Characterization of MSC Growth, Differentiation, and EV Production in CNF Hydrogels Under Static and Dynamic Cultures in Hypoxic and Normoxic Conditions
Abstract
1. Introduction
2. Materials and Methods
2.1. Cells
2.1.1. adMSCs
2.1.2. PBMCs
2.2. Preparation of to-CNFs
2.3. Cell Encapsulation in to-CNF Hydrogels Spheres for In Vitro Cultivation
2.4. to-CNF Hydrogel Sphere Cultivation in Bioreactors
2.5. Live/Dead Cell Visualization
2.6. Cell Metabolic Activity Assessment
2.7. DNA Quantification
2.8. Isolation of Extracellular Vesicles
2.9. Protein Concentration Assay
2.10. Nanoparticle Tracking Analysis
2.11. PBMC Co-Culture with adMSCs or adMSC-EVs
2.12. Flow Cytometry Analysis
2.13. Extracellular Vesicle Surface Marker Analysis
2.14. Adipogenic Differentiation
2.15. Intracellular Lipid Droplet Staining
2.16. Mechanical Characterization of to-CNF Hydrogels
2.17. Statistical Analysis
3. Results
3.1. Effect of CaCl2-Induced Crosslinking on Mechanical Properties of to-CNF Hydrogel Spheres and Growth Dynamics of Encapsulated adMSCs
3.2. Dynamic Cultivation Enhances adMSC Proliferation and Protein Secretion in to-CNF Hydrogel Spheres
3.3. Hypoxia Enhances adMSC Proliferation and Protein Secretion in to-CNF Hydrogel Spheres
3.4. Enhanced Adipogenic Differentiation of adMSCs in to-CNF Hydrogel Spheres Under Hypoxic Conditions
3.5. Generation and Characterization of Immunomodulatory EVs by to-CNF Hydrogel Sphere-Encapsulated adMSCs
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Nikolits, I.; Chariyev-Prinz, F.; Egger, D.; Liebner, F.; Mytzka, N.; Kasper, C. Characterization of MSC Growth, Differentiation, and EV Production in CNF Hydrogels Under Static and Dynamic Cultures in Hypoxic and Normoxic Conditions. Bioengineering 2024, 11, 1050. https://doi.org/10.3390/bioengineering11101050
Nikolits I, Chariyev-Prinz F, Egger D, Liebner F, Mytzka N, Kasper C. Characterization of MSC Growth, Differentiation, and EV Production in CNF Hydrogels Under Static and Dynamic Cultures in Hypoxic and Normoxic Conditions. Bioengineering. 2024; 11(10):1050. https://doi.org/10.3390/bioengineering11101050
Chicago/Turabian StyleNikolits, Ilias, Farhad Chariyev-Prinz, Dominik Egger, Falk Liebner, Nicolas Mytzka, and Cornelia Kasper. 2024. "Characterization of MSC Growth, Differentiation, and EV Production in CNF Hydrogels Under Static and Dynamic Cultures in Hypoxic and Normoxic Conditions" Bioengineering 11, no. 10: 1050. https://doi.org/10.3390/bioengineering11101050
APA StyleNikolits, I., Chariyev-Prinz, F., Egger, D., Liebner, F., Mytzka, N., & Kasper, C. (2024). Characterization of MSC Growth, Differentiation, and EV Production in CNF Hydrogels Under Static and Dynamic Cultures in Hypoxic and Normoxic Conditions. Bioengineering, 11(10), 1050. https://doi.org/10.3390/bioengineering11101050