Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Skovronova, Renata, Grange, Cristina, Dimuccio, Veronica, Deregibus, Maria Chiara, Camussi, Giovanni, Bussolati, Benedetta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8616318/
https://www.ncbi.nlm.nih.gov/pubmed/34831170
http://dx.doi.org/10.3390/cells10112948
_version_ 1784604318435901440
author Skovronova, Renata
Grange, Cristina
Dimuccio, Veronica
Deregibus, Maria Chiara
Camussi, Giovanni
Bussolati, Benedetta
author_facet Skovronova, Renata
Grange, Cristina
Dimuccio, Veronica
Deregibus, Maria Chiara
Camussi, Giovanni
Bussolati, Benedetta
author_sort Skovronova, Renata
collection PubMed
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 Online
Article
Text
id pubmed-8616318
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-86163182021-11-26 Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques Skovronova, Renata Grange, Cristina Dimuccio, Veronica Deregibus, Maria Chiara Camussi, Giovanni Bussolati, Benedetta Cells Article 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. MDPI 2021-10-29 /pmc/articles/PMC8616318/ /pubmed/34831170 http://dx.doi.org/10.3390/cells10112948 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Skovronova, Renata
Grange, Cristina
Dimuccio, Veronica
Deregibus, Maria Chiara
Camussi, Giovanni
Bussolati, Benedetta
Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques
title 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_short 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8616318/
https://www.ncbi.nlm.nih.gov/pubmed/34831170
http://dx.doi.org/10.3390/cells10112948
work_keys_str_mv AT skovronovarenata surfacemarkerexpressioninsmallandmediumlargemesenchymalstromalcellderivedextracellularvesiclesinnaiveorapoptoticconditionusingorthogonaltechniques
AT grangecristina surfacemarkerexpressioninsmallandmediumlargemesenchymalstromalcellderivedextracellularvesiclesinnaiveorapoptoticconditionusingorthogonaltechniques
AT dimuccioveronica surfacemarkerexpressioninsmallandmediumlargemesenchymalstromalcellderivedextracellularvesiclesinnaiveorapoptoticconditionusingorthogonaltechniques
AT deregibusmariachiara surfacemarkerexpressioninsmallandmediumlargemesenchymalstromalcellderivedextracellularvesiclesinnaiveorapoptoticconditionusingorthogonaltechniques
AT camussigiovanni surfacemarkerexpressioninsmallandmediumlargemesenchymalstromalcellderivedextracellularvesiclesinnaiveorapoptoticconditionusingorthogonaltechniques
AT bussolatibenedetta surfacemarkerexpressioninsmallandmediumlargemesenchymalstromalcellderivedextracellularvesiclesinnaiveorapoptoticconditionusingorthogonaltechniques