Semipermeable Cellulose Beads Allow Selective and Continuous Release of Small Extracellular Vesicles (sEV) From Encapsulated Cells

Frontiers in Pharmacology
Gabriela ZavalaMaroun Khoury

Abstract

The clinical benefit of therapies using Mesenchymal Stem Cells (MSCs) is attributable to their pleiotropic effect over cells and tissues, mainly through their secretome. This paracrine effect is mediated by secreted growth factors and extracellular vesicles (EV) including small EV (sEV). sEV are extra-cellular, membrane encompassed vesicles of 40 to 200 nm diameter that can trigger and signal many cellular responses depending on their cargo protein and nucleic acid repertoire. sEV are purified from cell culture conditioned media using several kits and protocols available that can be tedious and time-consuming, involving sequences of ultracentrifugations and density gradient separations, making their production a major challenge under Good Manufacturing Practices (GMP) conditions. We have developed a method to efficiently enrich cell culture media with high concentrations of sEV by encapsulating cells in semipermeable cellulose beads that allows selectively the release of small particles while offering a 3D culture condition. This method is based on the pore size of the capsules, allowing the release of particles of ≤ 200 nm including sEV. As a proof-of-principle, MSCs were encapsulated and their sEV release rate (sEV-Cap) was m...Continue Reading

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Citations

Jan 9, 2021·Stem Cell Research & Therapy·Mesude BicerDarius Widera
Nov 27, 2020·Drug Discovery Today·Tania B Lopez-MendezGorka Orive
Jun 15, 2021·Frontiers in Bioengineering and Biotechnology·Aliosha I Figueroa-ValdésFrancisca Alcayaga-Miranda

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Methods Mentioned

BETA
size exclusion chromatography
Assay
FACS
flow cytometry
electron microscopy
density-gradient centrifugation

Software Mentioned

ImageJ Analyze Particles
Olympus iTEM
WimTube
sEV
GraphPad Prism
ImageJ
FlowJo
Cap

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