Bojmar Linda, Kim Han Sang, Tobias Gabriel C, Pelissier Vatter Fanny A, Lucotti Serena, Gyan Kofi Ennu, Kenific Candia M, Wan Zurong, Kim Kyung-A, Kim DooA, Hernandez Jonathan, Pascual Virginia, Heaton Todd E, La Quaglia Michael P, Kelsen David, Trippett Tanya M, Jones David R, Jarnagin William R, Matei Irina R, Zhang Haiying, Hoshino Ayuko, Lyden David
Children's Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children's Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY 10021, USA.
Department of Biomedical and Clinical Sciences, Linköping University, Linköping 58183, Sweden.
STAR Protoc. 2020 Dec 22;2(1):100225. doi: 10.1016/j.xpro.2020.100225. eCollection 2021 Mar 19.
We developed a modified protocol, based on differential ultracentrifugation (dUC), to isolate extracellular vesicles and particles (specifically exomeres) (EVPs) from various human and murine sources, including cell lines, surgically resected tumors and adjacent tissues, and bodily fluids, such as blood, lymphatic fluid, and bile. The diversity of these samples requires robust and highly reproducible protocols and refined isolation technology, such as asymmetric-flow field-flow fractionation (AF4). Our isolation protocol allows for preparation of EVPs for various downstream applications, including proteomic profiling. For complete details on the use and execution of this protocol, please refer to Hoshino et al. (2020).
我们开发了一种基于差速超速离心（dUC）的改良方案，用于从各种人类和小鼠来源中分离细胞外囊泡和颗粒（特别是外泌子）（EVPs），这些来源包括细胞系、手术切除的肿瘤及相邻组织，以及血液、淋巴液和胆汁等体液。这些样本的多样性需要稳健且高度可重复的方案以及完善的分离技术，如不对称流场流分离法（AF4）。我们的分离方案能够制备用于各种下游应用（包括蛋白质组分析）的EVPs。有关该方案的使用和执行的完整详细信息，请参考星野等人（2020年）的文献。
