Minicircle mediated gene delivery to canine and equine mesenchymal stem cells

Research output: Contribution to journalArticle

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4 Downloads (Pure)

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.
Original languageEnglish
Article number819
Pages (from-to)1-14
Number of pages14
JournalInternational Journal of Molecular Sciences
Volume18
Issue number4
DOIs
Publication statusPublished - 12 Apr 2017

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stem cells
Stem cells
Mesenchymal Stromal Cells
genes
Horses
Transfection
Canidae
delivery
Genes
Cartilage
Plasmids
cartilage
plasmids
Repair
Green Fluorescent Proteins
Regeneration
Rodentia
therapy
Animal Models
cells

Cite this

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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.",
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Minicircle mediated gene delivery to canine and equine mesenchymal stem cells. / Tidd, Naomie; Michelsen, Jacob; Hilbert, Bryan; Quinn, Jane C.

In: International Journal of Molecular Sciences, Vol. 18, No. 4, 819, 12.04.2017, p. 1-14.

Research output: Contribution to journalArticle

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