Long-term expression of foreign genes in normal human epidermal keratinocytes after transfection with lipid/DNA complexes

Histochem Cell Biol. 2001 Jan;115(1):41-7. doi: 10.1007/s004180000225.

Abstract

Normal human epidermal keratinocytes were isolated and cultivated in serum-free medium. The expression of the integrin subunits alpha6 and beta1 indicated that a high number of keratinocytes from the stem cell system was present. These cells were transfected with complexes made of different cationic lipids and marker genes. Effectene showed a 20-fold higher transfection efficiency, compared to Lipofectin and Lipofectamine, and a similar low toxicity. The transfection protocol was optimised. A DNA/lipid ratio of 0.133 showed the highest transfection efficiency. Keratinocytes expressed the marker gene luciferase for 20 days. The maximum expression occurred after 3-4 days, where individual patches of fluorescent keratinocytes were detected. Transfected keratinocytes, cultivated at the air-liquid interface, expressed the marker gene beta-galactosidase for at least 7 weeks.

Publication types

  • Comparative Study

MeSH terms

  • Cation Exchange Resins / metabolism
  • Cation Exchange Resins / standards
  • Cell Culture Techniques / methods*
  • DNA / metabolism
  • Gene Expression
  • Gene Transfer Techniques*
  • Genes, Reporter / genetics*
  • Humans
  • Indicators and Reagents / metabolism
  • Indicators and Reagents / standards
  • Integrin alpha6beta1
  • Integrin beta1 / genetics
  • Integrin beta1 / metabolism
  • Integrins / genetics
  • Integrins / metabolism
  • Keratinocytes / cytology
  • Keratinocytes / metabolism*
  • Lipid Metabolism
  • Lipids / standards
  • Liposomes / metabolism
  • Luciferases / genetics
  • Luciferases / standards
  • Phosphatidylethanolamines / metabolism
  • Phosphatidylethanolamines / standards
  • Stem Cells / cytology
  • Time Factors
  • Transfection / methods
  • Transfection / standards*
  • beta-Galactosidase / genetics
  • beta-Galactosidase / standards

Substances

  • Cation Exchange Resins
  • Indicators and Reagents
  • Integrin alpha6beta1
  • Integrin beta1
  • Integrins
  • Lipids
  • Lipofectamine
  • Liposomes
  • Phosphatidylethanolamines
  • 1,2-dielaidoylphosphatidylethanolamine
  • DNA
  • Luciferases
  • beta-Galactosidase