Conditions for gene transfection into the HL-60 human leukaemia cell line by electroporation.
Electroporation represents a powerful technique for cell transfection; however, its efficiency in haemopoietic cells (approximately 1%) 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 microF) followed by separation of living cells on a Ficoll-Paque discontinuous gradient. Using 10-20 micrograms of plasmid, we routinely achieve 12-14% of transfectants.
Laboratory of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic.
J
J
Pacherník
R
R
Janík
J
J
Hofmanová
V
V
Bryja
A
A
Kozubík
D002470:Cell Survival / Q000502:physiology; D018274:Electroporation; D049452:Green Fluorescent Proteins; D018922:HL-60 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins