China Biotechnology

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Mammalian Exosomes: Isolation and Purification, Clinical Applications and Prospects

JIANG Shanshan, YIN Yuqi, LI Qianyu, ZHANG Liangyu, YANG Mei, WANG Yonghua

China Biotechnology, 2024, 44(12): 51-63. DOI: 10.13523/j.cb.2404028

Fig.1 Schematic diagram of exosome biogenesis (by Figdraw) Exosomes are small membrane vesicles. They are formed as intraluminal vesicles by inward budding of the cell membrane into early endosomes, subsequently undergo migration and sorting to produce multivesicular body (MVB) and form intraluminal vesicles (ILVs). Upon fusion of thus arising MVB with the plasma membrane, these vesicles are released as exosomes

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Table 1 Comparison of common exosomes isolation methods and their advantages /disadvantages^[18⇓-20]
Fig.2 Schematic diagram of the routine technique of exosome isolation (modified from the references[18], by Figdraw) A: Ultracentrifugation. Exosomes were obtained by multiple centrifugation, centrifugal forces from 300 g up to 100 000 g, after each centrifugation step, the supernatant including cells and cellular debris was removed and collected for subsequent centrifugation, the centrifugation is performed at 4℃ B: Density gradient ultracentrifugation. Preparation of an isometric density gradient ultracentrifugation by gradually decreasing the density layer from bottom to top. After prolonged centrifugation, exosomes and other extracellular components reach a static position in a medium of similar density to each component C: Size-exclusion chromatography. When passing a solution through a stationary phase consisting of porous resin particles, molecules can be separated according to size D: Immunoaffinity. After incubating exosome-containing liquids with exosome-specific antibody coupled to the solid substrates, exosomes can be enriched onto solid substrates
Table 2 Summary of exosome isolation on different body fluid types^[57]
Table 3 Exosomal non-coding RNAs as diagnostic/prognostic markers^[74⇓-76]

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