In Vitro Conditioning of Adipose-Derived Mesenchymal Stem Cells by the Endothelial Microenvironment: Modeling Cell Responsiveness towards Non-Genetic Correction of Haemophilia A
Abstract
:1. Introduction
2. Results
2.1. Characterization of Ad-MSC Cultures
2.2. Cell Migration Capacity
2.3. Endothelial Differentiation
2.4. FVIII Release
2.5. Coagulation Assay
3. Discussion
4. Materials and Methods
4.1. In Vitro Culture of Human Adipose-Derived Mesenchymal Stem Cells (Ad-MSCs)
4.2. Morphological Evaluation by Optical Microscopy
4.3. Immunophenotype Characterization by Flow Cytometry
4.4. Endothelial Differentiation
4.5. Cell Migration Assay
4.6. Acquisition of Endothelial-like Characteristics
4.6.1. Gene Expression Study by qPCR
4.6.2. Protein Expression Study by Flow Cytometry
4.7. FVIII Protein Study
4.8. Coagulation Assay
4.9. Statistics
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Target Gene | TaqMan® Probe ID | Species | Amplicon Length (bp) |
---|---|---|---|
CD34 | Hs02576480_m1 | Human | 63 |
PDGFRα | Hs00998026_m1 | Human | 60 |
VE-cadherin | Hs00170986_m1 | Human | 70 |
CD31 | Hs01065282_m1 | Human | 67 |
vWF | Hs01109446_m1 | Human | 56 |
FVIII | Hs00252034_m1 | Human | 127 |
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Barbon, S.; Stocco, E.; Rajendran, S.; Zardo, L.; Macchi, V.; Grandi, C.; Tagariello, G.; Porzionato, A.; Radossi, P.; De Caro, R.; et al. In Vitro Conditioning of Adipose-Derived Mesenchymal Stem Cells by the Endothelial Microenvironment: Modeling Cell Responsiveness towards Non-Genetic Correction of Haemophilia A. Int. J. Mol. Sci. 2022, 23, 7282. https://doi.org/10.3390/ijms23137282
Barbon S, Stocco E, Rajendran S, Zardo L, Macchi V, Grandi C, Tagariello G, Porzionato A, Radossi P, De Caro R, et al. In Vitro Conditioning of Adipose-Derived Mesenchymal Stem Cells by the Endothelial Microenvironment: Modeling Cell Responsiveness towards Non-Genetic Correction of Haemophilia A. International Journal of Molecular Sciences. 2022; 23(13):7282. https://doi.org/10.3390/ijms23137282
Chicago/Turabian StyleBarbon, Silvia, Elena Stocco, Senthilkumar Rajendran, Lorena Zardo, Veronica Macchi, Claudio Grandi, Giuseppe Tagariello, Andrea Porzionato, Paolo Radossi, Raffaele De Caro, and et al. 2022. "In Vitro Conditioning of Adipose-Derived Mesenchymal Stem Cells by the Endothelial Microenvironment: Modeling Cell Responsiveness towards Non-Genetic Correction of Haemophilia A" International Journal of Molecular Sciences 23, no. 13: 7282. https://doi.org/10.3390/ijms23137282