Improvement of stem cell-derived exosome release efficiency by surface-modified nanoparticles

Abstract

Background: Mesenchymal 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. Results: The 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. Conclusions: It 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.

Comparative Analysis of MSC-Derived Exosomes Depending on Cell Culture Media for Regenerative Bioactivity

“…For this reason, it is crucial to isolate highly purified exosomes secreted from MSCs without contaminants. For clinical applications, various attempts to standardize on the method of separating exosome and culturing the cells have been reported for increasing production yield and purity of exosomes [47][48][49][50][51]. Especially, the types of media used for cell culture play an important role not only in determining the state of cells, but also in determining the properties of exosomes secreted from the cells.…”

Engineering of Extracellular Vesicles Based on Payload Changes for Tissue Regeneration

“…However, previously reports showed excellent therapeutic potential efficacy with application in EVs through bioengineering techniques such as cargo loading and surface modification [84,85]. The low EV yield and the lack of the ability to target specific organs or molecules are two disadvantages associated with the use of EV; however, these limitations can be overcome by international regulation [77,78]. In addition, clinical research using EVs will be accelerated by the development of 3D culture technology and increasing the yield of EVs by the use of bioreactors [93,94].…”

“…Recently, our research team has developed a technology that can increase the number of EVs generated from stem cells by approximately 100 times by using iron oxide nanoparticles with a positive surface charge [78]. It was found that the components of the obtained exosomes contained miRNA factors related to proliferation, anti-inflammatory, and ROS reduction.…”

Technological advances towards extracellular vesicles mass production

“…Finally, exposition of cells to several types of exogenous particles have also been reported to increase EV secretion. Internalization of positively charged iron oxide nanoparticles encapsulated in PLGA-PEI polymer by MSCs increased secretion of extracellular vesicles, which appeared to be linked to increased autophagosome formation 85 . In Emam et al 86 , four cancerous cell lines were incubated for 48 h with different concentration of neutral, cationic-bare or PEGylated liposomes solutions.…”