A comparative study of serum exosome isolation using differential ultracentrifugation and three commercial reagents

PLoS ONE

Volumn 12, Issue 1, 2017, Pages

  Helwa, Inas ^a   Cai, Jingwen ^a   Drewry, Michelle D ^a   Zimmerman, Arthur ^a   Dinkins, Michael B ^a   Khaled, Mariam Lotfy ^a   Seremwe, Mutsa ^a   Dismuke, W Michael ^b   Bieberich, Erhard ^a   Stamer, W Daniel ^b,c   Hamrick, Mark W ^a   Liu, Yutao ^a  

^a MEDICAL COLLEGE OF GEORGIA   (United States)

^b DUKE UNIVERSITY   (United States)

^c Duke University   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CD63 ANTIGEN; CD9 ANTIGEN; EXOSOMAL RNA; MICRORNA; MICRORNA 16; MICRORNA 451; RNA; UNCLASSIFIED DRUG; DYES, REAGENTS, INDICATORS, MARKERS AND BUFFERS; NANOPARTICLE;

ANTIGEN EXPRESSION; ARTICLE; CELL ISOLATION; CELL SIZE; CHEMICAL ANALYZER; CONTROLLED STUDY; EXOSOME; HUMAN; HUMAN CELL; SERUM; ULTRACENTRIFUGATION; ZETA POTENTIAL; CHEMISTRY; COMPARATIVE STUDY; METABOLISM; POLYMERASE CHAIN REACTION; PROCEDURES; PROTEOMICS; TRANSMISSION ELECTRON MICROSCOPY; WESTERN BLOTTING;

BLOTTING, WESTERN; EXOSOMES; HUMANS; INDICATORS AND REAGENTS; MICROSCOPY, ELECTRON, TRANSMISSION; NANOPARTICLES; POLYMERASE CHAIN REACTION; PROTEOMICS; RNA; ULTRACENTRIFUGATION;

EID: 85010295300     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0170628     Document Type: Article

References (81)

1. Admyre C, Johansson SM, Qazi KR, Filen JJ, Lahesmaa R, Norman M, et al. Exosomes with immune modulatory features are present in human breast milk. Journal of immunology. 2007; 179(3):1969-78.
2. Alegre E, Sanmamed MF, Rodriguez C, Carranza O, Martin-Algarra S, Gonzalez A. Study of circulating microRNA-125b levels in serum exosomes in advanced melanoma. Archives of pathology & laboratory medicine. 2014; 138(6):828-32.
3. Conde-Vancells J, Rodriguez-Suarez E, Embade N, Gil D, Matthiesen R, Valle M, et al. Characterization and Comprehensive Proteome Profiling of Exosomes Secreted by Hepatocytes. J Proteome Res. 2008; 7(12):5157-66. PMID: 19367702
4. Hegmans JPJJ, Bard MPL, Hemmes A, Luider TM, Kleijmeer MJ, Prins J-B, et al. Proteomic Analysis of Exosomes Secreted by Human Mesothelioma Cells. Am J Pathol. 2004; 164(5):1807-15. doi: 10.1016/ S0002-9440(10)63739-X PMID: 15111327
5. Nilsson J, Skog J, Nordstrand A, Baranov V, Mincheva-Nilsson L, Breakefield XO, et al. Prostate cancer-derived urine exosomes: a novel approach to biomarkers for prostate cancer. British journal of cancer. 2009; 100(10):1603-7. PubMed Central PMCID: PMC2696767. doi: 10.1038/sj.bjc.6605058 PMID: 19401683
6. Rabinowits G, Gercel-Taylor C, Day JM, Taylor DD, Kloecker GH. Exosomal microRNA: a diagnostic marker for lung cancer. Clinical lung cancer. 2009; 10(1):42-6. doi: 10.3816/CLC.2009.n.006 PMID: 19289371
7. Zhang W, Zhou X, Zhang H, Yao Q, Liu Y, Dong Z. Extracellular Vesicles in Diagnosis and Therapy of Kidney Diseases. Am J Physiol Renal Physiol. 2016:ajprenal 00429 2016.
8. Thery C, Amigorena S, Raposo G, Clayton A. Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Current protocols in cell biology / editorial board, Juan S Bonifacino [et al]. 2006;Chapter 3:Unit 3 22.
9. Mathivanan S, Ji H, Simpson RJ. Exosomes: extracellular organelles important in intercellular communication. Journal of proteomics. 2010; 73(10):1907-20. doi: 10.1016/j.jprot.2010.06.006 PMID: 20601276
10. van der Pol E, Boing AN, Harrison P, Sturk A, Nieuwland R. Classification, functions, and clinical relevance of extracellular vesicles. Pharmacological reviews. 2012; 64(3):676-705. doi: 10.1124/pr.112. 005983 PMID: 22722893
11. Camussi G, Deregibus MC, Bruno S, Grange C, Fonsato V, Tetta C. Exosome/microvesicle-mediated epigenetic reprogramming of cells. American journal of cancer research. 2011; 1(1):98-110. PubMed Central PMCID: PMC3180104. PMID: 21969178
12. Rak J, Guha A. Extracellular vesicles-vehicles that spread cancer genes. BioEssays: news and reviews in molecular, cellular and developmental biology. 2012; 34(6):489-97.
13. Dinkins MB, Enasko J, Hernandez C, Wang G, Kong J, Helwa I, et al. Neutral Sphingomyelinase-2 Deficiency Ameliorates Alzheimer's Disease Pathology and Improves Cognition in the 5XFAD Mouse. J Neurosci. 2016; 36(33):8653-67. doi: 10.1523/JNEUROSCI.1429-16.2016 PMID: 27535912
14. Ibrahim A, Marban E. Exosomes: Fundamental Biology and Roles in Cardiovascular Physiology. Annu Rev Physiol. 2015.
15. Vader P, Breakefield XO, Wood MJ. Extracellular vesicles: emerging targets for cancer therapy. Trends in molecular medicine. 2014; 20(7):385-93. PubMed Central PMCID: PMC4082760. doi: 10.1016/j. molmed.2014.03.002 PMID: 24703619
16. Lawson C, Vicencio JM, Yellon DM, Davidson SM. Microvesicles and exosomes: new players in metabolic and cardiovascular disease. The Journal of endocrinology. 2016; 228(2):R57-71. doi: 10.1530/ JOE-15-0201 PMID: 26743452
17. Yanez-Mo M, Siljander PR, Andreu Z, Zavec AB, Borras FE, Buzas EI, et al. Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles. 2015; 4:27066. PubMed Central PMCID: PMCPMC4433489. doi: 10.3402/jev.v4.27066 PMID: 25979354
18. Emanueli C, Shearn AI, Angelini GD, Sahoo S. Exosomes and exosomal miRNAs in cardiovascular protection and repair. Vascular pharmacology. 2015; 71:24-30. PubMed Central PMCID: PMC4838026. doi: 10.1016/j.vph.2015.02.008 PMID: 25869502
19. An T, Qin S, Xu Y, Tang Y, Huang Y, Situ B, et al. Exosomes serve as tumour markers for personalized diagnostics owing to their important role in cancer metastasis. J Extracell Vesicles. 2015; 4:27522. PubMed Central PMCID: PMC4475684. doi: 10.3402/jev.v4.27522 PMID: 26095380
20. Ailawadi S, Wang X, Gu H, Fan GC. Pathologic function and therapeutic potential of exosomes in cardiovascular disease. Biochimica et biophysica acta. 2015; 1852(1):1-11. PubMed Central PMCID: PMC4268281. doi: 10.1016/j.bbadis.2014.10.008 PMID: 25463630
21. Zhao L, Liu W, Xiao J, Cao B. The role of exosomes and "exosomal shuttle microRNA" in tumorigenesis and drug resistance. Cancer letters. 2015; 356(2 Pt B):339-46.
22. Lotvall J, Hill AF, Hochberg F, Buzas EI, Di Vizio D, Gardiner C, et al. Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J Extracell Vesicles. 2014; 3:26913. PubMed Central PMCID: PMC4275645. doi: 10.3402/jev.v3.26913 PMID: 25536934
23. Szatanek R, Baran J, Siedlar M, Baj-Krzyworzeka M. Isolation of extracellular vesicles: Determining the correct approach (Review). International journal of molecular medicine. 2015; 36(1):11-7. PubMed Central PMCID: PMC4494580. doi: 10.3892/ijmm.2015.2194 PMID: 25902369
24. Jeppesen DK, Hvam ML, Primdahl-Bengtson B, Boysen AT, Whitehead B, Dyrskjot L, et al. Comparative analysis of discrete exosome fractions obtained by differential centrifugation. J Extracell Vesicles. 2014; 3:25011. PubMed Central PMCID: PMC4224706. doi: 10.3402/jev.v3.25011 PMID: 25396408
25. Sohel MMH, Hoelker M, Noferesti SS, Salilew-Wondim D, Tholen E, Looft C, et al. Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence. Plos One. 2013; 8(11).
26. Hartman ZC, Wei J, Glass OK, Guo H, Lei G, Yang XY, et al. Increasing vaccine potency through exosome antigen targeting. Vaccine. 2011; 29(50):9361-7. PubMed Central PMCID: PMC3350974. doi: 10.1016/j.vaccine.2011.09.133 PMID: 22001882
27. da Silveira JC, Veeramachaneni DN, Winger QA, Carnevale EM, Bouma GJ. Cell-secreted vesicles in equine ovarian follicular fluid contain miRNAs and proteins: a possible new form of cell communication within the ovarian follicle. Biology of reproduction. 2012; 86(3):71. doi: 10.1095/biolreprod.111.093252 PMID: 22116803
28. Franzen CA, Blackwell RH, Foreman KE, Kuo PC, Flanigan RC, Gupta GN. Urinary Exosomes: The Potential for Biomarker Utility, Intercellular Signaling and Therapeutics in Urological Malignancy. The Journal of urology. 2016; 195(5):1331-9. doi: 10.1016/j.juro.2015.08.115 PMID: 26714199
29. Hoshino A, Costa-Silva B, Shen TL, Rodrigues G, Hashimoto A, Tesic Mark M, et al. Tumour exosome integrins determine organotropic metastasis. Nature. 2015; 527(7578):329-35. PubMed Central PMCID: PMC4788391. doi: 10.1038/nature15756 PMID: 26524530
30. Filipe V, Hawe A, Jiskoot W. Critical evaluation of Nanoparticle Tracking Analysis (NTA) by NanoSight for the measurement of nanoparticles and protein aggregates. Pharmaceutical research. 2010; 27 (5):796-810. PubMed Central PMCID: PMC2852530. doi: 10.1007/s11095-010-0073-2 PMID: 20204471
31. Mehdiani A, Maier A, Pinto A, Barth M, Akhyari P, Lichtenberg A. An innovative method for exosome quantification and size measurement. Journal of visualized experiments: JoVE. 2015;(95):50974. PubMed Central PMCID: PMC4354536. doi: 10.3791/50974 PMID: 25650897
32. Wang Y, Zhang L, Li Y, Chen L, Wang X, Guo W, et al. Exosomes/microvesicles from induced pluripotent stem cells deliver cardioprotective miRNAs and prevent cardiomyocyte apoptosis in the ischemic myocardium. International journal of cardiology. 2015; 192:61-9. PubMed Central PMCID: PMC4469495. doi: 10.1016/j.ijcard.2015.05.020 PMID: 26000464
33. Gallego-Urrea JA, Tuoriniemi J, Hassellöv M. Applications of particle-tracking analysis to the determination of size distributions and concentrations of nanoparticles in environmental, biological and food samples. Trends Analyt Chem. 2011; 30(3):473-83.
34. Seader JD, Henley E. J. & Roper D. K. Separation process principles (Chemical and biochemical operations). 3 ed: John Wiley and Sons; 2011.
35. Sokolova V, Ludwig AK, Hornung S, Rotan O, Horn PA, Epple M, et al. Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. Colloids and surfaces B, Biointerfaces. 2011; 87(1):146-50. doi: 10.1016/j.colsurfb.2011.05.013 PMID: 21640565
36. Hanaor D, Michelazzi M, Leonelli C, Sorrell CC. The effects of carboxylic acids on the aqueous dispersion and electrophoretic deposition of ZrO2. J Eur Ceram Soc. 2012; 32(1):235-44.
37. Alvarez ML, Khosroheidari M, Kanchi Ravi R, DiStefano JK. Comparison of protein, microRNA, and mRNA yields using different methods of urinary exosome isolation for the discovery of kidney disease biomarkers. Kidney international. 2012; 82(9):1024-32. doi: 10.1038/ki.2012.256 PMID: 22785172
38. Lane RE, Korbie D, Anderson W, Vaidyanathan R, Trau M. Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing. Scientific reports. 2015; 5:7639. PubMed Central PMCID: PMC4648344. doi: 10.1038/srep07639 PMID: 25559219
39. Rekker K, Saare M, Roost AM, Kubo AL, Zarovni N, Chiesi A, et al. Comparison of serum exosome isolation methods for microRNA profiling. Clinical biochemistry. 2014; 47(1-2):135-8. doi: 10.1016/j. clinbiochem.2013.10.020 PMID: 24183884
40. Webber J, Clayton A. How pure are your vesicles? J Extracell Vesicles. 2013; 2. PubMed Central PMCID: PMC3760653.
41. Andreu Z, Rivas E, Sanguino-Pascual A, Lamana A, Marazuela M, Gonzalez-Alvaro I, et al. Comparative analysis of EV isolation procedures for miRNAs detection in serum samples. J Extracell Vesicles. 2016; 5:31655. PubMed Central PMCID: PMC4916259. doi: 10.3402/jev.v5.31655 PMID: 27330048
42. Crossland RE, Norden J, Bibby LA, Davis J, Dickinson AM. Evaluation of optimal extracellular vesicle small RNA isolation and qRT-PCR normalisation for serum and urine. Journal of immunological methods. 2016; 429:39-49. doi: 10.1016/j.jim.2015.12.011 PMID: 26723490
43. Kalra H, Adda CG, Liem M, Ang CS, Mechler A, Simpson RJ, et al. Comparative proteomics evaluation of plasma exosome isolation techniques and assessment of the stability of exosomes in normal human blood plasma. Proteomics. 2013; 13(22):3354-64. doi: 10.1002/pmic.201300282 PMID: 24115447
44. Tauro BJ, Greening DW, Mathias RA, Ji H, Mathivanan S, Scott AM, et al. Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes. Methods. 2012; 56(2):293-304. doi: 10.1016/j.ymeth.2012.01. 002 PMID: 22285593
45. Baranyai T, Herczeg K, Onodi Z, Voszka I, Modos K, Marton N, et al. Isolation of Exosomes from Blood Plasma: Qualitative and Quantitative Comparison of Ultracentrifugation and Size Exclusion Chromatography Methods. Plos One. 2015; 10(12).
46. Lobb RJ, Becker M, Wen SW, Wong CS, Wiegmans AP, Leimgruber A, et al. Optimized exosome isolation protocol for cell culture supernatant and human plasma. J Extracell Vesicles. 2015; 4:27031. ubMed Central PMCID: PMC4507751. doi: 10.3402/jev.v4.27031 PMID: 26194179
47. Kowal J, Arras G, Colombo M, Jouve M, Morath JP, Primdal-Bengtson B, et al. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proceedings of the National Academy of Sciences of the United States of America. 2016; 113(8):E968-77. ubMed Central PMCID: PMC4776515. doi: 10.1073/pnas.1521230113 PMID: 26858453
48. Shalaby SM, Khater MK, Perucho AM, Mohamed SA, Helwa I, Laknaur A, et al. Magnetic nanoparticles as a new approach to improve the efficacy of gene therapy against differentiated human uterine fibroid cells and tumor-initiating stem cells. Fertility and sterility. 2016; 105(6):1638-48 e8. PubMed Central PMCID: PMC4971775. doi: 10.1016/j.fertnstert.2016.03.001 PMID: 27020169
49. Dismuke WM, Challa P, Navarro I, Stamer WD, Liu Y. Human aqueous humor exosomes. Experimental eye research. 2015; 132:73-7. PubMed Central PMCID: PMC4352394. doi: 10.1016/j.exer.2015.01. 19 PMID: 25619138
50. Liu Y, Bailey JC, Helwa I, Dismuke WM, Cai J, Drewry M, et al. A Common Variant in MIR182 Is Associated With Primary Open-Angle Glaucoma in the NEIGHBORHOOD Consortium. Invest Ophthalmol Vis Sci. 2016; 57(10):3974-81.
51. El-Khoury V, Pierson S, Kaoma T, Bernardin F, Berchem G. Assessing cellular and circulating miRNA recovery: the impact of the RNA isolation method and the quantity of input material. Scientific reports. 2016; 6:19529. PubMed Central PMCID: PMC4726450. doi: 10.1038/srep19529 PMID: 26787294
52. Lasser C, Eldh M, Lotvall J. Isolation and characterization of RNA-containing exosomes. Journal of visualized experiments: JoVE. 2012;(59):e3037. Epub 2012/01/20. PubMed Central PMCID: PMC3369768. doi: 10.3791/3037 PMID: 22257828
53. Cai J, Perkumas KM, Qin X, Hauser MA, Stamer WD, Liu Y. Expression Profiling of Human Schlemm's Canal Endothelial Cells From Eyes With and Without Glaucoma. Invest Ophthalmol Vis Sci. 2015; 56 (11):6747-53. PubMed Central PMCID: PMC4614909. doi: 10.1167/iovs.15-17720 PMID: 26567786
54. Chevillet JR, Kang Q, Ruf IK, Briggs HA, Vojtech LN, Hughes SM, et al. Quantitative and stoichiometric analysis of the microRNA content of exosomes. Proc Natl Acad Sci USA. 2014; 111(41):14888-93. doi: 10.1073/pnas.1408301111 PMID: 25267620
55. Li M, Rai AJ, DeCastro GJ, Zeringer E, Barta T, Magdaleno S, et al. An optimized procedure for exosome isolation and analysis using serum samples: Application to cancer biomarker discovery. Methods. 2015; 87:26-30. doi: 10.1016/j.ymeth.2015.03.009 PMID: 25814440
56. Ruxton GD. The unequal variance t-test is an underused alternative to Student's t-test and the Mann-Whitney U test. Behavioral Ecology. 2006; 17(4):688-90.
57. Akers JC, Ramakrishnan V, Nolan JP, Duggan E, Fu CC, Hochberg FH, et al. Comparative Analysis of Technologies for Quantifying Extracellular Vesicles (EVs) in Clinical Cerebrospinal Fluids (CSF). PLoS One. 2016; 11(2):e0149866. PubMed Central PMCID: PMC4763994. doi: 10.1371/journal.pone. 149866 PMID: 26901428
58. Van Deun J, Mestdagh P, Sormunen R, Cocquyt V, Vermaelen K, Vandesompele J, et al. The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling. J Extracell Vesicles. 2014; 3. PubMed Central PMCID: PMC4169610.
59. EL Andaloussi S, Mager I, Breakefield XO, Wood MJ. Extracellular vesicles: biology and emerging therapeutic opportunities. Nature reviews Drug discovery. 2013; 12(5):347-57. doi: 10.1038/nrd3978 MID: 23584393
60. Properzi F, Logozzi M, Fais S. Exosomes: the future of biomarkers in medicine. Biomarkers in medicine. 2013; 7(5):769-78. doi: 10.2217/bmm.13.63 PMID: 24044569
61. Zarovni N, Corrado A, Guazzi P, Zocco D, Lari E, Radano G, et al. Integrated isolation and quantitative analysis of exosome shuttled proteins and nucleic acids using immunocapture approaches. Methods. 2015; 87:46-58. doi: 10.1016/j.ymeth.2015.05.028 PMID: 26044649
62. Rider MA, Hurwitz SN, Meckes DG Jr. ExtraPEG: A Polyethylene Glycol-Based Method for Enrichment of Extracellular Vesicles. Scientific reports. 2016; 6:23978. PubMed Central PMCID: PMC4828635. doi: 10.1038/srep23978 PMID: 27068479
63. Rood IM, Deegens JK, Merchant ML, Tamboer WP, Wilkey DW, Wetzels JF, et al. Comparison of three methods for isolation of urinary microvesicles to identify biomarkers of nephrotic syndrome. Kidney international. 2010; 78(8):810-6. doi: 10.1038/ki.2010.262 PMID: 20686450
64. Linares R, Tan S, Gounou C, Arraud N, Brisson AR. High-speed centrifugation induces aggregation of extracellular vesicles. J Extracell Vesicles. 2015; 4:29509. PubMed Central PMCID: PMCPMC4689953. doi: 10.3402/jev.v4.29509 PMID: 26700615
65. Caradec J, Kharmate G, Hosseini-Beheshti E, Adomat H, Gleave M, Guns E. Reproducibility and efficiency of serum-derived exosome extraction methods. Clinical biochemistry. 2014; 47(13-14):1286-92. doi: 10.1016/j.clinbiochem.2014.06.011 PMID: 24956264
66. Deregibus MC, Figliolini F, D'Antico S, Manzini PM, Pasquino C, De Lena M, et al. Charge-based precipitation of extracellular vesicles. International journal of molecular medicine. 2016; 38(5):1359-66. doi: 10.3892/ijmm.2016.2759 PMID: 28025988
67. Lorincz AM, Timar CI, Marosvari KA, Veres DS, Otrokocsi L, Kittel A, et al. Effect of storage on physical and functional properties of extracellular vesicles derived from neutrophilic granulocytes. J Extracell Vesicles. 2014; 3:25465. PubMed Central PMCID: PMC4275651. doi: 10.3402/jev.v3.25465 PMID: 25536933
68. Schageman J, Zeringer E, Li M, Barta T, Lea K, Gu J, et al. The complete exosome workflow solution: from isolation to characterization of RNA cargo. BioMed research international. 2013; 2013:253957. ubMed Central PMCID: PMC3800616. doi: 10.1155/2013/253957 PMID: 24205503
69. Lee M, Ban J-J, Im W, Kim M. Influence of storage condition on exosome recovery. Biotechnology and Bioprocess Engineering. 2016; 21(2):299-304.
70. Zhou H, Yuen PS, Pisitkun T, Gonzales PA, Yasuda H, Dear JW, et al. Collection, storage, preservation, and normalization of human urinary exosomes for biomarker discovery. Kidney international. 2006; 69(8):1471-6. PubMed Central PMCID: PMC2276656. doi: 10.1038/sj.ki.5000273 PMID: 16501490
71. Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nature cell biology. 2007; 9(6):654-9. doi: 10.1038/ncb1596 PMID: 17486113
72. Zhang J, Li S, Li L, Li M, Guo C, Yao J, et al. Exosome and exosomal microRNA: trafficking, sorting, and function. Genomics, proteomics & bioinformatics. 2015; 13(1):17-24. PubMed Central PMCID: PMC4411500.
73. Alvarez-Garcia I, Miska EA. MicroRNA functions in animal development and human disease. Development. 2005; 132(21):4653-62. doi: 10.1242/dev.02073 PMID: 16224045
74. Mikaelian I, Scicchitano M, Mendes O, Thomas RA, Leroy BE. Frontiers in preclinical safety biomarkers: microRNAs and messenger RNAs. Toxicologic pathology. 2013; 41(1):18-31. Epub 2012/06/05. doi: 10.1177/0192623312448939 PMID: 22659243
75. Vickers KC, Palmisano BT, Shoucri BM, Shamburek RD, Remaley AT. MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins. Nature cell biology. 2011; 13 (4):423-33. Epub 2011/03/23. PubMed Central PMCID: PMCPmc3074610. doi: 10.1038/ncb2210 MID: 21423178
76. Tabet F, Vickers KC, Cuesta Torres LF, Wiese CB, Shoucri BM, Lambert G, et al. HDL-transferred microRNA-223 regulates ICAM-1 expression in endothelial cells. Nature communications. 2014; 5:3292. Epub 2014/03/01. PubMed Central PMCID: PMCPmc4189962. doi: 10.1038/ncomms4292 MID: 24576947
77. Peterson MF, Otoc N, Sethi JK, Gupta A, Antes TJ. Integrated systems for exosome investigation. Methods. 2015; 87:31-45. doi: 10.1016/j.ymeth.2015.04.015 PMID: 25916618
78. Sodar BW, Kittel A, Paloczi K, Vukman KV, Osteikoetxea X, Szabo-Taylor K, et al. Low-density lipoprotein mimics blood plasma-derived exosomes and microvesicles during isolation and detection. Scientific reports. 2016; 6:24306-16. Epub 2016/04/19. PubMed Central PMCID: PMCPmc4834552.
79. Patel RS, Carter G, El Bassit G, Patel AA, Cooper DR, Murr M, et al. Adipose-derived stem cells from lean and obese humans show depot specific differences in their stem cell markers, exosome contents and senescence: role of protein kinase C delta (PKCdelta) in adipose stem cell niche. Stem cell investigation. 2016; 3:2. Epub 2016/07/01. PubMed Central PMCID: PMCPmc4923648. doi: 10.3978/j.issn. 2306-9759.2016.01.02 PMID: 27358894
80. Laurent LC, Abdel-Mageed AB, Adelson PD, Arango J, Balaj L, Breakefield X, et al. Meeting report: discussions and preliminary findings on extracellular RNA measurement methods from laboratories in the NIH Extracellular RNA Communication Consortium. J Extracell Vesicles. 2015; 4:26533. Epub 2015/ 09/01. PubMed Central PMCID: PMCPmc4553263. doi: 10.3402/jev.v4.26533 PMID: 26320937
81. Ueda K, Ishikawa N, Tatsuguchi A, Saichi N, Fujii R, Nakagawa H. Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes. Scientific reports. 2014; 4:6232. Epub 2014/08/30. PubMed Central PMCID: PMCPmc4148700. doi: 10.1038/srep06232 PMID: 25167841