JPS604714B2 - Tissue culture carrier particles - Google Patents
Tissue culture carrier particlesInfo
- Publication number
- JPS604714B2 JPS604714B2 JP9848680A JP9848680A JPS604714B2 JP S604714 B2 JPS604714 B2 JP S604714B2 JP 9848680 A JP9848680 A JP 9848680A JP 9848680 A JP9848680 A JP 9848680A JP S604714 B2 JPS604714 B2 JP S604714B2
- Authority
- JP
- Japan
- Prior art keywords
- carrier particles
- tissue culture
- particles
- culture carrier
- culture
- 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.)
- Expired
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
【発明の詳細な説明】 本発明は組織培養用担体粒子に関する。[Detailed description of the invention] The present invention relates to carrier particles for tissue culture.
細胞組織を大量に培養するために、近年、ＤＥＡＥ−デ
キストランからなる微小球状担体粒子を培養液中で浮遊
させ、細胞を粒子表面に接着させて増殖させる方法が行
なわれている。In order to culture cell tissues in large quantities, a method has recently been used in which microspherical carrier particles made of DEAE-dextran are suspended in a culture medium and cells are allowed to adhere to the particle surface and proliferate.
この方法によれば、装置容積に比べて細胞の接着、増殖
する培養面積を著しく大きくすることができるからであ
るが、しかし、一方において、従来から知られているよ
うな上記担体粒子は多孔質で水膨潤性であって、機械的
強度に劣るため、例えば担体粒子を培養液中で蝿杵、浮
遊させる際に担体粒子が破損したり、粒子相互に付着し
たりし、一般に耐久性に乏しい。さらに、従来の担体粒
子はＤＥＡＥーデキストラン重合体をェマルジョン化し
て製造されるため、その粒径を均一にそろえることが困
難であり「均一な粒径の担体粒子を得るには煩雑な分級
操作を必要とする。本発明は上記した問題を解決するた
めになされたものであって、機械的強度、耐久性にすぐ
れると共に、均一な粒径を有し、さらに細胞生育性、増
殖性にもすぐれる組織培養用担体粒子を提供することを
目的とする。According to this method, the culture area for cell adhesion and proliferation can be significantly increased compared to the device volume. However, on the other hand, the previously known carrier particles are porous. Because it is water-swellable and has poor mechanical strength, for example, when carrier particles are suspended in a culture medium, they may be damaged or adhere to each other, and generally have poor durability. . Furthermore, because conventional carrier particles are manufactured by emulsifying DEAE-dextran polymers, it is difficult to make the particle size uniform, and ``complicated classification operations are required to obtain carrier particles with a uniform particle size.'' The present invention has been made to solve the above-mentioned problems, and has excellent mechanical strength and durability, uniform particle size, and quick cell growth and proliferation. The purpose of the present invention is to provide carrier particles for tissue culture.
本発明の組織培養用担体粒子は、合成樹脂粒子からなり
、その表面がＢ線照射されていることを特徴とする。The tissue culture carrier particles of the present invention are made of synthetic resin particles, and are characterized in that their surfaces are irradiated with B-rays.
本発明においては担体粒子素材としての合成樹脂には種
々のものが用いられるが、普通、ポリスチレン、ポリ塩
化ビニル、スチレンーアクリロニトリル共重合体、ポリ
メチルメタクリレート、スチレンーメチルメタクリレー
ト共重合体、スチレンー無水マレィン酸共重合体、ポリ
エチレン、ポリプロピレン、ポリメチルベソテン−１等
が好ましく用いられる。In the present invention, various synthetic resins are used as carrier particle materials, but usually polystyrene, polyvinyl chloride, styrene-acrylonitrile copolymer, polymethyl methacrylate, styrene-methyl methacrylate copolymer, styrene-anhydrous Maleic acid copolymers, polyethylene, polypropylene, polymethylbesothene-1, etc. are preferably used.
このような合成樹脂を素材として、担体粒子は懸濁重合
又は乳化重合により製造される。重合法は何ら限定され
ず、従来から知られている方法に従えばよい。例えば懸
濁重合は、ポリメタクリル酸ナトリウム等の水溶性重合
体を懸濁剤として用いて重合を行なうのがよく、この方
法は通常、粒径１０〃以上の担体粒子の製造に適する。
また、乳化重合の場合にはドデシルベンゼンスルホン酸
ナトＩＪウム等のアニオン系界面活性剤及び／又はポリ
オキシェチレンィソオクチルフェニルェーテル等の非イ
オン系界面活性剤を乳化剤として用いて重合するのがよ
く、この方法は通常、５０〆以下の粒径の担体粒子の製
造に適する。本発明においては粒蓬１０〜５００仏の粒
子が好ましく用いられる。本発明の組織培養用担体粒子
は上記のような合成樹脂粒子表面にＢ線照射して得られ
、８線源としては通常の電子線加速装置を用いることが
できる。Using such a synthetic resin as a raw material, carrier particles are manufactured by suspension polymerization or emulsion polymerization. The polymerization method is not limited at all, and any conventionally known method may be followed. For example, suspension polymerization is preferably carried out using a water-soluble polymer such as sodium polymethacrylate as a suspending agent, and this method is usually suitable for producing carrier particles having a particle size of 10 mm or more.
In addition, in the case of emulsion polymerization, anionic surfactants such as sodium dodecylbenzenesulfonate and/or nonionic surfactants such as polyoxyethyleneisooctylphenyl ether are used as emulsifiers for polymerization. This method is generally suitable for producing carrier particles with a particle size of 50 mm or less. In the present invention, particles of 10 to 500 grains are preferably used. The carrier particles for tissue culture of the present invention are obtained by irradiating the surface of the synthetic resin particles as described above with B rays, and an ordinary electron beam accelerator can be used as the 8-ray source.
担体粒子による８線の吸収線量は７Ｍｒａｄ以上が好ま
しい。吸収線量が少なすぎるときは、担体粒子表面への
細胞の付着性、増殖性が十分ではない。一方、数百Ｍｒ
ａｄの吸収線量を有するように照射しても、数十Ｍｒａ
ｄの吸収線量の場合に比べて細胞の付着性や増殖性に目
立った効果がないので、通常は１００ＭＭｄ以下の吸収
線量で十分である。８線照射を行なう際の雰囲気ガスは
特に限定されないが、普通は空気、酸素、窒素、炭酸ガ
ス、一酸化炭素等が用いられる。The absorbed dose of 8 lines by the carrier particles is preferably 7 Mrad or more. If the absorbed dose is too small, the adhesion and proliferation of cells to the surface of the carrier particles will not be sufficient. On the other hand, several hundred Mr.
Even if irradiated with an absorbed dose of ad, the
An absorbed dose of 100 MMd or less is usually sufficient since there is no noticeable effect on cell adhesion or proliferation compared to the case of an absorbed dose of 100 MMd. The atmospheric gas used in the 8-ray irradiation is not particularly limited, but usually air, oxygen, nitrogen, carbon dioxide, carbon monoxide, etc. are used.
また、６線照射に際して、担体粒子は静層されていても
よく、凝拝されていてもよい。走行するベルト上に散布
され、連続的に照射されていてもよい。本発明の組織培
養用担体粒子は、一般に組織培養試験の対象となるすべ
ての細胞に対して適用することができるが、。Further, during the 6-ray irradiation, the carrier particles may be in a static layer or may be in a fixed layer. The light may be scattered on a running belt and irradiated continuously. The tissue culture carrier particles of the present invention can generally be applied to all cells that are the subject of tissue culture tests.
例えば代表的な株細胞としてＨｅＬａ（ヒト子宮頚部癌
）、Ｌ（マウス正常皮下結合組織）、ＷＩ−３８（ヒト
正常胎児肺）、ＫＢ（ヒト口底癌）、ＣｈａｎｇＬｉｖ
ｅｒ（ヒト正常肝臓）、ＦＬ（ヒト正常羊膜）、Ｄｏｎ
（チャイニーズハムスター正常肺）、ＢＨＫ−２１（シ
リアンハムスター正常賢）、ＩＭＲ−９０（ヒト肺由来
正二倍体の繊維芽細胞）等を例示することができる。本
発明の組織培養用担体粒子は、以上のように合成樹脂粒
子に８線照射されてなり、機械的強度にすぐれているの
で、カラムに充填して培養液を流動化させたり、また、
培養液中で燈拝して浮遊させる際に破損や粒子相互の付
着が起こらず、極めて耐久性にすぐれており、さらに合
成樹脂粒子は懸濁重合又は乳化重合によって粒蓬の均一
なものが容易に得られ、従って煩雑な分級操作を要せず
して、粒径のそろった担体粒子を得ることができる。For example, typical cell lines include HeLa (human cervical cancer), L (mouse normal subcutaneous connective tissue), WI-38 (human normal fetal lung), KB (human floor of mouth cancer), and ChangLiv.
er (human normal liver), FL (human normal amniotic membrane), Don
(Chinese hamster normal lung), BHK-21 (Syrian hamster normal lung), IMR-90 (eudiploid fibroblast derived from human lung), and the like. The tissue culture carrier particles of the present invention are made by irradiating synthetic resin particles with 8 rays as described above and have excellent mechanical strength, so they can be packed into columns to fluidize the culture medium, and
When floating in a culture solution, there is no breakage or adhesion of particles to each other, and the particles are extremely durable.Furthermore, synthetic resin particles can be easily made into uniform particles by suspension polymerization or emulsion polymerization. Therefore, carrier particles of uniform particle size can be obtained without the need for complicated classification operations.
また、その表面が８線処理されて良好な親水性を有する
ために、細胞の付着性、増殖性にもすぐれている。実施
例
通常の懸濁重合により得られた平均粒径１９０仏のポリ
スチレン粒子を分級せずに、加速電圧２×１ぴＶ、出力
１皿Ｗの電子線加速装置を用いて空気中で吸収線量が１
９Ｍｒａｄとなるように６線照射した。In addition, since its surface is 8-line treated and has good hydrophilicity, it has excellent cell adhesion and proliferation properties. Example Polystyrene particles with an average particle size of 190 mm obtained by ordinary suspension polymerization were measured in air using an electron beam accelerator with an accelerating voltage of 2 × 1 piV and an output of 1 plate W, without classifying them. is 1
Six rays were irradiated at 9 Mrad.
こうして得られた組織培養用担体粒子を酸化エチレンガ
スにて滅菌後、その０．６夕（乾燥重量）を蝿洋器付き
容器に入れ、さらに培養液ｌｏｏの‘を加えた。After sterilizing the tissue culture carrier particles thus obtained with ethylene oxide gas, 0.6 liters (dry weight) of the carrier particles were placed in a container with a container, and a culture solution LOOO was added thereto.
培養液はＥａｇｅＭ旧Ｍ培地７１．４の‘に成一ｖｉｎ
ｅｓｅｍｍ１４．３の‘を加え、７．５％炭酸水素ナト
リウムにてｐＨ調整したものを用いた。この培養液にＨ
ｅＬａＳ−３細胞１×１ぴ個を含む液１０泌を投与した
後、３７ｏ０の温度で５％炭酸ガス雰囲気中、櫨拝しな
がら培養を行なった。培養を開始して７日後に０．２５
％トリプシン液を加え、担体粒子表面に生育した細胞を
離脱させた。The culture medium is Seiichi vin in EageM old M medium 71.4'.
esemm14.3' was added and the pH was adjusted with 7.5% sodium hydrogen carbonate. This culture solution has H
After administering 10 volumes of a solution containing 1 x 1 cells of eLaS-3 cells, the cells were cultured at a temperature of 37°C in a 5% carbon dioxide atmosphere with shaking. 0.25 7 days after starting culture
% trypsin solution was added to detach cells grown on the surface of the carrier particles.
Claims (1)
いることを特徴とする組織培養用担体粒子。 ２ β線の吸収線量が７Ｍｒａｄ以上であることを特徴
とする特許請求の範囲第１項記載の組織培養用担体粒子
。[Scope of Claims] 1. A tissue culture carrier particle comprising synthetic resin particles, the surface of which is irradiated with β-rays. 2. The tissue culture carrier particles according to claim 1, wherein the absorbed dose of β-rays is 7 Mrad or more.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP9848680A JPS604714B2 (en) | 1980-07-17 | 1980-07-17 | Tissue culture carrier particles |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP9848680A JPS604714B2 (en) | 1980-07-17 | 1980-07-17 | Tissue culture carrier particles |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5722690A JPS5722690A (en) | 1982-02-05 |
| JPS604714B2 true JPS604714B2 (en) | 1985-02-06 |
Family
ID=14220974
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP9848680A Expired JPS604714B2 (en) | 1980-07-17 | 1980-07-17 | Tissue culture carrier particles |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS604714B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02139123A (en) * | 1988-11-18 | 1990-05-29 | Shizuoka Seiki Co Ltd | Short circuit foreseeing method in electrolytic finishing |
| JPH02152722A (en) * | 1988-12-01 | 1990-06-12 | Shizuoka Seiki Co Ltd | Short-circuit foreseeing method in electrolytic finishing |
-
1980
- 1980-07-17 JP JP9848680A patent/JPS604714B2/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5722690A (en) | 1982-02-05 |
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