Dataset Information

Human and mouse brain, and human cerebrospinal fluid extracellular vesicles LC-MSMS

ABSTRACT: Extracellular vesicles (EVs) are a unified terminology of membrane-enclosed vesicular species ubiquitously secreted by almost every cell type and present in all body fluids. They carry a cargo of lipids, metabolites, nucleic acids and proteins for their clearance from cells as well as for cell-to-cell communications. The exact composition of EVs and their specific functions are not well understood due to the underdevelopment of the separation protocols, especially those from the central nervous system including animal and human brain tissues as well as cerebrospinal fluids. To understand their exact molecular composition and their functional roles, development of the reliable protocols for EV separation is necessary. Known EV separation methods include differential centrifugation-ultracentrifugation, sucrose gradient ultracentrifugation, size exclusion chromatography, ultra-filtration, microfluidics, high-resolution flow cytometric sorting, polymer-based precipitation, immunoaffinity capture, affinity capture, and asymmetric-flow field-flow fractionation. In this report, we describe our EV separation protocols from human biospecimens, such as unfixed frozen brain tissues and cerebrospinal fluids. We also discuss the quality control measures of the above separation protocols by biophysical and proteomic characterizations.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Brain, Cerebrospinal Fluid

SUBMITTER: Satoshi Muraoka

LAB HEAD: Tsuneya Ikezu

PROVIDER: PXD017273 | Pride | 2020-02-17

REPOSITORIES: Pride

ACCESS DATA

Publications

Assessment of separation methods for extracellular vesicles from human and mouse brain tissues and human cerebrospinal fluids.

Muraoka Satoshi S Lin Weiwei W Chen Mei M Hersh Samuel W SW Emili Andrew A Xia Weiming W Ikezu Tsuneya T

Methods (San Diego, Calif.) 20200205

Extracellular vesicle (EV) is a unified terminology of membrane-enclosed vesicular species ubiquitously secreted by almost every cell type and present in all body fluids. They carry a cargo of lipids, metabolites, nucleic acids and proteins for their clearance from cells as well as for cell-to-cell communications. The exact composition of EVs and their specific functions are not well understood due to the underdevelopment of the separation protocols, especially those from the central nervous sys ...[more]

PMID: 32035230

Similar Datasets

Project description:Extracellular vesicles (EVs) are involved in a wide range of physiological and pathological processes by shuttling material out of and between cells. Tissue EVs may thus lend insights into disease mechanisms and also betray disease when released into easily accessed biological fluids. Since brain-derived EVs (bdEVs) and their cargo may serve as biomarkers of neurodegenerative diseases, we evaluated modifications to a published, rigorous protocol for separation of EVs from brain tissue and studied effects of processing variables on quantitative and qualitative outcomes. To this end, size exclusion chromatography (SEC) and sucrose density gradient ultracentrifugation were compared as final separation steps in protocols involving stepped ultracentrifugation. bdEVs were separated from brain tissues of human, macaque. Effects of post-mortem interval (PMI) before final bdEV separation were probed. MISEV2018-compliant EV characterization was performed, and both small RNA and protein profiling were done. We conclude that the modified, SEC-employing protocol achieves EV separation efficiency roughly similar to a protocol using gradient density ultracentrifugation, while decreasing operator time and, potentially, variability. The protocol appears to yield bdEVs of higher purity for human tissues compared with macaque, suggesting opportunities for optimization. The interval between death/tissue storage/processing and final bdEV separation can also affect bdEV populations and composition and should thus be recorded for rigorous reporting. Different populations of EVs obtained through the modified method reported herein display characteristic RNA and protein content that hint at biomarker potential. To conclude, this study finds that the automatable and increasingly employed technique of SEC can be applied to tissue EV separation, and also reveals more about the importance of species-specific and technical considerations when working with tissue EVs. These results are expected to enhance the use of bdEVs in revealing and understanding brain disease.

Dataset Information

Human and mouse brain, and human cerebrospinal fluid extracellular vesicles LC-MSMS

Publications

Assessment of separation methods for extracellular vesicles from human and mouse brain tissues and human cerebrospinal fluids.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure