Multiple glycerol shocks increase the calcium phosphate transfection of non-synchronized CHO cells.
The exposure of CHO DG44 cells to an osmotic shock, after DNA uptake, results in a cellular volume decrease of approx. 55%. Repetitive osmotic shocks targeted different sub-populations of cells as was demonstrated using two different fluorescent reporter genes. Also the exposure of a calcium phosphate-DNA coprecipitate to high osmolarity in vitro caused the release of the DNA from the precipitate. The results demonstrate the importance of the osmotic shock on the efficient delivery of plasmid DNA to the nucleus of CHO cells following calcium phosphate-mediated transfection.
Grosjean
Frederic
F
Institute of Biochemistry, Chemin des Boveresses 155, 1066, Epalinges, Switzerland. Frederic.Grosjean@unil.ch
Bertschinger
Martin
M
Hacker
David L
DL
Wurm
Florian M
FM
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D002130:Calcium Phosphates / Q000737:chemistry*; D018929:Cell Culture Techniques / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry; D005990:Glycerol / Q000737:chemistry*; D008860:Microscopy, Ultraviolet; D009995:Osmosis; D009997:Osmotic Pressure; D010957:Plasmids / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
D002130:Calcium Phosphates; D004247:DNA; C020243:calcium phosphate; D005990:Glycerol