Physical characterization and profiling of airway epithelial derived exosomes using light scattering.
Physical characterization and profiling of airway epithelial derived exosomes using light scattering.
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DOI：
10.1016/j.ymeth.2015.03.013
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发表时间：
2015-10-01
期刊：
影响因子：
--
通讯作者：
Gupta R
中科院分区：
文献类型：
--
作者：
Kesimer M;Gupta R
外泌体和其他细胞外囊泡由于其在生物学、疾病发病机制和生物标志物方面的新兴作用，在过去十年中引起了人们的兴趣。几乎所有发表的有关外泌体和其他细胞外囊泡的研究都包括某种形式的物理表征。因此，在研究这些囊泡作为细胞间信使、生物标志物或治疗工具的生物学作用之前，应该对其进行精确的描绘和物理表征。利用光散射技术，包括动态光散射（DLS）和多角度激光光散射结合尺寸排除分离（SEC-MALLS），我们对两种不同培养的气道上皮细胞的根尖分泌物进行了物理表征并比较了不同的细胞外囊泡。结果表明，上皮细胞释放的囊泡具有不同的物理性质和大小。人原代气管支气管细胞培养（HTBE）衍生囊泡的流体动力学半径（Rh）约为340 nm，而其旋转半径（Rg）约为200 nm。然而，电子显微镜分析显示，它们的球形成分的尺寸为40-100纳米，它们的表面携带丝状的、纠缠的膜粘蛋白，这增加了它们的整体半径。用光散射技术测量，表面的粘蛋白装饰决定了它们的大小和电荷。它们的表面特性反映了它们来源于的细胞的特性。这可能为研究人员提供一个独特的工具来阐明正常气道生物学和先天和适应性防御中未解决的问题，包括炎症、肿瘤发生和转移期间气道的重塑。
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.