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Int. J. Mol. Sci. 2017, 18(4), 819;

Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells

School of Animal and Veterinary Science, Charles Sturt University, Boorooma Street, Locked Bag 588, Wagga Wagga, NSW 2678, Australia
Graham Centre for Agricultural Innovation (NSW Department of Primary Industries and Charles Sturt University), Wagga Wagga, NSW 2678 Australia
Author to whom correspondence should be addressed.
Academic Editors: Adriano Piatelli and Barbara Zavan
Received: 25 February 2017 / Revised: 3 April 2017 / Accepted: 10 April 2017 / Published: 12 April 2017
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Gene-directed tissue repair offers the clinician, human or veterinary, the chance to enhance cartilage regeneration and repair at a molecular level. Non-viral plasmid vectors have key biosafety advantages over viral vector systems for regenerative therapies due to their episomal integration however, conventional non-viral vectors can suffer from low transfection efficiency. Our objective was to identify and validate in vitro a novel non-viral gene expression vector that could be utilized for ex vivo and in vivo delivery to stromal-derived mesenchymal stem cells (MSCs). Minicircle plasmid DNA vector containing green fluorescent protein (GFP) was generated and transfected into adipose-derived MSCs from three species: canine, equine and rodent and transfection efficiency was determined. Both canine and rat cells showed transfection efficiencies of approximately 40% using minicircle vectors with equine cells exhibiting lower transfection efficiency. A Sox9-expressing minicircle vector was generated and transfected into canine MSCs. Successful transfection of the minicircle-Sox9 vector was confirmed in canine cells by Sox9 immunostaining. This study demonstrate the application and efficacy of a novel non-viral expression vector in canine and equine MSCs. Minicircle vectors have potential use in gene-directed regenerative therapies in non-rodent animal models for treatment of cartilage injury and repair. View Full-Text
Keywords: canine; equine; mesenchymal stem cell; minicircle; transfection; Sox9 canine; equine; mesenchymal stem cell; minicircle; transfection; Sox9

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Tidd, N.; Michelsen, J.; Hilbert, B.; Quinn, J.C. Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells. Int. J. Mol. Sci. 2017, 18, 819.

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