JPH05252933A - Method for supplying oxygen and device for culturing organismic cell by the method - Google Patents
Method for supplying oxygen and device for culturing organismic cell by the methodInfo
- Publication number
- JPH05252933A JPH05252933A JP5477892A JP5477892A JPH05252933A JP H05252933 A JPH05252933 A JP H05252933A JP 5477892 A JP5477892 A JP 5477892A JP 5477892 A JP5477892 A JP 5477892A JP H05252933 A JPH05252933 A JP H05252933A
- Authority
- JP
- Japan
- Prior art keywords
- oxygen supply
- culture
- supply zone
- partition wall
- oxygen
- 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.)
- Pending
Links
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
- C12M23/34—Internal compartments or partitions
-
- 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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
-
- 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
- C12M29/08—Air lift
Landscapes
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Clinical Laboratory Science (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
【0001】[0001]
【産業上の利用分野】本発明は、生体の細胞を生体外で
増殖させるための細胞への酸素供給方法及びその培養装
置に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for supplying oxygen to cells for in vitro growth of cells in the living body, and a culture apparatus therefor.
【0002】[0002]
【従来の技術】液中通気方式による細胞への酸素供給方
法及び培養装置に関しては、ケミカルエンジニアリン
グ、1987年1月16日号121〜129ペ−ジ(Ch
emicalEngineering,February,16,1987,p1
21〜129)、及びサイトテクノロジ、1990年第
3巻39〜42ページ(Cytotechnology,3,p39〜
42,1990)、及び特開昭63−164879号公報に記載
されている。また、細胞は培養槽に留めたまま使用済の
培地のみを抜き出し、新鮮な培地を供給する、いわゆ
る、灌流培養を実施するに当り、微細孔を有するスクリ
ーンによって培養液中に分散している細胞と液とを分離
する方法については、デベロップ.バイオル1.スタン
ダード、66,169〜175ページ、1987年(De
velop. Biol.Standard,66,p169〜175、19
87)に記載されている。また、デベロップ.バイオ
ル.スタンダード、66,227〜240ページ、19
87年(Develop.Biol. Standard,66,p227〜2
40、1987)には細胞等の付着を防止するため酸素
供給用チューブユニットを振動させることが記載されて
いる。2. Description of the Related Art Regarding the method of supplying oxygen to cells by a submerged aeration system and a culture apparatus, Chemical Engineering, January 16, 1987, pages 121-129 (Ch.
emicalEngineering, February, 16, 1987, p1
21-129), and Site Technology, 1990 Volume 3, pages 39-42 (Cytotechnology, 3, p39-
42, 1990) and JP-A-63-164879. In addition, when cells are kept in the culture tank, only the used medium is extracted and fresh medium is supplied, that is, cells that are dispersed in the culture medium by a screen having fine pores when performing so-called perfusion culture. For the method of separating liquid and liquid, see Development. Biol 1. Standard, 66, 169-175 pages, 1987 (De
velop. Biol. Standard, 66, p169-175, 19
87). Also, development. Biol. Standard, pages 66, 227-240, 19
1987 (Develop. Biol. Standard, 66, p227-2
40, 1987), vibrating the tube unit for oxygen supply to prevent attachment of cells and the like.
【0003】[0003]
【発明が解決しようとする課題】液中通気方式は酸素供
給能力に優れているが、細胞の増殖や物質生産に影響を
与える場合や、細胞が多数凝集して細胞塊を形成した
り、マイクロキャリアで足場依存性細胞を培養する場合
に適用できない場合があった。従来技術では、これらの
培養に適用可能とするため、培養槽内にスクリーンで囲
った細胞やマイクロキャリアの存在しない領域を設けて
その領域内で液中通気する方式をとっている。これらの
従来技術では、細胞やマイクロキャリアの浸入を防ぐス
クリーンの目詰まりによる酸素供給能力の低下が大きな
問題であり、目詰まりを防止するため、スクリーンに回
転や上下振動を与えることが講じられているがその効果
は十分とは言えない。Although the submerged aeration method is excellent in oxygen supply capacity, it may affect cell growth or substance production, or a large number of cells may aggregate to form a cell mass, or a micro It may not be applicable when culturing anchorage-dependent cells in a carrier. In the prior art, in order to be applicable to these cultures, a region surrounded by a screen where cells and microcarriers do not exist is provided in the culture tank, and liquid is aerated in the region. In these conventional techniques, the decrease in oxygen supply capacity due to the clogging of the screen that prevents the infiltration of cells and microcarriers is a major problem, and in order to prevent the clogging, rotation and vertical vibration are applied to the screen. However, the effect is not sufficient.
【0004】本発明は、細胞やマイクロキャリアの存在
しない区画を形成するための隔壁の目詰まりを防止し、
液中通気による効率的な酸素供給を実施する方法及び培
養装置を提供することにある。The present invention prevents clogging of partition walls for forming compartments in which cells and microcarriers do not exist,
It is an object of the present invention to provide a method and a culture device for efficiently supplying oxygen by submerged aeration.
【0005】[0005]
【課題を解決するための手段】本発明は、生体の細胞を
培養するにあたり、培養液中の細胞への酸素の供給を液
中通気方法により行なう方法において、液中通気を培養
槽内に設けた培養細胞の凝集塊および/またはマイクロ
キャリアを実質的に透過しない微細孔を有し、かつ、位
置を変化しうる酸素供給ゾーン隔壁により区別された酸
素供給ゾーン内で行なうこと、前記酸素供給ゾーン隔壁
上端は培養液面より上に下端部は培養槽内に開口されて
おり、かつ、酸素供給ゾーン隔壁は弾性を有する部材に
よって培養槽に固定されてその位置の変化が可能なこ
と、前記隔壁下端部開口部近傍に酸素供給ゾーン内部の
溶液を培養槽内に排出するための手段を設けたこと、酸
素供給ゾーン隔壁微細孔に圧力変動を生じさせる手段を
設けたこと、培養液面かつ酸素供給ゾーン隔壁上端より
上に泡沫破泡手段を設けたことを特徴とする酸素の供給
方法である。The present invention provides a method for supplying oxygen to cells in a culture medium by submerged aeration when culturing cells of a living body, wherein submerged aeration is provided in a culture tank. Oxygen supply zone having micropores that are substantially impermeable to aggregates of cultured cells and / or microcarriers and capable of changing position, performed in an oxygen supply zone separated by partition walls, said oxygen supply zone The upper end of the partition wall is opened above the culture liquid surface and the lower end is opened in the culture tank, and the oxygen supply zone partition wall is fixed to the culture tank by an elastic member so that its position can be changed. A means for discharging the solution inside the oxygen supply zone into the culture tank was provided in the vicinity of the lower end opening, a means for causing pressure fluctuation in the oxygen supply zone partition micropores, and a culture solution. And an oxygen supply method which is characterized in that a foam-breaking means above the oxygen supply zone bulkhead upper.
【0006】前記酸素供給ゾーン内部の溶液を培養槽内
に排出するための手段として、前記酸素供給ゾーン隔壁
下端部開口部の近傍に回転軸と直行する整流板を有する
撹拌翼を設置して吐出流を生じさせることを特徴とする
酸素の供給方法である。As a means for discharging the solution in the oxygen supply zone into the culture tank, a stirring blade having a straightening plate perpendicular to the rotary shaft is installed near the lower end opening of the partition wall of the oxygen supply zone for discharge. A method of supplying oxygen characterized by generating a flow.
【0007】また、前記培養槽内に設けた酸素供給ゾー
ン隔壁の外部に整流隔壁を設け、両隔壁間に培養液の流
通部分を設けたことを特徴とする。Further, the present invention is characterized in that a rectifying partition wall is provided outside the oxygen supply zone partition wall provided in the culture tank, and a culture liquid circulation portion is provided between both partition walls.
【0008】さらに、前記培養槽内に設けた酸素供給ゾ
ーン隔壁が細孔径を変化させる機構を有することを特徴
とする。Further, the oxygen supply zone partition wall provided in the culture tank has a mechanism for changing the pore diameter.
【0009】本発明は、生体の細胞を培養する装置にお
いて、培養液中の細胞に酸素を供給する手段として酸素
含有ガスを直接液中に通気する液中通気方法を用い、か
つ、培養細胞の凝集塊および/またはマイクロキャリア
を実質的に透過しない微細孔を有する酸素供給ゾーン隔
壁により区別された酸素供給ゾーン内で液中通気を行な
うこと、酸素供給ゾーン隔壁上端は培養液面より上に下
端部は培養液内に開口されており、かつ、酸素供給ゾー
ン隔壁は弾性を有する部材によって培養槽に固定されて
その位置の変化が可能なこと、前記酸素供給ゾーン下端
部開口の近傍に酸素供給ゾーン内部の溶液を培養槽内に
排出するための手段として撹拌翼を有すること、培養液
面上に泡沫破泡手段を有することを特徴とする生体の細
胞を培養する装置である。The present invention, in a device for culturing cells of a living body, uses a submerged aeration method in which an oxygen-containing gas is directly aerated in the liquid as a means for supplying oxygen to the cells in the culture liquid, and Aeration in the liquid is carried out in the oxygen supply zone separated by the oxygen supply zone partition having micropores that do not substantially permeate aggregates and / or microcarriers. Part is opened in the culture solution, and the oxygen supply zone partition wall is fixed to the culture tank by an elastic member so that its position can be changed, and oxygen is supplied near the opening of the lower end of the oxygen supply zone. A device for culturing cells of a living body, characterized by having a stirring blade as a means for discharging the solution in the zone into the culture tank and having a foam breaking means on the surface of the culture liquid. A.
【0010】前記培養槽内に設けた酸素供給ゾーン隔壁
の外部に、上端部が液面下に開口し、かつ下端部が撹拌
翼の近傍の上部に開口している整流隔壁を設け、両隔壁
間に培養液の流通部分を設けたことを特徴とする細胞を
培養する装置である。Outside the oxygen supply zone partition provided in the culture tank, a rectifying partition having an upper end opening below the liquid surface and a lower end opening above the stirring blade is provided. It is an apparatus for culturing cells, which is characterized in that a circulation portion of a culture solution is provided therebetween.
【0011】また、前記培養槽内に設けた酸素供給ゾー
ンを区別するための隔壁が細孔径を変化せしめる機構を
有することを特徴とする細胞を培養する装置である。[0012] Further, the apparatus for culturing cells is characterized in that the partition for distinguishing the oxygen supply zone provided in the culture tank has a mechanism for changing the pore diameter.
【0012】さらに、培養槽内に設けた撹拌翼を駆動す
るための軸の一部に軸芯より偏芯させた偏芯部材を設置
し、前記偏芯部材の回転に追従してしゅう動するしゅう
動部材を設置し、前記しゅう動部材を酸素供給ゾーン隔
壁と結合させたことにより、撹拌軸の回転により前記隔
壁に外力を加えてその位置を変化させることを特徴とす
る細胞を培養する装置である。Further, an eccentric member eccentric from the shaft center is installed on a part of a shaft for driving a stirring blade provided in the culture tank, and slides following the rotation of the eccentric member. An apparatus for culturing cells characterized in that a sliding member is installed and the sliding member is connected to an oxygen supply zone partition so that an external force is applied to the partition by rotation of a stirring shaft to change its position. Is.
【0013】[0013]
【作用】本発明の液中通気方法により培養液中の細胞へ
酸素の供給を行なう方法は、培養槽内に設けた培養細胞
の凝集塊および/またはマイクロキャリアを実質的に透
過しない微細孔をもち、かつ、位置を変化しうる酸素供
給ゾーン隔壁により区別された酸素供給ゾーン内で液中
通気を行なう。酸素供給ゾーン隔壁上端は培養液面より
上に下端部は培養槽内に開口されており、かつ、酸素供
給ゾーン隔壁は弾性を有する部材によって培養槽に固定
されてその位置の変化が可能な状態にある。隔壁下端部
開口部の近傍には、酸素供給ゾーン内部の溶液を培養槽
内に排出するための手段として撹拌翼を設置して吐出流
を生じさせる。また、酸素供給ゾーン隔壁の外部に整流
隔壁を設け、両隔壁間に培養液の流通部分を設けたこと
により酸素供給ゾーン隔壁の近傍での培養液の流動を効
率良く生じさせる。その結果、酸素供給ゾーン隔壁の微
細孔内への目詰まりを効果的に防止することができる。
さらに、酸素供給ゾーン隔壁を振動させる機構により、
酸素供給ゾーン隔壁微細孔に圧力変動を生じさせること
により、より一層、微細孔内への目詰まりを効果的に防
ぐことができる。また、目詰まりを生じた微細孔内の物
質を排除する手段として、細孔径を変化させる機構を有
する。液中通気により泡沫層を形成する場合には、培養
液面かつ酸素供給ゾーン隔壁上端より上に泡沫破泡手段
を設ける。The method of supplying oxygen to the cells in the culture solution by the submerged aeration method of the present invention is a method of forming agglomerates of cultured cells and / or micropores that are substantially impermeable to microcarriers, provided in the culture tank. The submerged aeration is performed in the oxygen supply zone which is divided by the oxygen supply zone partition wall which has a variable position. Oxygen supply zone partition wall The upper end is opened above the culture solution surface and the lower end is opened inside the culture tank, and the oxygen supply zone partition wall is fixed to the culture tank by an elastic member and its position can be changed. It is in. In the vicinity of the opening at the lower end of the partition wall, a stirring blade is installed as a means for discharging the solution inside the oxygen supply zone into the culture tank to generate a discharge flow. Further, by providing a rectifying partition wall outside the oxygen supply zone partition wall and providing a culture solution flow portion between both partition walls, the flow of the culture solution in the vicinity of the oxygen supply zone partition wall is efficiently generated. As a result, it is possible to effectively prevent clogging of the oxygen supply zone partition wall into the fine holes.
Furthermore, by the mechanism that vibrates the oxygen supply zone partition,
By causing a pressure fluctuation in the oxygen supply zone partition wall fine holes, it is possible to further effectively prevent clogging of the fine holes. Further, it has a mechanism for changing the pore diameter as a means for eliminating the substance in the fine pores that have been clogged. When a foam layer is formed by submerged aeration, foam breaking means is provided above the culture solution surface and above the upper end of the oxygen supply zone partition wall.
【0014】本発明の生体細胞の培養装置は、培養槽,
培地供給系として新鮮培地貯留槽,培養液貯留槽,酸素
供給系として酸素源,炭酸ガス源,窒素源、空気源、及
び、培養系の無菌状態を維持するための除菌フィルタ、
及び、培養状況を監視し、制御を行なうためのコンピュ
ータより構成されている。培養槽は、培養液中の細胞に
酸素を供給する手段として酸素含有ガスを、直接、液中
に通気する液中通気法を用い、培養細胞を実質的に透過
しない微細孔を有する酸素供給ゾーン隔壁により区別さ
れた酸素供給ゾーン内に液中通気する。酸素供給ゾーン
隔壁上端は培養液面より上に下端部は培養液内に開口さ
れており、かつ、酸素供給ゾーン隔壁は弾性を有する部
材によって培養槽に固定されてその位置の変化が可能で
ある。酸素供給ゾーン下端部開口の近傍には酸素供給ゾ
ーン内部の溶液を培養槽内に排出するための手段として
撹拌翼を有する。酸素供給ゾーン隔壁の外部に、上端部
が液面下に開口し、かつ、下端部が撹拌翼の近傍上部に
開口している整流隔壁を設け、両隔壁間に培養液の流通
部分を設けている。撹拌翼を駆動するための軸の一部に
軸芯より偏芯させた偏芯部材を設置し、偏芯部材の回転
に追従してしゅう動するしゅう動部材を設置し、しゅう
動部材を酸素供給ゾーン隔壁と結合させたことにより、
撹拌軸の回転により隔壁に外力を加えてその位置を変化
させる。酸素供給ゾーンを区別するための隔壁には細孔
径を変化させる機構をもつ。培養液面上には泡沫破泡手
段を有する。The biological cell culturing apparatus of the present invention comprises a culture tank,
A fresh medium storage tank as a medium supply system, a culture solution storage tank, an oxygen source, a carbon dioxide gas source, a nitrogen source, an air source as an oxygen supply system, and a sterilization filter for maintaining a sterile state of the culture system,
And a computer for monitoring and controlling the culture condition. The culture tank uses an in-liquid aeration method in which an oxygen-containing gas is directly aerated in the liquid as a means for supplying oxygen to the cells in the culture liquid, and an oxygen supply zone having fine pores that does not substantially permeate the cultured cells. Aeration is submerged in the oxygen supply zone separated by the partition. Oxygen supply zone partition wall The upper end is opened above the culture solution surface and the lower end is opened in the culture solution, and the oxygen supply zone partition wall is fixed to the culture tank by an elastic member and its position can be changed. .. A stirring blade is provided in the vicinity of the lower end opening of the oxygen supply zone as a means for discharging the solution inside the oxygen supply zone into the culture tank. Outside the oxygen supply zone partition, a rectifying partition with an upper end opening below the liquid surface and a lower end opening near the agitating blade in the upper part was provided, and a culture fluid flow section was provided between both partition walls. There is. An eccentric member that is eccentric from the shaft center is installed on a part of the shaft for driving the stirring blade, and a sliding member that slides following the rotation of the eccentric member is installed. By combining with the supply zone partition,
An external force is applied to the partition wall by the rotation of the stirring shaft to change its position. The partition for distinguishing the oxygen supply zone has a mechanism for changing the pore size. A foam breaking means is provided on the culture liquid surface.
【0015】[0015]
【実施例】以下、本発明の実施例を詳細に説明する。EXAMPLES Examples of the present invention will be described in detail below.
【0016】〈実施例1〉図1は本発明の培養液中の細
胞への酸素供給方法の1実施例を示す図である。培養槽
1内には酸素供給ゾーン隔壁2が設けられている。酸素
供給ゾーン隔壁2は、培養細胞の凝集塊および/または
マイクロキャリアを実質的に透過しない微細孔を有して
いる。酸素供給ゾーン隔壁上端は培養液面より上に下端
部は培養槽内に開口し、かつ、酸素供給ゾーン隔壁は弾
性部材3をその一部に用いた支持体4によって培養槽に
固定されており、その位置の変化が可能な構造をしてい
る。酸素の供給は、酸素供給ゾーン隔壁により区別され
た酸素供給ゾーン内に酸素含有ガスをガス供給管20よ
り撹拌軸8に設けた管路13を経てノズル14より、直
接、培養液中に通気することにより行なう。隔壁下端部
開口部の近傍には、酸素供給ゾーン内部の液を培養槽内
に排出するための手段として撹拌翼5を設置して吐出流
を生じさせる。撹拌翼5には、整流板6を設けており、
酸素供給ゾーン内部の液を効率良く培養槽内に排出する
とともに、酸素供給ゾーン内部への細胞の凝集塊および
/またはマイクロキャリアの侵入を防止する。また、酸
素供給ゾーン隔壁の外部に整流隔壁7を設け、両隔壁間
に培養液の流通部分を設けて酸素供給ゾーン隔壁の近傍
での培養液の流動を効率良く生じさせる。その結果、酸
素供給ゾーン隔壁2の微細孔内への目詰まりを効果的に
防ぐことができる。さらに、撹拌軸8の一部に軸芯より
偏芯させた偏芯部材9を設置し、偏芯部材9の回転に追
従してしゅう動するしゅう動部材10を設置し、しゅう
動部材10を酸素供給ゾーン隔壁2と結合させ、撹拌軸
の回転により酸素供給ゾーン隔壁2を振動させ、酸素供
給ゾーン隔壁微細孔に圧力変動を加えることにより、よ
り一層、微細孔内への目詰まりを効果的に防止すること
ができる。液中通気により培養液面上に泡沫層11を形
成する場合には、培養液面かつ酸素供給ゾーン隔壁上端
より上に泡沫破泡層12を設ける。泡沫破泡層12に
は、表面をポリシロキサンで疎水化したステンレス製金
網(約10メッシュ)を用いる。液面上に形成された泡
沫は、破泡層に接触した時点で破裂消滅して液滴となり
培養液中に戻る。通気したガスは、排気管15より培養
槽外に排出される。Example 1 FIG. 1 is a diagram showing an example of the method for supplying oxygen to cells in a culture medium according to the present invention. An oxygen supply zone partition wall 2 is provided in the culture tank 1. The oxygen supply zone partition wall 2 has micropores that are substantially impermeable to aggregates of cultured cells and / or microcarriers. The oxygen supply zone partition has an upper end opened above the culture solution surface and a lower end opened into the culture tank, and the oxygen supply zone partition is fixed to the culture tank by a support 4 using an elastic member 3 as a part thereof. , It has a structure that can change its position. Oxygen is supplied to the culture solution directly from the nozzle 14 through the gas supply pipe 20 through the pipe line 13 provided on the stirring shaft 8 in the oxygen supply zone separated by the oxygen supply zone partition wall. By doing. In the vicinity of the lower end opening of the partition wall, a stirring blade 5 is installed as a means for discharging the liquid inside the oxygen supply zone into the culture tank, and a discharge flow is generated. The stirring blade 5 is provided with a straightening plate 6,
The liquid inside the oxygen supply zone is efficiently discharged into the culture tank, and the invasion of cell aggregates and / or microcarriers into the oxygen supply zone is prevented. Further, a rectifying partition wall 7 is provided outside the oxygen supply zone partition wall, and a culture solution flow portion is provided between both partition walls to efficiently cause the flow of the culture solution near the oxygen supply zone partition wall. As a result, it is possible to effectively prevent clogging of the oxygen supply zone partition wall 2 into the fine holes. Further, an eccentric member 9 which is eccentric from the shaft center is installed on a part of the stirring shaft 8, and a sliding member 10 that slides following the rotation of the eccentric member 9 is installed. By combining with the oxygen supply zone partition wall 2, vibrating the oxygen supply zone partition wall 2 by rotating the stirring shaft, and applying pressure fluctuation to the oxygen supply zone partition wall micropores, the clogging into the micropores is further effective. Can be prevented. When the foam layer 11 is formed on the surface of the culture solution by submerged aeration, the foam breaking layer 12 is provided on the surface of the culture solution and above the upper end of the partition wall of the oxygen supply zone. For the foam breaking layer 12, a stainless wire mesh (about 10 mesh) whose surface is hydrophobized with polysiloxane is used. The foam formed on the liquid surface bursts and disappears when it comes into contact with the foam-breaking layer, and returns to the culture solution as droplets. The aerated gas is discharged from the culture tank through the exhaust pipe 15.
【0017】〈実施例2〉図2は本発明の他の実施例を
示す。本実施例の特徴は、実施例1において培養槽内撹
拌翼用駆動軸と酸素供給ゾーン隔壁2を振動させるため
の加振軸とを分離したことにある。すなわち、槽内の撹
拌および酸素供給ゾーン内部の液を排出するための撹拌
翼は培養槽底部に設けた撹拌軸8により駆動され、酸素
供給ゾーン隔壁2の加振は培養槽の上部に設けた加振軸
16により行なわれる。<Embodiment 2> FIG. 2 shows another embodiment of the present invention. The feature of this embodiment is that the drive shaft for stirring blades in the culture tank and the vibrating shaft for vibrating the oxygen supply zone partition wall 2 are separated from each other in the first embodiment. That is, the stirring blades for stirring in the tank and discharging the liquid in the oxygen supply zone were driven by the stirring shaft 8 provided at the bottom of the culture tank, and the vibration of the oxygen supply zone partition wall 2 was provided at the top of the culture tank. It is performed by the vibrating shaft 16.
【0018】本実施例によれば、培養液の撹拌強度と、
酸素供給ゾーン隔壁を振動を別個に調節することがで
き、培養中の細胞濃度の変化や酸素供給ゾーン隔壁微細
孔の目詰まり状況に応じた培養条件を設定することがで
きる。According to this embodiment, the stirring strength of the culture solution,
The vibration of the partition walls of the oxygen supply zone can be adjusted separately, and the culture conditions can be set according to changes in the cell concentration during culture and the clogging of the micropores of the partition walls of the oxygen supply zone.
【0019】〈実施例3〉図3は本発明の他の実施例を
示す。本実施例の特徴は、実施例2において、酸素供給
ゾーン隔壁2が培養槽底部に設けた撹拌軸8により振動
されることにある。すなわち、撹拌軸8に軸芯より偏芯
させた偏芯部材9を設置し、偏芯部材9の回転に追従し
てしゅう動するしゅう動部材10を設置し、しゅう動部
材10を酸素供給ゾーン隔壁2と結合させ、撹拌軸の回
転により酸素供給ゾーン隔壁2を振動させる。<Third Embodiment> FIG. 3 shows another embodiment of the present invention. The feature of this embodiment is that in the second embodiment, the oxygen supply zone partition wall 2 is vibrated by the stirring shaft 8 provided at the bottom of the culture tank. That is, an eccentric member 9 which is eccentric from the shaft center is installed on the stirring shaft 8, and a sliding member 10 which slides following the rotation of the eccentric member 9 is installed. The oxygen supply zone partition wall 2 is vibrated by being combined with the partition wall 2 and rotating the stirring shaft.
【0020】本実施例によれば、撹拌軸の軸受部と酸素
供給ゾーン隔壁加振用偏芯部材との間隔を小さくするこ
とができ、その結果として、酸素供給ゾーン隔壁の振動
による撹拌軸及び軸受に加わる応力が軽減され、より簡
易な構造にすることができる。According to this embodiment, the distance between the bearing portion of the stirring shaft and the eccentric member for exciting the oxygen supply zone partition wall can be reduced, and as a result, the stirring shaft and The stress applied to the bearing is reduced, and a simpler structure can be obtained.
【0021】〈実施例4〉図4は本発明になる酸素供給
方法を用いた培養装置の概略を示す図である。<Embodiment 4> FIG. 4 is a schematic view of a culture apparatus using the oxygen supply method according to the present invention.
【0022】本発明になる生体の細胞を培養する装置
は、内部の熱殺菌が可能なようにステンレススチール製
の培養槽1,撹拌翼駆動用モータ31,培地供給系とし
て新鮮培地貯留槽33,培養液貯留槽32,酸素供給系
として酸素源34,炭酸ガス源35,窒素源36,空気
源37、及び、培養系の無菌状態を維持するための除菌
フィルタ38、及び、培養状況を監視し、制御を行なう
ためのコンピュータ50より構成されている。The apparatus for culturing cells of a living body according to the present invention comprises a stainless steel culture tank 1, a stirring blade driving motor 31, a fresh medium storage tank 33 as a medium supply system, so as to enable heat sterilization of the inside. Culture solution storage tank 32, oxygen source 34, carbon dioxide gas source 35, nitrogen source 36, air source 37 as an oxygen supply system, sterilization filter 38 for maintaining the sterility of the culture system, and the culture condition are monitored. In addition, it is composed of a computer 50 for controlling.
【0023】培養槽1内には本発明の酸素供給方法が使
用されている。すなわち、培養細胞の凝集塊および/ま
たはマイクロキャリアを実質的に透過しない微細孔を有
する酸素供給ゾーン隔壁2が設けられ、隔壁上端は培養
液面より上に下端部は培養槽内に開口している。酸素の
供給は、酸素供給ゾーン隔壁により区別された酸素供給
ゾーン内に酸素含有ガスをガス供給管20より撹拌軸8
に設けた管路を経て、直接、培養液中に通気することに
より行なう。隔壁下端部の開口部近傍には、酸素供給ゾ
ーン内部の液を培養槽内に排出する手段として撹拌翼5
を設置して吐出流を生じさせる。撹拌翼5には、整流板
6を設けており、酸素供給ゾーン内部の液を効率良く培
養槽内に排出するとともに、酸素供給ゾーン内部への細
胞の凝集塊および/またはマイクロキャリアの侵入を防
止する。また、酸素供給ゾーン隔壁の外部に整流隔壁7
を設け、両隔壁間に培養液の流通部分を設けたことによ
り酸素供給ゾーン隔壁の近傍での培養液の流動を効率良
く生じさせる。撹拌軸8の一部に軸芯より偏芯させた偏
芯部材、該偏芯部材の回転に追従するしゅう動部材、し
ゅう動部材を酸素供給ゾーン隔壁2と結合させ、撹拌軸
の回転により酸素供給ゾーン隔壁2を振動させ、酸素供
給ゾーン隔壁微細孔に圧力変動を加えることにより、微
細孔内への目詰まりを防止する。液中通気により培養液
面上に泡沫11を形成する場合には、培養液面かつ酸素
供給ゾーン隔壁上端より上に泡沫破泡層12を設ける。
泡沫破泡層12には、表面をポリシロキサンで疎水化し
たステンレス製金網(約10メッシュ)を用いた。液面
上に形成された泡沫は、破泡層に接触した時点で破裂消
滅して液滴となり培養液中に戻る。通気したガスは、除
菌フィルタ38を経て培養槽外に排出される。The oxygen supply method of the present invention is used in the culture tank 1. That is, an oxygen supply zone partition wall 2 having micropores that do not substantially permeate aggregates of cultured cells and / or microcarriers is provided, and the partition wall upper end is above the culture liquid surface and the lower end portion is opened in the culture tank. There is. The oxygen is supplied by supplying the oxygen-containing gas from the gas supply pipe 20 into the oxygen supply zone, which is distinguished by the oxygen supply zone partition wall.
It is carried out by directly ventilating the culture solution through the conduit provided in. In the vicinity of the opening at the lower end of the partition wall, a stirring blade 5 is provided as a means for discharging the liquid inside the oxygen supply zone into the culture tank.
To generate a discharge flow. The stirring blade 5 is provided with a straightening plate 6 to efficiently discharge the liquid inside the oxygen supply zone into the culture tank and prevent the infiltration of cell aggregates and / or microcarriers into the oxygen supply zone. To do. The rectifying partition 7 is provided outside the oxygen supply zone partition.
Is provided and the flow portion of the culture solution is provided between both partition walls, so that the flow of the culture solution in the vicinity of the oxygen supply zone partition wall is efficiently generated. An eccentric member that is eccentric from the shaft center on a part of the stirring shaft 8, a sliding member that follows the rotation of the eccentric member, and a sliding member are connected to the oxygen supply zone partition wall 2 and the oxygen is generated by rotating the stirring shaft. By vibrating the supply zone partition walls 2 and applying pressure fluctuations to the oxygen supply zone partition wall fine holes, clogging of the fine holes is prevented. When the foam 11 is formed on the surface of the culture solution by aeration in the liquid, the foam breaking layer 12 is provided on the surface of the culture solution and above the upper end of the partition wall of the oxygen supply zone.
For the foam-breaking layer 12, a stainless wire mesh (about 10 mesh) whose surface was made hydrophobic with polysiloxane was used. The foam formed on the liquid surface bursts and disappears when it comes into contact with the foam-breaking layer, and returns to the culture solution as droplets. The aerated gas is discharged to the outside of the culture tank through the sterilization filter 38.
【0024】本培養装置で培養される生体細胞は、動物
の細胞,微生物,植物細胞等である。特に、培養液中に
100〜1000μm程度の粒子が浮遊している培養液
が適している。Living cells cultured in the present culture device are animal cells, microorganisms, plant cells and the like. Particularly, a culture solution in which particles of about 100 to 1000 μm are suspended in the culture solution is suitable.
【0025】まず、あらかじめ培養槽1及び新鮮培地貯
留槽33,培養液貯留槽32、及び配管内を高圧蒸気滅
菌する。ついで、培養を目的とする生体細胞の成育に必
要な栄養成分を溶かし込んだ培地を調製し殺菌処理して
新鮮培地貯留槽に保管する。次いで、その一部を培養槽
1内に移送し、これに種細胞を無菌的に接種して培養を
開始する。培養は、培養槽外部に設けたジャケット(図
示せず)に温水を通じて培養液を加温して行なう。培養
温度は、培養する細胞の至適温度にコントロールする。
細胞の増殖により培養液の物理化学的性質が変化し、栄
養成分の枯渇や、乳酸等の老廃成分の蓄積が起こる。こ
のため、培養槽1には温度センサ39,pHセンサ4
0,DOセンサ41、及びDCO2 センサ42を設置
し、温度、pH,DO(溶存酸素濃度)、DCO2 (溶
存炭酸ガス濃度)を計測する。これらの計測信号はコン
ピュータ50に伝送され、培養状況の判定がなされる。
また、必要に応じて、培養液を一部抜き出し、液の濁度
や、乳酸濃度,グルコース濃度,細胞濃度、及び形状,
大きさ,輝度等の細胞の画像情報等をコンピュータに入
力する(図示せず)。コンピュータ50はこれらの情報
と内部に蓄積した過去の培養情報とをもとにその時点で
の培養状況を推論,判定し、ガス通気量,通気ガス中の
酸素濃度,炭酸ガス濃度を調節する指令を各調節弁に与
える。さらに、培養液中の栄養成分の枯渇や乳酸の蓄積
に対応して新鮮培地供給及び使用済みの培養液抜き出し
を各ポンプに指令し、培地の灌流を行なわせる。なお、
使用済みの培養液抜き出しにあたっては、培養液中に浮
遊する固形物を分離した上で行なうのが望ましい。First, the culture tank 1, the fresh medium storage tank 33, the culture solution storage tank 32, and the inside of the pipe are sterilized by high-pressure steam in advance. Then, a medium in which the nutrient components necessary for the growth of living cells for culture are dissolved is prepared, sterilized, and stored in a fresh medium storage tank. Then, a part thereof is transferred into the culture tank 1, and seed cells are aseptically inoculated into the culture tank 1 to start the culture. The culture is performed by warming the culture solution through warm water through a jacket (not shown) provided outside the culture tank. The culture temperature is controlled to the optimum temperature of the cells to be cultured.
The physicochemical properties of the culture solution change due to cell growth, causing depletion of nutrients and accumulation of waste components such as lactic acid. Therefore, the temperature sensor 39 and the pH sensor 4 are provided in the culture tank 1.
0, DO sensor 41, and DCO 2 sensor 42 are installed, and temperature, pH, DO (dissolved oxygen concentration), and DCO 2 (dissolved carbon dioxide concentration) are measured. These measurement signals are transmitted to the computer 50, and the culture status is determined.
In addition, if necessary, a part of the culture solution may be extracted to obtain turbidity of the solution, lactate concentration, glucose concentration, cell concentration, and shape,
Image information of cells such as size and brightness is input to a computer (not shown). The computer 50 infers and judges the culture state at that time based on these information and the past culture information accumulated therein, and commands to adjust the gas ventilation amount, the oxygen concentration in the ventilation gas, and the carbon dioxide concentration. Is given to each control valve. Further, in response to the depletion of nutrients in the culture solution and the accumulation of lactic acid, each pump is instructed to supply the fresh medium and withdraw the used culture solution to perfuse the medium. In addition,
When extracting the used culture solution, it is desirable to separate the solid matter floating in the culture solution.
【0026】本培養装置により、培養液中に細胞凝集塊
やマイクロキャリア等の固形物を含有する場合にも泡沫
の発生を懸念することなく所要量の通気量で液中通気に
よる酸素供給が可能となり、生体細胞の高密度培養が可
能となる。With the main culturing device, oxygen can be supplied by submerged aeration with a required amount of aeration without concern about the occurrence of foam even when the culture contains solid matter such as cell aggregates and microcarriers. Therefore, high density culture of living cells becomes possible.
【0027】[0027]
【発明の効果】本発明によれば、培養液中に細胞凝集塊
やマイクロキャリア等の固形物を含有する培養でも液中
通気による酸素供給が可能となり、泡沫の発生を懸念す
ることなく所要量の通気量で通気することができるた
め、高密度培養が可能となる。EFFECTS OF THE INVENTION According to the present invention, oxygen can be supplied by submerged aeration even in a culture containing solid matters such as cell aggregates and microcarriers in the culture liquid, and the required amount can be obtained without fear of foam formation. Since it can be aerated with the aeration amount of, a high-density culture becomes possible.
【図1】本発明の酸素供給方法を用いた1実施例の断面
図。FIG. 1 is a cross-sectional view of one embodiment using the oxygen supply method of the present invention.
【図2】本発明の酸素供給方法を用いた1実施例の断面
図。FIG. 2 is a sectional view of one embodiment using the oxygen supply method of the present invention.
【図3】本発明の酸素供給方法を用いた1実施例の断面
図。FIG. 3 is a cross-sectional view of one embodiment using the oxygen supply method of the present invention.
【図4】本発明の酸素供給方法を用いた培養装置の1実
施例の系統図。FIG. 4 is a system diagram of one example of a culture device using the oxygen supply method of the present invention.
1…培養槽、2…酸素供給ゾーン隔壁、5…撹拌翼、6
…整流板、7…整流隔壁、8…撹拌軸、9…偏芯部材、
10…しゅう動部材、12…泡沫破泡層、14…ノズ
ル。1 ... Culture tank, 2 ... Oxygen supply zone partition wall, 5 ... Stirring blade, 6
... rectifying plate, 7 ... rectifying partition wall, 8 ... stirring shaft, 9 ... eccentric member,
10 ... Sliding member, 12 ... Foam breaking layer, 14 ... Nozzle.
Claims (8)
の細胞への酸素の供給を液中通気方法により行なう方法
において、液中通気を培養槽内に設けた培養細胞の凝集
塊および/またはマイクロキャリアを実質的に透過しな
い微細孔を有し、位置を変化しうる酸素供給ゾーンの隔
壁により区別された前記酸素供給ゾーン内で行なうこ
と、前記酸素供給ゾーンの隔壁の上端は前記培養液面よ
り上に下端部は前記培養槽内に開口されており、前記酸
素供給ゾーン隔壁は弾性を有する部材によって前記培養
槽に固定されてその位置の変化が可能なこと、前記隔壁
の下端部開口部の近傍に前記酸素供給ゾーン内部の溶液
を前記培養槽内に排出するための手段を設けたこと、前
記酸素供給ゾーン隔壁の微細孔に圧力変動を生じさせる
手段を設けたこと、前記培養液面でかつ前記酸素供給ゾ
ーンの隔壁上端より上に泡沫破泡手段を設けたことを特
徴とする酸素の供給方法。1. In culturing cells of a living body, in a method of supplying oxygen to cells in a culture solution by a submerged aeration method, submerged aeration is provided in a culture tank, and aggregates of cultured cells and / or Alternatively, it has micropores that are substantially impermeable to microcarriers, and is carried out in the oxygen supply zone distinguished by the partition walls of the oxygen supply zone whose position can be changed. The upper end of the partition wall of the oxygen supply zone is the culture medium. The lower end above the surface is opened in the culture tank, the oxygen supply zone partition wall is fixed to the culture tank by a member having elasticity and its position can be changed, and the lower end opening of the partition wall A means for discharging the solution inside the oxygen supply zone into the culture tank in the vicinity of the section, and a means for causing a pressure fluctuation in the micropores of the oxygen supply zone partition wall, Oxygen supply method, wherein a serial provided foam-breaking means above the partition wall upper end of the culture liquid surface a and the oxygen supply zone.
内部の溶液を前記培養槽内の培養部に排出するための手
段として、前記酸素供給ゾーンの隔壁下端部開口部近傍
に回転軸と直行する整流板を有する撹拌翼を設置して吐
出流を生じさせる酸素の供給方法。2. The means for discharging the solution inside the oxygen supply zone to the culture section in the culture tank according to claim 1, wherein the oxygen supply zone is orthogonal to the rotary shaft near the opening at the lower end of the partition wall of the oxygen supply zone. A method of supplying oxygen to generate a discharge flow by installing a stirring blade having a current plate.
に設けた前記酸素供給ゾーン隔壁の外部に整流隔壁を設
け、両隔壁間に前記培養液の流通部分を設けた酸素の供
給方法。3. The oxygen supply method according to claim 1, wherein a rectifying partition is provided outside the oxygen supply zone partition provided in the culture tank, and a portion for circulating the culture solution is provided between the partition walls.
槽内に設けた前記酸素供給ゾーン隔壁が細孔径を変化せ
しめる機構を有する酸素の供給方法。4. The oxygen supply method according to claim 1, 2, or 3, wherein the oxygen supply zone partition wall provided in the culture tank has a mechanism for changing the pore diameter.
槽,培地供給系として新鮮培地貯留槽,培養液貯留槽,
酸素供給系として酸素源,炭酸ガス源,窒素源,空気
源、及び培養系の無菌状態を維持するための除菌フィル
タ、及び培養状況を監視し、制御を行なうためのコンピ
ュータより構成され、培養液中の細胞に酸素を供給する
手段として酸素含有ガスを直接液中に通気する液中通気
方法を用い、かつ、培養細胞の凝集塊および/またはマ
イクロキャリアを実質的に透過しない微細孔を有する酸
素供給ゾーン隔壁により区別された酸素供給ゾーン内で
液中通気を行なうこと、前記酸素供給ゾーン隔壁の上端
は培養液面より上に下端部は培養液内に開口されてお
り、前記酸素供給ゾーン隔壁は弾性を有する部材によっ
て前記培養槽に固定されてその位置の変化が可能なこ
と、前記酸素供給ゾーンの下端部開口近傍に前記酸素供
給ゾーン内部の溶液を前記培養槽内に排出するための手
段として撹拌翼を有すること、前記培養液面上に泡沫破
泡手段を有することを特徴とする生体細胞の培養装置。5. A device for culturing living cells, comprising a culture tank, a fresh medium storage tank as a medium supply system, a culture solution storage tank,
An oxygen source, a carbon dioxide gas source, a nitrogen source, an air source as an oxygen supply system, and a sterilization filter for maintaining the sterility of the culture system, and a computer for monitoring and controlling the culture condition, An in-liquid aeration method in which an oxygen-containing gas is directly aerated is used as a means for supplying oxygen to cells in the liquid, and it has fine pores that do not substantially permeate aggregates of cultured cells and / or microcarriers. Aeration in the liquid is carried out in the oxygen supply zone separated by the oxygen supply zone partition, the upper end of the oxygen supply zone partition is opened above the culture solution surface, and the lower end is opened in the culture solution, the oxygen supply zone The partition wall is fixed to the culture tank by an elastic member and its position can be changed, and the solution inside the oxygen supply zone is placed in the vicinity of the lower end opening of the oxygen supply zone. Having a stirring blade as a means for discharging the culture tank, a culture apparatus for biological cells, characterized by having a foam-breaking unit on the culture surface.
前記酸素供給ゾーン隔壁の外部に、上端部が液面下に開
口し、かつ下端部が前記撹拌翼の近傍上部に開口してい
る整流隔壁を設け、両隔壁間に前記培養液の流通部分を
設けた生体細胞の培養装置。6. The method according to claim 5, wherein an upper end is opened below the liquid surface and a lower end is opened above the stirring blade outside the oxygen supply zone partition wall provided in the culture tank. An apparatus for culturing living cells, in which a rectifying partition is provided, and a portion where the culture solution flows is provided between both partitions.
に設けた酸素供給ゾーンを区別するための隔壁が細孔径
を変化させる機構を有する生体細胞の培養装置。7. The apparatus for culturing living cells according to claim 5, wherein the partition wall for distinguishing the oxygen supply zone provided in the culture tank has a mechanism for changing the pore diameter.
槽内に設けた前記撹拌翼を駆動するための軸の一部に軸
芯より偏芯させた偏芯部材を設置し、前記偏芯部材の回
転に追従してしゅう動するしゅう動部材を設置し、前記
しゅう動部材を前記酸素供給ゾーン隔壁と結合させたこ
とにより、前記撹拌軸の回転により前記隔壁に外力を加
えてその位置を変化させる生体細胞の培養装置。8. The eccentric member eccentric to the shaft center is installed on a part of a shaft for driving the stirring blade provided in the culture tank according to claim 5, 6 or 7. By installing a sliding member that slides following the rotation of the core member and connecting the sliding member with the oxygen supply zone partition wall, an external force is applied to the partition wall by the rotation of the stirring shaft and its position A device for culturing living cells that changes temperature.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP5477892A JPH05252933A (en) | 1992-03-13 | 1992-03-13 | Method for supplying oxygen and device for culturing organismic cell by the method |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP5477892A JPH05252933A (en) | 1992-03-13 | 1992-03-13 | Method for supplying oxygen and device for culturing organismic cell by the method |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH05252933A true JPH05252933A (en) | 1993-10-05 |
Family
ID=12980230
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP5477892A Pending JPH05252933A (en) | 1992-03-13 | 1992-03-13 | Method for supplying oxygen and device for culturing organismic cell by the method |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH05252933A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105820952A (en) * | 2016-03-31 | 2016-08-03 | 上海拜高乐生物技术有限公司 | Bioreactor for cell perfusion culture |
WO2018048181A3 (en) * | 2016-09-06 | 2018-05-03 | 한국화학연구원 | Cell culture receptacle |
-
1992
- 1992-03-13 JP JP5477892A patent/JPH05252933A/en active Pending
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105820952A (en) * | 2016-03-31 | 2016-08-03 | 上海拜高乐生物技术有限公司 | Bioreactor for cell perfusion culture |
WO2018048181A3 (en) * | 2016-09-06 | 2018-05-03 | 한국화학연구원 | Cell culture receptacle |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0353893B1 (en) | Cell culture bioreactor | |
JP5271083B2 (en) | Cell culture method and apparatus for carrying out the same | |
EP0422149B1 (en) | Static oxygenator for suspension culture of animal cells | |
JPH0321149B2 (en) | ||
US4727040A (en) | Sparger for fermentation and tissue culturing vessels | |
US20030054544A1 (en) | Oxygen enriched bioreactor and method of culturing cells | |
EP0414887A1 (en) | Spin filter for removing substantially cell-free culture medium from suspension cell culture system. | |
IL179311A (en) | Membrane bioreactor | |
DK174748B1 (en) | Fermentor for growing cell cultures | |
IL104385A (en) | Method and apparatus for growing biomass particles | |
US3969190A (en) | Apparatus and method for microbial fermentation in a zero gravity environment | |
JP4561192B2 (en) | Cell culture device and cell culture method | |
JPH0595778A (en) | Porous separation membrane integrated type incubator equipped with stirrer | |
JPH07203945A (en) | Apparatus for culturing cell of living body | |
JP2011177046A (en) | Method and apparatus for culturing cell | |
JPH05252933A (en) | Method for supplying oxygen and device for culturing organismic cell by the method | |
JP3289984B2 (en) | Apparatus and method for culturing living cells | |
CN110734858A (en) | multi-mode three-dimensional perfusion type cell culture instrument | |
CN102272285A (en) | Method for reducing deposits during the cultivation of organisms | |
JPH06133765A (en) | Method for supplying oxygen and culture device used therefor | |
JP2613892B2 (en) | Animal cell culture method and culture device | |
US5151362A (en) | Apparatus containing a septum which impedes cell permeation for cell culture and method of use | |
JPH0775549A (en) | Living body cell culture arrangement | |
JPH04158781A (en) | Culture device | |
JPH0420240Y2 (en) |