JPS62275679A - Preparation of cell containing fine magnetic particle - Google Patents
Preparation of cell containing fine magnetic particleInfo
- Publication number
- JPS62275679A JPS62275679A JP61119909A JP11990986A JPS62275679A JP S62275679 A JPS62275679 A JP S62275679A JP 61119909 A JP61119909 A JP 61119909A JP 11990986 A JP11990986 A JP 11990986A JP S62275679 A JPS62275679 A JP S62275679A
- Authority
- JP
- Japan
- Prior art keywords
- cell
- magnetic particle
- fine magnetic
- cells
- magnetotactic
- 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
- 239000006249 magnetic particle Substances 0.000 title claims abstract description 13
- 241000894006 Bacteria Species 0.000 claims description 15
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 210000004027 cell Anatomy 0.000 abstract description 40
- 230000001580 bacterial effect Effects 0.000 abstract description 6
- 230000007910 cell fusion Effects 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 210000000265 leukocyte Anatomy 0.000 abstract description 3
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 abstract description 2
- 210000002540 macrophage Anatomy 0.000 abstract description 2
- 210000002244 magnetosome Anatomy 0.000 abstract description 2
- 241000193830 Bacillus <bacterium> Species 0.000 abstract 1
- 241001478240 Coccus Species 0.000 abstract 1
- 230000005381 magnetic domain Effects 0.000 abstract 1
- 210000003743 erythrocyte Anatomy 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 4
- 239000011859 microparticle Substances 0.000 description 4
- 102000016943 Muramidase Human genes 0.000 description 3
- 108010014251 Muramidase Proteins 0.000 description 3
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 3
- 239000010419 fine particle Substances 0.000 description 3
- 229960000274 lysozyme Drugs 0.000 description 3
- 235000010335 lysozyme Nutrition 0.000 description 3
- 239000004325 lysozyme Substances 0.000 description 3
- 210000001938 protoplast Anatomy 0.000 description 3
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000035931 haemagglutination Effects 0.000 description 2
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 102000003996 Interferon-beta Human genes 0.000 description 1
- 108090000467 Interferon-beta Proteins 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 210000000224 granular leucocyte Anatomy 0.000 description 1
- 230000003067 hemagglutinative effect Effects 0.000 description 1
- 210000004754 hybrid cell Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- -1 polyethylene Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 108010082737 zymolyase Proteins 0.000 description 1
Abstract
Description
【発明の詳細な説明】
３、発明の詳細な説明
〔産業上の利用分野〕
本発明は、医学的治療、有用物質の生産、分析等の手段
として有用である、磁気微粒子含有細胞の製法に関する
。Detailed Description of the Invention 3. Detailed Description of the Invention [Field of Industrial Application] The present invention relates to a method for producing cells containing magnetic particles, which are useful as a means for medical treatment, production of useful substances, analysis, etc. .
走磁性細菌が、磁鉄鉱からなる磁気微粒子を鎖条に配列
した状態で含んでおり、地磁気に悪心することが、Ｒｌ
Ｐ、Ｂｌａｋｅｍｏｒｅにより発見され、報告されてい
る。しかしながら、通常の細胞には上記のような性質を
持ち合せたものは発見されていない。Rl
It was discovered and reported by P. Blakemore. However, no normal cells have been found that have the above properties.
例えば、赤血球、白血球のような特別な生理機能を有す
る細胞や有用物質生産能を有する細胞などに磁気感受性
を付与することができれば、これら細胞の運動を人為的
に制御することができるため、医学的治療、分析等に有
用である。しかし、従来、走磁性細菌以外の細胞に磁気
感受性を持たせる試みはまったく行なわれていない。For example, if it is possible to impart magnetic sensitivity to cells with special physiological functions such as red blood cells and white blood cells, or cells with the ability to produce useful substances, it is possible to artificially control the movement of these cells. It is useful for medical treatment, analysis, etc. However, no attempt has been made to make cells other than magnetotactic bacteria magnetically sensitive.
そこで、本発明の目的は、磁気感受性を有する磁気微粒
子含有細胞の製法を提供することにある。Therefore, an object of the present invention is to provide a method for producing magnetic microparticle-containing cells having magnetic susceptibility.
本発明は、かかる磁気感受性細胞の製法として、走磁性
細菌と他の細胞とを細胞融合させることにより、磁気微
粒子を前記他の細胞中へ導入することからなる磁気微粒
子含有細胞の製法を提供する。The present invention provides a method for producing magnetically sensitive cells, which comprises introducing magnetic microparticles into other cells by fusing magnetotactic bacteria with other cells. .
本発明に用いられる走磁性細菌は、菌体内に５００人程
度の磁鉄鉱から成る磁気微粒子が１０〜２０個程連なっ
たマグネトソームと呼ばれるチェーンを、１〜数本有す
る桿菌、球菌またはらせん菌である。具体例としてはＡ
ＴＣＣ３１６３２として寄託されている菌株＜ＭＳ−１
）を挙げることができる。The magnetotactic bacteria used in the present invention are rods, cocci, or spiral bacteria that have one to several chains called magnetosomes in which about 10 to 20 magnetic microparticles made of about 500 magnetites are connected in the bacterial body. . A specific example is A
Strain deposited as TCC31632<MS-1
) can be mentioned.
このような走磁性細菌は、例えば、本願出願人による特
願昭６０−２０３１２９号に提案された採取器により淡
水および海水から容易に採取することができる。Such magnetotactic bacteria can be easily collected from freshwater and seawater using, for example, a collector proposed in Japanese Patent Application No. 60-203129 by the applicant of the present invention.
また、本発明の方法により走磁性細菌中に存在する磁気
微粒子を導入することができる他の細胞は、特に制限さ
れず、次のものを例示することができる。Further, other cells into which magnetic microparticles present in magnetotactic bacteria can be introduced by the method of the present invention are not particularly limited, and the following may be exemplified.
多形核白血球。Polymorphonuclear leukocytes.
マクロファージ。macrophage.
クツパー細胞。Kzupah cell.
Ｔリンパ球。T lymphocytes.
Ｂリンパ球 等の免疫担当細胞； モノクローナル抗体生産性リンパ球。Immune-competent cells such as B lymphocytes; Monoclonal antibody-producing lymphocytes.
α−インターフェロン生産性白血球。α-interferon-producing leukocytes.
β−インターフェロン生産性繊維芽細胞等の有用物質生
産細胞；
、 赤血球等の血球凝集反応関連細胞。Useful substance-producing cells such as β-interferon-producing fibroblasts; and hemagglutination-related cells such as red blood cells.
さて、走磁性細菌と他方の細胞との細胞融合は、公知の
方法により行なうことができ、走磁性細菌および必要に
応じ他方の細胞をもプロトプラスト化またはスフェロプ
ラスト化した後、適当な細胞融合活性を有する物質を用
いて、あるいは電気パルスを作用させて細胞融合を生起
させる。Now, cell fusion between magnetotactic bacteria and the other cell can be performed by a known method. After converting the magnetotactic bacteria and, if necessary, the other cell into protoplasts or spheroplasts, appropriate cell fusion is performed. Cell fusion is caused using active substances or by applying electrical pulses.
走磁性細菌のプロトプラスト化あるいはスフェロプラス
ト化は、例えば、リゾチーム（ｉ ｙｓｏｚｙｍｅ）等
を用いて行なうことができる。また、他方の細胞のプロ
トプラスト化が必要である場合には、例えば、リゾチー
ム、サイモリアーゼ（ｚｙｍｏｌｙａｓｅ）、セルラー
ゼ（ｃｅｌｌｕｌａｓｅ）、マセロサイム（ｍａｃｅｒ
ｏｚｙｍｅ）等を使用することができる。また、使用す
ることができる細胞融合活性物質としては、例えば、ポ
リエチレングリコール、１（ＶＪウィルス等を挙げるこ
とができる。Protoplast formation or spheroplast formation of magnetotactic bacteria can be carried out using, for example, lysozyme. In addition, if it is necessary to convert the other cell into a protoplast, for example, lysozyme, zymolyase, cellulase, macerocyme, etc.
ozyme) etc. can be used. Furthermore, examples of cell fusion active substances that can be used include polyethylene glycol, 1 (VJ virus, etc.).
本発明の方法により得られる細胞は、用いられた走磁性
細菌と他方の細胞の雑種細胞であるが、後者の細胞が本
来備えている、例えば、前述の免疫に関する作用、機能
、有用物質生産性、血球凝集機能等を有し、かつ走磁性
細菌から磁鉄鉱の磁気微粒子を受継いだもので磁気感受
性を有している。The cells obtained by the method of the present invention are hybrid cells of the magnetotactic bacteria used and the other cells, and the latter cells inherently have, for example, the above-mentioned immune-related effects, functions, and useful substance productivity. , has a hemagglutinating function, etc., and has magnetic susceptibility because it has inherited magnetic fine particles of magnetite from magnetotactic bacteria.
以下、本発明を実施例により具体的に説明する。 Hereinafter, the present invention will be specifically explained with reference to Examples.
実施例１
捕集した走磁性細菌を微量遠心機で′ａ縮、分離後、洗
浄した。Example 1 The collected magnetotactic bacteria were collapsed using a microcentrifuge, separated, and then washed.
遠心から分離までの操作を数回繰り返した。得られた走
磁性細凹を、０．５Ｍシｇ糖、０．２Ｍ塩化マグネシウ
ム、および０．０２Ｍマレイン酸塩からなるマレイン酸
緩衝液（ｐＨ６，５）の３ＭＭ溶液にＱ＄５し、リゾチ
ーム、ＥＤＴＡをそれぞれ０．１％（ｗ／ｖ）、０．２
％（ｗ／ｖ） となるように加えて、３７℃２時間イ
ンキニーベートしスフェロプラストを行なった。この条
件下でのスフェロプラスト率は、低張処理し、浸透圧シ
ョックで細胞を破裂させることで測定し、６０％以上で
あることを確ｔ＝した。The operations from centrifugation to separation were repeated several times. The obtained magnetotactic fine grooves were placed in a 3MM solution of maleic acid buffer (pH 6.5) consisting of 0.5M sig sugar, 0.2M magnesium chloride, and 0.02M maleate for Q$5, and lysozyme was added. , 0.1% (w/v) and 0.2% EDTA, respectively.
% (w/v) and incubate at 37°C for 2 hours to perform spheroplasty. The spheroplast rate under these conditions was determined by subjecting the cells to hypotonic treatment and rupturing the cells with osmotic shock, and was confirmed to be 60% or more.
このように調製した走磁性細菌のスフェロプラストと洗
浄したヒツジの赤血球を混合し、４０％（ｗ／ｖ）のポ
リエチレン（分子！１ｌｔ６０００）の３ＭＭ溶液中で
２５゛Ｃ１１０分インキュベートすることで走磁性細菌
を赤血球に融合させた。磁気微粒子が赤血球中に導入さ
れたことを透過型電子顕微鏡で確認した。Magnetotactic bacterial spheroplasts prepared in this manner were mixed with washed sheep red blood cells and incubated in a 3MM solution of 40% (w/v) polyethylene (molecules! 1lt6000) at 25°C for 110 minutes. Magnetic bacteria were fused to red blood cells. The introduction of magnetic particles into red blood cells was confirmed using a transmission electron microscope.
この磁気微粒子含有赤血球は、光学顕微鏡による観察に
より磁石を近づけるとその向きが変わることがわかり、
磁気感受性を持ったことが確認された。Observation using an optical microscope revealed that the orientation of these red blood cells containing magnetic particles changes when a magnet is brought close to them.
It was confirmed that he had magnetic susceptibility.
天】〔生ｌ
赤血球の代りに大腸菌を用いた以外は、実施例１と同様
にして走磁性細菌を大腸菌に融合させた。Heaven] [Live l Magnetotactic bacteria were fused to E. coli in the same manner as in Example 1, except that E. coli was used instead of red blood cells.
大腸菌に磁気微粒子が導入され、磁気感受性を示す大腸
菌が得られた。Magnetic particles were introduced into E. coli, resulting in E. coli that exhibits magnetic susceptibility.
本発明の製法により得られる磁気微粒子含有細胞は、磁
気微粒子が単は区構造を有するものであるため磁気モー
メントが大きくて比較的弱い磁場で移動を制御すること
ができる。この細胞は、融合に用いられた一方の細胞が
本来有していた能力を保持するものであるが、磁場の適
用により所望箇所に局部的に集中させることができるた
めその機能、能力を一層強く発揮させることができる上
に、回収なども容易である。例えば、免疫担当細胞の場
合には、体内の患部に人為的に集中させることが可能と
なり、また有用物質生産細胞や血球凝集試験における赤
血球の磁気誘導が可能となり、凝集度が向上し、回収が
容易となる。The magnetic fine particle-containing cells obtained by the production method of the present invention have a large magnetic moment because the magnetic fine particles have a monogonal structure, and their movement can be controlled with a relatively weak magnetic field. These cells retain the original abilities of one of the cells used in the fusion, but by applying a magnetic field they can be locally concentrated at a desired location, making their functions and abilities even stronger. Not only can it be exploited, but it can also be easily recovered. For example, in the case of immunocompetent cells, it is possible to artificially concentrate them in the affected area of the body, and it is also possible to magnetically guide useful substance-producing cells and red blood cells in hemagglutination tests, improving the degree of agglutination and making recovery easier. It becomes easier.
Claims (1)
より、磁気微粒子を前記他の細胞中へ導入することから
なる磁気微粒子含有細胞の製法。(1) A method for producing cells containing magnetic particles, which comprises introducing magnetic particles into the other cells by fusing magnetotactic bacteria with the other cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61119909A JPS62275679A (en) | 1986-05-23 | 1986-05-23 | Preparation of cell containing fine magnetic particle |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61119909A JPS62275679A (en) | 1986-05-23 | 1986-05-23 | Preparation of cell containing fine magnetic particle |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS62275679A true JPS62275679A (en) | 1987-11-30 |
Family
ID=14773198
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61119909A Pending JPS62275679A (en) | 1986-05-23 | 1986-05-23 | Preparation of cell containing fine magnetic particle |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62275679A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007140617A1 (en) * | 2006-06-08 | 2007-12-13 | Multi-Magnetics Incorporated | Magnetosome gene expression in eukaryotic cells |
| JP2015504680A (en) * | 2012-01-13 | 2015-02-16 | ベル・バイオシステムズ，インコーポレーテッド | Host cells containing artificial endosymbiosis |
| JP2017522022A (en) * | 2014-07-15 | 2017-08-10 | ベル・バイオシステムズ，インコーポレーテッド | Eukaryotic cells containing artificial endosymbiosis for multimodal detection |
| US9827333B2 (en) | 2012-01-13 | 2017-11-28 | Bell Biosystems, Inc. | Host cells with artificial endosymbionts |
| US10076579B2 (en) | 2012-01-13 | 2018-09-18 | Bell Biosystems, Inc. | Host cells with artificial endosymbionts |
| US10184114B2 (en) | 2013-09-03 | 2019-01-22 | Bell Biosystems, Inc. | Host cell modification with artificial endosymbionts |
-
1986
- 1986-05-23 JP JP61119909A patent/JPS62275679A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007140617A1 (en) * | 2006-06-08 | 2007-12-13 | Multi-Magnetics Incorporated | Magnetosome gene expression in eukaryotic cells |
| US9556238B2 (en) | 2006-06-08 | 2017-01-31 | Multi-Magnetics Inc. | Magnetosome gene expression in eukaryotic cells |
| JP2015504680A (en) * | 2012-01-13 | 2015-02-16 | ベル・バイオシステムズ，インコーポレーテッド | Host cells containing artificial endosymbiosis |
| US9827333B2 (en) | 2012-01-13 | 2017-11-28 | Bell Biosystems, Inc. | Host cells with artificial endosymbionts |
| EP3263696A1 (en) * | 2012-01-13 | 2018-01-03 | Bell Biosystems, Inc. | Host cells with artificial endosymbionts |
| EP2802334B1 (en) * | 2012-01-13 | 2018-03-07 | Bell Biosystems, Inc. | Host cells with artificial endosymbionts |
| JP2018108107A (en) * | 2012-01-13 | 2018-07-12 | ベル・バイオシステムズ，インコーポレーテッド | Host cells with artificial endosymbionts |
| US10076579B2 (en) | 2012-01-13 | 2018-09-18 | Bell Biosystems, Inc. | Host cells with artificial endosymbionts |
| US10280403B2 (en) | 2012-01-13 | 2019-05-07 | Bell Biosystems, Inc. | Host cells with artificial endosymbionts |
| US10184114B2 (en) | 2013-09-03 | 2019-01-22 | Bell Biosystems, Inc. | Host cell modification with artificial endosymbionts |
| JP2017522022A (en) * | 2014-07-15 | 2017-08-10 | ベル・バイオシステムズ，インコーポレーテッド | Eukaryotic cells containing artificial endosymbiosis for multimodal detection |
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