The article provides a comprehensive review of the evolution of Minimal Information for Studies of Extracellular Vesicles (MISEV) from 2014 to 2023. MISEV has been updated in three editions—MISEV2014, MISEV2018, and MISEV2023—to refine standards and improve the rigor, reproducibility, and transparency of extracellular vesicle (EV) research. The evolution of MISEV reflects the growing interest and advancements in the field of EVs, with a focus on refining experimental design, data reporting, and sample treatment. Key changes in MISEV2023 include: 1. **Nomenclature**: The 2023 edition clarifies the definition of EVs and emphasizes the distinction between "naturally" produced EVs and those produced in various cell culture conditions. It also highlights the importance of non-vesicular extracellular particles (NVEPs). 2. **Collection and Pre-Processing**: The 2023 edition adds detailed guidelines for collecting and pre-processing samples from various sources, such as blood, milk, and urine, emphasizing the need for comprehensive documentation of experimental conditions. 3. **EV Separation and Concentration**: The 2023 edition refines traditional separation methods and introduces new techniques like fluid flow-based separation. It provides detailed recommendations for various separation and concentration methods, including their application in different sample types. 4. **EV Characterization**: The 2023 edition updates the five-component framework for EV protein characterization, emphasizing the importance of evaluating both EV marker proteins and NVEP to ensure sample purity and specificity. 5. **EV Release and Uptake**: The 2023 edition discusses methods for observing and modulating EV release and uptake, which are crucial for understanding EV functions and their potential applications in drug delivery. 6. **Functional Studies**: The 2023 edition recommends dose-response and time-course assays for comprehensive assessment of EV biological effects and the use of appropriate negative controls to ensure the accuracy of experiments. 7. **In Vivo Analysis**: The 2023 edition summarizes in vivo EV studies, providing insights into the advantages and genetic similarities of different in vivo models, and detailing the reporting requirements for exogenous EV administration parameters. The article concludes by highlighting the ongoing challenges and future directions in EV research, including the need for more transparent experimental processes, improved isolation techniques, and the integration of machine learning in EV analysis. The continuous updates to MISEV aim to enhance the quality and reliability of EV research, fostering a more robust and comprehensive understanding of these particles.The article provides a comprehensive review of the evolution of Minimal Information for Studies of Extracellular Vesicles (MISEV) from 2014 to 2023. MISEV has been updated in three editions—MISEV2014, MISEV2018, and MISEV2023—to refine standards and improve the rigor, reproducibility, and transparency of extracellular vesicle (EV) research. The evolution of MISEV reflects the growing interest and advancements in the field of EVs, with a focus on refining experimental design, data reporting, and sample treatment. Key changes in MISEV2023 include: 1. **Nomenclature**: The 2023 edition clarifies the definition of EVs and emphasizes the distinction between "naturally" produced EVs and those produced in various cell culture conditions. It also highlights the importance of non-vesicular extracellular particles (NVEPs). 2. **Collection and Pre-Processing**: The 2023 edition adds detailed guidelines for collecting and pre-processing samples from various sources, such as blood, milk, and urine, emphasizing the need for comprehensive documentation of experimental conditions. 3. **EV Separation and Concentration**: The 2023 edition refines traditional separation methods and introduces new techniques like fluid flow-based separation. It provides detailed recommendations for various separation and concentration methods, including their application in different sample types. 4. **EV Characterization**: The 2023 edition updates the five-component framework for EV protein characterization, emphasizing the importance of evaluating both EV marker proteins and NVEP to ensure sample purity and specificity. 5. **EV Release and Uptake**: The 2023 edition discusses methods for observing and modulating EV release and uptake, which are crucial for understanding EV functions and their potential applications in drug delivery. 6. **Functional Studies**: The 2023 edition recommends dose-response and time-course assays for comprehensive assessment of EV biological effects and the use of appropriate negative controls to ensure the accuracy of experiments. 7. **In Vivo Analysis**: The 2023 edition summarizes in vivo EV studies, providing insights into the advantages and genetic similarities of different in vivo models, and detailing the reporting requirements for exogenous EV administration parameters. The article concludes by highlighting the ongoing challenges and future directions in EV research, including the need for more transparent experimental processes, improved isolation techniques, and the integration of machine learning in EV analysis. The continuous updates to MISEV aim to enhance the quality and reliability of EV research, fostering a more robust and comprehensive understanding of these particles.