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 degrees 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. Transformed cells were stable and different numbers of plasmid DNA copies were detected.
Ogawa
T
T
Cosmetics Laboratory, Kanebo, Ltd., Kanagawa, Japan.
Hyodo
M
M
Ihara
S
S
Takekoshi
M
M
Minato
K
K
Suzuki
K
K
D000818:Animals; D002463:Cell Membrane Permeability; D002470:Cell Survival; D004247:DNA / Q000235:genetics*; D005260:Female; D008325:Mammary Neoplasms, Experimental; D051379:Mice; D009693:Nucleic Acid Hybridization; D009997:Osmotic Pressure; D010957:Plasmids; D014162:Transfection; D014407:Tumor Cells, Cultured
D004247:DNA