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

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.