title: Physical characterization and profiling of airway epithelial derived exosomes using light scattering.
abstract: Exosomes and other extracellular vesicles have been gaining interest during the last decade due to their emerging role in biology and, disease pathogenesis and their biomarker potential. Almost all published research related to exosomes and other extracellular vesicles include some form of physical characterization. Therefore, these vesicles should be precisely profiled and characterized physically before studying their biological role as intercellular messengers, biomarkers or therapeutic tools. Using a combination of light scattering techniques, including dynamic light scattering (DLS) and multi-angle laser light scattering combined with size exclusion separation (SEC-MALLS), we physically characterized and compared distinct extracellular vesicles derived from the apical secretions of two different cultured airway epithelial cells. The results indicated that epithelial cells release vesicles with distinct physical properties and sizes. Human primary tracheobronchial cell culture (HTBE) derived vesicles have a hydrodynamic radius (Rh) of approximately 340 nm while their radius of gyration (Rg) is approximately 200 nm. Electron microscopy analysis, however, revealed that their spherical component is 40-100 nm in size, and they carry filamentous, entangled membrane mucins on their surface that increases their overall radius. The mucin decoration on the surface defines their size and charge as measured using light scattering techniques. Their surface properties mirror the properties of the cells from which they are derived. This may provide a unique tool for researchers to elucidate the unanswered questions in normal airway biology and innate and adaptive defense, including the remodeling of airways during inflammation, tumorigenesis and metastasis.
authors.affiliation: Department of Pathology and Laboratory Medicine, Cystic Fibrosis/Pulmonary Research and Treatment Center, Marsico Lung Institute, University of North Carolina, Chapel Hill, NC 27599-7248, United States. Electronic address: kesimer@med.unc.edu.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Kesimer
authors.affiliation: Department of Pathology and Laboratory Medicine, Cystic Fibrosis/Pulmonary Research and Treatment Center, Marsico Lung Institute, University of North Carolina, Chapel Hill, NC 27599-7248, United States.
authors.forename: Richa
authors.initials: R
authors.lastname: Gupta
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D002460:Cell Line; D004847:Epithelial Cells / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D055354:Exosomes / Q000737:chemistry*; D006801:Humans; D008027:Light; D009077:Mucins / Q000737:chemistry / Q000378:metabolism; D010316:Particle Size; D061251:Primary Cell Culture; D020545:Respiratory Mucosa / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D012542:Scattering, Radiation; D055672:Static Electricity
chemical_list: D015415:Biomarkers; D009077:Mucins
keywords: Airway mucus; Dynamic light scattering; Exosomes; Membrane tethered mucins; Static light scattering
references.citation: Genes Chromosomes Cancer. 2014 Sep;53(9):713-24
references.pmid: 24764226
references.citation: Hum Pathol. 2010 Sep;41(9):1265-75
references.pmid: 20447677
references.citation: Cell Tissue Res. 2013 Apr;352(1):33-47
references.pmid: 22610588
references.citation: Expert Rev Proteomics. 2014 Oct;11(5):621-39
references.pmid: 25115191
references.citation: Colloids Surf B Biointerfaces. 2011 Oct 1;87(1):146-50
references.pmid: 21640565
references.citation: J Biol Chem. 1987 Jul 5;262(19):9412-20
references.pmid: 3597417
references.citation: J Cell Biol. 2007 May 21;177(4):671-81
references.pmid: 17502425
references.citation: J Immunol Methods. 2014 Sep;411:55-65
references.pmid: 24952243
references.citation: PLoS One. 2014;9(4):e92921
references.pmid: 24705249
references.citation: Curr Protoc Cell Biol. 2006 Apr;Chapter 3:Unit 3.22
references.pmid: 18228490
references.citation: Nanomedicine. 2011 Dec;7(6):780-8
references.pmid: 21601655
references.citation: J Proteome Res. 2012 Aug 3;11(8):4013-23
references.pmid: 22663354
references.citation: Proteomics. 2013 May;13(10-11):1672-86
references.pmid: 23585443
references.citation: PLoS One. 2013;8(12):e82589
references.pmid: 24391718
references.citation: J Physiol. 2007 Oct 1;584(Pt 1):245-59
references.pmid: 17656429
references.citation: Blood. 2013 Sep 12;122(11):1873-80
references.pmid: 23798713
references.citation: Anal Biochem. 2012 Sep 1;428(1):44-53
references.pmid: 22691960
references.citation: Clin Immunol. 2013 Nov;149(2):236-43
references.pmid: 23706172
references.citation: Allergy. 2012 Jul;67(7):911-9
references.pmid: 22620679
references.citation: Nat Rev Immunol. 2002 Aug;2(8):569-79
references.pmid: 12154376
references.citation: Nat Rev Mol Cell Biol. 2012 May;13(5):328-35
references.pmid: 22510790
references.citation: J Reprod Immunol. 2008 Oct;79(1):12-7
references.pmid: 18715652
references.citation: J Membr Biol. 2014 Nov;247(11):1129-36
references.pmid: 25135166
references.citation: FASEB J. 2009 Jun;23(6):1858-68
references.pmid: 19190083
references.citation: Methods Mol Med. 2005;107:183-206
references.pmid: 15492373
references.citation: Nat Cell Biol. 2012 Jul;14(7):654-5
references.pmid: 22743708
references.citation: J Immunol. 2014 Aug 1;193(3):1006-11
references.pmid: 25049431
references.citation: Respir Res. 2010;11:94
references.pmid: 20618958
references.citation: Front Physiol. 2012 Sep 07;3:354
references.pmid: 22973237
references.citation: Nat Immunol. 2013 Aug;14(8):793-803
references.pmid: 23832071
references.citation: Sci Rep. 2014;4:5750
references.pmid: 25034888
references.citation: Biol Pharm Bull. 2013;36(1):66-75
references.pmid: 23302638
references.citation: J Am Soc Nephrol. 2014 Sep;25(9):2017-27
references.pmid: 24700864
references.citation: Oral Dis. 2010 Jan;16(1):34-8
references.pmid: 19627513
references.citation: Am J Physiol Lung Cell Mol Physiol. 2009 Jan;296(1):L92-L100
references.pmid: 18931053
references.citation: Gene. 2014 Nov 1;551(1):55-64
references.pmid: 25158130
references.citation: J Allergy Clin Immunol. 2013 Apr;131(4):1194-203, 1203.e1-14
references.pmid: 23414598
references.citation: PLoS One. 2012;7(10):e47480
references.pmid: 23094054
references.citation: Diabetes Metab Syndr Obes. 2012;5:247-82
references.pmid: 22924003
references.citation: Alzheimers Dement. 2015 Jun;11(6):600-7.e1
references.pmid: 25130657
references.citation: Fertil Steril. 2007 Oct;88(4 Suppl):1212-9
references.pmid: 17562335