Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma
Abstract Small extracellular vesicles (sEV) purified from blood have great potential clinically as biomarkers for systemic disease; however interpretation is complicated by release of sEV ex vivo after blood taking. To quantify the problem and devise ways to minimise it, we characterised sEV in pair...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-05-01
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| Series: | Journal of Extracellular Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jex2.88 |
| _version_ | 1827888324309680128 |
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| author | Małgorzata S. Małys Maximilian C. Köller Kristin Papp Christof Aigner Daffodil Dioso Patrick Mucher Helga Schachner Michael Bonelli Helmuth Haslacher Andrew J. Rees Renate Kain |
| author_facet | Małgorzata S. Małys Maximilian C. Köller Kristin Papp Christof Aigner Daffodil Dioso Patrick Mucher Helga Schachner Michael Bonelli Helmuth Haslacher Andrew J. Rees Renate Kain |
| author_sort | Małgorzata S. Małys |
| collection | DOAJ |
| description | Abstract Small extracellular vesicles (sEV) purified from blood have great potential clinically as biomarkers for systemic disease; however interpretation is complicated by release of sEV ex vivo after blood taking. To quantify the problem and devise ways to minimise it, we characterised sEV in paired serum, plasma and platelet poor plasma (PPP) samples from healthy donors. Immunoblotting showed twofold greater abundance of CD9 in sEV fractions from fresh serum than from fresh plasma or PPP. MACSPlex confirmed this, and showed that proteins expressed on platelet sEV, either exclusively (CD41b, CD42a and CD62P) or more widely (HLA‐ABC, CD24, CD29 and CD31) were also twofold more abundant; by contrast non‐platelet proteins (including CD81) were no different. Storage of plasma (but not serum) increased abundance of platelet and selected leukocyte sEV proteins to at least that of serum, and this could be recapitulated by activating cells in fresh plasma by Ca2+, an effect abrogated in PPP. This suggests that a substantial proportion of sEV in serum and stored plasma were generated ex vivo, which is not the case for fresh plasma or PPP. Thus we provide strategies to minimise ex vivo sEV generation and criteria for identifying those that were present in vivo. |
| first_indexed | 2024-03-12T20:32:05Z |
| format | Article |
| id | doaj.art-35b7093b746b459cacb6441565783024 |
| institution | Directory Open Access Journal |
| issn | 2768-2811 |
| language | English |
| last_indexed | 2024-03-12T20:32:05Z |
| publishDate | 2023-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Biology |
| spelling | doaj.art-35b7093b746b459cacb64415657830242023-08-01T19:00:30ZengWileyJournal of Extracellular Biology2768-28112023-05-0125n/an/a10.1002/jex2.88Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasmaMałgorzata S. Małys0Maximilian C. Köller1Kristin Papp2Christof Aigner3Daffodil Dioso4Patrick Mucher5Helga Schachner6Michael Bonelli7Helmuth Haslacher8Andrew J. Rees9Renate Kain10Department of Pathology Medical University Vienna Vienna AustriaDepartment of Pathology Medical University Vienna Vienna AustriaDepartment of Pathology Medical University Vienna Vienna AustriaDepartment of Pathology Medical University Vienna Vienna AustriaDepartment of Internal Medicine III, Division of Rheumatology Medical University of Vienna Vienna AustriaDepartment of Laboratory Medicine Medical University of Vienna Vienna AustriaDepartment of Pathology Medical University Vienna Vienna AustriaDepartment of Internal Medicine III, Division of Rheumatology Medical University of Vienna Vienna AustriaDepartment of Laboratory Medicine Medical University of Vienna Vienna AustriaDepartment of Pathology Medical University Vienna Vienna AustriaDepartment of Pathology Medical University Vienna Vienna AustriaAbstract Small extracellular vesicles (sEV) purified from blood have great potential clinically as biomarkers for systemic disease; however interpretation is complicated by release of sEV ex vivo after blood taking. To quantify the problem and devise ways to minimise it, we characterised sEV in paired serum, plasma and platelet poor plasma (PPP) samples from healthy donors. Immunoblotting showed twofold greater abundance of CD9 in sEV fractions from fresh serum than from fresh plasma or PPP. MACSPlex confirmed this, and showed that proteins expressed on platelet sEV, either exclusively (CD41b, CD42a and CD62P) or more widely (HLA‐ABC, CD24, CD29 and CD31) were also twofold more abundant; by contrast non‐platelet proteins (including CD81) were no different. Storage of plasma (but not serum) increased abundance of platelet and selected leukocyte sEV proteins to at least that of serum, and this could be recapitulated by activating cells in fresh plasma by Ca2+, an effect abrogated in PPP. This suggests that a substantial proportion of sEV in serum and stored plasma were generated ex vivo, which is not the case for fresh plasma or PPP. Thus we provide strategies to minimise ex vivo sEV generation and criteria for identifying those that were present in vivo.https://doi.org/10.1002/jex2.88biobankingexosomesextracellular vesiclesex vivo generationMACSPlexplatelets |
| spellingShingle | Małgorzata S. Małys Maximilian C. Köller Kristin Papp Christof Aigner Daffodil Dioso Patrick Mucher Helga Schachner Michael Bonelli Helmuth Haslacher Andrew J. Rees Renate Kain Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma Journal of Extracellular Biology biobanking exosomes extracellular vesicles ex vivo generation MACSPlex platelets |
| title | Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma |
| title_full | Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma |
| title_fullStr | Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma |
| title_full_unstemmed | Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma |
| title_short | Small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma |
| title_sort | small extracellular vesicles are released ex vivo from platelets into serum and from residual blood cells into stored plasma |
| topic | biobanking exosomes extracellular vesicles ex vivo generation MACSPlex platelets |
| url | https://doi.org/10.1002/jex2.88 |
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