Conditions for gene transfection into the HL-60 human leukaemia cell line by electroporation
Publikace: FOLIA BIOLOGICA 48, 154-156 Autoři: Pachernik, J., Janik, R., Hofmanova, J., Bryja, V., Kozubik, A. Rok: 2002
Electroporation represents a powerful technique for cell transfection; however, its efficiency in haemopoietic cells (similar to1%) is largely unsatisfactory. Biological processes in haemopoietic cells are often studied using leukaemia cell line HL-60. For this reason we developed conditions for efficiently introducing plasmids to HL-60 cells by electroporation, as an alternative to other techniques. This technique employs the electric pulse (250 270 V; 1000 muF) followed by separation of living cells on a Ficoll-Paque discontinuous gradient. Using 10-20 mug of plasmid, we routinely achieve 12-14% of transfectants.