Fol. Biol. 2002, 48, 154-156
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 (~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 μF) followed by separation of living cells on a Ficoll-Paque discontinuous gradient. Using 10-20 μg of plasmid, we routinely achieve 12-14% of transfectants.
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Funding
This work was supported by the Grant Agency of the Czech Republic No. 524/99/0694 and by the Grant Agency of the Academy of Sciences of the Czech Republic Nos. S5004009, Z50040920 and KSK5011112.
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Copyright
This is an open-access article distributed under the terms of the Creative Commons Attribution License.