Human Platelet-derived Extracellular Vesicles (EVs): A new EVs reference Material from Blood Components
Our method of isolation of platelet EVs ensure scalable manufacturing, reproducibility, and a consistent lipoprotein reduction.
Platelet Extracellular Vesicles
Platelet-derived Extracellular Vesicles (EVs) are involved in various physiological processes, including immune response, inflammation, and wound healing. In the last years, they have gained attention for their potential roles in diseases like cardiovascular disorders and cancer, as well as their therapeutic potential. Indeed, these EVs have shown promise in regenerative medicine, aiding in tissue repair, wound healing, and promoting angiogenesis [1,2]. Additionally, their ability to deliver therapeutic molecules, such as drugs or genetic material, makes them valuable for application in personalized and precision medicine.
In this technical note, we present the method used for the isolation of Platelet EVs, which ensures scalable manufacturing, reproducibility, and a consistent reduction in lipoprotein content compared to other commonly used methods.
Platelet EVs: Purification Workflow
Platelet EVs: Nanoparticle Characterization
Nanoparticle quality control was performed accordingly:
- Particle size distribution and concentration: Nanoparticle tracking analysis (NTA) and flow cytometry
- Marker expression: Flow cytometry, ELISA, Western blot
- Morphology: Transmission electron microscopy (TEM)
a) Nanoparticle Tracking Analysis
| Particle concentration | 2.2E+11 +/- 2.1E+10 part/ml |
|---|---|
| Particle size | 100.7 +/- 5.7 nm |
Table 1: Particle concentration and size measured by NTA
b) Nanoparticle Morphology
c) Marker Expression
Platelet EVs: Final Format
- Format: 100 μl vials, each one containing 2 - 2.5x10E+10 particles, in PBS 1x buffer.
- Certificate of analysis: including particle size distribution and concetration, measuread by NTA (Zetaview, Particle Metrix), assessement of 4 membrane markers (CD9, CD81,CD63,CD41), 1 internal marker (ALIX), 1 negative marker (Calnexin).
- Lyophilized (shipping and storing at 4°C) or frozen (-80°C) on request.
References
[1] Torres, N. P., Moradi, Z., Martini, A. C., Bischoff , S. R., & Ran, H. (2020). Exosomes derived from activated platelets as novel therapeutics in tissue repair and regeneration. Journal of Extracellular Vesicles, 9(1), 1750205. doi:10.1080/20013078.2020.1750205.
[2] Aatonen, M. T., Ohman, T., Nyman, T. A., Laitinen, S., Grönholm, M., & Siljander, P. R. M. (2014). Isolation and characterization of platelet-derived extracellular vesicles. Journal of Extracellular Vesicles, 3, 24692. doi:10.3402/jev.v3.24692