High-Frequency Transfection of Mouse FM3A Mammary Carcinoma Cells in Suspension Culture with Plasmid DNA

High-Frequency Transfection of Mouse FM3A Mammary Carcinoma Cells in Suspension Culture with Plasmid DNA

High-frequency transfection of mouse FM3A cells, grown in suspension, with plasmid pSV2neo DNA was achieved by incubation of the cells with DNA plus polybrene for 6 h followed by an osmotic shock with a hypertonic NaCl solution. When incubated for 20 min at 34°C, FM3A cells showed resistance to the...

Full description

Saved in:
Bibliographic Details
Published in:Cell Structure and Function Vol. 12; no. 6; pp. 567 - 574
Main Authors: Ogawa, Tadatomo, Hyodo, Masao, Ihara, Seiji, Takekoshi, Masataka, Minato, Kaori, Suzuki, Kenshi
Format: Journal Article
Language:English
Published: Kyoto Japan Society for Cell Biology 1987
Japan Science and Technology Agency
Subjects:
Animals
Biological and medical sciences
Biotechnology
Cell Membrane Permeability
Cell Survival
Diverse techniques
DNA - genetics
Female
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Mammary Neoplasms, Experimental
Methods. Procedures. Technologies
Mice
Molecular and cellular biology
Nucleic Acid Hybridization
Osmotic Pressure
Plasmids
Transfection
Tumor Cells, Cultured
Vectors (cloning, transfer, expression). Insertion sequences and transposons
Cell culture
Calcium
Osmolarity
Selection
Rodentia
Genetic transfer
Vertebrata
Mammalia
Transfection
Mouse
Tumor
Mammary gland
Vector
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:High-frequency transfection of mouse FM3A cells, grown in suspension, with plasmid pSV2neo DNA was achieved by incubation of the cells with DNA plus polybrene for 6 h followed by an osmotic shock with a hypertonic NaCl solution. When incubated for 20 min at 34°C, FM3A cells showed resistance to the osmolarity change from 0.1 to 9.0% NaCl in the medium. Within this concentration range, 5-7 % gave the highest efficiency of transfection. Both linear and circular forms of plasmid DNA produced transformants with equal efficiency. This method was simple, reproducible, and carrier DNA was not required. The efficiency was about 100 times higher than that of the method with DNA-calcium phosphate precipitates. Trans-formed cells were stable and different numbers of plasmid DNA copies were detected.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0386-7196
1347-3700
DOI:10.1247/csf.12.567