Extracellular Vesicles Generated by Mesenchymal Stem Cells in Stirred Suspension Bioreactors Promote Angiogenesis in Human-Brain-Derived Endothelial Cells
Abstract
:1. Introduction
2. Results
2.1. SSB Culture Affects MSC Proliferation and Gene Expression, with Dependence on Confluence
2.2. Dynamic Culture Conditions and Cell Confluency Alter EV Secretion and Composition
2.3. EVs Isolated from MSCs Cultured in SSBs Have Improved Angiogenic Properties
3. Discussion
4. Materials and Methods
4.1. MSC Culture
4.2. RT-qPCR Analysis of MSCs
4.3. EV Isolation
4.4. TEM
4.5. SP-IRIS
4.6. Luminex Multiplex Analysis
4.7. CMEC Culture
4.8. Proliferation Assay
4.9. Tube Formation Assay
4.10. Statistical Methods
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Primer Sequence (5′-3′) | Origin |
---|---|---|
18S | F: TGG TCG CTC GCT CCT CTC C R: CGC CTG CTG CCT TCC TTG G | NR_003286 |
ANG1 | F: CCT GAT CTT ACA CGG TGC R: GCT TTC ATA ATC GCT TCT | NM_001314051 |
BCL2 | F: GAT GAC TGA GTA CCT GAA CC R: AGT TCC ACA AAG GCA TCC | EU287875 |
FGF2 | F: CGC GGT TGC AAC GGG AT R: GGG TTC ACG GAT GGT TGT CT | NM_27968 |
HIF1 | F: CCA GTT ACG TTC CTT CGA TCA GT R: TTT GAG GAC TTG CGC TTT CA | NM_001243084 |
HMOX1 | F: ATG ACA CCA AGG ACC AGA GC R: GTG TAA GGA CCC ATC GGA GA | NM_002133 |
IL6 | F: TCA ATA TTA GAG TCT CAA CCC CCA R: TTC TCT TTC GTT CCC GGT GG | NM_000600 |
MCP1 | F: GCA ATC AAT GCC CCA GTC AC R: TCT TTG GGA CAC TTG CTG CT | S71513 |
SDF1 | F: GGA CTT TCC GCT AGA CCC AC R: GCC CGA TCC CAG ATC AAT GT | NM_199168 |
TGFB1 | F: GGG GAA ATT GAG GGC TTT CG R: CCA GGA CCT TGC TGT ACT GC | NM_000660 |
VEGF | F: ACG GTC CCT CTT GGA ATT GG R: GGC CGC GGT GTG TCT A | M32977 |
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Phelps, J.; Hart, D.A.; Mitha, A.P.; Duncan, N.A.; Sen, A. Extracellular Vesicles Generated by Mesenchymal Stem Cells in Stirred Suspension Bioreactors Promote Angiogenesis in Human-Brain-Derived Endothelial Cells. Int. J. Mol. Sci. 2024, 25, 5219. https://doi.org/10.3390/ijms25105219
Phelps J, Hart DA, Mitha AP, Duncan NA, Sen A. Extracellular Vesicles Generated by Mesenchymal Stem Cells in Stirred Suspension Bioreactors Promote Angiogenesis in Human-Brain-Derived Endothelial Cells. International Journal of Molecular Sciences. 2024; 25(10):5219. https://doi.org/10.3390/ijms25105219
Chicago/Turabian StylePhelps, Jolene, David A. Hart, Alim P. Mitha, Neil A. Duncan, and Arindom Sen. 2024. "Extracellular Vesicles Generated by Mesenchymal Stem Cells in Stirred Suspension Bioreactors Promote Angiogenesis in Human-Brain-Derived Endothelial Cells" International Journal of Molecular Sciences 25, no. 10: 5219. https://doi.org/10.3390/ijms25105219