NG2 as an Identity and Quality Marker of Mesenchymal Stem Cell Extracellular Vesicles

Barilani, Mario; Peli, Valeria; Cherubini, Alessandro; Dossena, Marta; Dolo, Vincenza; Lazzari, Lorenza

doi:10.3390/cells8121524

Cited by 19 publications

(23 citation statements)

References 38 publications

(55 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MSC-EV defining markers CD44, CD73 and CD90 were present ( Figure 1 e), as well as generic EV-markers CD63 and CD81 ( Figure 1 f). CD9, another postulated EV-marker, resulted expressed at very low levels ( Figure 1 f), as previously described for ASC-EVs [ 38 ] and fetal MSCs [ 39 ].…”

Section: Resultssupporting

confidence: 74%

In Vitro Study of Extracellular Vesicles Migration in Cartilage-Derived Osteoarthritis Samples Using Real-Time Quantitative Multimodal Nonlinear Optics Imaging

et al. 2020

View full text Add to dashboard Cite

Mesenchymal stromal cells (MSCs)-derived extracellular vesicles (EVs) are promising therapeutic nano-carriers for the treatment of osteoarthritis (OA). The assessment of their uptake in tissues is mandatory but, to date, available technology does not allow to track and quantify incorporation in real-time. To fill this knowledge gap, the present study was intended to develop an innovative technology to determine kinetics of fluorescent MSC-EV uptake by means of time-lapse quantitative microscopy techniques. Adipose-derived mesenchymal stromal cells (ASCs)-EVs were fluorescently labeled and tracked during their uptake into chondrocytes micromasses or cartilage explants, both derived from OA patients. Immunofluorescence and time-lapse coherent anti-Stokes Raman scattering, second harmonic generation and two-photon excited fluorescence were used to follow and quantify incorporation. EVs penetration appeared quickly after few minutes and reached 30–40 μm depth after 5 h in both explants and micromasses. In explants, uptake was slightly faster, with EVs signal overlapping both extracellular matrix and chondrocytes, whereas in micromasses a more homogenous diffusion was observed. The finding of this study demonstrates that this innovative technology is a powerful tool to monitor EVs migration in tissues characterized by a complex extracellular network, and to obtain data resembling in vivo conditions.

show abstract

Section: Resultssupporting

confidence: 74%

In Vitro Study of Extracellular Vesicles Migration in Cartilage-Derived Osteoarthritis Samples Using Real-Time Quantitative Multimodal Nonlinear Optics Imaging

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Additionally, hAMSC-EVs strongly expressed both CD63 and CD81, consistent with previously reported characteristics of EVs ( Figure 1E). Eventually, CD9, another EV marker, staining gave a weak signal of the entire population ( Figure 1E), similarly to vesicles derived from other MSC types [64]. EVs isolated from hAMSCs were first analyzed by nanoparticle tracking analysis (NTA).…”

Section: Hamscs and Evs Characterizationmentioning

confidence: 93%

miRNA Reference Genes in Extracellular Vesicles Released from Amniotic Membrane-Derived Mesenchymal Stromal Cells

et al. 2020

View full text Add to dashboard Cite

Human amniotic membrane and amniotic membrane-derived mesenchymal stromal cells (hAMSCs) have produced promising results in regenerative medicine, especially for the treatment of inflammatory-based diseases and for different injuries including those in the orthopedic field such as tendon disorders. hAMSCs have been proposed to exert their anti-inflammatory and healing potential via secreted factors, both free and conveyed within extracellular vesicles (EVs). In particular, EV miRNAs are considered privileged players due to their impact on target cells and tissues, and their future use as therapeutic molecules is being intensely investigated. In this view, EV-miRNA quantification in either research or future clinical products has emerged as a crucial paradigm, although, to date, largely unsolved due to lack of reliable reference genes (RGs). In this study, a panel of thirteen putative miRNA RGs (let-7a-5p, miR-16-5p, miR-22-5p, miR-23a-3p, miR-26a-5p, miR-29a-5p, miR-101-3p, miR-103a-3p, miR-221-3p, miR-423-5p, miR-425-5p, miR-660-5p and U6 snRNA) that were identified in different EV types was assessed in hAMSC-EVs. A validated experimental pipeline was followed, sifting the output of four largely accepted algorithms for RG prediction (geNorm, NormFinder, BestKeeper and ΔCt method). Out of nine RGs constitutively expressed across all EV isolates, miR-101-3p and miR-22-5p resulted in the most stable RGs, whereas miR-423-5p and U6 snRNA performed poorly. miR-22-5p was also previously reported to be a reliable RG in adipose-derived MSC-EVs, suggesting its suitability across samples isolated from different MSC types. Further, to shed light on the impact of incorrect RG choice, the level of five tendon-related miRNAs (miR-29a-3p, miR-135a-5p, miR-146a-5p, miR-337-3p, let-7d-5p) was compared among hAMSC-EVs isolates. The use of miR-423-5p and U6 snRNA did not allow a correct quantification of miRNA incorporation in EVs, leading to less accurate fingerprinting and, if used for potency prediction, misleading indication of the most appropriate clinical batch. These results emphasize the crucial importance of RG choice for EV-miRNAs in hAMSCs studies and contribute to the identification of reliable RGs such as miR-101-3p and miR-22-5p to be validated in other MSC-EVs related fields.

show abstract

“…A postulated mechanism of growth factor transport is through binding to transmembrane EV proteins [63], which demonstrates the capability of EVs to serve as a transport vehicle for secreted proteins and thereby contributing to long-distance delivery.…”

Section: Neovascularization and Vascular Integritymentioning

confidence: 99%

Extracellular Vesicle-Associated Proteins in Tissue Repair

Roefs

Sluijter

Vader

2020

Trends in Cell Biology

119

View full text Add to dashboard Cite

The administration of (stem) cell-derived extracellular vesicles (EVs) promotes tissue repair through management of different inflammatory, proliferative and remodeling processes in the body. Despite the widely observed biological and therapeutic roles of EVs in wound healing and tissue repair, knowledge on how EVs activate recipient cells and which EV cargo is responsible for the subsequent functional effects is limited. Recent studies hint toward an important role for proteins as functional EV cargo. Here, we provide an overview of how EV-associated proteins promote tissue repair processes and discuss current challenges in evaluating their contribution to EV function and future directions for translating fundamental insights into clinically relevant EV therapies. Extracellular Vesicles: Natural Vehicles of Biological Cargo in Tissue Homeostasis and RepairApart from conventional secretory pathways (see Glossary), cells secrete proteins, nucleic acids, and lipids that are delivered to other cells through vesicles generated via the endosomal pathway or upon direct budding of the plasma membrane. Collectively, these vesicles are called extracellular vesicles (EVs). Despite initial thoughts about being nothing more than the garbage disposal system of cells, it is now well established that these EVs also play important roles in tissue homeostasis through short-and long-distance intercellular communication. EVs isolated from multiple cell types, tissues, and body fluids promote endogenous tissue repair through the modulation of several physiological processes in the body. As a result, EVs bear potential as therapeutics in regenerative medicine, and have been demonstrated to be a functional component of the stem and progenitor cell secretome [1]. Indeed, EVs have been found to promote different reparative processes in the body. Although many studies have set out to identify the functional cargo and cell activation pathways of EVs in tissue repair, the exact mechanisms by which these EVs exert their effects remain elusive. In this review, we describe why proteins expressed on (or in) EVs might be important contributors to EV function, and address the role of specific EV proteins in tissue repair processes in the body. Challenges in the study of individual EV-associated proteins and the implication for the translation of these insights into EV therapeutics will be discussed. EV Biogenesis and Recipient Cell ActivationEVs may be classified based on a set of characteristics including their size, density, process of cellular release, and their protein/lipid composition [2,3]. Three such subgroups are exosomes, microvesicles, and apoptotic bodies (Box 1). Although distinct in biogenesis, it is hard to capture and assign the heterogeneous population of EVs upon isolation toward different classes as they share overlapping physiochemical characteristics and biological markers [4]. To encompass all different vesicle types, the general term EV is increasingly used, as also in this review. EVs are enriched in distinct proteins and...

show abstract

NG2 as an Identity and Quality Marker of Mesenchymal Stem Cell Extracellular Vesicles

Cited by 19 publications

References 38 publications

In Vitro Study of Extracellular Vesicles Migration in Cartilage-Derived Osteoarthritis Samples Using Real-Time Quantitative Multimodal Nonlinear Optics Imaging

In Vitro Study of Extracellular Vesicles Migration in Cartilage-Derived Osteoarthritis Samples Using Real-Time Quantitative Multimodal Nonlinear Optics Imaging

miRNA Reference Genes in Extracellular Vesicles Released from Amniotic Membrane-Derived Mesenchymal Stromal Cells

Extracellular Vesicle-Associated Proteins in Tissue Repair

Contact Info

Product

Resources

About