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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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/5/2861 |
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| author | Miquel Saumell-Esnaola Diego Delgado Gontzal García del Caño Maider Beitia Joan Sallés Imanol González-Burguera Pello Sánchez Maider López de Jesús Sergio Barrondo Mikel Sánchez |
| author_facet | Miquel Saumell-Esnaola Diego Delgado Gontzal García del Caño Maider Beitia Joan Sallés Imanol González-Burguera Pello Sánchez Maider López de Jesús Sergio Barrondo Mikel Sánchez |
| author_sort | Miquel Saumell-Esnaola |
| collection | DOAJ |
| description | 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 CaCl<sub>2</sub>-activated platelets (PLT-Exos-Ca<sup>2+</sup>) released more exosomes than non-activated ones (PLT-Exos). Moreover, PLT-Exos-Ca<sup>2+</sup> 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-Ca<sup>2+</sup> 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. |
| first_indexed | 2024-03-09T20:35:41Z |
| format | Article |
| id | doaj.art-25b1bb1c54d44b17a9711ac90fb08cc9 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T20:35:41Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-25b1bb1c54d44b17a9711ac90fb08cc92023-11-23T23:10:49ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-03-01235286110.3390/ijms23052861Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological CharacterizationMiquel Saumell-Esnaola0Diego Delgado1Gontzal García del Caño2Maider Beitia3Joan Sallés4Imanol González-Burguera5Pello Sánchez6Maider López de Jesús7Sergio Barrondo8Mikel Sánchez9Department of Pharmacology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, SpainAdvanced Biological Therapy Unit, Hospital Vithas Vitoria, 01008 Vitoria-Gasteiz, SpainBioaraba, Neurofarmacología Celular y Molecular, 01008 Vitoria-Gasteiz, SpainAdvanced Biological Therapy Unit, Hospital Vithas Vitoria, 01008 Vitoria-Gasteiz, SpainDepartment of Pharmacology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, SpainBioaraba, Neurofarmacología Celular y Molecular, 01008 Vitoria-Gasteiz, SpainAdvanced Biological Therapy Unit, Hospital Vithas Vitoria, 01008 Vitoria-Gasteiz, SpainDepartment of Pharmacology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, SpainDepartment of Pharmacology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, SpainAdvanced Biological Therapy Unit, Hospital Vithas Vitoria, 01008 Vitoria-Gasteiz, SpainPlatelet-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 CaCl<sub>2</sub>-activated platelets (PLT-Exos-Ca<sup>2+</sup>) released more exosomes than non-activated ones (PLT-Exos). Moreover, PLT-Exos-Ca<sup>2+</sup> 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-Ca<sup>2+</sup> 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.https://www.mdpi.com/1422-0067/23/5/2861platelet-rich plasmahuman plateletsplatelet-derived-exosomesexosome markerscytokinesgrowth factors |
| spellingShingle | Miquel Saumell-Esnaola Diego Delgado Gontzal García del Caño Maider Beitia Joan Sallés Imanol González-Burguera Pello Sánchez Maider López de Jesús Sergio Barrondo Mikel Sánchez Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization International Journal of Molecular Sciences platelet-rich plasma human platelets platelet-derived-exosomes exosome markers cytokines growth factors |
| title | Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization |
| title_full | Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization |
| title_fullStr | Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization |
| title_full_unstemmed | Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization |
| title_short | Isolation of Platelet-Derived Exosomes from Human Platelet-Rich Plasma: Biochemical and Morphological Characterization |
| title_sort | isolation of platelet derived exosomes from human platelet rich plasma biochemical and morphological characterization |
| topic | platelet-rich plasma human platelets platelet-derived-exosomes exosome markers cytokines growth factors |
| url | https://www.mdpi.com/1422-0067/23/5/2861 |
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