Enhanced transfection of cell lines from Atlantic salmon through nucoleofection and antibiotic selection
1 Department of Pharmaceutical Biosciences, School of Pharmacy, University of Oslo, Norway
2 Department of Microbiology, Faculty of Science, University of Malaga, Malaga, Spain
3 Institute of Pathology, Rikshospitalet-Radiumhospitalet Medical Center, University of Oslo, Norway
4 Department of Basic Science and Aquatic Medicine, Norwegian School of Veterinary Science, Oslo, Norway
5 Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, Oslo, Norway
6 Department of Bacteriology and Immunology, The Norwegian Institute of Public Health, P.O. Box 4404 Nydalen, N-0403 Oslo, Norway
7 National Veterinary Institute, Oslo, Norway
8 Centre for Ecology and Evolutionary Synthesis, Department of Biology, University of Oslo, Norway
BMC Research Notes 2011, 4:136 doi:10.1186/1756-0500-4-136Published: 6 May 2011
Cell lines from Atlantic salmon kidney have made it possible to culture and study infectious salmon anemia virus (ISAV), an aquatic orthomyxovirus affecting farmed Atlantic salmon. However, transfection of these cells using calcium phosphate precipitation or lipid-based reagents shows very low transfection efficiency. The Amaxa Nucleofector technology™ is an electroporation technique that has been shown to be efficient for gene transfer into primary cells and hard to transfect cell lines.
Here we demonstrate, enhanced transfection of the head kidney cell line, TO, from Atlantic salmon using nucleofection and subsequent flow cytometry. Depending on the plasmid promoter, TO cells could be transfected transiently with an efficiency ranging from 11.6% to 90.8% with good viability, using Amaxa's cell line nucleofector solution T and program T-20. A kill curve was performed to investigate the most potent antibiotic for selection of transformed cells, and we found that blasticidin and puromycin were the most efficient for selection of TO cells.
The results show that nucleofection is an efficient way of gene transfer into Atlantic salmon cells and that stably transfected cells can be selected with blasticidin or puromycin.