Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study
Since the discovery of the beneficial therapeutical effects of extracellular vesicles (EVs), these agents have been attracting great interest as next-generation therapies. EVs are nanosized membrane bodies secreted by all types of cells that mediate cell–cell communication. Although the classificati...
| Autores: | , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Fecha de publicación: | 2022 |
| País: | España |
| Institución: | Universidad de Navarra |
| Repositorio: | Dadun. Depósito Académico Digital de la Universidad de Navarra |
| Idioma: | inglés |
| OAI Identifier: | oai:dadun.unav.edu:10171/63709 |
| Acceso en línea: | https://hdl.handle.net/10171/63709 |
| Access Level: | acceso abierto |
| Palabra clave: | Large and small extracellular vesicles Cardiovascular progenitors Ultracentrifugation Size-exclusion chromatography |
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Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative studySaludas-Echauri, L. (Laura)|||/items/944a0427-e0fd-47ba-9388-10417608d96bGarbayo, E. (Elisa)|||/items/c0fa35e4-82c5-44f7-b300-740027106dbaRuiz-Villalba, A. (Adrián)|||/items/feebed90-4714-4668-93cc-0267fd191547Hernández, S.C. (Silvia C.)|||/items/3189736d-aa68-465f-80ed-f05ec2748af8Vader, P. (Pieter)|||/items/396919de-52fd-4f36-9a38-f42af61021b0Prosper-Cardoso, F. (Felipe)|||/items/3d1b0b82-06c3-4e63-8280-e903dc4dc0c1Blanco-Prieto, M.J. (María José)|||/items/93e177db-635f-456f-b672-b79ef8befc40Large and small extracellular vesiclesCardiovascular progenitorsUltracentrifugationSize-exclusion chromatographySince the discovery of the beneficial therapeutical effects of extracellular vesicles (EVs), these agents have been attracting great interest as next-generation therapies. EVs are nanosized membrane bodies secreted by all types of cells that mediate cell–cell communication. Although the classification of different subpopulations of EVs can be complex, they are broadly divided into microvesicles and exosomes based on their biogenesis and in large and small EVs based on their size. As this is an emerging field, current investigations are focused on basic aspects such as the more convenient method for EV isolation. In the present paper, we used cardiac progenitor cells (CPCs) to study and compare different cell culture conditions for EV isolation as well as two of the most commonly employed purification methods: ultracentrifugation (UC) and size-exclusion chromatography (SEC). Large and small EVs were separately analysed. We found that serum starvation of cells during the EV collecting period led to a dramatic decrease in EV secretion and major cell death. Regarding the isolation method, our findings suggest that UC and SEC gave similar EV recovery rates. Separation of large and small EV-enriched subpopulations was efficiently achieved with both purification protocols although certain difference in sample heterogeneity was observed. Noteworthy, while calnexin was abundant in large EVs, ALIX and CD63 were mainly found in small EVs. Finally, when the functionality of EVs was assessed on primary culture of adult murine cardiac fibroblasts, we found that EVs were taken up by these cells, which resulted in a pronounced reduction in the proliferative and migratory capacity of the cells. Specifically, a tendency towards a larger effect of SEC-related EVs was observed. No differences could be found between large and small EVs. Altogether, these results contribute to establish the basis for the use of EVs as therapeutic platforms, in particular in regenerative fields.ElsevierDadun. Depósito Académico Digital Universidad de Navarra20222022-06-2820222022-01-0120222022-01-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10171/63709reponame:Dadun. Depósito Académico Digital de la Universidad de Navarrainstname:Universidad de NavarraInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:dadun.unav.edu:10171/637092026-06-21T12:47:57Z |
| dc.title.none.fl_str_mv |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study |
| title |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study |
| spellingShingle |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study Saludas-Echauri, L. (Laura)|||/items/944a0427-e0fd-47ba-9388-10417608d96b Large and small extracellular vesicles Cardiovascular progenitors Ultracentrifugation Size-exclusion chromatography |
| title_short |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study |
| title_full |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study |
| title_fullStr |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study |
| title_full_unstemmed |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study |
| title_sort |
Isolation methods of large and small extracellular vesicles derived from cardiovascular progenitors: A comparative study |
| dc.creator.none.fl_str_mv |
Saludas-Echauri, L. (Laura)|||/items/944a0427-e0fd-47ba-9388-10417608d96b Garbayo, E. (Elisa)|||/items/c0fa35e4-82c5-44f7-b300-740027106dba Ruiz-Villalba, A. (Adrián)|||/items/feebed90-4714-4668-93cc-0267fd191547 Hernández, S.C. (Silvia C.)|||/items/3189736d-aa68-465f-80ed-f05ec2748af8 Vader, P. (Pieter)|||/items/396919de-52fd-4f36-9a38-f42af61021b0 Prosper-Cardoso, F. (Felipe)|||/items/3d1b0b82-06c3-4e63-8280-e903dc4dc0c1 Blanco-Prieto, M.J. (María José)|||/items/93e177db-635f-456f-b672-b79ef8befc40 |
| author |
Saludas-Echauri, L. (Laura)|||/items/944a0427-e0fd-47ba-9388-10417608d96b |
| author_facet |
Saludas-Echauri, L. (Laura)|||/items/944a0427-e0fd-47ba-9388-10417608d96b Garbayo, E. (Elisa)|||/items/c0fa35e4-82c5-44f7-b300-740027106dba Ruiz-Villalba, A. (Adrián)|||/items/feebed90-4714-4668-93cc-0267fd191547 Hernández, S.C. (Silvia C.)|||/items/3189736d-aa68-465f-80ed-f05ec2748af8 Vader, P. (Pieter)|||/items/396919de-52fd-4f36-9a38-f42af61021b0 Prosper-Cardoso, F. (Felipe)|||/items/3d1b0b82-06c3-4e63-8280-e903dc4dc0c1 Blanco-Prieto, M.J. (María José)|||/items/93e177db-635f-456f-b672-b79ef8befc40 |
| author_role |
author |
| author2 |
Garbayo, E. (Elisa)|||/items/c0fa35e4-82c5-44f7-b300-740027106dba Ruiz-Villalba, A. (Adrián)|||/items/feebed90-4714-4668-93cc-0267fd191547 Hernández, S.C. (Silvia C.)|||/items/3189736d-aa68-465f-80ed-f05ec2748af8 Vader, P. (Pieter)|||/items/396919de-52fd-4f36-9a38-f42af61021b0 Prosper-Cardoso, F. (Felipe)|||/items/3d1b0b82-06c3-4e63-8280-e903dc4dc0c1 Blanco-Prieto, M.J. (María José)|||/items/93e177db-635f-456f-b672-b79ef8befc40 |
| author2_role |
author author author author author author |
| dc.contributor.none.fl_str_mv |
Dadun. Depósito Académico Digital Universidad de Navarra |
| dc.subject.none.fl_str_mv |
Large and small extracellular vesicles Cardiovascular progenitors Ultracentrifugation Size-exclusion chromatography |
| topic |
Large and small extracellular vesicles Cardiovascular progenitors Ultracentrifugation Size-exclusion chromatography |
| description |
Since the discovery of the beneficial therapeutical effects of extracellular vesicles (EVs), these agents have been attracting great interest as next-generation therapies. EVs are nanosized membrane bodies secreted by all types of cells that mediate cell–cell communication. Although the classification of different subpopulations of EVs can be complex, they are broadly divided into microvesicles and exosomes based on their biogenesis and in large and small EVs based on their size. As this is an emerging field, current investigations are focused on basic aspects such as the more convenient method for EV isolation. In the present paper, we used cardiac progenitor cells (CPCs) to study and compare different cell culture conditions for EV isolation as well as two of the most commonly employed purification methods: ultracentrifugation (UC) and size-exclusion chromatography (SEC). Large and small EVs were separately analysed. We found that serum starvation of cells during the EV collecting period led to a dramatic decrease in EV secretion and major cell death. Regarding the isolation method, our findings suggest that UC and SEC gave similar EV recovery rates. Separation of large and small EV-enriched subpopulations was efficiently achieved with both purification protocols although certain difference in sample heterogeneity was observed. Noteworthy, while calnexin was abundant in large EVs, ALIX and CD63 were mainly found in small EVs. Finally, when the functionality of EVs was assessed on primary culture of adult murine cardiac fibroblasts, we found that EVs were taken up by these cells, which resulted in a pronounced reduction in the proliferative and migratory capacity of the cells. Specifically, a tendency towards a larger effect of SEC-related EVs was observed. No differences could be found between large and small EVs. Altogether, these results contribute to establish the basis for the use of EVs as therapeutic platforms, in particular in regenerative fields. |
| publishDate |
2022 |
| dc.date.none.fl_str_mv |
2022 2022-06-28 2022 2022-01-01 2022 2022-01-01 |
| dc.type.none.fl_str_mv |
journal article http://purl.org/coar/resource_type/c_6501 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
https://hdl.handle.net/10171/63709 |
| url |
https://hdl.handle.net/10171/63709 |
| dc.language.none.fl_str_mv |
Inglés eng |
| language_invalid_str_mv |
Inglés |
| language |
eng |
| dc.rights.none.fl_str_mv |
open access http://purl.org/coar/access_right/c_abf2 |
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info:eu-repo/semantics/openAccess |
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open access http://purl.org/coar/access_right/c_abf2 |
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openAccess |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Elsevier |
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Elsevier |
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reponame:Dadun. Depósito Académico Digital de la Universidad de Navarra instname:Universidad de Navarra |
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Universidad de Navarra |
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Dadun. Depósito Académico Digital de la Universidad de Navarra |
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Dadun. Depósito Académico Digital de la Universidad de Navarra |
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15.300719 |