Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture

Clerici, Marta; Camilla Ciardulli, Maria; Pavel Lamparelli, Erwin; Lovecchio, Joseph; Giordano, Emanuele; Dale, Tina P.; Forsyth, Nicholas R.; Maffulli, Nicola; Della Porta, Giovanna

Abstract

Tendon injuries significantly impact quality of life, prompting the exploration of innovative solutions beyond conventional surgery. Extracellular Vesicles (EVs) have emerged as a promising strategy to enhance tendon regeneration. In this study, human Tendon Stem/Progenitor Cells (TSPCs) were isolated from surgical biopsies and cultured in a Growth-Differentiation Factor-5-supplemented medium to promote tenogenic differentiation under static and dynamic conditions using a custom-made perfusion bioreactor. Once at 80% confluence, cells were transitioned to a serum-free medium for conditioned media collection. Ultracentrifugation revealed the presence of vesicles with a 106 particles/mL concentration and sub-200nm diameter size. Dynamic culture yielded a 3-fold increase in EV protein content compared to static culture, as confirmed by Western-blot analysis. Differences in surface marker expression were also shown by flow cytometric analysis. Data suggest that we efficiently developed a protocol for extracting EVs from human TSPCs, particularly under dynamic conditions. This approach enhances EV protein content, offering potential therapeutic benefits for tendon regeneration. However, further research is needed to fully understand the role of EVs in tendon regeneration.

Citation

Clerici, M., Camilla Ciardulli, M., Pavel Lamparelli, E., Lovecchio, J., Giordano, E., Dale, T. P., …Della Porta, G. (2025). Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture. Artificial Cells, Nanomedicine, and Biotechnology, 53(1), https://doi.org/10.1080/21691401.2025.2475099

• Journal Article Type: Article
• Acceptance Date: Feb 24, 2025
• Online Publication Date: Mar 10, 2025
• Publication Date: Dec 31, 2025
• Deposit Date: Mar 18, 2025
• Publicly Available Date: Mar 18, 2025
• Journal: Artificial Cells, Nanomedicine, and Biotechnology
• Publisher: Taylor and Francis Group
• Peer Reviewed: Peer Reviewed
• Volume: 53
• Issue: 1
• DOI: https://doi.org/10.1080/21691401.2025.2475099
• Keywords: Extracellular vesicles; tendon stem/progenitor cells; tendon tissue engineering; bioreactors
• Public URL: https://keele-repository.worktribe.com/output/1107968
• Publisher URL: https://www.tandfonline.com/doi/full/10.1080/21691401.2025.2475099#abstract