JPS63283A - Bed material for cell culture - Google Patents
Bed material for cell cultureInfo
- Publication number
- JPS63283A JPS63283A JP61144257A JP14425786A JPS63283A JP S63283 A JPS63283 A JP S63283A JP 61144257 A JP61144257 A JP 61144257A JP 14425786 A JP14425786 A JP 14425786A JP S63283 A JPS63283 A JP S63283A
- Authority
- JP
- Japan
- Prior art keywords
- glutamate
- cell culture
- bed material
- cells
- copolymer
- 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
- 239000000463 material Substances 0.000 title claims abstract description 20
- 238000004113 cell culture Methods 0.000 title claims abstract description 16
- DHQUQYYPAWHGAR-UHFFFAOYSA-N dibenzyl 2-aminopentanedioate Chemical compound C=1C=CC=CC=1COC(=O)C(N)CCC(=O)OCC1=CC=CC=C1 DHQUQYYPAWHGAR-UHFFFAOYSA-N 0.000 claims abstract description 14
- YEJSPQZHMWGIGP-UHFFFAOYSA-N dl-glutamic acid dimethyl ester Natural products COC(=O)CCC(N)C(=O)OC YEJSPQZHMWGIGP-UHFFFAOYSA-N 0.000 claims abstract description 13
- SEWIYICDCVPBEW-UHFFFAOYSA-N methyl glutamate Chemical compound COC(=O)C(N)CCC(O)=O SEWIYICDCVPBEW-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229920005604 random copolymer Polymers 0.000 claims abstract description 9
- 238000009408 flooring Methods 0.000 claims description 4
- 229920001577 copolymer Polymers 0.000 abstract description 10
- 239000011521 glass Substances 0.000 abstract description 5
- 239000011324 bead Substances 0.000 abstract description 3
- 239000011248 coating agent Substances 0.000 abstract description 3
- 238000000576 coating method Methods 0.000 abstract description 3
- 239000000758 substrate Substances 0.000 abstract 2
- 230000035755 proliferation Effects 0.000 abstract 1
- 229920003002 synthetic resin Polymers 0.000 abstract 1
- 239000000057 synthetic resin Substances 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 14
- 229920000642 polymer Polymers 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 229920001519 homopolymer Polymers 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 230000021164 cell adhesion Effects 0.000 description 3
- -1 polyethylene Polymers 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 229920000193 polymethacrylate Polymers 0.000 description 2
- HFZKKJHBHCZXTQ-JTQLQIEISA-N (4s)-4-azaniumyl-5-oxo-5-phenylmethoxypentanoate Chemical compound OC(=O)CC[C@H](N)C(=O)OCC1=CC=CC=C1 HFZKKJHBHCZXTQ-JTQLQIEISA-N 0.000 description 1
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 description 1
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 235000010582 Pisum sativum Nutrition 0.000 description 1
- 240000004713 Pisum sativum Species 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 238000000944 Soxhlet extraction Methods 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000012510 hollow fiber Substances 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000010445 mica Substances 0.000 description 1
- 229910052618 mica group Inorganic materials 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 238000012758 nuclear staining Methods 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920002239 polyacrylonitrile Polymers 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 229920005553 polystyrene-acrylate Polymers 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 229920000915 polyvinyl chloride Polymers 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Polyamides (AREA)
Abstract
Description
【発明の詳細な説明】
〔技術分野〕
本発明は、細胞を培養し、増殖させる際に用いられる細
胞培養用床材に関するものである。DETAILED DESCRIPTION OF THE INVENTION [Technical Field] The present invention relates to a cell culture bed material used for culturing and proliferating cells.
一般に、動物細胞は浮遊状態では増殖せず、固体表面に
付着してのみ増殖する性質を有している。In general, animal cells do not grow in suspension, but only grow when attached to solid surfaces.
従って、そのような細胞を培養し、増殖させるためには
細胞を多量に付着する固体表面、つまり床材が必要とな
る。Therefore, in order to culture and proliferate such cells, a solid surface, ie, a bed material, to which a large amount of cells can adhere is required.
従来の細胞培養用床材は、一般に、ポリスチレン、ポリ
メタクリレート等の高分子材料から形成されているが、
これらの材料から形成された床材は、細胞が付着しにく
く、効率的な細胞培養を与えない。Conventional cell culture bedding is generally made of polymeric materials such as polystyrene and polymethacrylate.
Bedding formed from these materials is difficult for cells to adhere to and does not provide efficient cell culture.
本発明の目的は、細胞付着性の高い固体表面を有する細
胞培養用床材を提供することにある６〔構 成〕
本発明者は、細胞の付着しやすい性質を有する材料の開
発について種々研究を重ねた結果、グルタミン酸メチル
３５〜２５モル％とグルタミン酸ベンジル６５〜７５モ
ル％とのランダム共重合体はｆ４Ｉｌ鉋を著しく多量に
付着させる性質を有し、細胞培養用床材として好適であ
ることを見出し、本発明を完成するに到った。An object of the present invention is to provide a bed material for cell culture having a solid surface with high cell adhesion.6 [Structure] The present inventor has conducted various research on the development of materials that have properties to which cells easily adhere. As a result of repeated studies, it was found that a random copolymer of 35 to 25 mol% of methyl glutamate and 65 to 75 mol% of benzyl glutamate has the property of attaching a significantly large amount of f4Il planes, and is suitable as a flooring material for cell culture. They discovered this and completed the present invention.
本発明は、グルタミン酸メチル３５〜２５モル％とグル
タミン酸ベンジル６５〜７５モル％とのランダム共重合
体を表面に有することを特徴とする細胞培養用床材であ
る。The present invention is a cell culture floor material characterized by having a random copolymer of 35 to 25 mol% of methyl glutamate and 65 to 75 mol% of benzyl glutamate on its surface.
本明細書における細胞培養用床材としては、細胞を培養
する際に細胞を付着させて増殖をはかるために用いられ
る固体表面を有する物質を意味し。As used herein, the term "bed material for cell culture" refers to a material having a solid surface that is used to allow cells to adhere and grow during cell culture.
シート状、ｉ＊維状、ビーズ状、中空糸状、プレート状
等の形状を有するものである。It has a shape such as a sheet shape, an i*fiber shape, a bead shape, a hollow fiber shape, or a plate shape.
本発明の細胞培養（組織培養）川床材は、グルタミン酸
メチルとグルタミン酸ベンジルとのランダム共重合体か
らなるが、この場合、共重合体中のグルタミン酸ベンジ
ルの含量を６５〜７５モル％、好ましくは約７０モル％
付近にする。この範囲を逸脱すると、細胞付着性が悪化
する。The cell culture (tissue culture) river bed material of the present invention is made of a random copolymer of methyl glutamate and benzyl glutamate, and in this case, the content of benzyl glutamate in the copolymer is 65 to 75 mol%, preferably about 70 mol%
Make it nearby. Outside this range, cell adhesion deteriorates.
本発明で用いる共重合体の分子量は、皮膜を形成し得る
程度であればよく、通常１万〜１００万、好ましくは２
万〜１０万の範囲である６共重合体を構成するグルタミ
ン酸成分は、０体、１体又はラセミ体であることができ
る。The molecular weight of the copolymer used in the present invention may be as long as it can form a film, and is usually 10,000 to 1,000,000, preferably 20,000 to 1,000,000.
The glutamic acid component constituting the 6-copolymer, which ranges from 10,000 to 100,000, can be 0, 1, or racemic.
本発明による細胞培養用床材は、前記共重合体を所望の
形状に成形するか、又は支持体高分子で形成された成形
体の表面に共重合体をコーティングすることによって製
造することができる。アミノ酸系重合体は比較的高価で
あることから、安価な支持体高分子で形成した成形体の
表面に、前記共重合体をコーティングするのが好ましい
。支持体ポリマーとしては、ポリエチレン、ポリプロピ
レン、ポリスチレン、ポリアクリロニトリル、ポリメタ
クリレート、エチレン／プロピレン共重合体、ポリ塩化
ビニル、ポリエステル、ポリアミド等がある。支持体ポ
リマーの成形体は１発泡体又は非発泡体であることがで
きる。また、共重合体コーティング用支持体としては、
ガラスピーズ、ガラスフレーク、マイカ等の無機物も使
用される。The cell culture floor material according to the present invention can be produced by molding the copolymer into a desired shape or by coating the copolymer on the surface of a molded body made of a support polymer. Since amino acid-based polymers are relatively expensive, it is preferable to coat the surface of a molded article made of an inexpensive support polymer with the copolymer. Support polymers include polyethylene, polypropylene, polystyrene, polyacrylonitrile, polymethacrylate, ethylene/propylene copolymer, polyvinyl chloride, polyester, polyamide, and the like. The shaped bodies of the support polymer can be foamed or non-foamed. In addition, as a support for copolymer coating,
Inorganic materials such as glass peas, glass flakes, and mica are also used.
本発明の床材を用いて細胞培養を行う場合、固体表面と
して前記共重合体表面を用いる以外は。When carrying out cell culture using the flooring material of the present invention, the copolymer surface is used as the solid surface.
従来公知の方法に従って実施される。培養対象となる細
胞は、液体又はゼラチン状の培地に添加し。This is carried out according to conventionally known methods. Cells to be cultured are added to a liquid or gelatinous medium.
湿度３０〜４０℃で培養される。この培養において、細
胞は、固体表面によく付着し、効率的に増殖される。Cultured at a humidity of 30-40°C. In this culture, cells adhere well to solid surfaces and are efficiently grown.
次に本発明を実施例により説明する。 Next, the present invention will be explained by examples.
実施例Ｉ
し−グルタミン酸メチルとＬ−グルタミン酸ベンジルと
をそれぞれＮ−カルボキシアミノ酸無水物に変換した後
、それぞれの無水物を混合した。この場合、グルタミン
酸メチルとグルタミン酸ベンジルとの混合モル比は２９
／７１であった。この混合物をジオキサンと１，２−ジ
クロロエタンとの混合溶媒に溶かし、開始剤として゛ト
リエチルアミンを加えて重合させた。重合完結後、その
重合溶液をガラス板上に流延し、風乾して皮膜（膜厚約
０．０５ｍｍ）を得た。この皮膜をエタノールを用いた
ソックスレー抽出を行った後、乾燥した。Example I Methyl glutamate and benzyl L-glutamate were each converted into N-carboxyamino acid anhydrides, and then the respective anhydrides were mixed. In this case, the mixing molar ratio of methyl glutamate and benzyl glutamate is 29
/71. This mixture was dissolved in a mixed solvent of dioxane and 1,2-dichloroethane, and triethylamine was added as an initiator for polymerization. After the polymerization was completed, the polymer solution was cast onto a glass plate and air-dried to obtain a film (film thickness: about 0.05 mm). This film was subjected to Soxhlet extraction using ethanol and then dried.
次に、この皮膜上に、人由来の上皮性細胞を含む培養液
（約ｌＯ万個／ｍＱ）を接触させたまま、炭酸ガス濃度
５％、湿度１００％、３７°Ｃの部屋に静置した。Next, a culture medium containing human-derived epithelial cells (approximately 10,000 cells/mQ) was left in contact with the film and left in a room at 37°C with a carbon dioxide concentration of 5% and humidity of 100%. did.
１７時間後、皮膜上に付着している細胞の量を核染色法
により定量した。After 17 hours, the amount of cells adhering to the membrane was quantified by nuclear staining.
比較のため、グルタミン酸メチル５０モル％とグルタミ
ン酸ベンジル５０モル％とからなるランダム共重合体の
皮膜、グルタミン酸メチル単独重合体の皮膜及びグルタ
ミン酸ベンジル単独重合体の皮膜を用いて、同様の細胞
付着試験を行った。For comparison, a similar cell adhesion test was conducted using a random copolymer film consisting of 50 mol% methyl glutamate and 50 mol% benzyl glutamate, a methyl glutamate homopolymer film, and a benzyl glutamate homopolymer film. went.
皮膜に付着した細胞の量を標準試料（和光純薬工業株式
会社製組織培養用プラスチックシート）に付着した細胞
の量で割ることにより、細胞付着率を求めた。その結果
を第１表に示す。The cell attachment rate was determined by dividing the amount of cells attached to the film by the amount of cells attached to a standard sample (plastic sheet for tissue culture manufactured by Wako Pure Chemical Industries, Ltd.). The results are shown in Table 1.
なお、第１表に示した符号は次のことを意味する。Note that the symbols shown in Table 1 have the following meanings.
ＰＧＭ・・・グルタミン酸メチルの単独重合体Ｐ（ＧＭ
／Ｇ［ｌ） （Ｉ ）・・・グルタミン酸メチル５０モ
ル％とグルタミン酸ベンジル５０モル％とのランダム共
重合体
Ｐ （ＧＭ／ＧＢ） （■）・・・グルタミン酸メチル
２９モル％とグルタミン酸ベンジル７１モル％とのラン
ダム共重合体
１’ＧＢ・・・グルタミン酸ベンジルの単独重合体ＧＢ
・・・グルタミン酸ベンジル
第１表
実施例２
実施例１のＰ （ＧＭ／ＣＢ） （ＩＩ ）溶液（濃友
釣０．５％）中にガラスピーズ（直径約０．５ｍｍ）を
浸漬し、その後溶媒を除去、乾燥して、Ｐ （ＧＭ／Ｇ
Ｂ） （ｎ ）を表面に有するビーズ状細胞培養用床材
を得た。PGM...homopolymer P of methyl glutamate (GM
/G[l) (I)... Random copolymer P of methyl glutamate 50 mol% and benzyl glutamate 50 mol% (GM/GB) (■)... Methyl glutamate 29 mol% and benzyl glutamate 71 mol Random copolymer 1'GB with %...Homopolymer GB of benzyl glutamate
... Benzyl glutamate Table 1 Example 2 Glass beads (about 0.5 mm in diameter) were immersed in the P (GM/CB) (II) solution of Example 1 (0.5% Noyutsuri), and then the solvent is removed, dried, and P (GM/G
B) A bead-shaped cell culture bed material having (n) on the surface was obtained.
以上説明したように、本発明の細胞培養用床材は、極め
て多量の細胞を付着させることができ、その産業的意義
は大きい。As explained above, the cell culture flooring material of the present invention can attach an extremely large amount of cells, and has great industrial significance.
指定代理人 工業技術院製品科学研究所長高橋危司Designated Agent: Keiji Takahashi, Director, Product Science Research Institute, Agency of Industrial Science and Technology
Claims (1)
ン酸ベンジル６５〜７５モル％とのランダム共重合体を
表面に有することを特徴とする細胞培養用床材。(1) A cell culture flooring material having on its surface a random copolymer of 35 to 25 mol% of methyl glutamate and 65 to 75 mol% of benzyl glutamate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61144257A JPS63283A (en) | 1986-06-20 | 1986-06-20 | Bed material for cell culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61144257A JPS63283A (en) | 1986-06-20 | 1986-06-20 | Bed material for cell culture |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63283A true JPS63283A (en) | 1988-01-05 |
| JPH047671B2 JPH047671B2 (en) | 1992-02-12 |
Family
ID=15357890
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61144257A Granted JPS63283A (en) | 1986-06-20 | 1986-06-20 | Bed material for cell culture |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63283A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106665562A (en) * | 2017-03-14 | 2017-05-17 | 南京九寿堂医药科技有限公司 | Umbilical cord blood stem cell freezing tube |
-
1986
- 1986-06-20 JP JP61144257A patent/JPS63283A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106665562A (en) * | 2017-03-14 | 2017-05-17 | 南京九寿堂医药科技有限公司 | Umbilical cord blood stem cell freezing tube |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH047671B2 (en) | 1992-02-12 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EXPY | Cancellation because of completion of term |