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Volume 23
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10.3390/ijms23052861
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Article

Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization

by
Miquel Saumell-Esnaola
1,2,†,
Diego Delgado
3,†,
Gontzal García del Caño
2,4,
Maider Beitia
3,
Joan Sallés
1,2,5,*,
Imanol González-Burguera
2,4,
Pello Sánchez
3,
Maider López de Jesús
1,2,
Sergio Barrondo
1,2,5 and
Mikel Sánchez
3,6,*
1
Department of Pharmacology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, Spain
2
Bioaraba, Neurofarmacología Celular y Molecular, 01008 Vitoria-Gasteiz, Spain
3
Advanced Biological Therapy Unit, Hospital Vithas Vitoria, 01008 Vitoria-Gasteiz, Spain
4
Department of Neurosciences, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, Spain
5
Centro de Investigación Biomédica en Red de Salud Mental (CIBERSAM), 28029 Madrid, Spain
6
Arthroscopic Surgery Unit, Hospital Vithas Vitoria, 01008 Vitoria-Gasteiz, Spain
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Int. J. Mol. Sci. 2022, 23(5), 2861; https://doi.org/10.3390/ijms23052861
Submission received: 31 January 2022 / Revised: 25 February 2022 / Accepted: 4 March 2022 / Published: 5 March 2022
(This article belongs to the Special Issue New Insights into the Roles of Platelet-Rich Plasma in Tissue Repair/Regeneration)
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Abstract

Platelet-Rich Plasma (PRP) is enriched in molecular messengers with restorative effects on altered tissue environments. Upon activation, platelets release a plethora of growth factors and cytokines, either in free form or encapsulated in exosomes, which have been proven to promote tissue repair and regeneration. Translational research on the potential of exosomes as a safe nanosystem for therapeutic cargo delivery requires standardizing exosome isolation methods along with their molecular and morphological characterization. With this aim, we isolated and characterized the exosomes released by human PRP platelets. Western blot analysis revealed that CaCl2-activated platelets (PLT-Exos-Ca2+) released more exosomes than non-activated ones (PLT-Exos). Moreover, PLT-Exos-Ca2+ exhibited a molecular signature that meets the most up-to-date biochemical criteria for platelet-derived exosomes and possessed morphological features typical of exosomes as assessed by transmission electron microscopy. Array analysis of 105 analytes including growth factors and cytokines showed that PLT-Exos-Ca2+ exhibited lower levels of most analytes compared to PLT-Exos, but relatively higher levels of those consistently validated as components of the protein cargo of platelet exosomes. In summary, the present study provides new insights into the molecular composition of human platelet-derived exosomes and validates a method for isolating highly pure platelet exosomes as a basis for future preclinical studies in regenerative medicine and drug delivery.
Keywords: platelet-rich plasma; human platelets; platelet-derived-exosomes; exosome markers; cytokines; growth factors platelet-rich plasma; human platelets; platelet-derived-exosomes; exosome markers; cytokines; growth factors

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MDPI and ACS Style

Saumell-Esnaola, M.; Delgado, D.; García del Caño, G.; Beitia, M.; Sallés, J.; González-Burguera, I.; Sánchez, P.; López de Jesús, M.; Barrondo, S.; Sánchez, M. Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization. Int. J. Mol. Sci. 2022, 23, 2861. https://doi.org/10.3390/ijms23052861

AMA Style

Saumell-Esnaola M, Delgado D, García del Caño G, Beitia M, Sallés J, González-Burguera I, Sánchez P, López de Jesús M, Barrondo S, Sánchez M. Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization. International Journal of Molecular Sciences. 2022; 23(5):2861. https://doi.org/10.3390/ijms23052861

Chicago/Turabian Style

Saumell-Esnaola, Miquel, Diego Delgado, Gontzal García del Caño, Maider Beitia, Joan Sallés, Imanol González-Burguera, Pello Sánchez, Maider López de Jesús, Sergio Barrondo, and Mikel Sánchez. 2022. "Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization" International Journal of Molecular Sciences 23, no. 5: 2861. https://doi.org/10.3390/ijms23052861

APA Style

Saumell-Esnaola, M., Delgado, D., García del Caño, G., Beitia, M., Sallés, J., González-Burguera, I., Sánchez, P., López de Jesús, M., Barrondo, S., & Sánchez, M. (2022). Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization. International Journal of Molecular Sciences, 23(5), 2861. https://doi.org/10.3390/ijms23052861

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