ORIGINAL RESEARCH article
Front. Cell Dev. Biol.
Sec. Stem Cell Research
Volume 13 - 2025 | doi: 10.3389/fcell.2025.1633912
GMP-compliant, serum-free cultures preserve therapeutic potential of extracellular vesicles from human mesenchymal stromal cells
Provisionally accepted
Filippo Calascibetta1
Annalisa Martorana1Margot Lo Pinto1
Claudia Carcione1Salvatore D'Arpa2
Giandomenico Amico1
Vitale Miceli3Nicola Cuscino3
Gioacchin Iannolo3Lorenzo Volpe3
Simone Dario Scilabra1
Pier Giulio Conaldi3
Cinzia Maria Chinnici1*- 1Ri.MED Foundation, Palermo, Italy
- 2La Maddalena Clinic for Cancer, Palermo, Italy
- 3Istituto Mediterraneo per i Trapianti e Terapie ad Alta Specializzazione, Palermo, Italy
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The therapeutic potential of extracellular vesicles (EVs) derived from human mesenchymal stromal cells (MSCs) is limited by the lack of standardized, Good Manufacturing Practice (GMP)-compliant production protocols. This study investigates the effects of MSC-Brew, a commercially available GMP-grade medium, on MSC-derived EVs in comparison to those produced in conventional cultures with DMEM supplemented with 10% fetal bovine serum (FBS). MSCs from adult dermis were successfully isolated and expanded in Brew medium while retaining their characteristic surface marker expression. MSC-EVs derived from Brew cultures met the Minimal Information for Studies of Extracellular Vesicles (MISEV) criteria, including particle size, concentration, marker expression, and minimal inflammatory cytokine content. Notably, Brew-EVs exhibited a significantly higher particleto-protein ratio compared to EVs produced in FBS-containing cultures, indicating improved purity. Proteomic analysis revealed a largely conserved composition between Brew-EVs and conventionally produced EVs, and microRNA (miRNA) profiling identified only four differentially expressed miRNAs. Brew-EVs were enriched in anti-fibrotic miRNAs and effectively reduced collagen secretion in transforming growth factor (TGF)-β1-activated LX-2 cells, a human hepatic stellate cell line used as a model of liver fibrosis. These findings support MSC-Brew medium as a standardized, serum-free platform for the consistent production of high-quality EVs suitable for therapeutic applications.
Keywords: Mesenchymal stromal cells1, Extracellular vesicles2, GMP-compliant medium3, liver fibrosis4, LX-2 cells5, collagen secretion6
Received: 23 May 2025; Accepted: 25 Jul 2025.
Copyright: © 2025 Calascibetta, Martorana, Lo Pinto, Carcione, D'Arpa, Amico, Miceli, Cuscino, Iannolo, Volpe, Scilabra, Conaldi and Chinnici. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Cinzia Maria Chinnici, Ri.MED Foundation, Palermo, Italy
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