Journal article
Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes
BJ Tauro, DW Greening, RA Mathias, H Ji, S Mathivanan, AM Scott, RJ Simpson
Methods | Published : 2012
Abstract
Exosomes are 40-100. nm extracellular vesicles that are released from a multitude of cell types, and perform diverse cellular functions including intercellular communication, antigen presentation, and transfer of oncogenic proteins as well as mRNA and miRNA. Exosomes have been purified from biological fluids and in vitro cell cultures using a variety of strategies and techniques. However, all preparations invariably contain varying proportions of other membranous vesicles that co-purify with exosomes such as shed microvesicles and apoptotic blebs. Using the colorectal cancer cell line LIM1863 as a cell model, in this study we performed a comprehensive evaluation of current methods used for e..
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Awarded by Australian Cancer Research Foundation
Funding Acknowledgements
This work was supported, in part, by the National Health & Medical Research Council of Australia (program Grant #487922 (R.J.S and A.M.S). The University of Melbourne Research Scholarship (B.J.T). Analysis of proteomic data described in this work was supported using the Australian Proteomics Computational Facility funded by the National Health & Medical Research Council of Australia Grant #381413. This work was supported by funds from the Operational Infrastructure Support Program provided by the Victorian Government Australia. We acknowledge the Australian Cancer Research Foundation for providing funds to purchase the Orbitrap (TM) mass spectrometer.