Nucleofection: A New Method for Cutaneous Gene Transfer?

Jacobsen, Frank; Mertens-Rill, Janine; Beller, Juergen; Hirsch, Tobias; Daigeler, Adrien; Langer, Stefan; Lehnhardt, Marcus; Steinau, Hans-Ulrich; Steinstraesser, Lars

doi:https://doi.org/10.1155/JBB/2006/26060

BioMed Research International

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Methods Report | Open Access

Volume 2006 | Article ID 026060 | https://doi.org/10.1155/JBB/2006/26060

Nucleofection: A New Method for Cutaneous Gene Transfer?

Frank Jacobsen,¹Janine Mertens-Rill,¹Juergen Beller,¹Tobias Hirsch,¹Adrien Daigeler,¹Stefan Langer,¹Marcus Lehnhardt,¹Hans-Ulrich Steinau,¹and Lars Steinstraesser¹

Received22 May 2006

Revised03 Aug 2006

Accepted24 Aug 2006

Published20 Nov 2006

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, β-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. Conclusion. Related to the method-dependent increase of cytotoxicity, transgene expression after nucleofection was earlier and higher than after lipofection.

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Copyright © 2006 Frank Jacobsen et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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