Gualerzi Alice, Kooijmans Sander Alexander Antonius, Niada Stefania, Picciolini Silvia, Brini Anna Teresa, Camussi Giovanni, Bedoni Marzia
IRCCS Fondazione Don Carlo Gnocchi, Milano, Italy.
Bioindustry Park Silvano Fumero SpA, Colleretto Giacosa, Italy.
J Extracell Vesicles. 2019 Jan 27;8(1):1568780. doi: 10.1080/20013078.2019.1568780. eCollection 2019.
Extracellular vesicles (EVs) from a variety of stem cell sources are believed to harbour regenerative capacity, which may be exploited for therapeutic purposes. Because of EV interaction with other soluble secreted factors, EV activity may depend on the employed purification method, which limits cross-study comparisons and therapeutic development. Raman spectroscopy (RS) is a quick and easy method to assess EV purity and composition, giving in-depth biochemical overview on EV preparation. Hereby, we show how this method can be used to characterise EVs isolated from human liver stem cells and bone marrow mesenchymal stem/stromal cells by means of conventional ultracentrifugation (UC) and size exclusion chromatography (SEC) protocols. The obtained EV preparations were demonstrated to be characterised by different degrees of purity and a specific Raman fingerprint that represents both the cell source and the isolation procedure used. Moreover, RS provided useful hints to explore the factors underlying the functional diversity of EV preparations from the same cell source, thus representing a valuable tool to assess EV quality prior to functional assays or therapeutic application.
来自多种干细胞来源的细胞外囊泡（EVs）被认为具有再生能力，可用于治疗目的。由于EV与其他可溶性分泌因子相互作用，EV活性可能取决于所采用的纯化方法，这限制了跨研究比较和治疗开发。拉曼光谱（RS）是一种快速简便的评估EV纯度和组成的方法，能对EV制剂进行深入的生化概述。在此，我们展示了如何通过传统超速离心（UC）和尺寸排阻色谱（SEC）方案，使用该方法对从人肝干细胞和骨髓间充质干/基质细胞中分离出的EVs进行表征。所获得的EV制剂表现出不同程度的纯度以及代表细胞来源和所用分离程序的特定拉曼指纹。此外，RS为探索来自同一细胞来源的EV制剂功能多样性背后的因素提供了有用线索，因此是在功能测定或治疗应用之前评估EV质量的有价值工具。
