Polyethyleneimine-mediated gene delivery into rat pheochromocytoma PC-12 cells.
In this study, we examined the use of polyethyleneimine (PEI) as a non-viral gene carrier and lipofectamine(trade mark) 2000 as control for rat pheochromocytoma PC-12 cells. The complex formation of PEI and DNA or lipofectamine and DNA was characterized by gel electrophoresis and measurement of particle size and surface charge. A gradual increase in surface charge (from 0.7 to 43 mV) and a gradual decrease in particle size (from 900 to 130 nm) was observed in the PEI-DNA complex with higher PEI concentrations. The cytotoxicity of PC-12 cells for lipofectamine-DNA complex was similar to PEI-DNA complex at N:P charge ratios of 4 and 8. Transfection efficiency was 14% for lipofectamine and 15% for PEI. At low N:P ratio, DNA condenses poorly, so the particle size tends to be large and polydispersed, resulting in poor transfection efficiency. Meanwhile, a high N:P ratio results in high transfection efficiency and cytotoxicity. Transfected PC-12 cells showed the generation of neurites from transfected PC-12 cells in the presence of NGF, indicating the differentiation of PC-12 cells. NGF-differentiated PC-12 cells were transfected by PEI-DNA complex of N:P charge ratio 8. From real-time imaging for transfection, the enhanced green fluorescent protein (EGFP) started to localize in the nuclei of PC-12 cells at 5 h and localized in the cytoplasm from 15 h. Our study demonstrates that PEI or lipofectamine may be applied as an effective gene carrier for PC-12 cells.
Lee
Jung Hwa
JH
Fusion Biotechnology Research Centre, Korea Research Institute of Chemical Technology, PO Box 107, Yuseong, Daejeon 305-600, Korea.
Ahn
Hyun Hee
HH
Kim
Kyung Sook
KS
Lee
Ju Young
JY
Kim
Moon Suk
MS
Lee
Bong
B
Khang
Gilson
G
Lee
Hai Bang
HB
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D004247:DNA / Q000737:chemistry; D008055:Lipids / Q000737:chemistry; D020932:Nerve Growth Factor / Q000494:pharmacology; D016716:PC12 Cells; D010673:Pheochromocytoma / Q000235:genetics / Q000473:pathology; D011094:Polyethyleneimine / Q000008:administration & dosage*; D051381:Rats; D014162:Transfection
D008055:Lipids; C086724:Lipofectamine; D011094:Polyethyleneimine; D004247:DNA; D020932:Nerve Growth Factor