Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of...
Patent
1997-05-22
1998-10-20
Degen, Nancy
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
435243, 435374, 435395, 435402, C12N 502, C12N 510, C12N 516, C12N 522
Patent
active
058245472
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to a method for production of transfected cells, more particularly, a method which makes possible to transfer a foreign gene into target cells with high efficiency in the fields such as cell technology, genetic engineering, developmental engineering and the like.
BACKGROUND OF THE INVENTION
As a method for transferring a foreign gene into target cells, there are known a calcium phosphate method, a DEAE-dextran method, a liposome method, an electroporation method, a microinjection method, a particle gun method and the like. All of these methods have both advantages and disadvantages with respect to manipulation procedures, efficiency, damage on cells and the like. Among these methods, a perforation method such as an electroporation method, a microinjection method, a particle gun method and the like enables easy handling of the cells without using special reagents and provides good transfer efficiency. However, damage to cells by perforation cannot be avoided.
The object of the present invention is to provide a method for improving the transfer efficiency when a foreign gene is transferred into target cells by a perforation method to produce transfected cells.
SUMMARY OF THE INVENTION
The first aspect of the present invention relates to a method for the production of transfected cells, which is characterized by comprising a step of culturing the cells in the presence of a cell-adhesive substance, after injection of a foreign gene into target cells using a perforation method.
The second aspect of the present invention relates to transfected cells containing a foreign gene which are produced by the method of the present invention.
The third aspect of the present invention relates to a kit, which is used for a method for the production of transfected cells according to the first aspect of the present invention and is characterized by containing a cell-adhesive substance.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a graph showing the effect of cell-adhesive polypeptide treatment on gene transfer efficiency in the transfer of pCAT-control vector into human epidermoid cancer cell A-431.
FIG. 2 is a graph showing the effect of cell-adhesive polypeptide treatment on gene transfer efficiency in the transfer of pCAT-control vector into African green monkey kidney cell COS-7-7.
DETAILED DESCRIPTION OF THE INVENTION
The method of the present invention is characterized by culturing the cell in the presence of a substance having the cell-adhesive activity after a foreign gene is transferred into target cells using a perforation method.
As used herein, the perforation method means a method for injection of a gene by perforating a cell wall, including an electroporation method, a microinjection method, a particle gun method and the like. The electroporation method is as described in, for example, Tanpakushitsu, Kakusan, Koso, volume 31, pages 1591-1603 (1986). The microinjection method is as described in, for example, Cell, volume 22, pages 479-488 (1980). The particle gun method is as described in, for example, Technique, volume 3, pages 3-16 (1991). These methods include the known methods used for transferring a gene into cells.
As cells to be used in these perforation methods, for example, there are animal cells which are prepared according to a known method edited by Nippon Seikagakugakkai, published by Tokyo Kagakudojin! and cultured animal cells may be used.
As used herein, a cell-adhesive substance refers to a substance having the cell-adhesive, that is, the activity to make target cells adhere to a cell, to an extracellular matrix which is a substance filling a space between cells in the tissue, or to a material such as plastic, glass and the like. In the present invention, any substances having the activity can be used as long as they give no adverse effects on transfection of target cells. Such the activity is to fix cells, for example, to culture ware covered with a cell-adhesive substance while maintaining the cell in its form, or in a spreaded f
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Hashino Kimikazu
Kato Ikunoshin
Matsushita Hideyuki
Degen Nancy
Takara Shuzo Co. Ltd.
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