MISEV2023 is an updated guide to extracellular vesicle (EV) research and applications. It provides a comprehensive overview of EV studies, including nomenclature, collection and pre-processing, separation and concentration, characterization, release and uptake, functional studies, and in vivo studies. The document emphasizes the importance of standardization and reproducibility in EV research, while also acknowledging its limitations. MISEV2023 was developed through a collaborative process involving over 1051 authors from 53 countries, with extensive community input and feedback. The document highlights the need for continued efforts to improve EV research and applications, while also recognizing the challenges of keeping up with the rapidly growing EV literature. MISEV2023 is not a comprehensive review of the literature but rather a guide to help researchers understand and apply EV research principles. The authors acknowledge that MISEV is not a complete solution but a tool to enhance the rigor and reproducibility of EV studies. The document also encourages ongoing dialogue and critique to improve the field. The authors express their gratitude to all contributors and emphasize the importance of collaboration in advancing EV research. The authors also note that future MISEV updates may not be necessary if the field can benefit from more focused guidelines rather than a broad revision. The document concludes with a call to action for the EV community to continue working together to advance the field.MISEV2023 is an updated guide to extracellular vesicle (EV) research and applications. It provides a comprehensive overview of EV studies, including nomenclature, collection and pre-processing, separation and concentration, characterization, release and uptake, functional studies, and in vivo studies. The document emphasizes the importance of standardization and reproducibility in EV research, while also acknowledging its limitations. MISEV2023 was developed through a collaborative process involving over 1051 authors from 53 countries, with extensive community input and feedback. The document highlights the need for continued efforts to improve EV research and applications, while also recognizing the challenges of keeping up with the rapidly growing EV literature. MISEV2023 is not a comprehensive review of the literature but rather a guide to help researchers understand and apply EV research principles. The authors acknowledge that MISEV is not a complete solution but a tool to enhance the rigor and reproducibility of EV studies. The document also encourages ongoing dialogue and critique to improve the field. The authors express their gratitude to all contributors and emphasize the importance of collaboration in advancing EV research. The authors also note that future MISEV updates may not be necessary if the field can benefit from more focused guidelines rather than a broad revision. The document concludes with a call to action for the EV community to continue working together to advance the field.