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Isolation, Characterization, and Therapeutic Application of Extracellular Vesicles from Cultured Human Mesenchymal Stem Cells
Published on: September 23, 2022
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Cellular In Vitro Responses Induced by Human Mesenchymal Stem/Stromal Cell-Derived Extracellular Vesicles Obtained
Ingrid L M Souza1, Andreia A Suzukawa1, Raphaella Josino2
1Laboratory of Basic Biology of Stem Cells (Labcet), Carlos Chagas Institute, Fiocruz, Curitiba 81350-010, PR, Brazil.
International Journal of Molecular Sciences
|July 27, 2024
Summary
Extracellular vesicles (EVs) from adipose-derived mesenchymal stem cells (ADMSCs) cultured on microcarriers promote skin cell proliferation, migration, and blood vessel formation. These ADMSC-EVs also help balance inflammatory responses in macrophages, supporting tissue regeneration potential.
Area of Science:
- Regenerative Medicine
- Biomaterials Science
- Cell Biology
Background:
- Mesenchymal stem/stromal cells (MSCs) and their extracellular vesicles (MSC-EVs) are crucial for tissue regeneration, exhibiting roles in repair, inflammation control, angiogenesis, and extracellular matrix remodeling.
- MSC-EVs offer advantages for regenerative therapies, including ease of dosing, histocompatibility, and low immunogenicity, reducing rejection risk.
Purpose of the Study:
- To evaluate the potential activity of adipose-derived MSC-EVs (ADMSC-EVs) cultured on cross-linked dextran microcarriers for scalable and reproducible EV production.
- To assess the impact of ADMSC-EVs on cellular responses including proliferation, migration, angiogenesis, and macrophage phenotype switching.
Main Methods:
- ADMSC-EVs were isolated from cells cultured on microcarriers.
- EVs were characterized using nanoparticle tracking analysis, electron microscopy, and immunophenotyping.
- ADMSC-EVs were applied to cultures of human dermal fibroblasts, keratinocytes, endothelial cells, and THP-1 derived macrophages to evaluate cellular responses.
Main Results:
- ADMSC-EVs significantly enhanced keratinocyte proliferation and fibroblast/keratinocyte migration.
- Endothelial cell cultures showed increased tube formation, indicating angiogenesis induction.
- Treatment with ADMSC-EVs modulated macrophage inflammatory cytokine expression, with higher doses increasing TGF-β and IL-1β in M1 macrophages.
Conclusions:
- EVs derived from microcarrier-cultured ADMSCs effectively modulate cell behavior, promoting skin cell migration and proliferation.
- These ADMSC-EVs stimulate angiogenesis and influence macrophage inflammatory responses, suggesting potential for tissue regeneration.
- Scalable isolation of EVs from ADMSC suspension cultures enables in vitro cellular responses for in vivo tissue regeneration studies.
