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Anti-Inflammatory and Angiogenic Effects of Stem Cell Secretome

Title: Anti-Inflammatory and Angiogenic Effects of Stem Cell Secretome
Authors: Shawn P. Grogan; Grant Stinebaugh; Darryl D. D’Lima
Source: International Journal of Molecular Sciences ; Volume 27 ; Issue 5 ; Pages: 2325
Publisher Information: Multidisciplinary Digital Publishing Institute
Publication Year: 2026
Collection: MDPI Open Access Publishing
Subject Terms: mesenchymal stem cell secretome; extracellular vesicles (EV); exosomes; hypoxia-conditioned secretome; endothelial network formation and angiogenesis; osteoarthritis and human cartilage explants; cell-free regenerative therapy
Description: Mesenchymal stem cells (MSCs) exert biological effects in part through their secretome which includes extracellular vesicles. In this study, we isolated and characterized the secretome from clinically relevant stem cell lines: human embryonic stem cell–derived mesenchymal stem cell line (ES-MSCs) and Infrapatellar fat pad derived MSC (IPFP-MSC) cultured in xeno-free medium. We assessed the biological activity of concentrated cell secretome or isolated fractions of extracellular vesicles (EVs) on cell proliferation, microvascular formation, and cartilage degradation in a human osteoarthritic (OA) ex vivo model. Serum-free conditioned medium from ES-MSC (N = 1) or IPFP-MSC (N = 2) monolayer cultures were concentrated by ultrafiltration to generate concentrated conditioned medium (CCM). Size exclusion chromatography was used to fractionate extracellular vesicles (EVs). Vesicle size, concentration, morphology, and surface markers were characterized by nanoparticle tracking analysis, transmission electron microscopy, and flow cytometry. Biological activity was evaluated by treating human umbilical vein endothelial cells (HUVECs), IPFP-MSCs, and ES-MSCs with CCM and EVs at defined particle concentrations. Endothelial network formation was tested in fibrin gels with different cell and secretome combinations. For analysis of cartilage degradation, human cartilage explants (N = 4; 3.5 mm in diameter) were harvested from patients undergoing total knee arthroplasty and subjected to IL-1β stimulation to induce an OA phenotype. Explants were treated with varying doses from CCM or EVs. Release of glycosaminoglycan in the medium and RNA analysis of catabolic genes were used as readouts. Secretome preparations yielded on average approximately 50 billion vesicles per mL with a similar particle size distribution between 50–200 nm in ES-MSC and IPFP-MSC cultures. Transmission electron microscopy confirmed vesicle morphology and flow cytometry confirmed expression of exosomal surface markers (CD9, CD63, CD81). Functionally, CCM and ...
Document Type: text
File Description: application/pdf
Language: English
Relation: Molecular Biology; https://dx.doi.org/10.3390/ijms27052325
DOI: 10.3390/ijms27052325
Availability: https://doi.org/10.3390/ijms27052325
Rights: https://creativecommons.org/licenses/by/4.0/
Accession Number: edsbas.D47981F1
Database: BASE