K2 Transfection System Boosts the Adenoviral Transduction of Murine Mesenchymal Stromal Cells
Abstract
:1. Introduction
2. Results
2.1. Adenoviral Transduction of MSC and the Effect of Potential Boosters
2.1.1. Adenoviral Transduction of MSC Isolated from NOD, C57BL/6, and BALB/c Mice
2.1.2. Adenoviral Transduction of MSC in the Presence of Different Potential Enhancers of the Transduction
2.2. Optimization of the Adenoviral Transduction of MSC in the Presence of K2TS
2.2.1. K2TR Optimization for Efficient Transduction Boosting
2.2.2. K2M Optimization
2.2.3. The Expression of Adenoviral Receptors in MSC Derived from Different Mouse Strains
2.3. Evaluation of the MSC Properties Following Adenoviral Transduction in the Presence of K2TS
2.3.1. Exposure to K2TS Does Not Modify the Expression of MSC Markers
2.3.2. Exposure to K2TS Does Not Influence the Multipotency of MSC
2.3.3. The Immunosuppressive Properties of MSC Are Not Affected by the K2TS
3. Discussion
4. Materials and Methods
4.1. MSC Isolation and Culture
4.2. Adenovirus Packaging, Purification, and Titration
4.3. Adenoviral Transduction of MSC
4.4. Adenoviral Transduction of MSC in the Presence of Various Potential Boosters
4.5. Optimization of the K2TS Components for MSC Adenoviral Transduction
4.6. RT-PCR and Quantitative Real-Time PCR
4.7. MSC Differentiation
4.8. Immunosuppression Assay
4.9. Flow Cytometry
4.10. Statistical Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
AdV | Adenovirus |
CAR | Coxsackievirus and Adenovirus Receptors |
CFSE | Carboxy-fluorescein succinimidyl ester |
GFP | Green fluorescent protein |
K2M | K2 Multiplier |
K2TR | K2 Transfection Reagent |
K2TS | K2 Transfection System |
MOI | multiplicity of infection |
MSC | Mesenchymal Stromal Cell |
NOD | Non-Obese Diabetic |
PI | Propidium iodide |
Sca-1 | Stem Cells Antigen-1 |
TU | Transduction Units |
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Dumitrescu, M.; Vacaru, A.M.; Trusca, V.G.; Fenyo, I.M.; Ionita, R.; Gafencu, A.V. K2 Transfection System Boosts the Adenoviral Transduction of Murine Mesenchymal Stromal Cells. Int. J. Mol. Sci. 2021, 22, 598. https://doi.org/10.3390/ijms22020598
Dumitrescu M, Vacaru AM, Trusca VG, Fenyo IM, Ionita R, Gafencu AV. K2 Transfection System Boosts the Adenoviral Transduction of Murine Mesenchymal Stromal Cells. International Journal of Molecular Sciences. 2021; 22(2):598. https://doi.org/10.3390/ijms22020598
Chicago/Turabian StyleDumitrescu, Madalina, Ana Maria Vacaru, Violeta Georgeta Trusca, Ioana Madalina Fenyo, Radu Ionita, and Anca Violeta Gafencu. 2021. "K2 Transfection System Boosts the Adenoviral Transduction of Murine Mesenchymal Stromal Cells" International Journal of Molecular Sciences 22, no. 2: 598. https://doi.org/10.3390/ijms22020598