JPS60234580A - Method of cell culture of animal - Google Patents
Method of cell culture of animalInfo
- Publication number
- JPS60234580A JPS60234580A JP59090903A JP9090384A JPS60234580A JP S60234580 A JPS60234580 A JP S60234580A JP 59090903 A JP59090903 A JP 59090903A JP 9090384 A JP9090384 A JP 9090384A JP S60234580 A JPS60234580 A JP S60234580A
- Authority
- JP
- Japan
- Prior art keywords
- culture
- semipermeable membrane
- culture solution
- oxygen
- air
- 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
Links
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
〔発明の利用分野〕
本発明は、動物細胞の生体外培養方法に係り、特に担体
培養法等で大量培養を行なうのに好適な動物細胞培養方
法に関するものである。DETAILED DESCRIPTION OF THE INVENTION [Field of Application of the Invention] The present invention relates to a method for culturing animal cells in vitro, and particularly to a method for culturing animal cells suitable for carrying out large-scale culture using a carrier culture method or the like.
動物細胞を生体外で大量培養する方法として、例えばＩ
細胞培養マニュアルＪ １９８２年５月１日講談社発行
、宗村庚修編、６３〜７０頁に「細胞の大量培養法」と
して、小量培養に用いるルーびんを必要数だけ用いる方
法、複数の円筒びんなゆっくり回転しながら培養を行な
うローラーボトル法、多層平板法等が種々論じられてい
る。これらは、いずれも培養表面積を大き鳴するための
方法であるが、培養表面積の割に設置場所、労力を必要
とするため、大規模培養には不向きである。As a method for mass culturing animal cells in vitro, for example, I
Cell Culture Manual J, published by Kodansha on May 1, 1982, edited by Kosuke Munemura, pp. 63-70, describes the method of using the required number of Lou bottles used for small-scale culture, and the method of using multiple cylindrical bottles. Various methods have been discussed, such as a roller bottle method in which culture is carried out while slowly rotating, a multilayer plate method, etc. These are all methods for increasing the culture surface area, but they are unsuitable for large-scale culture because they require installation space and labor relative to the culture surface area.
更に培養表面積を飛躍的に増大させる方法として、スー
パービーズ法（あるいはミクロキャリア法）が論じられ
ているが、ミクロキャリア法では、培養表面積は増大し
たものの、細胞への酸素供給に関しては十分考慮されて
な曵、つぎのような問題がある。Furthermore, the superbead method (or microcarrier method) is being discussed as a method to dramatically increase the culture surface area, but although the microcarrier method increases the culture surface area, sufficient consideration is not given to oxygen supply to cells. Well, I have the following problem.
一般に微生物培養では、微生物の生育および増殖に必要
な酸素は、培養液中に空気を吹込むことによって供給し
ている。しかしながら、動物細胞の培養においては、培
地として用いる直前が極めて発泡し易々、わずかな空気
の吹込みでも培養液が発泡して、培養に支障を生ずる等
の理由により、培養槽の気相部にのみ空気を張込むこと
で培養液中に酸素を供給している。この方法では、実験
室段階の培養に対しては十分な酸素供給能力を有するが
、培養槽を工業規模まで大形化した場合、培養液の増加
に対して液表面の増加の割合が小さいため、酸素供給能
力が不足する。この酸素供給能力の不足を補うため、気
相部への空気張込みと空気の吹込みを共用すると、前述
した如く、発泡の問題のため、これにも限度がある。Generally, in microbial culture, the oxygen necessary for the growth and proliferation of microorganisms is supplied by blowing air into the culture solution. However, when culturing animal cells, the gas phase of the culture tank is extremely prone to foaming just before it is used as a culture medium, and even the slightest amount of air blowing can cause the culture solution to foam, causing problems with culture. Oxygen is supplied to the culture medium by injecting air only into the culture medium. This method has sufficient oxygen supply capacity for laboratory-stage culture, but when the culture tank is enlarged to an industrial scale, the ratio of increase in the liquid surface to the increase in culture liquid is small. , oxygen supply capacity is insufficient. In order to compensate for this lack of oxygen supply ability, if air is pumped into the gas phase and air is blown into the gas phase, there is a limit to this because of the problem of foaming, as described above.
本発明の目的は、動物細胞の生体外での大量培養におい
て、培養槽を大形化しても培養液が発泡することなく、
十分な酸素供給能力を有する動物細胞培養方法を提供す
ることにある。The purpose of the present invention is to eliminate foaming of the culture solution even when the culture tank is enlarged in large-scale culture of animal cells in vitro.
The object of the present invention is to provide an animal cell culture method having sufficient oxygen supply capacity.
本発明は、動物細胞を生体外で大量培養する動物細胞培
養方法において、半透膜を介して培養液中に酸素を供給
することにより、培養液の発泡を生ずることなく、培養
槽の大形化に対応した酸素供給能力を持たせるようにし
たものである。The present invention provides an animal cell culture method for culturing animal cells in large quantities in vitro, by supplying oxygen to the culture solution through a semipermeable membrane, without causing foaming of the culture solution, and by increasing the size of the culture tank. It is designed to have an oxygen supply capacity that corresponds to the current demand.
以下、本発明の一実施例を第１図、第２図により説明す
る。第１図、第２図において、ｌは培養液およびそれに
浮遊した動物細胞または細胞が付着した担体を収容した
培養槽本体、２は培養槽本体ｌ内の培養液を攪拌する回
転翼、３は培養液中に空気中の酸素を供給するため、培
養槽本体ｌ内に複数本設置された半透膜管で、両端を空
気人口４および空気出口５に接続されている。An embodiment of the present invention will be described below with reference to FIGS. 1 and 2. In FIGS. 1 and 2, 1 is a culture tank main body containing a culture solution and animal cells floating thereon or carriers to which cells are attached, 2 is a rotor that stirs the culture solution in the culture tank main body 1, and 3 is a rotary blade that stirs the culture solution in the culture tank main body 1. In order to supply oxygen from the air to the culture solution, a plurality of semipermeable membrane tubes are installed inside the culture tank body 1, and both ends thereof are connected to the air port 4 and the air outlet 5.
培養槽本体ｌ内の培養液は、回転翼２によって動物細胞
または細胞が付着した担体が培養液中に均一に浮遊する
ように攪拌されており、細胞の生育および増殖に必要な
酸素は、培養槽本体１内気相部に張込まれた空気より、
培養液自由表面から吸収されると共に、空気人口４より
半透膜管３内を流通して空気出口５より出て行く空気中
の酸素が、半透膜を通って培養液中に浸出し供給される
。The culture solution in the culture tank main body 1 is stirred by rotary blades 2 so that animal cells or carriers to which cells are attached are evenly suspended in the culture solution, and the oxygen necessary for cell growth and proliferation is supplied to the culture medium. From the air injected into the gas phase inside the tank body 1,
Oxygen in the air that is absorbed from the free surface of the culture solution, flows through the semipermeable membrane tube 3 through the air outlet 5, and exits from the air outlet 5 passes through the semipermeable membrane and is supplied to the culture solution. be done.
この場合、半透膜管３を形成する半透膜は、酸素分子が
容易に通過できるものであればよく、また、適当な半透
膜を選択することにより、酸素を単一分子の形で培養液
中に溶出させることができ、気泡による発泡を防止する
ことができる。In this case, the semipermeable membrane forming the semipermeable membrane tube 3 may be one that allows oxygen molecules to pass through easily, and by selecting an appropriate semipermeable membrane, oxygen can be transferred in the form of a single molecule. It can be eluted into the culture solution and can prevent foaming due to air bubbles.
上述した実施例によれば、培養槽本体ｌの大形化に対応
して半透膜管３の設置本数を増加することができるので
、液体槽に対する液自由表面の割合が減少した場合にお
いても、培養液中への酸素供給能力を任意に増加するこ
とができる。According to the embodiment described above, the number of semipermeable membrane tubes 3 installed can be increased in response to an increase in the size of the culture tank main body l, so even when the ratio of the liquid free surface to the liquid tank is reduced, , the oxygen supply capacity into the culture medium can be increased arbitrarily.
第３図および第４図は、本発明の他の実施例を示したも
ので、第３図は半透膜管３によって回転翼２を構成した
ものであり、第４図は円板型半透膜６によって回転翼２
を構成したものであって、それぞれ半透膜管３および円
板型半透膜６を回転させることにより、半透膜管３およ
び円板型半透膜６表面と培養液との表面更新が促進され
るので、酸素供給能力を増大させることができる。3 and 4 show other embodiments of the present invention, in which the rotor blade 2 is constructed by a semipermeable membrane tube 3, and in FIG. The rotor 2 is connected by the permeable membrane 6.
By rotating the semipermeable membrane tube 3 and disk-shaped semipermeable membrane 6, respectively, the surfaces of the semipermeable membrane tube 3 and disk-shaped semipermeable membrane 6 and the culture solution can be renewed. The oxygen supply capacity can be increased.
本発明は以上述べたように、半透膜を介して培養液中に
酸素を供給するようにしたものであるから、培養液の発
泡を生ずることなく、培養液の自由表面以外からの酸素
供給能力を増加させることができると共に、培養槽の大
形化に対応した酸素供給能力を持たせることができ、培
養槽の培養効率を向上させることができる。As described above, the present invention is designed to supply oxygen into the culture medium through a semipermeable membrane, so oxygen can be supplied from other than the free surface of the culture medium without causing foaming of the culture medium. The capacity can be increased, and the culture tank can be provided with an oxygen supply capacity that corresponds to the larger size of the culture tank, and the culture efficiency of the culture tank can be improved.
第１図は本発明の一実施例を示す培養槽の縦断面図、第
２図は半透膜管の拡大詳細図、第３図および第４図は本
発明の他の実施例を示す培養槽の縦断面図である。
ｌ・・・・・・培養槽本体、２・・・・・・回転翼、３
・・・・・・半透膜管、４・・・・・・空気入口、５・
・・・・空気出口、６・・・・・・円板型半透膜
才１０
才３図FIG. 1 is a vertical sectional view of a culture tank showing one embodiment of the present invention, FIG. 2 is an enlarged detailed view of a semipermeable membrane tube, and FIGS. 3 and 4 are culture tanks showing other embodiments of the present invention. It is a longitudinal cross-sectional view of a tank. l... Culture tank main body, 2... Rotor blade, 3
... Semipermeable membrane tube, 4 ... Air inlet, 5.
... Air outlet, 6 ... Disk-shaped semipermeable membrane 10 years old 3 figures
Claims (1)
法において、半透膜を介して培養液中に酸素を供給する
ことを特徴とする動物細胞培養方法。 ２、回転翼を形成した半透膜を介して培養液中に酸素を
供給するようにした特許請求の範囲第１項記載の動物細
胞培養方法。 ３、円板型回転翼を形成した半透膜を介して培養液中に
酸素を供給するようにした特許請求の範囲第１項記載の
動物細胞培養方法。[Scope of Claims] 1. An animal cell culture method for culturing animal cells in large quantities in vitro, which is characterized in that oxygen is supplied to the culture solution through a semipermeable membrane. 2. The method for culturing animal cells according to claim 1, wherein oxygen is supplied to the culture solution through a semipermeable membrane having rotating blades. 3. The animal cell culture method according to claim 1, wherein oxygen is supplied to the culture solution through a semipermeable membrane formed with a disk-shaped rotor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59090903A JPS60234580A (en) | 1984-05-09 | 1984-05-09 | Method of cell culture of animal |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59090903A JPS60234580A (en) | 1984-05-09 | 1984-05-09 | Method of cell culture of animal |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS60234580A true JPS60234580A (en) | 1985-11-21 |
| JPH0328187B2 JPH0328187B2 (en) | 1991-04-18 |
Family
ID=14011359
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59090903A Granted JPS60234580A (en) | 1984-05-09 | 1984-05-09 | Method of cell culture of animal |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS60234580A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4959322A (en) * | 1989-03-07 | 1990-09-25 | W. L. Gore & Associates, Inc. | Culturing apparatus |
| US5110741A (en) * | 1987-03-18 | 1992-05-05 | Toyo Boseki Kabushiki Kaisha | Aeration apparatus for the culture of mammalian cells |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS50155677A (en) * | 1973-05-31 | 1975-12-16 | ||
| US3941662A (en) * | 1971-06-09 | 1976-03-02 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Apparatus for culturing cells |
| JPS5661988A (en) * | 1979-10-05 | 1981-05-27 | Intermedicat Gmbh | Fermentation container |
| JPS57118787A (en) * | 1980-11-18 | 1982-07-23 | Chemap Ag | Method and apparatus for culturing bacteria culture in liquid |
| JPS58116678A (en) * | 1981-12-28 | 1983-07-11 | Ajinomoto Co Inc | Method and apparatus for cultivation of suspensible animal cell |
-
1984
- 1984-05-09 JP JP59090903A patent/JPS60234580A/en active Granted
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3941662A (en) * | 1971-06-09 | 1976-03-02 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Apparatus for culturing cells |
| JPS50155677A (en) * | 1973-05-31 | 1975-12-16 | ||
| JPS5661988A (en) * | 1979-10-05 | 1981-05-27 | Intermedicat Gmbh | Fermentation container |
| JPS57118787A (en) * | 1980-11-18 | 1982-07-23 | Chemap Ag | Method and apparatus for culturing bacteria culture in liquid |
| JPS58116678A (en) * | 1981-12-28 | 1983-07-11 | Ajinomoto Co Inc | Method and apparatus for cultivation of suspensible animal cell |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5110741A (en) * | 1987-03-18 | 1992-05-05 | Toyo Boseki Kabushiki Kaisha | Aeration apparatus for the culture of mammalian cells |
| US4959322A (en) * | 1989-03-07 | 1990-09-25 | W. L. Gore & Associates, Inc. | Culturing apparatus |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0328187B2 (en) | 1991-04-18 |
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