Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

Linda Bojmar; Han Sang Kim; Gabriel C Tobias; Fanny A Pelissier Vatter; Serena Lucotti; Kofi Ennu Gyan; Candia M Kenific; Zurong Wan; Kyung-A Kim; DooA Kim; Jonathan Hernandez; Virginia Pascual; Todd E Heaton; Michael P La Quaglia; David Kelsen; Tanya M Trippett; David R Jones; William R Jarnagin; Irina R Matei; Haiying Zhang; Ayuko Hoshino; David Lyden

doi:10.1016/j.xpro.2020.100225

Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

STAR Protoc. 2020 Dec 22;2(1):100225. doi: 10.1016/j.xpro.2020.100225. eCollection 2021 Mar 19.

Authors

Linda Bojmar^{1

2}, Han Sang Kim^{1

3}, Gabriel C Tobias^{1

4}, Fanny A Pelissier Vatter¹, Serena Lucotti¹, Kofi Ennu Gyan^{1

5}, Candia M Kenific¹, Zurong Wan^{1

6}, Kyung-A Kim³, DooA Kim³, Jonathan Hernandez^{7

8}, Virginia Pascual^{6

9}, Todd E Heaton^{10

11}, Michael P La Quaglia^{10

11}, David Kelsen¹², Tanya M Trippett¹⁰, David R Jones¹³, William R Jarnagin⁷, Irina R Matei¹, Haiying Zhang¹, Ayuko Hoshino^{1

14}, David Lyden¹

Affiliations

¹ 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.
² Department of Biomedical and Clinical Sciences, Linköping University, Linköping 58183, Sweden.
³ Yonsei Cancer Center, Division of Medical Oncology, Department of Internal Medicine, Graduate School of Medical Science, Brain Korea 21 Project, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Korea.
⁴ School of Physical Education and Sport, University of Sao Paulo, Sao Paulo, Brazil.
⁵ Tri-Institutional PhD Program in Computational Biology and Medicine, New York, NY, USA.
⁶ Department of Pediatrics, Weill Cornell Medicine, New York, NY, USA.
⁷ Hepatopancreatobiliary Service, Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
⁸ Surgical Oncology Program, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
⁹ Drukier Institute for Children's Health, Weill Cornell Medicine, New York, NY, USA.
¹⁰ Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
¹¹ Pediatric Surgical Service, Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
¹² Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
¹³ Thoracic Surgery Service, Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
¹⁴ School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.

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).

Keywords: Cell Membrane; Cell culture; Cell isolation; Exomeres; Exosomes; Extracellular vesicles and particles; Isolation.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Body Fluids / chemistry*
Cell Line
Centrifugation, Density Gradient*
Extracellular Vesicles / chemistry*
Fractionation, Field Flow*
Humans
Mice
Proteomics*

Abstract

Publication types

MeSH terms

Grants and funding