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Non-viral genetic transfection of rat Schwann cells with FuGENE HD© lipofection and AMAXA© nucleofection is feasible but impairs cell viability

Published online by Cambridge University Press:  07 June 2011

Armin Kraus ,
Joachim Täger ,
Konrad Kohler ,
Max Haerle ,
Frank Werdin ,
Hans-Eberhard Schaller  and
Nektarios Sinis
Armin Kraus*
Affiliation:
Department of Hand, Plastic, Reconstructive and Burn Surgery, Eberhard-Karls-University of Tübingen, BG-Trauma Center, Germany
Joachim Täger
Affiliation:
Center for Regenerative Biology and Regenerative Medicine, Eberhard-Karls-University of Tübingen, Germany
Konrad Kohler
Affiliation:
Center for Regenerative Biology and Regenerative Medicine, Eberhard-Karls-University of Tübingen, Germany
Max Haerle
Affiliation:
Department of Hand and Plastic Surgery, Orthopaedic Hospital Markgroeningen, Kurt-Lindemann-Weg 10, D-71706 Markgroeningen, Germany
Frank Werdin
Affiliation:
Department of Hand, Plastic, Reconstructive and Burn Surgery, Eberhard-Karls-University of Tübingen, BG-Trauma Center, Germany
Hans-Eberhard Schaller
Affiliation:
Department of Hand, Plastic, Reconstructive and Burn Surgery, Eberhard-Karls-University of Tübingen, BG-Trauma Center, Germany
Nektarios Sinis
Affiliation:
Department of Hand, Plastic, Reconstructive and Burn Surgery, Eberhard-Karls-University of Tübingen, BG-Trauma Center, Germany
*
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: [email protected]
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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.

Keywords

Schwann cellperipheral nervenon-viral transfectionmethodology
Type
Neurotechniques
Information
Neuron Glia Biology , Volume 6 , Issue 4 , November 2010 , pp. 225 - 230
Copyright
Copyright © Cambridge University Press 2011

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