Neuron Glia Biology
- Neuron Glia Biology / Volume 6 / Issue 04 / November 2010, pp 225-230
- Copyright © Cambridge University Press 2011
- DOI: http://dx.doi.org/10.1017/S1740925X11000056 (About DOI), Published online: 07 June 2011
Neurotechniques
Non-viral genetic transfection of rat Schwann cells with FuGENE HD© lipofection and AMAXA© nucleofection is feasible but impairs cell viability
Armin Krausa1 c1, Joachim Tägera2, Konrad Kohlera2, Max Haerlea3, Frank Werdina1, Hans-Eberhard Schallera1 and Nektarios Sinisa1
a1 Department of Hand, Plastic, Reconstructive and Burn Surgery, Eberhard-Karls-University of Tübingen, BG-Trauma Center, Germany
a2 Center for Regenerative Biology and Regenerative Medicine, Eberhard-Karls-University of Tübingen, Germany
a3 Department of Hand and Plastic Surgery, Orthopaedic Hospital Markgroeningen, Kurt-Lindemann-Weg 10, D-71706 Markgroeningen, Germany
Abstract
Purpose: To determine transfection efficiency of FuGENE HD© lipofection and AMAXA© nucleofection on rat Schwann cells (SC).
Methods: The ischiadic and median nerves of 6-8 week old Lewis rats were cultured in modified melanocyte-growth medium. SCs were genetically transfected with green fluorescent protein (GFP) as reporter gene using FuGENE HD© lipofection and AMAXA© nucleofection. Transfection rates were determined by visualization of GFP fluorescence under fluorescence microscopy and cell counting. Transfected cell to non-transfected cell relation was determined.
Results: Purity of Schwann cell culture was 88% as determined by immunohistologic staining. Transfection rate of FuGENE HD© lipofection was 2%, transfection rate of AMAXA© nucleofection was 10%. With both methods, Schwann cells showed pronounced aggregation behavior which made them unfeasible for further cultivation. Settling of Schwann cells on laminin and poly-l-ornithine coated plates was compromised by either method.
Conclusion: Non-viral transfection of rat SC with FuGENE HD© lipofection and AMAXA© nucleofection is basically possible with a higher transfection rate for nucleofection than for lipofection. As cell viability is compromised by either method however, viral transfection is to be considered if higher efficiency is required.
Correspondence:
c1 Correspondence should be addressed to: Dr. Armin Kraus, Department of Hand, Plastic, Reconstructive and Burn Surgery, Eberhard-Karls-Universität Tübingen, BG-Trauma Center, Schnarrenbergstrasse 95, 72076 Tübingen, Germany phone: + 49 7071 606 1036 fax: + 49 7071 606 1037 email: arminkraus@hotmail.com
