Extracellular vesicles (EVs) are categorized according to their formation mechanisms and come in three basic types: exosomes, microvesicles, and apoptotic bodies. Exosomes are a subtype of EVs, which are small, round sacs with a diameter of between 30 and 120 nm that are released by most cell types. Exosomes have the potential to be used as novel therapeutic agents as well as tools for the diagnosis and prognosis of various diseases. The main requirement for exosomal large-scale implementation in medical practice is rapid, easy, high purity, high yield isolation, and characterization processes. In this review, various isolation and characterization methods of exosomes are discussed together with their advantages and disadvantages.