Exosomes are phospholipid bilayer vesicles secreted by living cells that can carry a variety of signaling molecules, such as RNA, DNA, protein, and lipids. Exosomes play a role in the transmission of signaling molecules between cells, thus regulating many physiological and pathological processes. The methods of extracting exosomes include differential centrifugation, density gradient centrifugation, exclusion chromatography, ultrafiltration, coprecipitation, polymer immune affinity, microfluidic separation technology, etc. Each of these extraction technologies has advantages and disadvantages; however, there is no unified international standard. In addition, the expression of specific proteins and genetic material of exosomes from different cell sources are different; thus, their expression characteristics and functions are also distinctive. Based on this situation, research on exosomes is limited to preclinical studies, and difficulties and challenges still exist in clinical application. This paper summarizes the progress of research in the field of exosomes, to understand the characteristics, modification and application of exosomes from different cell sources, and to summarize their advantages and disadvantages as well as challenges, which can help researchers better understand and master the performance of exosomes. Furthermore, improvement of standard procedures in the extraction and manufacturing of exosomes is important, as it will provide a reference for researchers to carry out exosome-related translational clinical research.