Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells
Abstract
:1. Introduction
2. Results
2.1. Isolation and Characterisation of Canine and Equine Mesenchymal Stem Cells
2.2. Further Chondrogenic Characterisation of Equine and Canine MSCs
2.3. Transfection Using a Minicircle Vector Shows Increased Transgene Expression Compared to Parental Plasmids in Rat, Canine and Equine MSCs
2.4. Minicircle Plasmid Demonstrates Similar GFP Expression Intensity in Rat, Canine and Equine MSCs and Is Diluted with Passage
2.5. Minicircle Vector-Driven Expression of Sox9 in Canine MSCs
2.6. Discussion
3. Materials and Methods
3.1. Vectors
3.2. Plasmid Production and Purification
3.3. Isolation and Subculture of Canine and Equine Adipose Tissue-Derived MSCs
3.4. Adipogenic Induction of Canine and Equine MSCs
3.5. Osteogenic Induction
3.6. Chondrogenic Induction
3.7. COL2A1 and COL10A1 Immunohistochemistry
3.8. Transfection of Rat, Canine and Equine MSCs
3.9. Comparison of Transfection Efficiency between Parental and Minicircle Plasmids
3.10. Passage of MSCs Transfected with MC-CAG
3.11. Transfection of Canine MSCs with MC.CAG-Sox9
3.12. Immunohistochemistry
3.13. Statistical Design and Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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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. https://doi.org/10.3390/ijms18040819
Tidd N, Michelsen J, Hilbert B, Quinn JC. Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells. International Journal of Molecular Sciences. 2017; 18(4):819. https://doi.org/10.3390/ijms18040819
Chicago/Turabian StyleTidd, Naomie, Jacob Michelsen, Bryan Hilbert, and Jane C. Quinn. 2017. "Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells" International Journal of Molecular Sciences 18, no. 4: 819. https://doi.org/10.3390/ijms18040819