Lipospermine-mediated gene transfer technique into murine cultured cortical cells.
In order to transfer exogenous DNA into embryonic cortical cells, we have chosen a transfection technique using a synthetic lipospermine (dipalmitoylphosphatidylethanolamylspermine, DPPES) which complexes DNA molecules and allows their penetration into the intracellular compartment. The procedure was optimized after testing several parameters: DPPES/DNA ratio, incubation time, kinetics of transgene expression, and growth medium. The protocol was achieved by following the expression of the E. coli LacZ reporter gene under the control of the cytomegalovirus promoter. The lipopolyamine-mediated transfection is efficient for terminally differentiated cells, since we routinely obtained transfection efficiencies of 30% for neurons.
Sola
B
B
Université de Caen, CNRS URA 1829, Laboratoire de Neurosciences, Caen,France.
Staedel
C
C
Remy
J S
JS
Bahr
A
A
Behr
J P
JP
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D003470:Culture Media / Q000494:pharmacology; D003587:Cytomegalovirus / Q000235:genetics; D004279:DNA, Viral; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D005998:Glycine / Q000031:analogs & derivatives*; D007700:Kinetics; D007763:Lac Operon; D051379:Mice; D008815:Mice, Inbred Strains; D009474:Neurons / Q000166:cytology / Q000187:drug effects; D010957:Plasmids; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D013096:Spermine / Q000031:analogs & derivatives*; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics
D003470:Culture Media; D004279:DNA, Viral; C061134:dioctadecylamidoglycylspermine; D013096:Spermine; D005998:Glycine