JPS63230070A - Cell fusion chamber - Google Patents
Cell fusion chamberInfo
- Publication number
- JPS63230070A JPS63230070A JP62065371A JP6537187A JPS63230070A JP S63230070 A JPS63230070 A JP S63230070A JP 62065371 A JP62065371 A JP 62065371A JP 6537187 A JP6537187 A JP 6537187A JP S63230070 A JPS63230070 A JP S63230070A
- Authority
- JP
- Japan
- Prior art keywords
- filter
- cells
- electrodes
- cell
- diameter
- 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 description 10
- 239000011810 insulating material Substances 0.000 claims 1
- 230000004927 fusion Effects 0.000 abstract description 7
- 239000012212 insulator Substances 0.000 abstract description 2
- 238000000034 method Methods 0.000 abstract description 2
- 230000005684 electric field Effects 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- 239000006285 cell suspension Substances 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000007499 fusion processing Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 239000012780 transparent material Substances 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）
本発明はバイオテクノロジーの分野において細胞どおし
を融合させる細胞融合装置で使用する細胞融合チャンバ
に関するものである。DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a cell fusion chamber used in a cell fusion device for fusing cells together in the field of biotechnology.
（従来の技術）
従来の細胞融合チャンバでは一対の電極を対向して設け
、その電極間に細胞を懸濁した細胞Ｕ濁液を入れ、一対
の電極間に高周波電圧を印加して細胞どおしを細胞帯電
荷のクーロン力によって接触させ、その後一対の電極間
に高電圧パルスを印加することにより電気的に融合させ
ている。(Prior art) In a conventional cell fusion chamber, a pair of electrodes is provided facing each other, a cell suspension containing cells is placed between the electrodes, and a high-frequency voltage is applied between the pair of electrodes to inject the cells. The cells are brought into contact by the Coulomb force of the cell charge, and then a high voltage pulse is applied between the pair of electrodes to electrically fuse them.
（発明が解決しようとする問題点）
細胞によっては海水などのように低抵抗の溶媒中でしか
生息できない細胞もある。そのような細胞の場合、溶媒
の電気抵抗が低すぎて細胞に帯電させることができない
。したがって、これまでは低抵抗の溶媒中でしか生息で
きない細胞どおしを電気的に接触させて融合させること
はできなかった。(Problems to be solved by the invention) Some cells can only live in low-resistance solvents such as seawater. For such cells, the electrical resistance of the solvent is too low to charge the cells. Therefore, until now, it has not been possible to fuse cells that can only live in low-resistance solvents by bringing them into electrical contact.
本発明は細胞とおしをクーロン力によらずに機械的に接
触させ、接触させた細胞に有効な電界を印加することが
でき、また、融合の過程などを顕微鏡などで光学的にモ
ニターすることのできる細胞融合チャンバを提供するこ
とを目的とするものである。The present invention makes it possible to bring cells into mechanical contact without relying on Coulomb force, apply an effective electric field to the contacted cells, and optically monitor the fusion process using a microscope or the like. The purpose of this research is to provide a cell fusion chamber that allows for cell fusion.
（問題点を解決するための手段）
実施例を示す第１図を参照して説明すると、本発明の細
胞融合チャンバでは、絶縁物にてなるチャンバ本体（２
）の内側を、対象となる細胞径より小さい短幅と大きい
長幅の長穴（６）をもつ透明絶縁性フィルタ（４）で水
平方向に仕切り、長大（６）に接近して一対の電極（８
，１０）を設けた。(Means for Solving the Problems) To explain with reference to FIG. 1 showing an embodiment, the cell fusion chamber of the present invention has a chamber body (two
) is horizontally partitioned by a transparent insulating filter (4) having a long hole (6) with a short width smaller and a longer width larger than the target cell diameter, and a pair of electrodes placed close to the long hole (6). (8
, 10) were established.
（実施例） 第１図は一実施例を表わす。(Example) FIG. 1 represents one embodiment.
２は絶縁物にてなるチャンバ本体であり、筒状に形成さ
れている。チャンバ本体２の内側には水平方向に透明で
絶縁性のフィルタ４が設けられており、このフィルタ４
には対象となる細胞の径より小さい短幅と対象となる細
胞の径より大きい長幅の長穴６があけられている。フィ
ルタ４の両端には長穴６の長手方向を挟むように一対の
電極８゜１０が設けられており、電極８，１０には融合
電界を印加するための電源１２が接続されている。Reference numeral 2 denotes a chamber body made of an insulator, which is formed into a cylindrical shape. A transparent and insulating filter 4 is provided horizontally inside the chamber body 2.
A long hole 6 is formed in the hole 6, which has a short width smaller than the diameter of the target cell and a long width larger than the diameter of the target cell. A pair of electrodes 8 and 10 are provided at both ends of the filter 4 so as to sandwich the longitudinal direction of the elongated hole 6, and a power source 12 for applying a fusion electric field is connected to the electrodes 8 and 10.
フィルタ４の材質は用途にもよるが、例えば石英ガラス
などを用いることによって容易に顕微鏡でモニタできる
ようになる。The material of the filter 4 depends on the application, but by using quartz glass, for example, it can be easily monitored with a microscope.
チャンバ本体２の上側から細胞を懸濁した細胞懸濁液が
入れられ、チャンバ本体２の下側から溶媒が排出されて
細胞がフィルタ４の長穴６に捕えられる。A cell suspension in which cells are suspended is introduced from the upper side of the chamber body 2, a solvent is discharged from the lower side of the chamber body 2, and the cells are captured in the elongated holes 6 of the filter 4.
本実施例におけるフィルタ４と電極８，１０の部分を第
２図に拡大して示す。第２（Ａ）は平面図、同図（Ｂ）
は同図（Ａ）のＡ−Ａ線位置での断面図、同図（Ｃ）は
同図（Ａ）のＢ−Ｂ線位置での断面図である。FIG. 2 shows an enlarged view of the filter 4 and electrodes 8, 10 in this embodiment. The second (A) is a plan view, the same figure (B)
is a cross-sectional view taken along the line AA in FIG. 1 (A), and FIG.
チャンバ本体２の上方から細胞懸濁液を入れると長穴６
を通って溶媒が下側に通過し、細胞１４゜１６が長穴６
を目詰りさせ、フィルタ４に収集される。長穴６はその
短幅が細胞１４．１６の直径よりも小さく、長幅が細胞
１４．１６の直径よりも大きく設定されているので、長
穴６の長手方向に沿って複数個の細胞１４．１６が集め
られ、かつ、互いに接触するようになる。When the cell suspension is poured into the chamber body 2 from above, the elongated hole 6
Through which the solvent passes to the lower side, cells 14 and 16 enter the elongated hole 6.
is collected in the filter 4. Since the elongated hole 6 has a short width smaller than the diameter of the cell 14.16 and a long width larger than the diameter of the cell 14.16, a plurality of cells 14 are formed along the longitudinal direction of the elongated hole 6. .16 are collected and come into contact with each other.
第２図に示されるように複数個の細胞が長穴６に集めら
れて互いに接触した状態で一対の電極８゜１０に融合用
の電界を印加することによって、細胞融合を起させるこ
とができる。As shown in FIG. 2, cell fusion can be caused by applying an electric field for fusion to a pair of electrodes 8 and 10 while a plurality of cells are collected in the elongated hole 6 and in contact with each other. .
第３図は他の実施例を表わす。FIG. 3 represents another embodiment.
４ａは第１図のフィルタ４に対応するフィルタであり、
このフィルタ４ａではその長穴６ａの断面形状が上側で
対象となる細胞１４．１６の直径よりも広く、下側で細
胞１４．１６の直径よりも小さくなるようにテーパ形に
形成されている点で異なっている。そして、フィルタ４
ａの表側に一方の電極２０が設けられ、裏面に他方の電
極２２が設けられ１両電極２０．２２間に融合用の電界
を印加するための電源１２が接続されている。4a is a filter corresponding to filter 4 in FIG. 1;
In this filter 4a, the cross-sectional shape of the elongated hole 6a is tapered so that the upper side is wider than the diameter of the target cell 14.16 and the lower side is smaller than the diameter of the cell 14.16. It's different. And filter 4
One electrode 20 is provided on the front side of a, the other electrode 22 is provided on the back side, and a power source 12 for applying an electric field for fusion is connected between the two electrodes 20 and 22.
本実施例ではフィルタ４ａとしては例えば透明な石英ガ
ラスを使用し、ａ極２０，２２としては透明電極を用い
る。フィルタ４ａと電極２０，２２をともに透明な材質
で構成することにより、細胞融合過程を顕微鏡などでモ
ニターすることができる。In this embodiment, for example, transparent quartz glass is used as the filter 4a, and transparent electrodes are used as the a-poles 20 and 22. By constructing both the filter 4a and the electrodes 20 and 22 from transparent materials, the cell fusion process can be monitored using a microscope or the like.
本実施例で上側から細胞懸濁液を投入し下側から溶媒を
排出させると、細胞１４．１６はフィルタ４ａの長穴６
ａに集められ、フィルタ４ａの厚さ方向に沿って接触す
る。そのため、一対の電極２０．２２に電圧を印加する
ことによって電界がフィルタ４ａの厚さ方向に印加され
、接触した細胞１４．１６が融合する。In this example, when the cell suspension is introduced from the upper side and the solvent is discharged from the lower side, the cells 14 and 16 are removed from the long holes 6 of the filter 4a.
a and contacts the filter 4a along the thickness direction. Therefore, by applying a voltage to the pair of electrodes 20.22, an electric field is applied in the thickness direction of the filter 4a, and the cells 14.16 in contact are fused.
第４図はさらに他の実施例を表わす。FIG. 4 shows yet another embodiment.
チャンバ本体２には下側に第３図と同様に断面がテーバ
形の長穴６ａをもつフィルタ４ａを設け、その上側に細
胞１４．１６が十分通過できる大きさの穴をもつフィル
タ４ｂを設け、フィルタ４ａの裏側とフィルタ４ｂの表
側にそれぞれ電極２２゜２０を設ける。電極２０．２２
間には融合電界印加用の電源１２を接続する。The chamber body 2 is provided with a filter 4a having a long hole 6a with a tapered cross section on the lower side as shown in FIG. , electrodes 22 and 20 are provided on the back side of the filter 4a and the front side of the filter 4b, respectively. Electrode 20.22
A power source 12 for applying a fused electric field is connected between them.
本実施例で上側から細胞懸濁液を投入し下側のフィルタ
４ａの長穴６ａから溶媒を排出させると、下側のフィル
タ４ａの長穴６ａでフィルタ４ａの厚さ方向に沿って複
数の細胞１４．１６が集められて接触する。そこで電極
２０と電極２２の間に融合電圧を印加することにより、
細胞１４と細胞１６を融合させることができる。In this example, when the cell suspension is introduced from the upper side and the solvent is discharged from the long hole 6a of the lower filter 4a, a plurality of Cells 14.16 are collected and contacted. Therefore, by applying a fusion voltage between the electrodes 20 and 22,
Cell 14 and cell 16 can be fused.
（発明の効果）
本発明ではフィルタで細胞懸濁液を濾過し、フィルタの
長穴に細胞を集めて互いに接触させるようにし、その状
態で融合電界を印加するので、従来のようにクーロン力
によって細胞どおしを接触させるのに比べて帯電用電界
による細胞へのストレスがかからない効果がある。(Effects of the Invention) In the present invention, the cell suspension is filtered using a filter, and the cells are collected in the long holes of the filter so that they come into contact with each other, and a fusion electric field is applied in that state. Compared to bringing cells into contact with each other, this method has the effect of reducing stress on cells due to the charging electric field.
フィルタに設ける長穴のサイズによって接触させる細胞
の個数を制御することができる。The number of cells brought into contact can be controlled by the size of the elongated holes provided in the filter.
また、接触した細胞が存在する位置を特定することがで
きるため、融合状態のモニタや処置が容易になる。Furthermore, since the location of the contacted cells can be specified, the fusion state can be easily monitored and treated.
さらに、本発明では細胞どおしを機械的に接触させるの
で、電気抵抗の低い溶媒中でしか生息できない細胞の場
合も接触させて融合させることができる。Furthermore, since cells are brought into mechanical contact with each other in the present invention, even cells that can only live in a solvent with low electrical resistance can be brought into contact and fused.
第１図は一実施例を示す概略斜視図、第２図（Ａ）はフ
ィルタと電極の部分を示す平面図、同図（Ｂ）は同図（
Ａ）のＡ−Ａ線位置での断面図、同図（Ｃ）は同図（Ａ
）のＢ−Ｂ線位置での断面図、第３図は他の実施例にお
けるフィルタの穴部分を示す断面図、第４図はさらに他
の実施例を示す断面図である。
２・・・・・・チャンバ本体、
４、．４ａ、４ｂ・・・・・・フィルタ、６．６ａ・・
・・・・長穴、
８．１０．２０．２２・・・・・・電極、１２・・・・
・・電源、
１４．１６・・・・・・細胞。Fig. 1 is a schematic perspective view showing one embodiment, Fig. 2 (A) is a plan view showing the filter and electrode portion, and Fig. 2 (B) is the same figure (
A) is a cross-sectional view taken along line A-A;
), FIG. 3 is a cross-sectional view showing the hole portion of a filter in another embodiment, and FIG. 4 is a cross-sectional view showing still another embodiment. 2... Chamber body, 4. 4a, 4b...Filter, 6.6a...
...Elongated hole, 8.10.20.22...Electrode, 12...
...Power supply, 14.16...Cell.
Claims (1)
る細胞径より小さい短幅と大きい長幅の長穴をもつ透明
絶縁性フィルタで水平方向に仕切り、前記長穴に接近し
て一対の電極を設けた細胞融合チャンバ。(1) The inside of the chamber body made of an insulating material is horizontally partitioned with a transparent insulating filter having long holes with a short width smaller than the diameter of the target cell and a long width larger than the diameter of the target cell, and a pair of filters are placed close to the long hole. Cell fusion chamber equipped with electrodes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62065371A JPS63230070A (en) | 1987-03-18 | 1987-03-18 | Cell fusion chamber |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62065371A JPS63230070A (en) | 1987-03-18 | 1987-03-18 | Cell fusion chamber |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS63230070A true JPS63230070A (en) | 1988-09-26 |
Family
ID=13285041
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62065371A Pending JPS63230070A (en) | 1987-03-18 | 1987-03-18 | Cell fusion chamber |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63230070A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2659347A1 (en) * | 1990-03-12 | 1991-09-13 | Agronomique Inst Nat Rech | Device for culturing cells which ensures their immobilisation |
| WO1993002178A1 (en) * | 1991-07-22 | 1993-02-04 | Schmukler Robert E | Apparatus and methods for electroporation and electrofusion |
| JP2007295912A (en) * | 2005-06-13 | 2007-11-15 | Tosoh Corp | Cell fusion chamber, cell fusion device, and method for cell fusion using the same |
| JP2007295922A (en) * | 2006-04-03 | 2007-11-15 | Tosoh Corp | Cell fusion device and method for cell fusion using the same |
-
1987
- 1987-03-18 JP JP62065371A patent/JPS63230070A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2659347A1 (en) * | 1990-03-12 | 1991-09-13 | Agronomique Inst Nat Rech | Device for culturing cells which ensures their immobilisation |
| WO1993002178A1 (en) * | 1991-07-22 | 1993-02-04 | Schmukler Robert E | Apparatus and methods for electroporation and electrofusion |
| JP2007295912A (en) * | 2005-06-13 | 2007-11-15 | Tosoh Corp | Cell fusion chamber, cell fusion device, and method for cell fusion using the same |
| JP2007295922A (en) * | 2006-04-03 | 2007-11-15 | Tosoh Corp | Cell fusion device and method for cell fusion using the same |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zimmermann et al. | High frequency fusion of plant protoplasts by electric fields | |
| US4578168A (en) | Apparatus for fusing live cells with electric fields | |
| CA1207275A (en) | Electrophoresis using alternating transverse electric fields | |
| BR8405727A (en) | BIOMEDICAL ELECTRODE AND PROCESS FOR MANUFACTURING BIOMEDICAL ELECTRODES | |
| JPH02131569A (en) | Microchamber plate, particle discrimination, particle treating device and cell treating device | |
| Scheurich et al. | Membrane fusion and deformation of red blood cells by electric fields | |
| JPS63230070A (en) | Cell fusion chamber | |
| SU1048994A3 (en) | Electrochromes device | |
| WO1996023432A3 (en) | Rack and shelving system, and method and apparatus for manufacture thereof | |
| KR100463583B1 (en) | Method of processing a ceramic capacitor | |
| JPH01141583A (en) | Cell-treating chamber | |
| JPH0336508B2 (en) | ||
| JPS62171687A (en) | Apparatus for introducing gene for such | |
| FR2527756B1 (en) | METALLURGICAL FUSION PROCESS AND ARC FURNACE FOR ITS IMPLEMENTATION | |
| JPH04190781A (en) | Chamber for electrically treating cell | |
| JP2576477B2 (en) | Electrodes for cell fusion chambers | |
| JPS6349068A (en) | Apparatus for electrical introduction of gene | |
| JP4918811B2 (en) | Cell fusion chamber, cell fusion device, and cell fusion method using them | |
| JPH0329844A (en) | Electrophoretic apparatus | |
| JPS62171667A (en) | Cell fusion apparatus | |
| JPH074216B2 (en) | Electric cell fusion method | |
| JPS6384478A (en) | Cell-fusion chamber | |
| JPS6387969A (en) | Chamber for cell fusion | |
| JPH0324075Y2 (en) | ||
| JPH05336944A (en) | Electrode device for cell fusion |