This systematic review examines the landscape of extracellular vesicle (EV)-related clinical trials to elucidate trends in clinical applications and methodologies, with a focus on EV subpopulations. By analyzing data from public repositories, 471 EV-related clinical trials were catalogued, covering over 200 diseases. Diagnostics and companion diagnostics represented the bulk of these trials, with cancer being the most frequent application. Therapeutics trials primarily utilized mesenchymal stromal cell (MSC) EVs for respiratory illnesses. Ultracentrifugation and RNA-sequencing were the most common isolation and characterization techniques, but methodology reporting was often lacking. Most characterization relied on bulk EV isolates, with only 11% using EV subpopulations. The review highlights the need for meticulous methodological reporting and consideration of EV subpopulations to enhance the translational success of EV-based interventions, pointing towards a paradigm shift in personalized medicine.This systematic review examines the landscape of extracellular vesicle (EV)-related clinical trials to elucidate trends in clinical applications and methodologies, with a focus on EV subpopulations. By analyzing data from public repositories, 471 EV-related clinical trials were catalogued, covering over 200 diseases. Diagnostics and companion diagnostics represented the bulk of these trials, with cancer being the most frequent application. Therapeutics trials primarily utilized mesenchymal stromal cell (MSC) EVs for respiratory illnesses. Ultracentrifugation and RNA-sequencing were the most common isolation and characterization techniques, but methodology reporting was often lacking. Most characterization relied on bulk EV isolates, with only 11% using EV subpopulations. The review highlights the need for meticulous methodological reporting and consideration of EV subpopulations to enhance the translational success of EV-based interventions, pointing towards a paradigm shift in personalized medicine.