Progress in Exosome Isolation Techniques

Exosomes are one type of membrane vesicles secreted into extracellular space by most types of cells. In addition to performing many biological functions particularly in cell-cell communication, cumulative evidence has suggested that several biological entities in exosomes like proteins and microRNAs are closely associated with the pathogenesis of most human malignancies and they may serve as invaluable biomarkers for disease diagnosis, prognosis, and therapy. This provides a commanding impetus and growing demands for simple, efficient, and affordable techniques to isolate exosomes. Capitalizing on the physicochemical and biochemical properties of exosomes, a number of techniques have been developed for the isolation of exosomes. This article summarizes the advances in exosome isolation techniques with an emphasis on their isolation mechanism, performance, challenges, and prospects. We hope that this article will provide an overview of exosome isolation techniques, opening up new perspectives towards the development more innovative strategies and devices for more time saving, cost effective, and efficient isolations of exosomes from a wide range of biological matrices.

[1]  T. Ørntoft,et al.  Comparative analysis of discrete exosome fractions obtained by differential centrifugation , 2014, Journal of extracellular vesicles.

[2]  M. Amiji,et al.  Exosome mediated communication within the tumor microenvironment. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[3]  Yi Li,et al.  Exosome‐Mediated Transfer of miR‐133b from Multipotent Mesenchymal Stromal Cells to Neural Cells Contributes to Neurite Outgrowth , 2012, Stem cells.

[4]  C. Melief,et al.  B lymphocytes secrete antigen-presenting vesicles , 1996, The Journal of experimental medicine.

[5]  A. Shiras,et al.  Tumor exosomes: cellular postmen of cancer diagnosis and personalized therapy. , 2016, Nanomedicine.

[6]  Diane M Simeone,et al.  Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes. , 2014, Lab on a chip.

[7]  B. Breitner [On cancer diagnostic]. , 1952, Wiener medizinische Wochenschrift.

[8]  Richard J Simpson,et al.  Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes. , 2012, Methods.

[9]  Rune Matthiesen,et al.  Characterization and comprehensive proteome profiling of exosomes secreted by hepatocytes. , 2008, Journal of proteome research.

[10]  S. Thibodeau,et al.  Characterization of human plasma-derived exosomal RNAs by deep sequencing , 2013, BMC Genomics.

[11]  Lynne T. Bemis,et al.  Standardization of sample collection, isolation and analysis methods in extracellular vesicle research , 2013, Journal of extracellular vesicles.

[12]  Sung‐Min Ahn,et al.  Proteomic analysis of exosomes from human neural stem cells by flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry. , 2008, Journal of proteome research.

[13]  Aled Clayton,et al.  Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological Fluids , 2006, Current protocols in cell biology.

[14]  S. Gabrielsson,et al.  Exosomes Derived from Burkitt’s Lymphoma Cell Lines Induce Proliferation, Differentiation, and Class-Switch Recombination in B Cells , 2014, The Journal of Immunology.

[15]  V. Bond,et al.  Isolation of Exosomes from the Plasma of HIV-1 Positive Individuals. , 2016, Journal of visualized experiments : JoVE.

[16]  J. Le Pecq,et al.  A phase I study of dexosome immunotherapy in patients with advanced non-small cell lung cancer , 2005, Journal of Translational Medicine.

[17]  Sanchita Bhatnagar,et al.  Exosome Function: From Tumor Immunology to Pathogen Biology , 2008, Traffic.

[18]  J. Slot,et al.  Proteomic and Biochemical Analyses of Human B Cell-derived Exosomes , 2003, The Journal of Biological Chemistry.

[19]  Mahdieh Khosroheidari,et al.  Comparison of protein, microRNA, and mRNA yields using different methods of urinary exosome isolation for the discovery of kidney disease biomarkers. , 2012, Kidney international.

[20]  H. Girish,et al.  Risk of tumor cell seeding through biopsy and aspiration cytology , 2014, Journal of International Society of Preventive & Community Dentistry.

[21]  A. Sivachenko,et al.  Massively Parallel Sequencing of Human Urinary Exosome/Microvesicle RNA Reveals a Predominance of Non-Coding RNA , 2014, PloS one.

[22]  S. Jeune,et al.  Diagnostic , 1982, Thromboses Veineuses Distales et Superficielles.

[23]  F. Young Biochemistry , 1955, The Indian Medical Gazette.

[24]  O. Barreiro,et al.  Tetraspanin-enriched microdomains: a functional unit in cell plasma membranes. , 2009, Trends in cell biology.

[25]  J. Klein,et al.  Comparison of three methods for isolation of urinary microvesicles to identify biomarkers of nephrotic syndrome. , 2010, Kidney international.

[26]  C. Harding,et al.  Exosomes: Looking back three decades and into the future , 2013, The Journal of cell biology.

[27]  D. Taylor,et al.  Tumour-derived exosomes and their role in cancer-associated T-cell signalling defects , 2005, British Journal of Cancer.

[28]  J. Lötvall,et al.  Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells , 2007, Nature Cell Biology.

[29]  Lubert Stryer,et al.  Biochemistry 5th ed , 2002 .

[30]  S. Wickline,et al.  A review of exosome separation techniques and characterization of B16-F10 mouse melanoma exosomes with AF4-UV-MALS-DLS-TEM , 2014, Analytical and Bioanalytical Chemistry.

[31]  Diego Diez,et al.  A novel affinity-based method for the isolation of highly purified extracellular vesicles , 2016, Scientific Reports.

[32]  Peter P. Lee,et al.  Tumor-Derived Exosomes Contain microRNAs with Immunological Function: Implications for a Novel Immunosuppression Mechanism. , 2014, MicroRNA.

[33]  Andrew F. Hill,et al.  Exosomes: Vehicles for the Transfer of Toxic Proteins Associated with Neurodegenerative Diseases? , 2012, Front. Physio..

[34]  Olivier Lantz,et al.  Vaccination of metastatic melanoma patients with autologous dendritic cell (DC) derived-exosomes: results of thefirst phase I clinical trial , 2005, Journal of Translational Medicine.

[35]  Jacopo Meldolesi,et al.  Shedding microvesicles: artefacts no more. , 2009, Trends in cell biology.

[36]  Fedor V. Karginov,et al.  Cell contact-dependent acquisition of cellular and viral nonautonomously encoded small RNAs. , 2009, Genes & development.

[37]  Michael Liem,et al.  Comparative proteomics evaluation of plasma exosome isolation techniques and assessment of the stability of exosomes in normal human blood plasma , 2013, Proteomics.

[38]  A. Malek,et al.  Lectin‐induced agglutination method of urinary exosomes isolation followed by mi‐RNA analysis: Application for prostate cancer diagnostic , 2016, The Prostate.

[39]  P. Low,et al.  Choroid plexus transcytosis and exosome shuttling deliver folate into brain parenchyma , 2013, Nature Communications.

[40]  M. Adam,et al.  Electron microscopic evidence for externalization of the transferrin receptor in vesicular form in sheep reticulocytes , 1985, The Journal of cell biology.

[41]  I. Kurochkin,et al.  Characterization of RNA in exosomes secreted by human breast cancer cell lines using next-generation sequencing , 2013, PeerJ.

[42]  P. Stahl,et al.  Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes , 1983, The Journal of cell biology.

[43]  A. Booth,et al.  Exosomes and HIV Gag bud from endosome-like domains of the T cell plasma membrane , 2006, The Journal of cell biology.

[44]  S. Raimondo,et al.  Exosomes as Intercellular Signaling Organelles Involved in Health and Disease: Basic Science and Clinical Applications , 2013, International journal of molecular sciences.

[45]  Trairak Pisitkun,et al.  Rapid isolation of urinary exosomal biomarkers using a nanomembrane ultrafiltration concentrator. , 2007, American journal of physiology. Renal physiology.

[46]  M Kersaudy-Kerhoas,et al.  Exosome isolation: a microfluidic road-map. , 2015, Lab on a chip.

[47]  Gerard Pasterkamp,et al.  Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. , 2010, Stem cell research.

[48]  Christian Pilarsky,et al.  Glypican-1 identifies cancer exosomes and detects early pancreatic cancer , 2015, Nature.

[49]  D. Prockop,et al.  Chromatographically isolated CD63+CD81+ extracellular vesicles from mesenchymal stromal cells rescue cognitive impairments after TBI , 2015, Proceedings of the National Academy of Sciences.

[50]  Y. Takakura,et al.  Effect of exosome isolation methods on physicochemical properties of exosomes and clearance of exosomes from the blood circulation. , 2016, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[51]  Tao Jiang,et al.  Exosomes derived from Rab27a‑overexpressing tumor cells elicit efficient induction of antitumor immunity. , 2013, Molecular medicine reports.

[52]  Anikó Szalai,et al.  Detection and isolation of cell-derived microparticles are compromised by protein complexes resulting from shared biophysical parameters. , 2011, Blood.

[53]  Paolo Guazzi,et al.  Integrated isolation and quantitative analysis of exosome shuttled proteins and nucleic acids using immunocapture approaches. , 2015, Methods.

[54]  I. Alevizos,et al.  Isolation of circulating microRNA in saliva. , 2013, Methods in molecular biology.

[55]  Fátima Sánchez-Cabo,et al.  Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells , 2011, Nature communications.

[56]  Emily Zeringer,et al.  Strategies for isolation of exosomes. , 2015, Cold Spring Harbor protocols.

[57]  J. Lötvall,et al.  Isolation and Characterization of RNA-Containing Exosomes , 2012, Journal of visualized experiments : JoVE.

[58]  H. Tahara,et al.  The role of exosomes and microRNAs in senescence and aging. , 2013, Advanced drug delivery reviews.

[59]  H. Geuze,et al.  Exosome: from internal vesicle of the multivesicular body to intercellular signaling device. , 2000, Journal of cell science.

[60]  O. Olivieri,et al.  Optimizing the purification and analysis of miRNAs from urinary exosomes , 2013, Clinical chemistry and laboratory medicine.

[61]  Thomas Würdinger,et al.  Endothelial cells require miR-214 to secrete exosomes that suppress senescence and induce angiogenesis in human and mouse endothelial cells. , 2013, Blood.

[62]  Kit Lam,et al.  Single exosome study reveals subpopulations distributed among cell lines with variability related to membrane content , 2015, Journal of extracellular vesicles.

[63]  Wei Huang,et al.  Ischemic Preconditioning Potentiates the Protective Effect of Stem Cells through Secretion of Exosomes by Targeting Mecp2 via miR-22 , 2014, PloS one.

[64]  V. Kralj-Iglič,et al.  The role of extracellular vesicles in phenotypic cancer transformation , 2013, Radiology and oncology.

[65]  A. Möller,et al.  Optimized exosome isolation protocol for cell culture supernatant and human plasma , 2015, Journal of extracellular vesicles.

[66]  Shinobu Ueda,et al.  Systemically Injected Exosomes Targeted to EGFR Deliver Antitumor MicroRNA to Breast Cancer Cells. , 2013, Molecular therapy : the journal of the American Society of Gene Therapy.

[67]  P. Stahl,et al.  Multivesicular Bodies and Multivesicular Endosomes: The "Ins and Outs" of Endosomal Traffic , 2002, Science's STKE.

[68]  Biana Godin,et al.  Ciliated micropillars for the microfluidic-based isolation of nanoscale lipid vesicles. , 2013, Lab on a chip.

[69]  Myung Soo Kim,et al.  Using exosomes, naturally-equipped nanocarriers, for drug delivery. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[70]  Laurence Zitvogel,et al.  Eradication of established murine tumors using a novel cell-free vaccine: dendritic cell derived exosomes , 1998, Nature Medicine.

[71]  R. Johnstone,et al.  Reticulocyte maturation and exosome release: transferrin receptor containing exosomes shows multiple plasma membrane functions. , 1989, Blood.

[72]  M. Record,et al.  Characterization of exosome subpopulations from RBL-2H3 cells using fluorescent lipids. , 2005, Blood cells, molecules & diseases.

[73]  Laurence Zitvogel,et al.  Exosomes: composition, biogenesis and function , 2002, Nature Reviews Immunology.

[74]  J. Meldolesi,et al.  Ectosomes and exosomes: shedding the confusion between extracellular vesicles. , 2015, Trends in cell biology.

[75]  J. Pérez-Gracia,et al.  Circulating melanoma exosomes as diagnostic and prognosis biomarkers. , 2016, Clinica chimica acta; international journal of clinical chemistry.

[76]  B. Pan,et al.  Fate of the transferrin receptor during maturation of sheep reticulocytes in vitro: Selective externalization of the receptor , 1983, Cell.

[77]  Richard J. Simpson,et al.  Proteomics Analysis of A33 Immunoaffinity-purified Exosomes Released from the Human Colon Tumor Cell Line LIM1215 Reveals a Tissue-specific Protein Signature* , 2009, Molecular & Cellular Proteomics.

[78]  Mark Blaxter,et al.  Extracellular Onchocerca-derived small RNAs in host nodules and blood , 2015, Parasites & Vectors.

[79]  Sascha Keller,et al.  Exosomes: from biogenesis and secretion to biological function. , 2006, Immunology letters.

[80]  S. Mathivanan,et al.  Exosomes: extracellular organelles important in intercellular communication. , 2010, Journal of proteomics.

[81]  T. Wurdinger,et al.  Microfluidic isolation and transcriptome analysis of serum microvesicles. , 2010, Lab on a chip.

[82]  Aled Clayton,et al.  Cancer exosomes trigger fibroblast to myofibroblast differentiation. , 2010, Cancer research.

[83]  Sébastien Roy,et al.  Mast cell- and dendritic cell-derived exosomes display a specific lipid composition and an unusual membrane organization. , 2004, The Biochemical journal.

[84]  T. Whiteside,et al.  Isolation and Characterization of CD34+ Blast-Derived Exosomes in Acute Myeloid Leukemia , 2014, PloS one.

[85]  Shivani Sharma,et al.  Elucidating diversity of exosomes: biophysical and molecular characterization methods. , 2016, Nanomedicine.

[86]  D. Hawke,et al.  Benchtop isolation and characterization of functional exosomes by sequential filtration. , 2014, Journal of chromatography. A.

[87]  Hamid Cheshmi Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers , 2011 .

[88]  Douglas D. Taylor,et al.  Exosomal microRNA: a diagnostic marker for lung cancer. , 2008, Clinical lung cancer.

[89]  Anna Janowska-Wieczorek,et al.  Microvesicles derived from activated platelets induce metastasis and angiogenesis in lung cancer , 2005, International journal of cancer.

[90]  Jaesung Park,et al.  Microfluidic filtration system to isolate extracellular vesicles from blood. , 2012, Lab on a chip.

[91]  Graça Raposo,et al.  Extracellular vesicles: Exosomes, microvesicles, and friends , 2013, The Journal of cell biology.

[92]  Hakho Lee,et al.  Acoustic purification of extracellular microvesicles. , 2015, ACS nano.

[93]  H. Nakagawa,et al.  Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes , 2014, Scientific Reports.

[94]  D. Gu,et al.  A Simplified Method to Recover Urinary Vesicles for Clinical Applications, and Sample Banking , 2014, Scientific Reports.

[95]  M. Wood,et al.  Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes , 2011, Nature Biotechnology.

[96]  Clotilde Théry,et al.  Exosome Secretion: Molecular Mechanisms and Roles in Immune Responses , 2011, Traffic.

[97]  Bernard Monsarrat,et al.  Exosomes account for vesicle-mediated transcellular transport of activatable phospholipases and prostaglandins[S] , 2010, Journal of Lipid Research.