Procoagulant Activity of Umbilical Cord-Derived Mesenchymal Stromal Cells’ Extracellular Vesicles (MSC-EVs)
Abstract
:1. Introduction
2. Results
2.1. EV Characterization
2.2. Nanoparticle Concentration via Nanoparticle Tracking Analysis (NTA)
2.3. Nanoparticle Size
2.4. EV Characterization via Dynamic Light Scattering (DLS)—Polydispersity Index (PDI) and Surface Charge (Zeta Potential)
2.5. Protein Content of EV Samples
2.6. Dot Blot
2.7. EV Procoagulant Activity Assay
3. Discussion
3.1. EV Characterization
3.2. Assaying Hemocompatibility
3.3. Limitations of the Present Work
3.4. Impact of Our Findings on Clinical Translation
4. Materials and Methods
4.1. Preparation of Conditioned Media from Canine Umbilical Cord-Derived Mesenchymal Stromal Cells
4.2. Design of Experiments Approach
4.3. EV Isolation via a Combination of Ultrafiltration and Size-Exclusion Chromatography (SEC)
4.4. EV Isolation via Ultracentrifugation (UC)
4.5. Lyophilization of EVs
4.6. Nanoparticle Tracking Analysis (NTA)
4.7. Dynamic Light Scattering (DLS)
4.8. Transmission Electron Microscopy (TEM)
4.9. Protein
4.10. Immunocytochemistry
4.11. Dot Blot
4.12. Procoagulant Assay
4.13. Statistics
4.14. Data Visualization
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Line | Cell Passage | Isolation Method | EV Number (per well) | FX-a Activity (ng/mL) | FX-a Generated (nM) | FX-a Generated per 1 × 106 EVs (nM) | FX-a Generated per 1 × 106 Cells |
---|---|---|---|---|---|---|---|
CUC 20 | Early | SEC | 5.00 × 107 | 37.7 | 0.82 | 0.016 | 260.114 |
CUC 20 | Late | SEC | 5.00 × 107 | 111.4 | 2.42 | 0.048 | 533.752 |
CUC 21 | Early | SEC | 5.00 × 107 | 52.8 | 1.15 | 0.023 | 735.681 |
CUC 21 | Late | SEC | 5.00 × 107 | 5.2 | 0.11 | 0.002 | 40.688 |
CUC 23 | Early | UC | 5.00 × 107 | 103.4 | 2.25 | 0.045 | 1928.998 |
CUC 23 | Late | UC | 5.00 × 107 | 95.7 | 2.08 | 0.042 | 1109.115 |
CUC 27 | Early | SEC | 5.00 × 107 | 8.2 | 0.18 | 0.004 | 59.294 |
CUC 27 | Late | SEC | 5.00 × 107 | 20.8 | 0.45 | 0.009 | 154.836 |
CUC 28 | Early | UC | 5.00 × 107 | 35.4 | 0.77 | 0.015 | 198.870 |
CUC 28 | Late | UC | 5.00 × 107 | 89.9 | 1.95 | 0.039 | 405.535 |
CUC 30 | Early | UC | 5.00 × 107 | 13.8 | 0.30 | 0.006 | 202.521 |
CUC 30 | Late | UC | 5.00 × 107 | 74.2 | 1.61 | 0.032 | 597.879 |
Mean ± SD | 0.023 ± 0.017 | 518.940 ± 543.138 |
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Wright, A.; Snyder, O.; He, H.; Christenson, L.K.; Fleming, S.; Weiss, M.L. Procoagulant Activity of Umbilical Cord-Derived Mesenchymal Stromal Cells’ Extracellular Vesicles (MSC-EVs). Int. J. Mol. Sci. 2023, 24, 9216. https://doi.org/10.3390/ijms24119216
Wright A, Snyder O, He H, Christenson LK, Fleming S, Weiss ML. Procoagulant Activity of Umbilical Cord-Derived Mesenchymal Stromal Cells’ Extracellular Vesicles (MSC-EVs). International Journal of Molecular Sciences. 2023; 24(11):9216. https://doi.org/10.3390/ijms24119216
Chicago/Turabian StyleWright, Adrienne, Orman (Larry) Snyder, Hong He, Lane K. Christenson, Sherry Fleming, and Mark L. Weiss. 2023. "Procoagulant Activity of Umbilical Cord-Derived Mesenchymal Stromal Cells’ Extracellular Vesicles (MSC-EVs)" International Journal of Molecular Sciences 24, no. 11: 9216. https://doi.org/10.3390/ijms24119216