Department of Otorhinolaryngology, Yonsei University Wonju College of Medicine, 20 Ilsan-ro, Wonju, Gangwon-do, 26426, South Korea.
Research Institute of Hearing Enhancement, Yonsei University Wonju College of Medicine, Wonju, South Korea.
J Nanobiotechnology. 2020 Dec 7;18(1):178. doi: 10.1186/s12951-020-00739-7.
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.
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.
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.
间充质干细胞(MSCs)是多能基质细胞,会释放细胞外囊泡(EVs)。EVs 包含各种生长因子和抗氧化剂,能对周围细胞产生积极影响。纳米级 MSC 来源的 EV,如外泌体,已被开发为生物稳定的纳米型材料。然而,产量低和定量困难等问题限制了它们的应用。我们假设使用纳米颗粒增强外泌体的产生会刺激细胞内分子的释放。
本研究旨在通过比较表面修饰的正电荷纳米颗粒的内化和 MSC 中外泌体的产生,阐明外泌体产生的分子机制。我们发现,Rab7,晚期内涵体和自噬体的标志物,在表达外泌体时增加,并与自噬体的形成有关。
我们开发的纳米颗粒通过网格蛋白介导的内吞作用被转运到溶酶体。此外,进入的纳米颗粒刺激自噬相关因子从 MSC 中释放外泌体。使用纳米颗粒的 MSC 来源的外泌体可以增加外泌体的产量,并能够发现纳米颗粒诱导的遗传因素。