Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells
| Autor: | Naomie Tidd, Bryan Hilbert, Jane Quinn, Jacob Michelsen |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
animal structures viruses Genetic Vectors Gene Expression canine Gene delivery Biology Minicircle Transfection Catalysis Article Viral vector Cell Line Inorganic Chemistry 03 medical and health sciences Dogs Genes Reporter Animals Humans Vector (molecular biology) Horses Transgenes Physical and Theoretical Chemistry Molecular Biology Spectroscopy mesenchymal stem cell minicircle equine Expression vector Organic Chemistry Mesenchymal stem cell fungi Gene Transfer Techniques Cell Differentiation Mesenchymal Stem Cells SOX9 Transcription Factor General Medicine Molecular biology Computer Science Applications Rats 030104 developmental biology embryonic structures transfection Sox9 Chondrogenesis Ex vivo Biomarkers |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences; Volume 18; Issue 4; Pages: 819 |
| ISSN: | 1422-0067 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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