Optimizing liposome-mediated gene transfer in primary rat septo-hippocampal cell cultures.
Although liposomes have been widely employed to transfect DNA into a variety of cell types, no previous studies have systematically examined conditions producing optimal liposomal-mediated transfection of DNA into central nervous system (CNS) cells. Thus, we used the beta-galactosidase (beta-gal) reporter gene to examine factors influencing the efficiency of liposome-mediated gene transfection in CNS cell cultures. Our results indicate that without increasing the amounts of DNA, increased liposome concentrations within certain limits enhanced transfection efficiency. However, higher liposome levels could produce cell lysis. Without increasing liposome concentrations, increased amounts of DNA did not improve transfection efficiency. Employing the optimal concentration (1 microgram DNA/3 microliters liposomes/well), beta-gal gene expression was sustained for at least two weeks after transfection in primary septo-hippocampal cultures.
Yang
K
K
Department of Neurosurgery, University of Texas Health Sciences Center at Houston 77030.
Faustinella
F
F
Xue
J J
JJ
Whitson
J
J
Kampfl
A
A
Mu
X S
XS
Zhao
X
X
Taglialatela
G
G
Perez-Polo
J R
JR
Clifton
G
G
D000818:Animals; D002478:Cells, Cultured; D005696:Galactosidases / Q000235:genetics; D018014:Gene Transfer Techniques; D006624:Hippocampus / Q000502:physiology*; D008081:Liposomes; D051381:Rats; D017207:Rats, Sprague-Dawley; D012686:Septal Nuclei / Q000502:physiology; D013997:Time Factors
D008081:Liposomes; D005696:Galactosidases