Production of Extracellular Vesicles Loaded with Therapeutic Cargo.
Production of Extracellular Vesicles Loaded with Therapeutic Cargo.
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DOI：
10.1007/978-1-4939-8661-3_4
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Jay SM
中科院分区：
文献类型：
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作者：
Lamichhane TN;Jay SM
细胞外囊泡（EV）是包含外泌体、微囊泡和由大多数细胞类型产生的其他异质纳米级囊泡群体的生物纳米颗粒。除了它们作为细胞间通讯的关键介质的假定作用外，EV已经开始被用作药物递送载体，早期证据表明它们可能比合成纳米颗粒递送系统在特定应用中具有显着优势。EV包封的货物的靶向递送已经实现，并且可能具有广泛的适用性;然而，用于生产和纯化EV以及用治疗分子装载它们的方法尚未标准化。在本章中，我们概述了EV分离和表征的步骤，并比较了目前使用短干扰RNA（siRNA）或小分子有效载荷进行EV主动和被动加载的方法，结果表明，通过电穿孔进行主动加载可以提高siRNA的加载效率，但不能提高HEK 293 T衍生EV中罗丹明B（一种小分子药物模型）的加载效率。本文所述的方法可以为将来设计通过EV靶向递送核酸或小分子提供信息。
Extracellular vesicles (EVs) are biological nanoparticles comprising exosomes, microvesicles, and other heterogeneous nanoscopic vesicle populations that are produced by most cell types. In addition to their putative roles as critical mediators of intercellular communication, EVs have begun to be harnessed as drug delivery vehicles, with early evidence indicating they may have significant advantages over synthetic nanoparticle delivery systems for particular applications. Targeted delivery of EV-encapsulated cargo has already been realized and may have broad applicability; however, methods for producing and purifying EVs and loading them with therapeutic molecules have yet to be standardized. In this chapter, we outline steps for EV isolation and characterization and compare current methods for active and passive loading of EVs with payloads of short interfering RNA (siRNA) or small molecules, with the results revealing that active loading via electroporation increases loading efficiency of siRNA but not of Rhodamine B, a model for a small molecule drug, in HEK293T-derived EVs. The methods described here may inform future design of targeted delivery of nucleic acids or small molecules via EVs.