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

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...

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Vydáno v:International journal of molecular sciences Ročník 23; číslo 5; s. 2861
Hlavní autoři: Saumell-Esnaola, Miquel, Delgado, Diego, García del Caño, Gontzal, Beitia, Maider, Sallés, Joan, González-Burguera, Imanol, Sánchez, Pello, López de Jesús, Maider, Barrondo, Sergio, Sánchez, Mikel
Médium: Journal Article
Jazyk:angličtina
Vydáno: Switzerland MDPI AG 05.03.2022
MDPI
Témata:
Antigens
Apoptosis
Biological products
Blood platelets
Blood Platelets - metabolism
Chemokines
Cytokines
Cytokines - metabolism
Endoplasmic reticulum
Exosomes - metabolism
Growth factors
Humans
Melanoma
Morphology
Plasma
Platelet-Rich Plasma - metabolism
Proteins
Regenerative medicine
Wound Healing
platelet-derived-exosomes
cytokines
growth factors
platelet-rich plasma
exosome markers
human platelets
ISSN:1422-0067, 1661-6596, 1422-0067
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Shrnutí: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.
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These authors contributed equally to this work.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms23052861