Updated project metadata. Fewer than 50% of patients develop vascular and valvular calcification, implying differential pathogenesis. Tissue-entrapped extracellular vesicles (EVs) are found in mineralization but their contents and functions are unstudied. Tissue EVs were isolated from normal and diseased human carotid arteries and aortic valves by enzymatic digestion, serial (ultra)centrifugation and density-gradient separation. Mass spectrometry characterized the density-gradient separation vs. ultracentrifugation.