Nucleofection: a new method for cutaneous gene transfer?
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.
Jacobsen
Frank
F
Department of Plastic Surgery, BG University Hospital Bergmannsheil, Ruhr University Bochum, Bochum 44789, Germany.
Mertens-Rill
Janine
J
Beller
Juergen
J
Hirsch
Tobias
T
Daigeler
Adrien
A
Langer
Stefan
S
Lehnhardt
Marcus
M
Steinau
Hans-Ulrich
HU
Steinstraesser
Lars
L