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Purification of High Yield Extracellular Vesicle Preparations Away from Virus
Published on: September 12, 2019
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Extracellular vesicles have variable dose-dependent effects on cultured draining cells in the eye
Saray Tabak1, Sofia Schreiber-Avissar1, Elie Beit-Yannai1
1Clinical Biochemistry and Pharmacology Department, The Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Journal of Cellular and Molecular Medicine
|February 8, 2018
Summary
Extracellular vesicles (EVs) exhibit concentration-dependent effects on Wnt signaling and MMP activity in ocular cells. Low EV doses decreased Wnt signaling proteins, while high doses enhanced MMP activity, revealing crucial dose-response relationships.
Area of Science:
- Biochemistry
- Cell Biology
- Ophthalmology
Background:
- Extracellular vesicles (EVs) mediate biological signals, but their effective dosage remains unknown.
- Understanding EV concentration effects is vital for their therapeutic applications.
Purpose of the Study:
- To investigate the dose-response effects of non-pigmented ciliary epithelium (NPCE)-derived EVs on Wnt signaling in normal trabecular meshwork (NTM) cells.
- To determine how varying EV concentrations influence gene and protein expression related to Wnt signaling and matrix metalloproteinases (MMPs).
Main Methods:
- EVs were isolated from NPCE and RPE cells using the PEG 8000 method.
- EV concentrations were quantified using Tunable Resistive Pulse Sensing.
- NTM cells were exposed to different EV concentrations, followed by analysis of Wnt pathway gene/protein expression (qPCR, Western blot) and MMP activity.
Main Results:
- Low NPCE-EV concentrations reduced β-Catenin and GSK-3β expression in NTM cells.
- High NPCE-EV concentrations significantly increased Pro-MMP9 and MMP9 activity.
- A concentration-dependent effect was observed for gene expression and MMP activity, distinct from Wnt protein responses.
Conclusions:
- EV biological effects are concentration-dependent in target cells.
- This study elucidates specific dose-response patterns for EVs in the human ocular drainage system model, impacting Wnt signaling and MMP activity.
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