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[1]. Vanderboom Patrick M,, et al. A size-exclusion-based approach for purifying extracellular vesicles from human plasma. Cell Rep Methods, 2021, 1(3):100055.£¨Ã·°ÂÕïËùҽѧ²¿£©

[2].Freitas D, et al. Different isolation approaches lead to diverse glycosylated extracellular vesicle populations.J Extracell Vesicles. 2019 Jun 3;8(1):1621131.£¨²¨¶ûͼ´óѧ½¡¿µÑо¿Óë´´ÐÂÑо¿Ëù£©IF=16.0

[3]. Xing F, et al. Loss of XIST in Breast Cancer Activates MSN-c-Met and Reprograms Microglia via Exosomal miRNA to Promote Brain Metastasis. Cancer Res. 2018 Aug 1; 78(15): 4316-4330.£¨Î¬¿ËÉ­ÁÖҽѧԺ°©Ö¢ÉúÎïѧϵ£©IF=11.2

[4]. Bao L, et al. Metastasis-associated miR-23a from nasopharyngeal carcinomaderived exosomes mediates angiogenesis by repressing a novel target gene TSGA10. Oncogene. 2018 May; 37(21): 2873-2889.£¨ÄÏͨ´óѧ¸½ÊôÒ½Ôº£©IF=8.0

[5]. Ramirez M, et al. Technical challenges of working with extracellular vesicles. Nanoscale. 2018 Jan 18; 10(3): 881-906.£¨A.C.¿¨Âí¸ê°©Ö¢ÖÐÐÄ£©

[6]. Takov K,et al. Comparison of small extracellular vesicles isolated from plasma by ultracentrifugation or size-exclusion chromatography: yield, purity and functional potential. J Extracell Vesicles.2018 Dec 28;8(1):1560809.£¨Â׶شóѧѧԺ¹þÌØÐÄѪ¹ÜÑо¿Ëù£©IF=16.0

[7]. Mol EA,et al. Higher functionality of extracellular vesicles isolated using size-exclusion chromatography compared to ultracentrifugation.Nanomedicine 2017 Aug;13(6):2061-2065.£¨ÎÚµÃÀÕÖ§´óѧҽѧÖÐÐÄ£©

[8]. Jurkoshek KS, et al. Interspecies Communication between Pathogens and Immune Cells via Bacterial Membrane Vesicles. Front Cell Dev Biol. 2016 Nov 11; 4: 125.£¨¿­Ë¹Î÷´¢´óѧ²¡Àíѧϵ£©

[9]. Lobb RJ, et al. Optimized exosome isolation protocol for cell culture supernatant and human plasma. J Extracell Vesicles. 2015 Jul 17; 4: 27031.£¨QIMR Berghofer ҽѧÑо¿Ëù£©IF=16.0