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Multiple glycerol shocks increase the calcium phosphate transfection of non-synchronized CHO cells |
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| Authors: | Frederic Grosjean Martin Bertschinger David L. Hacker Florian M. Wurm |
| Affiliation: | (1) Institute of Biochemistry, Chemin des Boveresses 155, 1066 Epalinges, Switzerland;(2) Glenmark Pharmaceuticals SA., Chemin de la Combeta 5, 2300 La Chaux-de-Fonds, Switzerland;(3) Laboratory of Cellular Biotechnology, école Polytechnique Fédérale de Lausanne (EPFL), 1015 Lausanne, Switzerland |
| Abstract: | 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. |
| Keywords: | Calcium phosphate Cellular volume decrease Glycerol shock Osmotic effect Plasmid release Transfection |
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