Preparation of cell membrane-derived nanovesicles by nitrogen cavitation. (A) The process to generate the nanovesicles via nitrogen cavitation, centrifugation and extrusion. Differentiated HL-60 cells were collected and disrupted by nitrogen cavitation under a pressure of approximately 350 psi, and membrane-formed vesicles, intracellular molecules, and the nucleus exist in the resulting solution. Afterwards, differential centrifugation was used to purify and obtain the needed vesicles. Copyright 2016, Elsevier.[16] (B) Cryo-TEM of cell membrane-formed vesicles, which are approximately 150 nm in diameter. (C) Proteomics of nanovesicles and their source cells. The total proteins (left) and membrane proteins (right) in nitrogen cavitation-generated EVs (NC-EVs), naturally secreted EVs (NS-EVs) and their parent cell analyzed by mass spectrometry. The shared proteins between EVs and their parent cells suggested that EVs were derived from their parent cells. (D) Quantification analysis of the yield of NC-EVs and NS-EVs indicated the reproductively of NC-EVs. Copyright 2017, Elsevier.[17]