbles B are blown out from the gas blowing means 14a (or 14b) by the aeration nozzle 23a (or 23b) along the inner periphery of the culture tube 12a (or 12b), and the bubbles B are cultured. It rises along the inner peripheral wall of the cylinder 12a (or 12b), enters the annular ring 27 of the upper convection promoting member 24a (or 24b), blows out from the gas discharge hole 29, and enters the gas phase portion Ga (or Gb). To be released. As a result, the culture solution 11 in the culture tube 12a (or 12b) moves to the other culture tube 12b (or 12a). In addition to the pressure of the gas phase portion Ga, the bubbles B are formed on the inner peripheral side. Since there are few bubbles B on the center side, there is a density difference between the inner peripheral side and the center side, a downward flow is generated on the center side, and the bubble B rises in the annular ring 27 of the convection promoting member 24a (or 24b). A part of the culture solution 11 is reversed at the outer periphery of the funnel-shaped nozzle 28 to become a downward flow. The culture solution 11 that enters the annular ring 27 together with the bubbles B and is released onto the convection promoting member 24a (or 24b) is reversed as shown by the arrow in the drawing, passes through the funnel-shaped nozzle 28, and then the culture tube. It will flow down the center of 12a (or 12b).

これにより、培養筒１２ａ（又は１２ｂ）の培養液１１の撹拌がさらに良好となる。   Thereby, stirring of the culture solution 11 of the culture cylinder 12a (or 12b) becomes still better.

１０ 培養槽
１１ 培養液
１２ａ、１２ｂ 培養筒
１４ａ、１４ｂ ガス吹込手段
１５ａ、１５ｂ 排気手段 DESCRIPTION OF SYMBOLS 10 Culture tank 11 Culture liquid 12a, 12b Culture cylinder 14a, 14b Gas blowing means 15a, 15b Exhaust means

Claims (4)

培養槽内に培養液を収容し、その培養槽にガスを通気して培養する細胞培養装置において、鉛直な一対の培養筒の下部を連結して反応槽を形成し、その両培養筒の下部にガス吹込手段をそれぞれ接続すると共に両培養筒の上部に排気手段をそれぞれ接続し、いずれか一方のガス吹込手段からその一方の培養筒の下部にガスを吹き込むと共に、他方の培養筒の上部から排気手段を介してガスを排出し、これを交互に繰り返して、両培養筒の培養液を交互に移動して撹拌することを特徴とする細胞培養装置。   In a cell culture apparatus that contains a culture solution in a culture tank and cultivates by venting gas to the culture tank, a lower part of a pair of vertical culture cylinders is connected to form a reaction tank, and the lower parts of both culture cylinders The gas blowing means are connected to each other and the exhaust means are respectively connected to the upper parts of both culture tubes, and the gas is blown from one of the gas blowing means to the lower part of one of the culture tubes and from the upper part of the other culture tube. A cell culturing apparatus characterized in that gas is discharged through an evacuation means, and this is alternately repeated to alternately move and agitate the culture solution in both culture tubes. 鉛直な一対の培養筒の下部を屈曲筒で接続してＵ字状の反応槽を形成し、ガス吹込手段と排気手段の切り換えで、屈曲筒を通して培養液を一方から他方、又はこの逆に培養液を移動させて撹拌する請求項１記載の細胞培養装置。   A U-shaped reaction tank is formed by connecting the lower part of a pair of vertical culture tubes with a bent tube, and the culture medium is cultured from one to the other or vice versa through the bent tube by switching between gas blowing means and exhaust means. The cell culture apparatus according to claim 1, wherein the liquid is moved and stirred. ガス吹込手段には、培養筒の下部の内周壁に沿って気泡を吹き出すエアレーションノズルが設けられる請求項１又は２記載の細胞培養装置。   The cell culture apparatus according to claim 1 or 2, wherein the gas blowing means is provided with an aeration nozzle that blows out bubbles along the inner peripheral wall of the lower part of the culture tube. 培養筒の上部には、その培養筒の内周側に沿って上昇した培養液を中心側に流下させる対流促進部材が設けられる請求項３記載の細胞培養装置。   4. The cell culture device according to claim 3, wherein a convection promoting member is provided at an upper part of the culture tube to cause the culture solution that has risen along the inner peripheral side of the culture tube to flow down to the center side.
JP2010270236A 2010-12-03 2010-12-03 Method of stirring culture medium in cell culture device Expired - Fee Related JP5768365B2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2010270236A JP5768365B2 (en) 2010-12-03 2010-12-03 Method of stirring culture medium in cell culture device

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP2010270236A JP5768365B2 (en) 2010-12-03 2010-12-03 Method of stirring culture medium in cell culture device

Publications (2)

Publication Number Publication Date
JP2012115232A true JP2012115232A (en) 2012-06-21
JP5768365B2 JP5768365B2 (en) 2015-08-26

Family

ID=46498885

Family Applications (1)

Application Number Title Priority Date Filing Date
JP2010270236A Expired - Fee Related JP5768365B2 (en) 2010-12-03 2010-12-03 Method of stirring culture medium in cell culture device

Country Status (1)

Country Link
JP (1) JP5768365B2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014058761A1 (en) * 2012-10-08 2014-04-17 Calysta Energy, Llc Gas-fed fermentation systems

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020037580A1 (en) * 2000-08-18 2002-03-28 Levitronix Llc Bioreactor and method for fluidly suspending a product
JP2003170189A (en) * 2001-12-11 2003-06-17 National Agricultural Research Organization Processing equipment
JP2006288244A (en) * 2005-04-07 2006-10-26 Espec Corp Culturing device and method for producing cell
JP2008182899A (en) * 2007-01-26 2008-08-14 Hitachi Plant Technologies Ltd Cell culture method and cell culture apparatus
JP2009505660A (en) * 2005-08-25 2009-02-12 エイ２ビーイー カーボン キャプチャー エルエルシー Method, apparatus and system for producing biodiesel from algae

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020037580A1 (en) * 2000-08-18 2002-03-28 Levitronix Llc Bioreactor and method for fluidly suspending a product
JP2003170189A (en) * 2001-12-11 2003-06-17 National Agricultural Research Organization Processing equipment
JP2006288244A (en) * 2005-04-07 2006-10-26 Espec Corp Culturing device and method for producing cell
JP2009505660A (en) * 2005-08-25 2009-02-12 エイ２ビーイー カーボン キャプチャー エルエルシー Method, apparatus and system for producing biodiesel from algae
JP2008182899A (en) * 2007-01-26 2008-08-14 Hitachi Plant Technologies Ltd Cell culture method and cell culture apparatus

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014058761A1 (en) * 2012-10-08 2014-04-17 Calysta Energy, Llc Gas-fed fermentation systems
CN104781387A (en) * 2012-10-08 2015-07-15 凯利斯塔公司 Gas-fed fermentation systems
US10501714B2 (en) 2012-10-08 2019-12-10 Calysta Energy, Inc. Gas-fed fermentation systems
US10889793B2 (en) 2012-10-08 2021-01-12 Calysta, Inc. C1 substrate-fed fermentation systems and methods for producing C4 compounds

Also Published As

Publication number Publication date
JP5768365B2 (en) 2015-08-26

Similar Documents

Publication Publication Date Title
US7819576B2 (en) Pneumatic bioreactor
US9499290B2 (en) Stationary bubble reactors
JP2021129590A (en) Culture method
US20160177248A1 (en) Pivoting pressurized single use bioreactor
US20170145361A1 (en) Bioreactor with separate co2 supply
JP2021521845A (en) Inflatable bioreactor and usage
JP2018500050A (en) Bioreactor
US7850365B1 (en) Toroidal convection mixing device
JP4561192B2 (en) Cell culture device and cell culture method
CN207628280U (en) A kind of waste water apparatus for kneading apparatus
JP5768365B2 (en) Method of stirring culture medium in cell culture device
KR20160064740A (en) Vertical microalgae culturing apparatus which is able to remove microalgae attached to photobioreactor by spiral flow generated by carbon dioxide and culturing medium
US10745305B2 (en) Methods and devices for performing reactions
US20090325282A1 (en) Vessels for mixing bioprocessing materials
CN115466661A (en) Bioreaction ventilation device, cell culture reactor and method of use thereof
JP2020503069A (en) Perfusion device
KR101516792B1 (en) Photo bioreactor
CN202988832U (en) Pulse aerating apparatus
CN205653229U (en) Bological aerated filter device
CN217895612U (en) Fermentation detoxification system
JP2008178389A (en) Small-scaled cell culture container
SU1551730A1 (en) Apparatus for cultivation of suspension cultures of cells or microorganisms
JP7173817B2 (en) Cultivation device having a bubble generator for removing carbon dioxide gas
CN103755115B (en) Fermentation tank for solid waste treatment process and solid waste treatment system
CN207294805U (en) A kind of cell co-cultures bottle

Legal Events

Date Code Title Description
A621 Written request for application examination

Free format text: JAPANESE INTERMEDIATE CODE: A621

Effective date: 20131029

A977 Report on retrieval

Free format text: JAPANESE INTERMEDIATE CODE: A971007

Effective date: 20141022

A131 Notification of reasons for refusal

Free format text: JAPANESE INTERMEDIATE CODE: A131

Effective date: 20141028

A521 Request for written amendment filed

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20141127

TRDD Decision of grant or rejection written
A01 Written decision to grant a patent or to grant a registration (utility model)

Free format text: JAPANESE INTERMEDIATE CODE: A01

Effective date: 20150526

A61 First payment of annual fees (during grant procedure)

Free format text: JAPANESE INTERMEDIATE CODE: A61

Effective date: 20150608

R151 Written notification of patent or utility model registration

Ref document number: 5768365

Country of ref document: JP

Free format text: JAPANESE INTERMEDIATE CODE: R151

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

LAPS Cancellation because of no payment of annual fees