JPS63173574A - Electrode for cell fusion - Google Patents
Electrode for cell fusionInfo
- Publication number
- JPS63173574A JPS63173574A JP62004040A JP404087A JPS63173574A JP S63173574 A JPS63173574 A JP S63173574A JP 62004040 A JP62004040 A JP 62004040A JP 404087 A JP404087 A JP 404087A JP S63173574 A JPS63173574 A JP S63173574A
- Authority
- JP
- Japan
- Prior art keywords
- electrode
- cell fusion
- fusion
- high polymer
- electrodes
- 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
- 230000007910 cell fusion Effects 0.000 title claims abstract description 21
- 239000011148 porous material Substances 0.000 claims abstract description 11
- 229920005597 polymer membrane Polymers 0.000 claims description 11
- 230000004927 fusion Effects 0.000 abstract description 15
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 abstract description 13
- 229910052697 platinum Inorganic materials 0.000 abstract description 6
- 229920002037 poly(vinyl butyral) polymer Polymers 0.000 abstract description 4
- 229920006254 polymer film Polymers 0.000 abstract description 4
- 239000007772 electrode material Substances 0.000 abstract description 3
- 239000007769 metal material Substances 0.000 abstract description 3
- 229910001220 stainless steel Inorganic materials 0.000 abstract description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 2
- 229920002284 Cellulose triacetate Polymers 0.000 abstract description 2
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 abstract description 2
- NNLVGZFZQQXQNW-ADJNRHBOSA-N [(2r,3r,4s,5r,6s)-4,5-diacetyloxy-3-[(2s,3r,4s,5r,6r)-3,4,5-triacetyloxy-6-(acetyloxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6s)-4,5,6-triacetyloxy-2-(acetyloxymethyl)oxan-3-yl]oxyoxan-2-yl]methyl acetate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](OC(C)=O)[C@H]1OC(C)=O)O[C@H]1[C@@H]([C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H](COC(C)=O)O1)OC(C)=O)COC(=O)C)[C@@H]1[C@@H](COC(C)=O)O[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@H]1OC(C)=O NNLVGZFZQQXQNW-ADJNRHBOSA-N 0.000 abstract description 2
- 229910052782 aluminium Inorganic materials 0.000 abstract description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 abstract description 2
- 229910052799 carbon Inorganic materials 0.000 abstract description 2
- 239000011651 chromium Substances 0.000 abstract description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 abstract description 2
- 229910052737 gold Inorganic materials 0.000 abstract description 2
- 239000010931 gold Substances 0.000 abstract description 2
- 239000000463 material Substances 0.000 abstract description 2
- 229920002492 poly(sulfone) Polymers 0.000 abstract description 2
- 229920002239 polyacrylonitrile Polymers 0.000 abstract description 2
- 229920000515 polycarbonate Polymers 0.000 abstract description 2
- 239000004417 polycarbonate Substances 0.000 abstract description 2
- 239000002861 polymer material Substances 0.000 abstract description 2
- 229920002635 polyurethane Polymers 0.000 abstract description 2
- 239000004814 polyurethane Substances 0.000 abstract description 2
- 239000010935 stainless steel Substances 0.000 abstract description 2
- 229910052804 chromium Inorganic materials 0.000 abstract 1
- 239000011248 coating agent Substances 0.000 abstract 1
- 238000000576 coating method Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 19
- 210000001938 protoplast Anatomy 0.000 description 9
- 238000000034 method Methods 0.000 description 7
- 239000006152 selective media Substances 0.000 description 7
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- -1 Conex Polymers 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 239000004695 Polyether sulfone Substances 0.000 description 1
- 240000005578 Rivina humilis Species 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 229920006217 cellulose acetate butyrate Polymers 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 150000004820 halides Chemical class 0.000 description 1
- 238000000464 low-speed centrifugation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 229920000058 polyacrylate Polymers 0.000 description 1
- 229920006393 polyether sulfone Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000004800 polyvinyl chloride Substances 0.000 description 1
- 229920000915 polyvinyl chloride Polymers 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000002747 voluntary effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- 108010082737 zymolyase Proteins 0.000 description 1
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
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Electromagnetism (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は細胞融合用電極に関し、より詳しくは細胞の融
合率を高め、融合の有無を効率よく判定することができ
る細胞融合用電極に関する。DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an electrode for cell fusion, and more particularly to an electrode for cell fusion that can increase the fusion rate of cells and efficiently determine the presence or absence of fusion.
本発明の細胞融合用電極は、食品、醗酵、農林水産業、
医薬品などの広範囲な分野で利用可能である。The cell fusion electrode of the present invention is applicable to food, fermentation, agriculture, forestry and fisheries industries,
It can be used in a wide range of fields such as pharmaceuticals.
細胞にパルスを印加することにより細胞融合させる方法
として、従来から均一電極と不均一電極とが使用されて
きた。均一電極としては、２枚の平板電極の向き合った
距離を調節する平行平板電極が代表的である。不均一電
極としては、微小電極法（マイクロマニピュレーター）
、白金線電極法などがある。Conventionally, uniform electrodes and non-uniform electrodes have been used as a method for cell fusion by applying pulses to cells. A typical example of the uniform electrode is a parallel plate electrode that adjusts the distance between two plate electrodes facing each other. As a non-uniform electrode, microelectrode method (micromanipulator)
, platinum wire electrode method, etc.
いずれの方法を採用しても、細胞を融合させる場合、融
合の有無及び融合率を判定するには、一般の光学顕微鏡
で観察する方法もあるが、この方法は光学顕微鏡で識別
可能な大きな細胞にしか適用できず、たまたま融合細胞
の存在する部位を鏡検したときにのみ細胞融合の判定を
下すことができるにすぎない。より確実な方法として、
融合処理を行った細胞液を選択培地を用いて増殖させ、
コロニーの出現の有無により判定している。No matter which method is used, when cells are fused, one way to determine the presence or absence of fusion and the fusion rate is to observe it with a general light microscope. Cell fusion can only be determined by microscopic examination of a site where fused cells happen to be present. As a more reliable method,
Proliferate the fusion-treated cell solution using a selective medium,
Judgment is made based on the presence or absence of colonies.
しかしながら、選択培地を使用する方法も融合率が低い
ときには、たまたま採取して選択培地に滴下した液の中
に融合細胞が含まれていなかった場合には陰性と判定さ
れる。そのため確実に判定するためには実験回数を増加
しなければならない欠点があった。したがって、融合率
を高め得る細胞融合用電極は望ましいものである。或い
は局所的に細胞融合率の高い部位が判明している細胞融
合用電極があれば、融合処理後その部位の細胞液をとっ
て選択培地で培養するならば効率よく融合の有無を判定
することができる。However, even with the method using a selective medium, when the fusion rate is low, a negative result is determined if the liquid sampled and dropped onto the selective medium does not contain fused cells. Therefore, there was a drawback that the number of experiments had to be increased in order to make a reliable determination. Therefore, a cell fusion electrode that can increase the fusion rate is desirable. Alternatively, if there is a cell fusion electrode in which a region with a locally high cell fusion rate is known, the presence or absence of fusion can be efficiently determined by taking the cell fluid from that region after fusion treatment and culturing it in a selective medium. Can be done.
本発明は、電気的細胞融合の操作において、細胞融合率
を高め、融合の有無を効率よく判定することができる細
胞融合用電極を提供することを目的とする。An object of the present invention is to provide a cell fusion electrode that can increase the cell fusion rate and efficiently determine the presence or absence of fusion in electrical cell fusion operations.
本発明は、表面が、多数の微小な孔を有する多孔質の高
分子膜で被覆されていることを特徴とする。The present invention is characterized in that the surface is coated with a porous polymer membrane having many minute pores.
本発明に係る細胞融合用電極の電極素材としては、白金
、金、アルミニウム、クロム、ステンレス鋼等の金属材
料或いは炭素等の非金属材料が挙げられる。形状は、第
１図に示すような平板状が好ましいが棒状でも差支えな
い。使用にあたっては、両極間に電流パルスを印加して
細胞を融合させる。印加する電流は直流でも交流でもよ
い。Examples of the electrode material for the cell fusion electrode according to the present invention include metal materials such as platinum, gold, aluminum, chromium, and stainless steel, and non-metallic materials such as carbon. The shape is preferably a flat plate as shown in FIG. 1, but it may also be rod-shaped. In use, a current pulse is applied between the poles to fuse the cells. The applied current may be direct current or alternating current.
本発明に使用する高分子膜材料としては、ポリスルホン
、酢酸セルロース、トリ酢酸セルロース、酢酸酪酸セル
ロース、ポリアクリロニトリル、ポリカーボネート、ポ
リウレタン、ポリ塩化ビニル、ポリビニルブチラール、
ナイロン、コーネックス、ポリアクリレート、ポリエー
テルスルホン、架橋ポリビニルピリジニウムハライドな
どからなる高分子多孔質膜が挙げられる。膜の孔径とし
ては、０．０５〜５０μ、好ましくは１〜３０μである
。Polymer membrane materials used in the present invention include polysulfone, cellulose acetate, cellulose triacetate, cellulose acetate butyrate, polyacrylonitrile, polycarbonate, polyurethane, polyvinyl chloride, polyvinyl butyral,
Examples include porous polymer membranes made of nylon, Conex, polyacrylate, polyether sulfone, crosslinked polyvinylpyridinium halide, and the like. The pore size of the membrane is 0.05 to 50μ, preferably 1 to 30μ.
このような多孔質の高分子膜を製造するにあたっては、
原料高分子素材を溶媒に溶解し、この中に電極素材を浸
漬すればよい。孔密度及び孔径は高分子溶液の濃度や温
度を制御することにより調節することができる。In manufacturing such porous polymer membranes,
The raw polymer material may be dissolved in a solvent, and the electrode material may be immersed in the solvent. The pore density and pore diameter can be adjusted by controlling the concentration and temperature of the polymer solution.
本発明に係る電極は、表面が多孔質の高分子膜で被覆さ
れているため、細胞は膜中の孔に優先的に捕捉され、電
極表面の細胞密度は特に高くなっている。更に、細胞融
合の頻度は電極界面で特に高いという事実が経験的に知
られている。このような事実から電極界面に最も近（、
かつ細胞が高密度で存在する高分子膜の孔中では、融合
率が高く、融合細胞が高頻度で存在する。しかもこの融
合細胞は高分子膜の孔中に捕捉されているため、細胞融
合用容器から選択培地に移す際に落下することなく電極
と共に移動させることができる。選択培地においては融
合細胞は電極表面で増殖し、孔から出てコロニーを形成
する。Since the electrode according to the present invention has a surface covered with a porous polymer membrane, cells are preferentially captured in the pores in the membrane, and the cell density on the electrode surface is particularly high. Furthermore, it is empirically known that the frequency of cell fusion is particularly high at the electrode interface. From this fact, the closest to the electrode interface (,
In addition, in the pores of a polymer membrane where cells exist at a high density, the fusion rate is high and fused cells are present at a high frequency. Furthermore, since the fused cells are captured in the pores of the polymer membrane, they can be moved together with the electrodes without falling when transferred from the cell fusion container to the selection medium. In the selective medium, the fused cells proliferate on the electrode surface and exit through the pores to form colonies.
第１図は本発明の１実施例の斜視図である。１）゛は白
金板からなる電極であり、２．２′は電極１の表面に設
けた高分子膜である。３はパルス発生装置である。本実
施例においては、高分子膜２は次のようにして作成した
。０．１ｇのポリビニルブチラールを１０ｍ１のジメヂ
ルホルムアミドに溶解した溶液に白金板を浸漬した。次
で、引上げて乾燥させ、白金板表面が多孔質のポリビニ
ルブチラールで覆われた本発明電極を作成した。FIG. 1 is a perspective view of one embodiment of the present invention. 1)' is an electrode made of a platinum plate, and 2.2' is a polymer film provided on the surface of the electrode 1. 3 is a pulse generator. In this example, the polymer membrane 2 was created as follows. A platinum plate was immersed in a solution in which 0.1 g of polyvinyl butyral was dissolved in 10 ml of dimedylformamide. Next, the platinum plate was pulled up and dried to produce an electrode of the present invention in which the surface of the platinum plate was covered with porous polyvinyl butyral.
別に、ｉｌｌとして鎖匹血皿肛朋り翌匹旦…匹（酵母）
の変異株であるロイシン要求株とトリプトファン要求株
とを用いた。各々の変異株をＹＰＤ完全培地（酵母エキ
ス１％、ポリペプトン２％、グルコース２％、残部蒸留
水）１００ｍｌにそれぞれ植菌し、３０℃で１晩振とう
培養した。菌体濃度１〜５ Ｘ １０’ ／ｍｌに達し
た時、遠心分離して各々集菌した。（３＋ＯＯＯｒｐｍ
、１０分）次いでこの酵母をＴＳ緩衝液（ソルビトール
１．２Ｍ、 Ｔｒｉｓ −ＨＣｌ １０ ｍＭ、ｐ １
）７．５）中に懸濁後、上記遠心条件と同一条件で遠心
して菌体を洗浄した。各菌体を２０ｍ ｌのＴＳｉｌ衝
液に再び懸濁してＺｙｍｏｌｙａｓｅ ６００００（
最８８濃度５０ ｐ ｇ／ｍｌ） 、２−メルカプトエ
タノール（最終濃度２０ｍＭ）を添加した。これを３０
℃で振とうしながら反応させ、プロトプラストを生成さ
せた。プロトプラストの確認は、少量をサンプリングし
、１０倍量の水に懸濁して顕微鏡下に観察し、バースト
の有無により確認した。Separately, Ill as a chain dog blood dish anus, the next day... (yeast)
A leucine auxotrophic strain and a tryptophan auxotrophic strain, which are mutant strains of , were used. Each mutant strain was inoculated into 100 ml of YPD complete medium (1% yeast extract, 2% polypeptone, 2% glucose, balance distilled water) and cultured with shaking at 30°C overnight. When the bacterial cell concentration reached 1 to 5 x 10'/ml, the cells were collected by centrifugation. (3+OOOrpm
, 10 min) then the yeast was incubated in TS buffer (Sorbitol 1.2 M, Tris-HCl 10 mM, p 1
) After suspending in 7.5), the cells were washed by centrifugation under the same centrifugation conditions as above. Resuspend each bacterial cell in 20 ml of TSil buffer and add Zymolyase 60000 (
2-mercaptoethanol (final concentration 20 mM) was added. 30 of this
The reaction was carried out with shaking at °C to generate protoplasts. Protoplasts were confirmed by sampling a small amount, suspending it in 10 times the volume of water, observing it under a microscope, and checking for the presence or absence of bursts.
次で低速遠心（２，００Ｏｒｐｍ 、５分）により各プ
ロトプラストを集め、Ｔ Ｓ ｉｌｌ液液２回洗浄し、
１０ｍＭＣａＣ１□を含むＴＳ緩衝液に懸濁させ、両者
を混合した。Next, each protoplast was collected by low-speed centrifugation (2,00 rpm, 5 minutes), washed twice with T Sill liquid,
It was suspended in a TS buffer containing 10 mM CaC1□, and both were mixed.
この混合液中に、第１図に示した２本の電極を挿入し、
パルス発生装置を用いて電気パルスを印加した。印加条
件は連続パルス（強度１００ Ｖ、パルス幅２０μ５ｅ
ｃ）を５秒印加後高電圧パルス（５００■）を印加した
。印加後この２本の電極を選択培地（Ｙｅａｓｔ Ｎｉ
ｔｒｏｇｅｎ ｂａｓｅ Ｏ，７％、グルコース２％、
ソルビトール２２％、寒天３％）上に置き、直ちに同組
成の寒天培地（４５℃）１０ｍｌを重層した。３０℃で
７日間培養したところコロニーが出現した。コロニーの
出現はロイシン要求株とトリプトファン要求株のプロト
プラスト同士が融合し、互いの要求性を相補しあったこ
とを意味する。Inserting the two electrodes shown in Figure 1 into this mixed solution,
Electrical pulses were applied using a pulse generator. The application conditions were continuous pulses (intensity 100 V, pulse width 20μ5e
After applying c) for 5 seconds, a high voltage pulse (500 μ) was applied. After application, these two electrodes were coated with selective medium (Yeast Ni
trogen base O, 7%, glucose 2%,
sorbitol 22%, agar 3%), and immediately overlaid with 10 ml of an agar medium (45°C) of the same composition. Colonies appeared after culturing at 30°C for 7 days. The appearance of a colony means that the protoplasts of the leucine auxotroph and the tryptophan auxotroph have fused with each other and complemented each other's auxotrophies.
本発明によれば、細胞の融合率を高め、更に融合率の高
い部位が電極表面であるため、電極をそのまま選択培地
に移すことにより、効率よく融合の有無を判定すること
ができる。According to the present invention, the fusion rate of cells is increased, and since the region with a high fusion rate is the electrode surface, the presence or absence of fusion can be efficiently determined by transferring the electrode as it is to a selective medium.
第１図は本発明の１実施例の斜視図である。
図面中、符号
１．１”は電極、２．２゛は高分子膜、３はパルス発生
装置である。
特許出願人 エヌオーケー株式会社
代理人 弁理士 吉 １）俊 夫
（外１名）
手続補正書
昭和６２年７月２〆日
特許庁長官 小 川 邦 夫 殿
１、事件の表示
昭和６２年特許願第４０４０号
２、発明の名称
細胞融合用電極〜
３、補正をする者
事件との関係 特許出願人
住所 東京都港区芝大門１丁目１２番１５号名称 エヌ
オーケー株式会社
４、代理人 ■１５０
住所 東京都渋谷区松濤−丁目２９番２１号５、補正命
令の日付 自発
６、補正の対象
７、補正の内容
（１）明細書、４頁、１３行の「浸漬すればよい。」を
「浸漬し、乾燥させればよい。」に訂正する。
（２）同、６頁、１４〜１５行の「プロトプラストを生
成させた。」を「プロトプラスト化させた。」に訂正す
る。
（３）同、６頁、１８〜１９行の「プロトプラストを集
め、」を「プロトプラスト化体を集め、」に訂正する。
（４）同、７頁、１２行の「プロトプラスト同士」を「
プロトプラスト化体同士」に訂正する。
以上FIG. 1 is a perspective view of one embodiment of the present invention. In the drawing, 1.1" is an electrode, 2.2" is a polymer membrane, and 3 is a pulse generator. Patent applicant Yoshi, Patent attorney, N.OK. Co., Ltd. 1) Toshio (one other person) Procedure Amendment dated July 2, 1988 Kunio Ogawa, Commissioner of the Patent Office 1. Indication of the case 1988 Patent Application No. 4040 2. Name of the invention Electrode for cell fusion ~ 3. Person making the amendment Relationship Patent applicant address 1-12-15 Shiba Daimon, Minato-ku, Tokyo Name NOK Co., Ltd. 4, Agent ■150 Address 29-21-5 Shoto-chome, Shibuya-ku, Tokyo Date of amendment order Voluntary action 6, Amendment Target 7, Contents of amendment (1) In the specification, page 4, line 13, "It is sufficient to soak." is corrected to "It is sufficient to soak and dry." (2) Same, page 6, lines 14-15, "protoplasts were generated." is corrected to "protoplast formation." (3) ``Collect protoplasts'' on page 6, lines 18-19 of the same publication is corrected to ``collect protoplasts.'' (4) Same, page 7, line 12, “protoplasts” are changed to “
Corrected to "protoplast-formed bodies."that's all
Claims (2)
膜で被覆されていることを特徴とする細胞融合用電極。(1) An electrode for cell fusion, the surface of which is coated with a porous polymer membrane having many minute pores.
求の範囲第１項記載の細胞融合用電極。(2) The electrode for cell fusion according to claim 1, wherein the polymer membrane has a pore size of 0.05 to 50μ.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62004040A JPS63173574A (en) | 1987-01-13 | 1987-01-13 | Electrode for cell fusion |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62004040A JPS63173574A (en) | 1987-01-13 | 1987-01-13 | Electrode for cell fusion |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS63173574A true JPS63173574A (en) | 1988-07-18 |
Family
ID=11573834
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62004040A Pending JPS63173574A (en) | 1987-01-13 | 1987-01-13 | Electrode for cell fusion |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63173574A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993002178A1 (en) * | 1991-07-22 | 1993-02-04 | Schmukler Robert E | Apparatus and methods for electroporation and electrofusion |
| JP2008054630A (en) * | 2006-09-01 | 2008-03-13 | Tosoh Corp | Cell fusion device and cell fusion method using the same |
| CN108828037A (en) * | 2018-06-26 | 2018-11-16 | 长春工业大学 | A kind of gold nano electrode and preparation method thereof |
-
1987
- 1987-01-13 JP JP62004040A patent/JPS63173574A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993002178A1 (en) * | 1991-07-22 | 1993-02-04 | Schmukler Robert E | Apparatus and methods for electroporation and electrofusion |
| JP2008054630A (en) * | 2006-09-01 | 2008-03-13 | Tosoh Corp | Cell fusion device and cell fusion method using the same |
| CN108828037A (en) * | 2018-06-26 | 2018-11-16 | 长春工业大学 | A kind of gold nano electrode and preparation method thereof |
| CN108828037B (en) * | 2018-06-26 | 2020-04-24 | 长春工业大学 | Gold nano electrode and preparation method thereof |
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