Abstract
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 (< 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.
Original language | English (US) |
---|---|
Pages (from-to) | 198-203 |
Number of pages | 6 |
Journal | Pflügers Archiv European Journal of Physiology |
Volume | 415 |
Issue number | 2 |
DOIs | |
State | Published - Nov 1989 |
Keywords
- Cystic fibrosis
- Epithelial cells
- Genetic engineering
- Transfection, human
ASJC Scopus subject areas
- Physiology
- Clinical Biochemistry
- Physiology (medical)