Efficient in situ electroporation of mammalian cells grown on microporous membranes.
Electroporation is a common technique for the introduction of DNA molecules into living cells. The method is currently limited by the necessity of applying the electrical discharge to cells in suspension. Adherent cells must therefore be removed from their substratum, which can induce unwanted physiological effects. We report here a new procedure for in situ electroporation of cells grown on microporous membranes of polyethylene terephthalate (PET) or polyester (PE). We demonstrate that this method of in situ electroporation employs only readily available materials and standard electroporation devices without any modifications, is as efficient as conventional electroporation of cells in suspension, and is applicable to a wide range of cell types. Efficient electroporation can be achieved under conditions of minimal cell killing, and can be performed with quiescent cells as well as with confluent epithelial sheets. The method is a useful extension of electroporation technology, and will allow the application of electroporation to a wider spectrum of biological systems.
Yang
T A
TA
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06520, USA.
Heiser
W C
WC
Sedivy
J M
JM
D000818:Animals; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D018274:Electroporation / Q000379:methods*; D006801:Humans; D008851:Micropore Filters; D010957:Plasmids / Q000235:genetics; D011091:Polyesters; D011093:Polyethylene Terephthalates; D051381:Rats; D014162:Transfection / Q000379:methods*
D011091:Polyesters; D011093:Polyethylene Terephthalates; D004247:DNA