Artikel
Nucelofection of canine Olfactory ensheating cells and canine Schwann cells for stable expression of GPF and tdREd under control of an EF1 alpha promotor
Nucleofection von caninen Olfactory Ensheathing Cells zur stabilen Expression von GFP und tdRED unter Kontrolle eines EF1 alpha promotors
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Veröffentlicht: | 27. September 2016 |
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Cell transplantation strategies are a promising toll with regard to reparative and regenerative processes after spinal cord trauma. Especially the transplantation of OECs into traumatic spinal cord injury and models can promote axonal sprouting and regeneration and functional recovery. Transfection of canine Olfactory ensheathing cells (OECs) is difficult and resulted in minimal transfection rates. Lenti-X lentiviral vector systems with EF-1 alpha promoters allow to achieve long-term expression of genes of interest in cell types in which CMV promoters are often silenced, such as hematopoietic and stem cells. The EF-1 alpha promoter is derived from the human EEF1A1 gene that expresses the alpha subunit of eukaryotic elongation factor 1. The described nucleofection was not used for glia cells yet. Here, we describe the successful transfection of canine OECS with a plasmid encoding enhanced green fluorescent protein eGFP under control of an EF1alpha promoter (pcDNAEF1-eGFP) for stable expression of GFP as a prerequisite for in vivo cell transplantation studies.