title: Non-viral genetic transfection of rat Schwann cells with FuGENE HD© lipofection and AMAXA© nucleofection is feasible but impairs cell viability.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors: Department of Hand, Plastic, Reconstructive and Burn Surgery, Eberhard-Karls-University of Tübingen, BG-Trauma Center, Tübingen, Germany. arminkraus@hotmail.com
authors: Armin
authors: A
authors: Kraus
authors: Joachim
authors: J
authors: Täger
authors: Konrad
authors: K
authors: Kohler
authors: Max
authors: M
authors: Haerle
authors: Frank
authors: F
authors: Werdin
authors: Hans-Eberhard
authors: HE
authors: Schaller
authors: Nektarios
authors: N
authors: Sinis
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods; D005434:Flow Cytometry; D007211:Indoles; D008055:Lipids / Q000235:genetics / Q000502:physiology*; D008475:Median Nerve / Q000166:cytology; D009414:Nerve Growth Factors / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D010714:Phosphatidylethanolamines / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D020800:Receptor, Nerve Growth Factor / Q000378:metabolism; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000378:metabolism; D012583:Schwann Cells / Q000378:metabolism*; D012584:Sciatic Nerve / Q000166:cytology; D014162:Transfection / Q000379:methods*
chemical_list: C411955:FuGene; D007211:Indoles; D008055:Lipids; D009414:Nerve Growth Factors; D010714:Phosphatidylethanolamines; D020800:Receptor, Nerve Growth Factor; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; C007293:DAPI; C020888:1,2-dielaidoylphosphatidylethanolamine