Gene transfer by lipofection in rabbit and human secretory epithelial cells.
Lipofection, a recently-developed method for gene transfer, was tested in secretory epithelial cells. Lipofection facilitated both transient DNA transfection with plasmids containing the chloramphenicol acetyltransferase gene and stable transfection with a plasmid containing the neomycin resistance gene, which confers resistance to the antibiotic G418 (Geneticin). Gene transfer occurred efficiently in a rabbit kidney medullary thick ascending limb cell line and in primary cultures of rabbit tracheal epithelial cells. The method was also effective in Simian virus 40-transformed human airway cells isolated from a normal individual and from a patient with cystic fibrosis (CF). Cytotoxicity was minimal, particularly when the time of exposure to the lipofectin-DNA was limited to 3-5 h (less than 5% cell loss). Thus, the lipofection method is useful for gene transfer in a variety of secretory epithelial cells and should be ideal for studies of defective secretory epithelial cell function in CF.
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Department of Physiology, Johns Hopkins School of Medicine, Baltimore, MD 21205.
Zeitlin
P L
PL
Guggino
W B
WB
Craig
R W
RW
D000818:Animals; D001980:Bronchi / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003550:Cystic Fibrosis / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D004351:Drug Resistance / Q000235:genetics; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D009355:Neomycin; D010957:Plasmids; D000644:Quaternary Ammonium Compounds; D011817:Rabbits; D014132:Trachea / Q000378:metabolism*; D014162:Transfection
D008081:Liposomes; D000644:Quaternary Ammonium Compounds; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D009355:Neomycin