The effects of cell type and culture condition on the procoagulant activity of human mesenchymal stromal cell-derived extracellular vesicles
| Autor: | James A. Bynum, Andrew P. Cap, Robin M. Kamucheka, Christopher R. Rathbone, Grantham C. Peltier, Tiffani C. Chance |
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| Rok vydání: | 2019 |
| Předmět: |
Stromal cell
Cytological Techniques Thrombogenicity Critical Care and Intensive Care Medicine Flow cytometry Extracellular Vesicles 03 medical and health sciences chemistry.chemical_compound Tissue factor 0302 clinical medicine Humans Medicine Blood Coagulation Cells Cultured medicine.diagnostic_test business.industry Mesenchymal stem cell Mesenchymal Stem Cells 030208 emergency & critical care medicine Phosphatidylserine Extracellular vesicle Cell biology chemistry Surgery business Wound healing |
| Zdroj: | Journal of Trauma and Acute Care Surgery. 87:S74-S82 |
| ISSN: | 2163-0763 2163-0755 |
| DOI: | 10.1097/ta.0000000000002225 |
| Popis: | Background Mesenchymal stromal cell (MSC)-derived extracellular vesicles (EVs) have great potential as a cell-free therapy in wound healing applications. Because EV populations are not equivalent, rigorous characterization is needed before clinical use. Although there has been much focus on their RNA composition and regenerative capabilities, relatively less is known regarding the effects of MSC cell type (adipose tissue [Ad-MSCs] or bone marrow [BM-MSCs]) and culture condition (monolayer or spheroid) on MSC-EV performance, including characteristics related to their ability to promote coagulation, which could determine EV safety if administered intravenously. Methods The successful isolation of EVs derived from Ad-MSCs or BM-MSCs cultured in either monolayer or spheroid cultures was confirmed by NanoSight (particle size distribution) and Western blot (surface marker expression). Extracellular vesicle surface expression of procoagulant molecules (tissue factor and phosphatidylserine) was evaluated by flow cytometry. Extracellular vesicle thrombogenicity was tested using calibrated thrombogram, and clotting parameters were assessed using thromboelastography and a flow-based adhesion model simulating blood flow over a collagen-expressing surface. Results The MSC cell type and culture condition did not impact EV size distribution. Extracellular vesicles from all groups expressed phosphatidylserine and tissue factor on their surfaces were functionally thrombogenic and tended to increase clotting rates compared to the negative control of serum-free media without EVs. On average, EVs did not form significantly larger or stronger clots than the negative control, regardless of cell source or culture condition. Additionally, EVs interfered with platelet adhesion in an in vitro flow-based assay. Conclusion Adipose-derived EVs were more thrombogenic and expressed higher amounts of phosphatidylserine. Our findings suggest that, like intact MSCs, source variability among EVs is an important factor when considering EVs for potential therapeutic purposes. Level of evidence Therapeutic care management, level II. |
| Databáze: | OpenAIRE |
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