Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection
Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and purification from body fluids remains challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma sa...
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Published in | Scientific reports Vol. 6; no. 1; p. 24316 |
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Main Authors | , , , , , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
London
Nature Publishing Group UK
18.04.2016
Nature Publishing Group |
Subjects | |
Online Access | Get full text |
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Abstract | Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and purification from body fluids remains challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma samples as well as human platelet concentrates. Using flow cytometry, we found that the majority of circulating particles within the size range of extracellular vesicles lacked common vesicular markers. We identified most of these particles as lipoproteins (predominantly low-density lipoprotein, LDL) which mimicked the characteristics of extracellular vesicles and also co-purified with them. Based on biophysical properties of LDL this finding was highly unexpected. Current state-of-the-art extracellular vesicle isolation and purification methods did not result in lipoprotein-free vesicle preparations from blood plasma or from platelet concentrates. Furthermore, transmission electron microscopy showed an association of LDL with isolated vesicles upon
in vitro
mixing. This is the first study to show co-purification and
in vitro
association of LDL with extracellular vesicles and its interference with vesicle analysis. Our data point to the importance of careful study design and data interpretation in studies using blood-derived extracellular vesicles with special focus on potentially co-purified LDL. |
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AbstractList | Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and purification from body fluids remains challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma samples as well as human platelet concentrates. Using flow cytometry, we found that the majority of circulating particles within the size range of extracellular vesicles lacked common vesicular markers. We identified most of these particles as lipoproteins (predominantly low-density lipoprotein, LDL) which mimicked the characteristics of extracellular vesicles and also co-purified with them. Based on biophysical properties of LDL this finding was highly unexpected. Current state-of-the-art extracellular vesicle isolation and purification methods did not result in lipoprotein-free vesicle preparations from blood plasma or from platelet concentrates. Furthermore, transmission electron microscopy showed an association of LDL with isolated vesicles upon in vitro mixing. This is the first study to show co-purification and in vitro association of LDL with extracellular vesicles and its interference with vesicle analysis. Our data point to the importance of careful study design and data interpretation in studies using blood-derived extracellular vesicles with special focus on potentially co-purified LDL. Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and purification from body fluids remains challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma samples as well as human platelet concentrates. Using flow cytometry, we found that the majority of circulating particles within the size range of extracellular vesicles lacked common vesicular markers. We identified most of these particles as lipoproteins (predominantly low-density lipoprotein, LDL) which mimicked the characteristics of extracellular vesicles and also co-purified with them. Based on biophysical properties of LDL this finding was highly unexpected. Current state-of-the-art extracellular vesicle isolation and purification methods did not result in lipoprotein-free vesicle preparations from blood plasma or from platelet concentrates. Furthermore, transmission electron microscopy showed an association of LDL with isolated vesicles upon in vitro mixing. This is the first study to show co-purification and in vitro association of LDL with extracellular vesicles and its interference with vesicle analysis. Our data point to the importance of careful study design and data interpretation in studies using blood-derived extracellular vesicles with special focus on potentially co-purified LDL. Abstract Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and purification from body fluids remains challenging. Here we studied human pre-prandial and 4 hours postprandial platelet-free blood plasma samples as well as human platelet concentrates. Using flow cytometry, we found that the majority of circulating particles within the size range of extracellular vesicles lacked common vesicular markers. We identified most of these particles as lipoproteins (predominantly low-density lipoprotein, LDL) which mimicked the characteristics of extracellular vesicles and also co-purified with them. Based on biophysical properties of LDL this finding was highly unexpected. Current state-of-the-art extracellular vesicle isolation and purification methods did not result in lipoprotein-free vesicle preparations from blood plasma or from platelet concentrates. Furthermore, transmission electron microscopy showed an association of LDL with isolated vesicles upon in vitro mixing. This is the first study to show co-purification and in vitro association of LDL with extracellular vesicles and its interference with vesicle analysis. Our data point to the importance of careful study design and data interpretation in studies using blood-derived extracellular vesicles with special focus on potentially co-purified LDL. |
ArticleNumber | 24316 |
Author | Falus, András Pállinger, Éva Buzás, Edit Irén Vukman, Krisztina V Drahos, László Giricz, Zoltán Vékey, Károly Szabó-Taylor, Katalin Pálóczi, Krisztina Wiener, Zoltán Sperlágh, Beáta Ferdinandy, Péter Németh, Andrea Turiák, Lilla Kittel, Ágnes Sódar, Barbara W Osteikoetxea, Xabier Baranyai, Tamás |
Author_xml | – sequence: 1 givenname: Barbara W surname: Sódar fullname: Sódar, Barbara W organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 2 givenname: Ágnes surname: Kittel fullname: Kittel, Ágnes organization: Institute of Experimental Medicine, Hungarian Academy of Sciences – sequence: 3 givenname: Krisztina surname: Pálóczi fullname: Pálóczi, Krisztina organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 4 givenname: Krisztina V surname: Vukman fullname: Vukman, Krisztina V organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 5 givenname: Xabier surname: Osteikoetxea fullname: Osteikoetxea, Xabier organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 6 givenname: Katalin surname: Szabó-Taylor fullname: Szabó-Taylor, Katalin organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 7 givenname: Andrea surname: Németh fullname: Németh, Andrea organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 8 givenname: Beáta surname: Sperlágh fullname: Sperlágh, Beáta organization: Institute of Experimental Medicine, Hungarian Academy of Sciences – sequence: 9 givenname: Tamás surname: Baranyai fullname: Baranyai, Tamás organization: Department of Pharmacology and Pharmacotherapy, Semmelweis University – sequence: 10 givenname: Zoltán surname: Giricz fullname: Giricz, Zoltán organization: Department of Pharmacology and Pharmacotherapy, Semmelweis University – sequence: 11 givenname: Zoltán surname: Wiener fullname: Wiener, Zoltán organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 12 givenname: Lilla surname: Turiák fullname: Turiák, Lilla organization: Research Centre for Natural Sciences, Hungarian Academy of Sciences – sequence: 13 givenname: László surname: Drahos fullname: Drahos, László organization: Research Centre for Natural Sciences, Hungarian Academy of Sciences – sequence: 14 givenname: Éva surname: Pállinger fullname: Pállinger, Éva organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 15 givenname: Károly surname: Vékey fullname: Vékey, Károly organization: Research Centre for Natural Sciences, Hungarian Academy of Sciences – sequence: 16 givenname: Péter surname: Ferdinandy fullname: Ferdinandy, Péter organization: Department of Pharmacology and Pharmacotherapy, Semmelweis University – sequence: 17 givenname: András surname: Falus fullname: Falus, András organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University – sequence: 18 givenname: Edit Irén surname: Buzás fullname: Buzás, Edit Irén organization: Department of Genetics, Cell- and Immunobiology, Semmelweis University |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/27087061$$D View this record in MEDLINE/PubMed |
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Snippet | Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their isolation and... Abstract Circulating extracellular vesicles have emerged as potential new biomarkers in a wide variety of diseases. Despite the increasing interest, their... |
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SubjectTerms | 14/28 631/1647/1407/1492 631/1647/2230 82/16 82/58 Adult Biomarkers - blood Blood Blood platelets Blood Platelets - chemistry Body fluids Data interpretation Data processing Electron microscopy Exosomes Exosomes - chemistry Extracellular Vesicles - chemistry Female Flow cytometry Humanities and Social Sciences Humans Lipoproteins Lipoproteins, LDL - blood Low density lipoprotein Male multidisciplinary Postprandial Period Purification Science Transmission electron microscopy Vesicles |
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Title | Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection |
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