title: Nucleofection: a new method for cutaneous gene transfer?
abstract: BACKGROUND
  Transfection efficacy after nonviral gene transfer in primary epithelial cells is limited. The aim of this study was to compare transfection efficacy of the recently available method of nucleofection with the established transfection reagent FuGENE6.
  
  
  METHODS
  Primary human keratinocytes (HKC), primary human fibroblasts (HFB), and a human keratinocyte cell line (HaCaT) were transfected with reporter gene construct by FuGENE6 or Amaxa Nucleofector device. At corresponding time points, beta-galactosidase expression, cell proliferation (MTT-Test), transduction efficiency (X-gal staining), cell morphology, and cytotoxicity (CASY) were determined.
  
  
  RESULTS
  Transgene expression after nucleofection was significantly higher in HKC and HFB and detected earlier (3 h vs. 24 h) than in FuGENE6. After lipofection 80%-90% of the cells remained proliferative without any influence on cell morphology. In contrast, nucleofection led to a decrease in keratinocyte cell size, with only 20%-42% proliferative cells.
  
  
  CONCLUSIONS
  Related to the method-dependent increase of cytotoxicity, transgene expression after nucleofection was earlier and higher than after lipofection.
authors.affiliation: Department of Plastic Surgery, BG University Hospital Bergmannsheil, Ruhr University Bochum, Bochum 44789, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Jacobsen
authors.forename: Janine
authors.initials: J
authors.lastname: Mertens-Rill
authors.forename: Juergen
authors.initials: J
authors.lastname: Beller
authors.forename: Tobias
authors.initials: T
authors.lastname: Hirsch
authors.forename: Adrien
authors.initials: A
authors.lastname: Daigeler
authors.forename: Stefan
authors.initials: S
authors.lastname: Langer
authors.forename: Marcus
authors.initials: M
authors.lastname: Lehnhardt
authors.forename: Hans-Ulrich
authors.initials: HU
authors.lastname: Steinau
authors.forename: Lars
authors.initials: L
authors.lastname: Steinstraesser