Abstract
BackgroundMesenchymal stem cells (MSCs) are pluripotent stromal cells that release extracellular vesicles (EVs). EVs contain various growth factors and antioxidants that can positively affect the surrounding cells. Nanoscale MSC-derived EVs, such as exosomes, have been developed as bio-stable nano-type materials. However, some issues, such as low yield and difficulty in quantification, limit their use. We hypothesized that enhancing exosome production using nanoparticles would stimulate the release of intracellular molecules.ResultsThe aim of this study was to elucidate the molecular mechanisms of exosome generation by comparing the internalization of surface-modified, positively charged nanoparticles and exosome generation from MSCs. We determined that Rab7, a late endosome and auto-phagosome marker, was increased upon exosome expression and was associated with autophagosome formation.ConclusionsIt was concluded that the nanoparticles we developed were transported to the lysosome by clathrin-mediated endocytosis. additionally, entered nanoparticles stimulated that autophagy related factors to release exosome from the MSC. MSC-derived exosomes using nanoparticles may increase exosome yield and enable the discovery of nanoparticle-induced genetic factors.
Highlights
Mesenchymal stem cells (MSCs) are pluripotent stromal cells that release extracellular vesicles (EVs)
When cells were treated with 10 μg/mL and 20 μg/mL NPs, viability decreased to 86.2% after 6 h and to 86% after 1 h, respectively. These findings indicated that the concentration of 5 μg/mL was optimal for internalization and that higher concentrations of positively charged polymeric clustered superparamagnetic (PCS) NPs were increasingly internalized by MSCs, affecting their viability (Fig. 2c)
The findings indicated that the PLGA-PEI PCS NPs were not internalized by clathrin-mediated endocytosis, and the approximately 40% inhibition by Dynasore was related to receptor-mediated endocytosis involving another mechanism (Fig. 4a, b)
Summary
Mesenchymal stem cells (MSCs) are pluripotent stromal cells that release extracellular vesicles (EVs). Nanoscale MSCderived EVs, such as exosomes, have been developed as bio-stable nano-type materials. Some issues, such as low yield and difficulty in quantification, limit their use. Therapeutic clinical studies using mesenchymal stem cells (MSCs) have been recently reported, and techniques using MSC-derived exosomes are being actively developed. Several studies have explored the application of cell-derived exosomes for medical purposes. MSC-derived exosomes have been used to suppress immune responses or as a therapeutic treatment for cancer [1, 2]. The yield of exosomes derived from stem cells is very low, which warrants optimization of conditions to increase yields. Nanoscale EVs are difficult to maintain their morphology and function, and exosomes originating from multiple vesicle bodies
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