介绍细胞外囊泡分离、工程技术的最新进展、与不同分离技术相关的局限性、不同的生物医学应用以及工程化细胞外囊泡的挑战，以造福学术界、工业界等研究人员。 材料和方法：同行-收集了来自大多数知名期刊的评论文章，并对所提供的信息进行了分析，以讨论收集、化学、电穿孔、细胞和膜表面工程，以设计用于各种治疗应用的细胞外囊泡。此外，我们还介绍了细胞外囊泡收集技术的应用和局限性。需要具有金标准的分离技术。然而，先进的细胞外囊泡分离技术显示出细胞外囊泡的回收率和纯度有所提高。肿瘤治疗是治疗部分的一个主要部分，它说明了工程细胞外囊泡在协同治疗（如光疗、治疗诊断和遗传物质递送）中的作用。此外，细胞外囊泡在治疗视网膜疾病、神经退行性疾病、结核病、骨质疏松症、炎症性肠病、疫苗生产和伤口愈合方面也显示出潜力。工程细胞外囊泡可以将货物递送到特定细胞，引发免疫反应并未来可用于疫苗的研发。然而，进展还处于初级阶段。总的来说，这篇综述将为不同生物医学领域的工程化细胞外囊泡的临床转化研究人员提供全面的了解，并为研究人员提供参考。版权所有©2024。由Elsevier Inc.出版。

To present the recent update on the isolation, engineering techniques for extracellular vesicles, limitations associated with different isolation techniques, different biomedical applications, and challenges of engineered extracellular vesicles for the benefit of researchers from academic, industry, etc. MATERIALS AND METHODS: Peer-reviewed articles from most recognized journals were collected, and presented information was analyzed to discuss collection, chemical, electroporation, cellular, and membrane surface engineering to design extracellular vesicles for various therapeutic applications. In addition, we present the applications and limitations of techniques for the collection of extracellular vesicles.There is a need for isolation techniques with the gold standard. However, advanced extracellular vesicle isolation techniques showed improved recovery, and purity of extracellular vesicles. Tumor therapy is a major part of the therapy section that illustrates the role of engineered extracellular vesicles in synergetic therapy such as phototherapy, theragnostic, and delivery of genetic materials. In addition, extracellular vesicles have shown their potential in the treatment of retinal disorders, neurodegenerative disease, tuberculosis, osteoporosis, inflammatory bowel disease, vaccine production, and wound healing.Engineered extracellular vesicles can deliver cargo to the specific cells, elicit an immune response and could be used for the development of the vaccine in the future. However, the progress is at the initial stage. Overall, this review will provide a comprehensive understanding and could serve as a reference for researchers in the clinical translation of engineered extracellular vesicles in different biomedical fields.Copyright © 2024. Published by Elsevier Inc.