Efficient electroporation of DNA and protein into confluent and differentiated epithelial cells in culture.
Electroporation-mediated delivery of molecules is a procedure widely used for transfecting complementary DNA in bacteria, mammalian and plant cells. This technique has proven very efficient for the introduction of macromolecules into cells in suspension culture and even into cells in their native tissue environment, e.g. retina and embryonic tissues. However, in spite of several attempts to date, there are no well-established procedures to electroporate polarized epithelial cells adhering to a tissue culture substrate (glass, plastic or filter). We report here the development of a simple procedure that uses available commercial equipment and works efficiently and reproducibly for a variety of epithelial cell lines in culture.
Deora
Ami A
AA
Margaret M. Dyson Vision Research Institute, Department of Ophthalmology, Weill Medical College of Cornell University, New York, NY 10021, USA.
Diaz
Fernando
F
Schreiner
Ryan
R
Rodriguez-Boulan
Enrique
E
D000818:Animals; D018938:Caco-2 Cells; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D004285:Dogs; D018274:Electroporation; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D011506:Proteins / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D011506:Proteins; D004247:DNA