JP2667447B2 - Method for immobilizing cells on a solid surface - Google Patents
Method for immobilizing cells on a solid surfaceInfo
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- JP2667447B2 JP2667447B2 JP63160229A JP16022988A JP2667447B2 JP 2667447 B2 JP2667447 B2 JP 2667447B2 JP 63160229 A JP63160229 A JP 63160229A JP 16022988 A JP16022988 A JP 16022988A JP 2667447 B2 JP2667447 B2 JP 2667447B2
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- cells
- ethyleneimine polymer
- solid
- plate
- treated
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Description
【発明の詳細な説明】 （産業上の利用分野） 本発明は固相免疫測定の混合凝集法あるいはアフィニ
ティークロマトグラフィーやバイオリアクター等に用い
られる細胞の固体表面への固定化方法に関する。TECHNICAL FIELD The present invention relates to a mixed agglutination method for solid-phase immunoassay or a method for immobilizing cells on a solid surface used for affinity chromatography, bioreactor and the like.
（従来の技術） 混合凝集法（mixed agglutination）は、抗原抗体反
応を利用した免疫学的凝集反応の検出方法の一つで、そ
の原理は1939年ウイーナー（Wiener）とハーマン（Herm
an）（J.Immunol.,（1939）36,255）によって提唱さ
れ、クームス（Coombs）らによって発展確立された〔Co
ombs,R.R.A.and BEDFORD,D.（1955）‘The A and B ant
igens on human Platelets demonstrated by means of
mixed erythrocyte−platelet agglutination.'Vox San
g.,5,111. Coombs,R.R.A.BEDFORD,D.and ROUILLARD,L.M.（195
6）．‘A and B blood−group antigens on human epid
ermal cell demonstrated by mixed agglutination'.La
ncet,i,461〕． さらに、それ以後応用が進みファーレウス（Fagraeu
s），エスプマーク（Espmark），ジョンソン（Johnsso
n）らがガラス表面上で培養細胞上の抗原を標識赤血球
試薬を用いて検出している〔Immunology,V.9.PP.161〜1
75（1962）〕。(Prior Art) The mixed agglutination method is one of the immunological agglutination detection methods using the antigen-antibody reaction, and its principle is 1939 Wiener and Herman.
an) (J. Immunol., (1939) 36, 255) and developed and established by Coombs et al. [Co
ombs, RRAand BEDFORD, D. (1955) 'The A and B ant
igens on human Platelets demonstrated by means of
mixed erythrocyte-platelet agglutination.'Vox San
g., 5,111. Coombs, RRABEDFORD, D.and ROUILLARD, LM (195
6). 'A and B blood-group antigens on human epid
ermal cell demonstrated by mixed agglutination'.La
ncet, i, 461]. In addition, the application has progressed since then, and
s), Espmark, Johnson (Johnsso)
n) et al. detect antigens on cultured cells on a glass surface using a labeled erythrocyte reagent [Immunology, V.9.PP.
75 (1962)].
またジュージ（Juji），カノウ（Kano）らによって血
小板抗原の分析に応用された（Int.Arch.Attergy,V.42,
PP.474〜484 Juji Kano and Milgrem）。Also applied to platelet antigen analysis by Juji, Kano et al. (Int. Arch. Attergy, V.42,
PP.474-484 Juji Kano and Milgrem).
さらに米国特許第4275053号および米国特許第4328183
号明細書で各種の血液型分析等に応用されるに至った。
混合凝集法において細胞等を安定に固体表面に結合させ
ることは非常に重要であり、種々の方法が実施されてい
る。Further, U.S. Pat.Nos. 4,427,053 and 4,328,183
In the specification, it has been applied to various blood group analysis and the like.
It is very important to stably bind cells and the like to a solid surface in the mixed aggregation method, and various methods have been implemented.
ジュージらはガラス試験管に吸着させているし、米国
特許第4275053号、米国特許第4328183号明細書ではポリ
スチレン表面をフィブリノーゲンとポリリジンで処理し
たり、化学的に共有結合させることができることを述べ
ている。Juge et al. Adsorb to glass test tubes and in U.S. Pat.No. 4,275,053 and U.S. Pat. I have.
（発明が解決しようとする課題） 細胞、オルガネラ等を固体表面に結合させる技術は、
混合凝集法の場合に限らず、アフィニテークロマトグラ
フィーや、バイオリアクター等の分野でも用いられてお
り、一般に架橋法と包括法がある。架橋法には米国特許
第4275053号明細書にも述べられているように反応可能
な固体表面をグルタルアルデヒドや臭化シアンで処理し
て細胞を結合させることができる。(Problem to be Solved by the Invention) The technology for binding cells, organelles, etc. to a solid surface is as follows.
It is used not only in the case of the mixed agglutination method but also in the fields of affinity chromatography, bioreactor and the like, and generally includes a cross-linking method and an inclusive method. For the cross-linking method, the reactive solid surface can be treated with glutaraldehyde or cyanogen bromide to attach cells, as also described in US Pat. No. 4,275,033.
しかし、これら共有結合を形成する反応性の基は、強
固な結合を形成するものの、安定性に難点があるため、
冷蔵保存もしくはすみやかに使用しなければならない。
また求核性であるがために反応はpHに大きく左右され、
一定した細胞の結合量をコントロールしにくいと言う欠
点をもっている。However, these reactive groups forming a covalent bond form a strong bond, but have a problem in stability.
Must be refrigerated or used immediately.
Also, because of its nucleophilicity, the reaction is highly dependent on pH,
It has the drawback that it is difficult to control the amount of fixed cell binding.
また米国特許第4275053号および米国特許第4328183号
明細書で述べられているフィブリノーゲンとポリリジン
による処理も２段階の処理が必要であったり、フィブリ
ノーゲン、ポリリジン自体安定性に乏しいために、処理
後すみやかに使用しなければならないという欠点を持っ
ている。In addition, the treatment with fibrinogen and polylysine described in U.S. Pat.No. 4,275,053 and U.S. Pat. It has the disadvantage of having to be used.
（課題を解決するための手段） そこで発明者らは安定かつ安価にしかも効率よく高速
に細胞を結合させることができる固体表面の処理方法を
開発すべく鋭意研究を行った結果、エチレンイミンポリ
マー（CH2CH2NH）ｎを吸着させた固体表面に生理的条件
下で、細胞が表面の機能をそこなわず効率良く結合でき
ることを見い出した。(Means for Solving the Problems) Therefore, the inventors conducted extensive research to develop a method for treating a solid surface, which is capable of binding cells stably, inexpensively, and efficiently at high speed. As a result, ethyleneimine polymer ( Under physiological conditions, it has been found that cells can efficiently bind to a solid surface to which CH 2 CH 2 NH) n has been adsorbed without impairing the surface function.
エチレンイミンポリマーは種々の有機プラスチック材
料に吸着するが、発明者等の検討ではヌンク（NUNC）社
製のImmuronで作られたポリスチレンマイクロプレート
が好適である。Ethyleneimine polymer is adsorbed on various organic plastic materials, but a polystyrene microplate made of Immuron manufactured by NUNC is preferable in the study of the present inventors.
さらに特開昭62−298763号公報に開示されているよう
に、エチレンイミンポリマーは、ガラス等無機材料にも
吸着し、前記有機材料同様細胞が効率良く結合できるこ
とを見出し本発明を達成するに至った。Further, as disclosed in JP-A-62-298763, the ethyleneimine polymer is also adsorbed on an inorganic material such as glass, and it has been found that cells can bind efficiently as in the case of the organic material, thus achieving the present invention. Was.
すなわち本発明は、抗原としての反応性を有する多数
個の所定の細胞をそのまま所定の容器状またはディスク
状の固体の表面に吸着させるための固体表面への細胞固
定化方法であって、該固体表面にエチレンイミンポリマ
ーをそのままの状態で結合させたのち、上記容器状また
はディスク状の固体の表面に結合されたエチレンイミン
ポリマーに対し生理的条件下で直接的に接触させること
を特徴とする固体表面への細胞固定化方法に関するもの
である。That is, the present invention is a method for immobilizing cells on a solid surface for adsorbing a large number of predetermined cells having reactivity as an antigen to the surface of a predetermined container-shaped or disc-shaped solid as they are, A solid characterized by directly bonding an ethyleneimine polymer on the surface thereof, and then directly contacting the ethyleneimine polymer bonded to the surface of the container-shaped or disc-shaped solid under physiological conditions. The present invention relates to a method for immobilizing cells on a surface.
エチレンイミンポリマー処理は水溶液の状態で固体表
面と接触させるだけで良く、その後、乾燥させても良
く、その際、室温で保存しても何ら性能に変化のない安
定な処理方法である。The ethyleneimine polymer treatment is a stable treatment method in which it may be brought into contact with the solid surface in the state of an aqueous solution and then dried, and the performance is not changed even if it is stored at room temperature.
またエチレンイミンポリマーは各種材料表面に化学的
処理を施した後化学的反応により導入することも可能で
ある。The ethyleneimine polymer can be introduced by a chemical reaction after subjecting various materials to a chemical treatment.
（実施例） 本発明を次の実施例により説明する。(Examples) The present invention will be described by the following examples.
実施例１ ＜ガラスチューブへの赤血球の固定化＞ ガラス試験管直径12mm×長さ75mm（コーニング社製）
に0.1％エチレンイミンポリマー（分子量約70,000、半
井化学（株）製）を200μ添加し、室温で30分反応さ
せた。Example 1 <Immobilization of red blood cells in a glass tube> Glass test tube diameter 12 mm x length 75 mm (Corning)
200 μl of 0.1% ethyleneimine polymer (molecular weight: about 70,000, manufactured by Hanoi Chemical Co., Ltd.) was added thereto, and the mixture was reacted at room temperature for 30 minutes.
精製水2mlで３回洗浄した後、0.3％に調整した赤血球
の生理食塩水（以下生食と言う）浮遊液200μを添加
し、1000rpmで１分間遠心した。次いでpH7.0の0.01Mリ
ン酸緩衝溶液（PBS）2mlで３回洗浄し、試験管底面に吸
着固定化した赤血球を無処理ガラス試験管につき同様に
行った結果と比較した。得た結果を次の第１表に示す。After washing 3 times with 2 ml of purified water, 200 μl of a suspension of erythrocyte physiological saline (hereinafter referred to as saline) adjusted to 0.3% was added and centrifuged at 1000 rpm for 1 minute. Then, the cells were washed 3 times with 2 ml of 0.01M phosphate buffer solution (PBS) having a pH of 7.0, and erythrocytes adsorbed and immobilized on the bottom surface of the test tube were compared with the results obtained in the same manner for untreated glass test tubes. The results obtained are shown in Table 1 below.
実施例２ ＜赤血球のプレートへの固定化＞ ヌンク社製マイクロプレート（No.464394）に0.1％エ
チレンイミンポリマー（半井化学（株）製）を100μ
／ウエル分注し、室温で30分インキュベートした後に、
0.01M PBS（pH＝7.0）300μ／ウエルで５回洗浄し、
室温で乾燥させて、エチレンイミンポリマー処理プレー
トを作成した。 Example 2 <Immobilization of erythrocytes to plate> 100 μl of 0.1% ethyleneimine polymer (manufactured by Hanui Chemical Co., Ltd.) was applied to a microplate (No. 464394) manufactured by Nunc.
/ Well dispensing and incubation at room temperature for 30 minutes,
Wash 5 times with 300μ / well of 0.01M PBS (pH = 7.0)
Dried at room temperature to make an ethyleneimine polymer treated plate.
次に上記エチレンイミンポリマー処理プレートに赤血
球の固定化を試みた。市販の赤血球であるサージスクリ
ーン（Surgiscreen）（商標名、オーソ社製、ロットNo.
3SS837）３％懸濁液を生食で適宜希釈して３％の他0.75
％、0.3％、0.15％の希釈懸濁液を作成し、各、懸濁液
を25μ／ウエルずつ分注し、室温で10分間インキュベ
ートした後0.1％（牛血清アルブミン）（BSA）含有、0.
01M PBS（pH＝7.0）200μ／ウエルで３回ウエルを洗
浄した。Next, an attempt was made to immobilize erythrocytes on the plate treated with the ethyleneimine polymer. A commercially available red blood cell, Surgiscreen (trade name, manufactured by Ortho, Lot No.
3SS837) 3% suspension diluted appropriately with saline, 3% and 0.75
%, 0.3%, 0.15% diluted suspensions were prepared, each suspension was dispensed at 25 μ / well, incubated at room temperature for 10 minutes, and then containing 0.1% (bovine serum albumin) (BSA), 0 .
The wells were washed three times with 200 μl / well of 01M PBS (pH = 7.0).
また、エチレンイミンポリマー処理していないヌンク
社製マイクロプレートについても同様な操作を行った。Further, the same operation was performed on a microplate manufactured by Nunc Co., Ltd., which was not treated with ethyleneimine polymer.
尚上記サージスクリーンのアンチグラム（抗原表）は
下記の通りである： Donor No.41901、Geno Type R2R2、Ｄ（＋）,C（０）,E
（＋）,c（０）,e（０）,f（０）,K（０）,k（＋）,Fya
（＋）,Fyb（＋）,JKa（０）,JKb（＋）,Xga（＋）,Lea
（０）,leb（＋）,S（＋）,s（０）,M（＋）,N（０）,P
1（＋）（＋）含有，（０）非含有 エチレンイミンポリマー処理、非処理プレートについ
て、固定化された赤血球量を求めるため倍率200倍で鏡
検して、単位面積あたりの赤血球数を算出した。得た結
果を第２表に示す。The antigram (antigen table) of the above surge screen is as follows: Donor No. 41901, Geno Type R 2 R 2 , D (+), C (0), E
(+), C (0) , e (0), f (0), K (0), k (+), Fy a
(+), Fy b (+), JK a (0), JK b (+), Xg a (+), Le a
(0), le b (+), S (+), s (0), M (+), N (0), P
1 (+) (+)-containing, (0) -free Plates treated with ethyleneimine polymer and untreated were microscopically examined at a magnification of 200 to determine the amount of immobilized red blood cells, and the number of red blood cells per unit area was calculated. did. The results obtained are shown in Table 2.
赤血球は、作用血球濃度で差はあるがプレート表面に
密に結合してるがエチレンイミンポリマー非処理プレー
トとは、処理プレートに比べて、ほとんど結合していな
いと言える。従って、エチレンイミンポリマー処理によ
って、赤血球が効率よくプレート面に結合させることが
できると言える。エチレンイミンポリマー濃度は0.001
％〜１％が好適であった。 The erythrocytes are tightly bound to the plate surface although there is a difference in the working blood cell concentration, but it can be said that the erythrocytes are hardly bound to the plate not treated with the ethyleneimine polymer as compared with the treated plate. Therefore, it can be said that the erythrocytes can be efficiently bound to the plate surface by the ethyleneimine polymer treatment. Ethyleneimine polymer concentration 0.001
% To 1% was preferred.
実施例３ ＜血小板のプレートへの固定＞ 実施例２で作成したエチレンイミンポリマー処理プレ
ートに血小板の固定を試みた。ACD添加全血を軽遠心110
0g−６分行い、上清として多血小板血漿を得た。更にこ
の上清を強遠心1500g−15分行って血小板を沈澱させ、
生食で３回洗浄し、1.0mlの懸濁液とした（２×109個/m
l）。この懸濁液を生食で適宜希釈し各種濃度の血小板
懸濁液を作成し、エチレンイミンポリマー処理プレート
と非処理プレートに25μ／ウエルずつ分注し、室温で
30分インキュベートした後、0.1％BSA含有0.01M PBS（p
H＝7.0）200μ／ウエルで３回洗浄した。Example 3 <Fixation of platelets on plate> Fixation of platelets on the plate treated with ethyleneimine polymer prepared in Example 2 was attempted. Lightly centrifuge ACD-supplemented whole blood 110
The operation was performed at 0 g for 6 minutes to obtain platelet-rich plasma as a supernatant. The supernatant was further centrifuged at 1500 g for 15 minutes to precipitate platelets,
Washed three times with saline to make a 1.0 ml suspension (2 × 10 9 / m
l). This suspension was appropriately diluted with saline to prepare platelet suspensions of various concentrations, and dispensed at 25 μ / well into an ethyleneimine polymer-treated plate and a non-treated plate at room temperature.
After incubating for 30 minutes, 0.01M PBS containing 0.1% BSA (p
(H = 7.0) The plate was washed three times with 200 μ / well.
倍率200倍で鏡検して固定された単位面積あたりの血
小板数を求めた。得た結果を第３表に示す。The number of platelets per fixed unit area was determined by microscopic examination at a magnification of 200 times. Table 3 shows the obtained results.
上表に示すとおり、エチレンイミンポリマー処理した
プレートの方が明らかに多くの血小板が固定化されてお
り、血小板固定プレート作成にもエチレンイミンポリマ
ー処理は有効であった。 As shown in the above table, the plate treated with the ethyleneimine polymer had clearly more platelets immobilized thereon, and the treatment with the ethyleneimine polymer was also effective in preparing a platelet-fixed plate.
実施例４ ＜抗グロブリン試験（coombs法）による不規則抗体の検
出＞ ・Ｏ型セル（Cell）固相プレート 実施例２で実施したようにエチレンイミンポリマー処
理したマイクロプレートに0.3％サージスクリーン２を
吸着固定化させたものを用いた。Example 4 <Detection of irregular antibody by antiglobulin test (coombs method)> O type cell (solid phase plate) 0.3% surge screen 2 was added to the ethyleneimine polymer-treated microplate as in Example 2. The one immobilized by adsorption was used.
・抗ヒトIgG感作粒子 抗ヒトIgGを感作させた血球の抗ヒトIgGセルキット
（商品名、オリンパス光学工業（株）製）を使用した。-Anti-human IgG sensitized particles An anti-human IgG cell kit (trade name, manufactured by Olympus Optical Co., Ltd.) of blood cells sensitized with anti-human IgG was used.
・不規則抗体の検出 前記Ｏ型Cell固相プレートにLiss液60μ、サンプル
としてオーソ社の抗、抗Ｋ、抗Fya、抗ｓの各抗血清3
0μを添加し、37℃20min反応させた。0.01M PBS（pH
7.0）200μで４回洗浄し、次いで前記抗ヒトIgG感作
粒子を25μづつ添加し、室温で４時間静置し、マイク
ロプレート底面に形成される反応パターンを検出した。
得られた結果を第４表に示す。- irregular antibody detection the O-type Cell solid plate Liss liquid 60 microns, as a sample Ortho's anti, anti K, anti-Fy a, each of the anti-s antiserum 3
0 µ was added and reacted at 37 ° C for 20 minutes. 0.01M PBS (pH
7.0) Washed 4 times with 200μ, then added 25μ each of the anti-human IgG sensitized particles, allowed to stand at room temperature for 4 hours, and detected the reaction pattern formed on the bottom surface of the microplate.
Table 4 shows the obtained results.
前記抗ヒトIgG感作粒子が、前記Ｏ型Cell固相プレー
トの底面に一様に広がったものを＋、中心に集まったも
のを−と判定した。 When the anti-human IgG-sensitized particles uniformly spread on the bottom surface of the O-type Cell solid-phase plate, it was judged as +, and when they were collected at the center, as-.
結果は、サージスクリーンのアンチグラム（以下の第
５表）と一致し、全ての抗原型が抗原としての反応を安
定に維持していることが確認された。The results were consistent with the antigram of the surge screen (Table 5 below), and it was confirmed that all serotypes maintained a stable reaction as an antigen.
実施例５ ＜リンパ球のプレートへの固定＞ 血液10mlにACD液を1.5ml加えた。400g−10分遠心を行
い、バフィーコートをピペットで約1.5ml吸引分取し、
1.5mlの生食を加えて、撹拌混和した。この懸濁液に分
離用比重液（モノポリ・レゾルビング・メジューム、Ｍ
−BRM（商品名、フロー・ラボラトリー社製）を静かに
積層した。600g−10分遠心し、リンパ球層を分取し、60
00g−１分遠心後沈渣を再び生食に懸濁し、1000−ｇ−
１分遠心し上清を吸引除去した。ハンクス液を加え、ト
ロンビン（100単位/ml）を１滴加え、混和した。1000g
−３秒遠心し、上清を再び1000g−１分遠心し、沈渣を1
mlの生食に懸濁させた（２×106個/ml）これを適宜生食
で希釈し、実施例２で作成したエチレンイミンポリマー
処理プレートと、非処理プレート25μ／ウエルずつ分
注し、室温で30分インキュベートした後、0.1％BSA含有
0.01M PBS（pH＝7.0）200μ／ウエルで３回洗浄し、
倍率200でリンパ球のプレートへの固定を単位面積あた
りのリンパ球をはかって算出した。得た結果を第６表に
示す。 Example 5 <Immobilization of lymphocytes on plate> 1.5 ml of ACD solution was added to 10 ml of blood. Perform centrifugation at 400 g for 10 minutes, aspirate and collect about 1.5 ml of the buffy coat with a pipette,
1.5 ml of raw food was added and mixed by stirring. The suspension is mixed with a specific gravity liquid for separation (Monopoly Resolving Medium, M
-BRM (trade name, manufactured by Flow Laboratories) was gently laminated. Centrifuge at 600 g for 10 minutes to separate the lymphocyte layer,
After centrifugation at 00 g for 1 minute, the precipitate was suspended again in
After centrifugation for 1 minute, the supernatant was removed by suction. The Hanks solution was added, and one drop of thrombin (100 units / ml) was added and mixed. 1000g
-3 seconds, and the supernatant was centrifuged again at 1000 g for 1 minute.
Suspended in 2 ml of saline (2 × 10 6 cells / ml), appropriately diluted with saline, dispensed with ethyleneimine polymer-treated plate prepared in Example 2 and non-treated plate (25 μ / well) at room temperature After 30 minutes incubation with 0.1% BSA
Wash three times with 200μ / well of 0.01M PBS (pH = 7.0)
The fixation of the lymphocytes to the plate at a magnification of 200 was calculated by measuring the lymphocytes per unit area. The results obtained are shown in Table 6.
表に示すとおり、エチレンイミンポリマー処理プレー
トの方が明らかに多くのリンパ球を固定することができ
た。 As shown in the table, the plate treated with the ethyleneimine polymer was able to clearly fix more lymphocytes.
実施例６ ＜ナイロンディスクへの赤血球の固定化＞ ナイロンディスク（6.6ナイロン）を2N HCl60℃、５
時間加水分解した後、純水で洗浄し、次に0.1％グルタ
ルアルデヒドを含む0.1M炭素水素ナトリウム水溶液に25
℃で１時間反応させた。反応後、純水で洗浄し、0.1％
エチレンイミンポリマー水溶液を室温で２時間反応させ
た。反応後純水で洗浄し、0.1Mグリシン−NaOH緩衝液
（pH8.5）中にて室温で一夜放置し、残余アルデヒド基
をブロッキングした。Example 6 <Immobilization of erythrocytes on nylon disc> Nylon disc (6.6 nylon) was treated with 2N HCl at 60 ° C,
After hydrolyzing for hours, wash with pure water and then add 25% aqueous solution of 0.1M sodium hydrogencarbonate containing 0.1% glutaraldehyde.
The reaction was carried out at a temperature of 1 hour. After the reaction, wash with pure water, 0.1%
The ethyleneimine polymer aqueous solution was reacted at room temperature for 2 hours. After the reaction, the reaction solution was washed with pure water and left overnight at room temperature in a 0.1 M glycine-NaOH buffer solution (pH 8.5) to block residual aldehyde groups.
純水で洗浄し、エチレンイミンポリマー処理ナイロン
ディスクを得た。0.3％に調整した赤血球の生理食塩水
浮遊液をエチレンイミンポリマー処理ナイロンディスク
に滴下し、15分室温静置した後、0.01M PBS（pH9.0）で
洗浄、前記ディスク上に残った赤血球をエチレンイミン
ポリマー無処理ナイロンディスクと比較した。得た結果
を第７表に示す。After washing with pure water, an ethyleneimine polymer-treated nylon disk was obtained. A suspension of erythrocytes in physiological saline adjusted to 0.3% was dropped onto an ethyleneimine polymer-treated nylon disk, left to stand at room temperature for 15 minutes, and then washed with 0.01M PBS (pH 9.0) to remove red blood cells remaining on the disk. A comparison was made with an ethyleneimine polymer untreated nylon disc. The results obtained are shown in Table 7.
（発明の効果） 本発明においては、架橋剤を介さずにエチレンイミン
ポリマーのみの吸着力で細胞を固定することによって、
固体表面と細胞との間に生理条件下で高密度に細胞を固
定できることが判明したので、安定な固定処理を実現で
きるとともに細胞の免疫学的機能を損なうことがない。 (Effect of the Invention) In the present invention, by fixing cells with the adsorption force of only ethyleneimine polymer without using a crosslinking agent,
Since it has been found that cells can be fixed at a high density between the solid surface and the cells under physiological conditions, stable fixing treatment can be realized and the immunological function of the cells is not impaired.
Claims (1)
の細胞をそのまま所定の容器状またはディスク状の固体
の表面に吸着させるための固体表面への細胞の固定化方
法であって、該固体表面にエチレンイミンポリマーをそ
のままの状態で結合させた後、上記容器状またはディス
ク状の固体の表面に結合されたエチレンイミンポリマー
に対し生理条件下で直接的に接触させることを特徴とす
る固体表面への細胞固定化方法。1. A method of immobilizing cells on a solid surface for adsorbing a large number of predetermined cells having reactivity as an antigen on a surface of a predetermined container-shaped or disk-shaped solid as it is, A solid characterized by directly bonding an ethyleneimine polymer to the surface of the solid, and then directly contacting the ethyleneimine polymer bonded to the surface of the container-shaped or disc-shaped solid under physiological conditions. A method for immobilizing cells on a surface.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63160229A JP2667447B2 (en) | 1988-06-28 | 1988-06-28 | Method for immobilizing cells on a solid surface |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63160229A JP2667447B2 (en) | 1988-06-28 | 1988-06-28 | Method for immobilizing cells on a solid surface |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0210160A JPH0210160A (en) | 1990-01-12 |
| JP2667447B2 true JP2667447B2 (en) | 1997-10-27 |
Family
ID=15710497
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63160229A Expired - Lifetime JP2667447B2 (en) | 1988-06-28 | 1988-06-28 | Method for immobilizing cells on a solid surface |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2667447B2 (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3835203A1 (en) * | 1988-10-15 | 1990-04-19 | Bayer Ag | MOLDS FROM POLYCARBONATE MIXTURES OF HIGH DISPERSE SOLUBILITY |
| DK1800129T3 (en) * | 2004-09-23 | 2013-07-01 | Tripath Imaging Inc | Polycationic polymer coatings for immobilization of biological samples |
| JP6779483B2 (en) | 2016-09-29 | 2020-11-04 | 住友ゴム工業株式会社 | Medical testing equipment and cell testing method |
| JP7109719B2 (en) | 2018-02-14 | 2022-08-01 | 住友ゴム工業株式会社 | Specific cell capture method |
| JP7158671B2 (en) | 2018-02-14 | 2022-10-24 | 住友ゴム工業株式会社 | Specific cell capture method |
| JP7170254B2 (en) * | 2018-02-14 | 2022-11-14 | 住友ゴム工業株式会社 | Specific cell capture method |
| US11614440B2 (en) | 2019-01-24 | 2023-03-28 | Sumitomo Rubber Industries, Ltd. | Specific cell fractionating and capturing methods |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6015560A (en) * | 1983-07-07 | 1985-01-26 | Sumitomo Bakelite Co Ltd | Immunological measuring molded article and preparation thereof |
| JPS6281565A (en) * | 1985-09-30 | 1987-04-15 | マイルス・インコーポレーテッド | Method of identifying nucleic acid to modified nylon substrate, nucleic acid identified by said method and methodof measuring polynucleotide configuration in sample |
-
1988
- 1988-06-28 JP JP63160229A patent/JP2667447B2/en not_active Expired - Lifetime
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6015560A (en) * | 1983-07-07 | 1985-01-26 | Sumitomo Bakelite Co Ltd | Immunological measuring molded article and preparation thereof |
| JPS6281565A (en) * | 1985-09-30 | 1987-04-15 | マイルス・インコーポレーテッド | Method of identifying nucleic acid to modified nylon substrate, nucleic acid identified by said method and methodof measuring polynucleotide configuration in sample |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0210160A (en) | 1990-01-12 |
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