Preconditioned Mesenchymal Stromal Cell-Derived Extracellular Vesicles (EVs) Counteract Inflammaging
Abstract
:1. Introduction
2. Materials and Methods
2.1. Mice
2.2. Isolation and Culture of Human Adipose Tissue-Derived Mesenchymal Stromal Cells (hMSCs)
2.3. Isolation and Culture of Mouse Bone Marrow-MSCs (mBMSCs)
2.4. Collection of hMSC and mBMSC Conditioned Medium (CM)
2.5. Immunohistochemistry
2.6. Extracellular Vesicle (EV) Separation and Characterization
2.7. Macrophage Isolation and Culture
2.8. Macrophage Polarization and Treatment with EVs
2.9. RNA Extraction and Quantitative Real Time-PCR
2.10. Proteome Profiler Array
2.11. Evaluation of Oxygen Consumption Rate
2.12. FoF1 ATP-Synthase Activity Assay
2.13. P/O Ratio
2.14. Cell Homogenate Preparation
2.15. ATP and AMP Intracellular Content Evaluation
2.16. Lactate Dehydrogenase Activity Assay
2.17. Malondialdehyde Evaluation
2.18. Statistical Analysis
3. Results
3.1. The Bone Marrow (BM) of Aged Mice Contains a Higher Number of Pro-Inflammatory Macrophages Compared to the Young Counterpart
3.2. The Aged Microenvironment Induces an Accumulation of CD38POS BM-Macrophages
3.3. Characterization of HypINF-hMSC-Derived EVs
3.4. EVs Released by HypINF-hMSCs Exert an Anti-Inflammatory Role on Aged Macrophages
3.5. HypINF hMSCs-Derived EVs Counteract Age-Associated Mitochondrial Dysfunction on Macrophages
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gorgun, C.; Africano, C.; Ciferri, M.C.; Bertola, N.; Reverberi, D.; Quarto, R.; Ravera, S.; Tasso, R. Preconditioned Mesenchymal Stromal Cell-Derived Extracellular Vesicles (EVs) Counteract Inflammaging. Cells 2022, 11, 3695. https://doi.org/10.3390/cells11223695
Gorgun C, Africano C, Ciferri MC, Bertola N, Reverberi D, Quarto R, Ravera S, Tasso R. Preconditioned Mesenchymal Stromal Cell-Derived Extracellular Vesicles (EVs) Counteract Inflammaging. Cells. 2022; 11(22):3695. https://doi.org/10.3390/cells11223695
Chicago/Turabian StyleGorgun, Cansu, Chiara Africano, Maria Chiara Ciferri, Nadia Bertola, Daniele Reverberi, Rodolfo Quarto, Silvia Ravera, and Roberta Tasso. 2022. "Preconditioned Mesenchymal Stromal Cell-Derived Extracellular Vesicles (EVs) Counteract Inflammaging" Cells 11, no. 22: 3695. https://doi.org/10.3390/cells11223695
APA StyleGorgun, C., Africano, C., Ciferri, M. C., Bertola, N., Reverberi, D., Quarto, R., Ravera, S., & Tasso, R. (2022). Preconditioned Mesenchymal Stromal Cell-Derived Extracellular Vesicles (EVs) Counteract Inflammaging. Cells, 11(22), 3695. https://doi.org/10.3390/cells11223695