Extracellular vesicles (EVs) are a heterogeneous group of cell-derived nanoscale membranous vesicles rich in lipids, proteins and nucleic acids. They are now considered as important carriers for intercellular communication. Extracellular vesicles derived from mesenchymal stem cell (MSC-EVs) have been widely concerned in recent years not only for their repair and regeneration properties similar to mesenchymal stem cells, but also for their advantages of stability and potential for large-scale preparation. Thus, MSC-EVs are expected to be used in clinic as an acellular therapeutic approach. This review summarizes the molecular composition, functional mechanism, isolation and identification methods of MSC-EVs. Moreover, their application in repairing tissue damage and their potential use as the drug delivery system are highlighted, which provide references for basic and translational researches of MSC-EVs.