LIPOSPERMINE-MEDIATED GENE-TRANSFER TECHNIQUE INTO MURINE CULTURED CORTICAL-CELLS

In order to transfer exogenous DNA into embryonic cortical cells, we h ave chosen a transfection technique using a synthetic lipospermine (di palmitoylphosphatidylethanolamylspermine, DPPES) which complexes DNA m olecules and allows their penetration into the intracellular compartme nt. The procedure was optimized after testing several parameters: DPPE S/DNA ratio, incubation time, kinetics of transgene expression, and gr owth medium. The protocol was achieved by following the expression of the E. coli LacZ reporter gene under the control of the cytomegaloviru s promoter. The lipopolyamine-mediated transfection is efficient for t erminally differentiated cells, since we routinely obtained transfecti on efficiencies of 30%, for neurons. (C) 1997 Elsevier Science B.V.