scholarly journals Chemical characterization of extracellular vesicles of mesenchymal stromal cells: TOF-SIMS and BCARS approach

2021 ◽  
Vol 2086 (1) ◽  
pp. 012107
Author(s):  
A V Aybush ◽  
A A Gulin ◽  
A A Kuzoiatova ◽  
M V Gubina ◽  
F E Gostev ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Stromal Cells ◽  
Chemical Characterization ◽  
Time Frame ◽  
Signaling Proteins ◽  
Label Free ◽  
Reasonable Time Frame ◽  
Membrane Bound ◽  
Scale Data

Abstract Paracrine functions of mesenchymal stem (stromal) cells (MSCs) rely, at least partly, on membrane-bound extracellular vesicles (EVs) with rich composition of lipids, nucleic acids and signaling proteins. Elucidation the underlying chemistry could potentially lead to MSCs-free therapy. However, the secretome of MSCs (EVs’ composition) is non-static and depends on many other factors including surrounding cells and medium. Thus, the research techniques must be able to provide not only bulk but microscopy-scale data within a reasonable time frame. Two of these label-free techniques are subject of this work toward the question of chemical composition of the EVs.

scholarly journals Label-Free Prostate Cancer Detection by Characterization of Extracellular Vesicles Using Raman Spectroscopy

2018 ◽  
Vol 90 (19) ◽  
pp. 11290-11296 ◽  
Author(s):  
Wooje Lee ◽  
Afroditi Nanou ◽  
Linda Rikkert ◽  
Frank A. W. Coumans ◽  
Cees Otto ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Raman Spectroscopy ◽  
Extracellular Vesicles ◽  
Cancer Detection ◽  
Label Free ◽  
Prostate Cancer Detection

Characterization of extracellular vesicles derived from two populations of human placenta derived mesenchymal stem/stromal cells

Cytotherapy ◽  
2020 ◽  
Vol 22 (5) ◽  
pp. S50
Author(s):  
R. Khanabdali ◽  
M. Shojaee ◽  
J. Johnson ◽  
S. Law ◽  
M. Whitmore ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Human Placenta ◽  
Stromal Cells ◽  
Two Populations

scholarly journals Mechanical Characterization of Extracellular Vesicles Derived from Immortalized Adipose Stromal Cells

2020 ◽  
Vol 118 (3) ◽  
pp. 252a
Author(s):  
Melissa C. Piontek ◽  
Sourav Maity ◽  
Linda A. Brouwer ◽  
Martin C. Harmsen ◽  
Wouter H. Roos
Keyword(s):  
Extracellular Vesicles ◽  
Stromal Cells ◽  
Mechanical Characterization ◽  
Adipose Stromal Cells

scholarly journals Comprehensive label-free characterization of extracellular vesicles and their surface proteins

2020 ◽  
Author(s):  
E. Priglinger ◽  
J. Strasser ◽  
B. Buchroithner ◽  
F. Weber ◽  
S. Wolbank ◽  
...  
Keyword(s):  
Quality Control ◽  
Extracellular Vesicles ◽  
Impedance Measurement ◽  
Surface Proteins ◽  
Label Free ◽  
Neuroprotective Effects ◽  
Characterization Methods ◽  
Minimal Sample ◽  
Flow Filtration

AbstractInterest in mesenchymal stem cell derived extracellular vesicles (MSC-EVs) as therapeutic agents has dramatically increased over the last decade. Preclinical studies show that MSC-EVs have anti-apoptotic and neuroprotective effects, boost wound healing, and improve the integration of allogeneic grafts through immunomodulation. Current approaches to the characterization and quality control of EV-based therapeutics include particle tracking techniques, Western blotting, and advanced cytometry, but standardized methods are lacking. In this study, we established and verified quartz crystal microbalance (QCM) as highly sensitive label-free immunosensing technique for characterizing clinically approved umbilical cord MSC-EVs enriched by tangential flow filtration and ultracentrifugation. Using QCM in conjunction with common characterization methods, we were able to specifically detect EVs via EV (CD9, CD63, CD81) and MSC (CD44, CD49e, CD73) markers and gauge their prevalence. Additionally, we characterized the topography and elasticity of these EVs by atomic force microscopy (AFM), enabling us to distinguish between EVs and non-vesicular particles (NVPs) in a therapeutic formulation. This measurement modality makes it possible to identify EV sub-fractions, discriminate between EVs and NVPs, and to characterize EV surface proteins, all with minimal sample preparation and using label-free measurement devices with low barriers of entry for labs looking to widen their spectrum of characterization techniques. Our combination of QCM with impedance measurement (QCM-I) and AFM measurements provides a robust multi-marker approach to the characterization of clinically approved EV formulations and opens the door to improved quality control.

Isolation and Characterization of Extracellular Vesicles from Mesenchymal Stromal Cells

2019 ◽  
pp. 15-23 ◽  
Author(s):  
Sally Yunsun Kim ◽  
Thanh Huyen Phan ◽  
Christina Limantoro ◽  
Bill Kalionis ◽  
Wojciech Chrzanowski
Keyword(s):  
Extracellular Vesicles ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Isolation And Characterization

scholarly journals Raman Fingerprint of Extracellular Vesicles and Conditioned Media for the Reproducibility Assessment of Cell-Free Therapeutics

2021 ◽  
Vol 9 ◽  
Author(s):  
Cristiano Carlomagno ◽  
Chiara Giannasi ◽  
Stefania Niada ◽  
Marzia Bedoni ◽  
Alice Gualerzi ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Stromal Cells ◽  
Cell Types ◽  
Dermal Fibroblasts ◽  
Cell Secretion ◽  
Free Products ◽  
Regenerative Effect ◽  
Different Cell Types ◽  
Non Destructive

Extracellular Vesicles (EVs) and Conditioned Medium (CM) are promising cell-free approaches to repair damaged and diseased tissues for regenerative rehabilitation purposes. They both entail several advantages, mostly in terms of safety and handling, compared to the cell-based treatment. Despite the growing interest in both EVs and CM preparations, in the light of a clinical translation, a number of aspects still need to be addressed mainly because of limits in the reproducibility and reliability of the proposed protocols. Raman spectroscopy (RS) is a non-destructive vibrational investigation method that provides detailed information about the biochemical composition of a sample, with reported ability in bulk characterization of clusters of EVs from different cell types. In the present brief report, we acquired and compared the Raman spectra of the two most promising cell-free therapeutics, i.e., EVs and CM, derived from two cytotypes with a history in the field of regenerative medicine, adipose-derived mesenchymal stem/stromal cells (ASCs) and dermal fibroblasts (DFs). Our results show how RS can verify the reproducibility not only of EV isolation, but also of the whole CM, thus accounting for both the soluble and the vesicular components of cell secretion. RS can provide hints for the identification of the soluble factors that synergistically cooperate with EVs in the regenerative effect of CM. Still, we believe that the application of RS in the pipeline of cell-free products preparation for therapeutic purposes could help in accelerating translation to clinics and regulatory approval.

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