Transfection of normal human epidermal keratinocytes with lipid/DNA complexes in vitro

Highly proliferative normal human epidermal keratinocytes (NHK) were isolat ed from human foreskin biopsies, cultivated in serum-free medium and charac terized by flow cytometry. The expression of cytokeratin 19, cytokeratin 14 and vimentin indicated that the suspension contained a high percentage of undifferentiated cells of the basal epidermal layer. The NHK were transfect ed in vitro with lipid/DNA complexes made of Effectene or Lipofectamine and different reporter genes. The transfection efficiency of Effectene/DNA com plexes was 20fold higher compared to Lipofectamine. Transfected keratinocyt es continued to grow and developed within 2 weeks a cellular multilayer (3- D culture). Areas of transfected cells were detected within this layer.