OPTIMIZED CONDITIONS FOR GENE TRANSFECTION INTO THE HUMAN EOSINOPHILIC CELL-LINE EOL-1 BY ELECTROPORATION

Eosinophils are emerging as an increasingly important cell in the immu noregulatory network of normal and pathological processes. No studies has yet described optimized experimental strategies to transfect DNA i nto human eosinophils. Using a frequently employed in vitro model of h uman eosinophil, the EoL-1 cells, we now described the optimal transfe ction of DNA into these cells by electroporation. Our results indicate that electroporation can efficiently and reproducibly transfect DNA i nto EoL-1 cells. Optimal electroporation conditions consist of the use of 1 X RPMI medium 1640 with 10% FBS. voltage setting at 275 V, 1150 mu F capacitance, 40 mg of DNA and 4.0 x 10(7) cells/ml per electropor ation in a total volume of 0.5 mi in 0.4 cm gap cuvettes. These condit ions may be a useful protocol for transfecting eosinophil cell lines. (C) 1998 Elsevier Science B.V. All rights reserved.