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

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 isolat...

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Main Authors: Marta Clerici, Maria Camilla Ciardulli, Erwin Pavel Lamparelli, Joseph Lovecchio, Emanuele Giordano, Tina P. Dale, Nicholas R. Forsyth, Nicola Maffulli, Giovanna Della Porta
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Artificial Cells, Nanomedicine, and Biotechnology
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Online Access:https://www.tandfonline.com/doi/10.1080/21691401.2025.2475099
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author Marta Clerici
Maria Camilla Ciardulli
Erwin Pavel Lamparelli
Joseph Lovecchio
Emanuele Giordano
Tina P. Dale
Nicholas R. Forsyth
Nicola Maffulli
Giovanna Della Porta
author_facet Marta Clerici
Maria Camilla Ciardulli
Erwin Pavel Lamparelli
Joseph Lovecchio
Emanuele Giordano
Tina P. Dale
Nicholas R. Forsyth
Nicola Maffulli
Giovanna Della Porta
author_sort Marta Clerici
collection DOAJ
description 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.
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spelling doaj-art-9ea70ce6d15e4776a27611c53bab81bc2025-08-20T03:05:25ZengTaylor & Francis GroupArtificial Cells, Nanomedicine, and Biotechnology2169-14012169-141X2025-12-0153111610.1080/21691401.2025.2475099Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic cultureMarta Clerici0Maria Camilla Ciardulli1Erwin Pavel Lamparelli2Joseph Lovecchio3Emanuele Giordano4Tina P. Dale5Nicholas R. Forsyth6Nicola Maffulli7Giovanna Della Porta8Department of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, Baronissi, Salerno, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, Baronissi, Salerno, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, Baronissi, Salerno, ItalySchool of Science and Engineering, Reykjavík University, Reykjavík, IcelandDepartment of Electrical, Electronic and Information Engineering “Guglielmo Marconi” (DEI), University of Bologna, Cesena, ItalySchool of Pharmacy and Bioengineering, Keele University, Stoke-on-Trent, UKSchool of Pharmacy and Bioengineering, Keele University, Stoke-on-Trent, UKSchool of Pharmacy and Bioengineering, Keele University, Stoke-on-Trent, UKDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, Baronissi, Salerno, ItalyTendon 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.https://www.tandfonline.com/doi/10.1080/21691401.2025.2475099Extracellular vesiclestendon stem/progenitor cellstendon tissue engineeringbioreactors
spellingShingle Marta Clerici
Maria Camilla Ciardulli
Erwin Pavel Lamparelli
Joseph Lovecchio
Emanuele Giordano
Tina P. Dale
Nicholas R. Forsyth
Nicola Maffulli
Giovanna Della Porta
Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture
Artificial Cells, Nanomedicine, and Biotechnology
Extracellular vesicles
tendon stem/progenitor cells
tendon tissue engineering
bioreactors
title Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture
title_full Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture
title_fullStr Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture
title_full_unstemmed Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture
title_short Human tendon stem/progenitor cell-derived extracellular vesicle production promoted by dynamic culture
title_sort human tendon stem progenitor cell derived extracellular vesicle production promoted by dynamic culture
topic Extracellular vesicles
tendon stem/progenitor cells
tendon tissue engineering
bioreactors
url https://www.tandfonline.com/doi/10.1080/21691401.2025.2475099
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