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Int. J. Mol. Sci. 2017, 18(4), 819; doi:10.3390/ijms18040819

Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells

1
School of Animal and Veterinary Science, Charles Sturt University, Boorooma Street, Locked Bag 588, Wagga Wagga, NSW 2678, Australia
2
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
View Full-Text   |   Download PDF [4111 KB, uploaded 13 April 2017]   |  

Abstract

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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MDPI and ACS Style

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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