Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids
Linda Bojmar, Han Sang Kim, Gabriel Cardial Tobias, Fanny A. Pelissier Vatter, Serena Lucotti, Kofi Ennu Gyan, Candia M. Kenific, Zurong Wan, Kyung‐A Kim, DooA Kim, Jonathan M. Hernandez, Virginia Pascual, Todd E. Heaton, Michael P. LaQuaglia, David P. Kelsen, Tanya Trippett, David R. Jones, William R. Jarnagin, Irina Matei, Haiying Zhang, Ayuko Hoshino, David Lyden
Abstract
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).