Gene transfer by lipofection in rabbit and human secretory epithelial cells

Luo Lu, Pamela L. Zeitlin, William B. Guggino, Ruth W. Craig

Research output: Contribution to journalArticlepeer-review

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 languageEnglish (US)
Pages (from-to)198-203
Number of pages6
JournalPflügers Archiv European Journal of Physiology
Volume415
Issue number2
DOIs
StatePublished - Nov 1989

Keywords

  • Cystic fibrosis
  • Epithelial cells
  • Genetic engineering
  • Transfection, human

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Physiology (medical)

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