Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells

Apoptotic vesicles (apoVs) are specific extracellular vesicles generated during apoptosis and play important roles in multiple physiological and pathophysiological settings. Here, we present a protocol using differential centrifugation to separate apoVs from human mesenchymal stem cells (MSCs) after...

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Detalles Bibliográficos
Autores principales: Chen, Da, Zhao, Zixuan, Zhang, Kai, Jin, Fang, Zheng, Chenxi, Jin, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9494288/
https://www.ncbi.nlm.nih.gov/pubmed/36129822
http://dx.doi.org/10.1016/j.xpro.2022.101695
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author Chen, Da
Zhao, Zixuan
Zhang, Kai
Jin, Fang
Zheng, Chenxi
Jin, Yan
author_facet Chen, Da
Zhao, Zixuan
Zhang, Kai
Jin, Fang
Zheng, Chenxi
Jin, Yan
author_sort Chen, Da
collection PubMed
description Apoptotic vesicles (apoVs) are specific extracellular vesicles generated during apoptosis and play important roles in multiple physiological and pathophysiological settings. Here, we present a protocol using differential centrifugation to separate apoVs from human mesenchymal stem cells (MSCs) after induction of apoptosis. We describe how to characterize apoV size and morphology by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM), and determination of specific biomarker expression by immunoblotting. Our protocol will be useful for preparing apoVs for further functional analysis. For complete details on the use and execution of this protocol, please refer to Zheng et al. (2021).
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spelling pubmed-94942882022-09-23 Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells Chen, Da Zhao, Zixuan Zhang, Kai Jin, Fang Zheng, Chenxi Jin, Yan STAR Protoc Protocol Apoptotic vesicles (apoVs) are specific extracellular vesicles generated during apoptosis and play important roles in multiple physiological and pathophysiological settings. Here, we present a protocol using differential centrifugation to separate apoVs from human mesenchymal stem cells (MSCs) after induction of apoptosis. We describe how to characterize apoV size and morphology by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM), and determination of specific biomarker expression by immunoblotting. Our protocol will be useful for preparing apoVs for further functional analysis. For complete details on the use and execution of this protocol, please refer to Zheng et al. (2021). Elsevier 2022-09-20 /pmc/articles/PMC9494288/ /pubmed/36129822 http://dx.doi.org/10.1016/j.xpro.2022.101695 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Chen, Da
Zhao, Zixuan
Zhang, Kai
Jin, Fang
Zheng, Chenxi
Jin, Yan
Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells
title Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells
title_full Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells
title_fullStr Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells
title_full_unstemmed Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells
title_short Protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells
title_sort protocol for differential centrifugation-based separation and characterization of apoptotic vesicles derived from human mesenchymal stem cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9494288/
https://www.ncbi.nlm.nih.gov/pubmed/36129822
http://dx.doi.org/10.1016/j.xpro.2022.101695
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