Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques

Extracellular vesicles released by mesenchymal stromal cells (MSC-EVs) are a promising resource for regenerative medicine. Small MSC-EVs represent the active EV fraction. A bulk analysis was applied to characterise MSC-EVs’ identity and purity, with the assessment of single EV morphology, size and i...

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Autores principales:	Renata Skovronova, Cristina Grange, Veronica Dimuccio, Maria Chiara Deregibus, Giovanni Camussi, Benedetta Bussolati
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	tetraspanins 10k MSC EVs 100k MSC EVs Nanosight MACSPlex ExoView Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/b01389360faf461eb299e566e041847e
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spelling	oai:doaj.org-article:b01389360faf461eb299e566e041847e2021-11-25T17:09:37ZSurface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques10.3390/cells101129482073-4409https://doaj.org/article/b01389360faf461eb299e566e041847e2021-10-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/2948https://doaj.org/toc/2073-4409Extracellular vesicles released by mesenchymal stromal cells (MSC-EVs) are a promising resource for regenerative medicine. Small MSC-EVs represent the active EV fraction. A bulk analysis was applied to characterise MSC-EVs’ identity and purity, with the assessment of single EV morphology, size and integrity using electron microscopy. We applied different methods to quantitatively analyse the size and surface marker expression in medium/large and small fractions, namely 10k and 100k fractions, of MSC-EVs obtained using sequential ultracentrifugation. Bone marrow, adipose tissue and umbilical cord MSC-EVs were compared in naive and apoptotic conditions. As detected by electron microscopy, the 100k EV size < 100 nm was confirmed by super-resolution microscopy and ExoView. Single-vesicle imaging using super-resolution microscopy revealed heterogeneous patterns of tetraspanins. ExoView allowed a comparative screening of single MSC-EV tetraspanin and mesenchymal markers. A semiquantitative bead-based cytofluorimetric analysis showed the segregation of immunological and pro-coagulative markers on the 10k MSC-EVs. Apoptotic MSC-EVs were released in higher numbers, without significant differences in the naive fractions in surface marker expression. These results show a consistent profile of MSC-EV fractions among the different sources and a safer profile of the 100k MSC-EV population for clinical application. Our study identified suitable applications for EV analytical techniques.Renata SkovronovaCristina GrangeVeronica DimuccioMaria Chiara DeregibusGiovanni CamussiBenedetta BussolatiMDPI AGarticletetraspanins10k MSC EVs100k MSC EVsNanosightMACSPlexExoViewBiology (General)QH301-705.5ENCells, Vol 10, Iss 2948, p 2948 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	tetraspanins 10k MSC EVs 100k MSC EVs Nanosight MACSPlex ExoView Biology (General) QH301-705.5
spellingShingle	tetraspanins 10k MSC EVs 100k MSC EVs Nanosight MACSPlex ExoView Biology (General) QH301-705.5 Renata Skovronova Cristina Grange Veronica Dimuccio Maria Chiara Deregibus Giovanni Camussi Benedetta Bussolati Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques
description	Extracellular vesicles released by mesenchymal stromal cells (MSC-EVs) are a promising resource for regenerative medicine. Small MSC-EVs represent the active EV fraction. A bulk analysis was applied to characterise MSC-EVs’ identity and purity, with the assessment of single EV morphology, size and integrity using electron microscopy. We applied different methods to quantitatively analyse the size and surface marker expression in medium/large and small fractions, namely 10k and 100k fractions, of MSC-EVs obtained using sequential ultracentrifugation. Bone marrow, adipose tissue and umbilical cord MSC-EVs were compared in naive and apoptotic conditions. As detected by electron microscopy, the 100k EV size < 100 nm was confirmed by super-resolution microscopy and ExoView. Single-vesicle imaging using super-resolution microscopy revealed heterogeneous patterns of tetraspanins. ExoView allowed a comparative screening of single MSC-EV tetraspanin and mesenchymal markers. A semiquantitative bead-based cytofluorimetric analysis showed the segregation of immunological and pro-coagulative markers on the 10k MSC-EVs. Apoptotic MSC-EVs were released in higher numbers, without significant differences in the naive fractions in surface marker expression. These results show a consistent profile of MSC-EV fractions among the different sources and a safer profile of the 100k MSC-EV population for clinical application. Our study identified suitable applications for EV analytical techniques.
format	article
author	Renata Skovronova Cristina Grange Veronica Dimuccio Maria Chiara Deregibus Giovanni Camussi Benedetta Bussolati
author_facet	Renata Skovronova Cristina Grange Veronica Dimuccio Maria Chiara Deregibus Giovanni Camussi Benedetta Bussolati
author_sort	Renata Skovronova
title	Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques
title_short	Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques
title_full	Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques
title_fullStr	Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques
title_full_unstemmed	Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques
title_sort	surface marker expression in small and medium/large mesenchymal stromal cell-derived extracellular vesicles in naive or apoptotic condition using orthogonal techniques
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/b01389360faf461eb299e566e041847e
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Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques

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