Differential detergent sensitivity of extracellular vesicle subpopulations
Differential detergent sensitivity of extracellular vesicle subpopulations
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DOI：
10.1039/c5ob01451d
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发表时间：
2015-01-01
影响因子：
3.2
通讯作者：
Buzas, Edit Iren
中科院分区：
文献类型：
--
作者：
Osteikoetxea, Xabier;Sodar, Barbara;Buzas, Edit Iren
细胞外囊泡（包括外泌体、微囊泡和凋亡小体）由于其携带复杂信息的能力以及作为自分泌、旁分泌甚至内分泌细胞间信使的作用，目前正吸引着生物学各个领域越来越多的关注。在本研究中，我们调查了不同浓度的洗涤剂，包括十二烷基硫酸钠，Triton X-100，吐温20和脱氧胆酸盐的细胞外囊泡的大小为基础的亚群的敏感性。我们确定了裂解Jurkat、THP-1、MiaPaCa和U937人细胞系分泌的每个囊泡亚群所需的去污剂浓度。我们通过可调电阻脉冲传感，流式细胞术和透射电子显微镜表征囊泡。发现微泡和凋亡小体比外泌体对去污剂裂解更敏感。此外，我们发现证据表明，十二烷基硫酸钠和Triton X-100在低浓度下比脱氧胆酸盐或吐温20更有效地溶解囊泡。综上所述，我们的数据表明，差异洗涤剂裂解与可调电阻脉冲传感或流式细胞术的组合可以证明对于外来体和其他细胞外囊泡亚群之间以及囊泡和非囊泡结构之间的简单和快速区分是有用的。
Extracellular vesicles (including exosomes, microvesicles and apoptotic bodies) are currently attracting rapidly increasing attention from various fields of biology due to their ability to carry complex information and act as autocrine, paracrine and even endocrine intercellular messengers. In the present study we investigated the sensitivity of size-based subpopulations of extracellular vesicles to different concentrations of detergents including sodium dodecyl sulphate, Triton X-100, Tween 20 and deoxycholate. We determined the required detergent concentration that lysed each of the vesicle subpopulations secreted by Jurkat, THP-1, MiaPaCa and U937 human cell lines. We characterized the vesicles by tunable resistive pulse sensing, flow cytometry and transmission electron microscopy. Microvesicles and apoptotic bodies were found to be more sensitive to detergent lysis than exosomes. Furthermore, we found evidence that sodium dodecyl sulphate and Triton X-100 were more effective in vesicle lysis at low concentrations than deoxycholate or Tween 20. Taken together, our data suggest that a combination of differential detergent lysis with tunable resistive pulse sensing or flow cytometry may prove useful for simple and fast differentiation between exosomes and other extracellular vesicle subpopulations as well as between vesicular and non-vesicular structures.