Coincubation as miR-Loading Strategy to Improve the Anti-Tumor Effect of Stem Cell-Derived EVs
Abstract
:1. Introduction
2. Materials and Methods
2.1. Renal Cancer Stem Cells Isolation and Culture
2.2. Human Liver Stem Cells Isolation and Culture
2.3. HLSC-EVs Isolation and Characterization
2.4. Electroporation Protocol
2.5. Coincubation Protocol
2.6. Cytofluorimetric EV Analysis
2.7. Apoptosis
2.8. Invasion
2.9. Super-Resolution Microscopy
2.10. EV Incorporation in Target Cells
2.11. miRNA Isolation and Real Time PCR
2.12. Protein Extraction and Western Blot
2.13. Statistical Analysis
3. Results
3.1. Comparison between Electroporation and Coincubation to Increase the EV Antitumor Effect
3.2. RNAse Treatment of Coincubated EVs
3.3. Anti-Tumor Effect and miR145 Transfer of RNase Treated Coincubated EVs
3.4. Protection of Surface Loaded miRNAs by RNA Binding Proteins
3.5. Coincubation Protocol Using Antitumor miRNAs
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Brossa, A.; Tapparo, M.; Fonsato, V.; Papadimitriou, E.; Delena, M.; Camussi, G.; Bussolati, B. Coincubation as miR-Loading Strategy to Improve the Anti-Tumor Effect of Stem Cell-Derived EVs. Pharmaceutics 2021, 13, 76. https://doi.org/10.3390/pharmaceutics13010076
Brossa A, Tapparo M, Fonsato V, Papadimitriou E, Delena M, Camussi G, Bussolati B. Coincubation as miR-Loading Strategy to Improve the Anti-Tumor Effect of Stem Cell-Derived EVs. Pharmaceutics. 2021; 13(1):76. https://doi.org/10.3390/pharmaceutics13010076
Chicago/Turabian StyleBrossa, Alessia, Marta Tapparo, Valentina Fonsato, Elli Papadimitriou, Michela Delena, Giovanni Camussi, and Benedetta Bussolati. 2021. "Coincubation as miR-Loading Strategy to Improve the Anti-Tumor Effect of Stem Cell-Derived EVs" Pharmaceutics 13, no. 1: 76. https://doi.org/10.3390/pharmaceutics13010076
APA StyleBrossa, A., Tapparo, M., Fonsato, V., Papadimitriou, E., Delena, M., Camussi, G., & Bussolati, B. (2021). Coincubation as miR-Loading Strategy to Improve the Anti-Tumor Effect of Stem Cell-Derived EVs. Pharmaceutics, 13(1), 76. https://doi.org/10.3390/pharmaceutics13010076