Multiple glycerol shocks increase the calcium phosphate transfection of non-synchronized CHO cells

Authors: Grosjean, Frederic; Bertschinger, Martin; Hacker, David; Wurm, Florian

Source: Biotechnology Letters, Volume 28, Number 22, November 2006 , pp. 1827-1833(7)

Publisher: Springer

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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

Document Type: Research article

DOI: http://dx.doi.org/10.1007/s10529-006-9167-7

Affiliations: 1: Email: Frederic.Grosjean@unil.ch

Publication date: 2006-11-01