 Exosomes are nano-sized subtypes of extracellular vesicles that play an important role in prognostic, diagnostic, and therapeutic aspects. However, common exosomal separation techniques suffer from operation complexity, time consumption, large sample volumes, and low purity, posing significant challenges for downstream analysis. Emerging technologies such as microfluidic chips have proposed superior strategies for exosome isolation and exhibited excellent performances. A comprehensive review of exosomal properties, biogenesis, contents, and functions is followed by an overview of conventional technologies and their challenges in clinical applications. Finally, emerging exosomal isolation strategies and large-scale GMP production of engineered exosomes are discussed to open up future perspectives for next-generation exodevices in cancer diagnosis and treatment. This article was authored by Jetsie Chen, Peilong Li, Tai Zhang, and others.