TRANSFECTION OF NORMAL HUMAN EPIDERMAL KERATINOCYTES WITH LIPID/DNA COMPLEXES IN VITRO

Sebastian Zellmer Institute of Biochemistry, Medical Faculty, University Leipzig, Leipzig, D-04103, Germany

Andrej Surovoy Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, Moscow, 117871, Russia

Frank Gaunitz Institute of Biochemistry, Medical Faculty, University Leipzig, Leipzig, D-04103, Germany

Jürgen Salvetter Institute of Anatomy, Medical Faculty, University Leipzig, Leipzig, D-04103, Germany

Rolf Gebhardt Institute of Biochemistry, Medical Faculty, University Leipzig, Leipzig, D-04103, Germany

Dieter Reissig Institute of Anatomy, Medical Faculty, University Leipzig, Leipzig, D-04103, Germany

Abstract

Highly proliferative normal human epidermal keratinocytes (NHK) were isolated from human foreskin biopsies, cultivated in serum-free medium and characterized 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 transfected in vitro with lipid/DNA complexes made of Effectene or Lipofectamine and different reporter genes. The transfection efficiency of Effectene/DNA complexes was 20fold higher compared to Lipofectamine. Transfected keratinocytes continued to grow and developed within 2 weeks a cellular multilayer (3-D culture). Areas of transfected cells were detected within this layer.

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TRANSFECTION OF NORMAL HUMAN EPIDERMAL KERATINOCYTES WITH LIPID/DNA COMPLEXES IN VITRO

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TRANSFECTION OF NORMAL HUMAN EPIDERMAL KERATINOCYTES WITH LIPID/DNA COMPLEXES IN VITRO

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