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细胞外囊泡亚群的不同去污剂敏感性。

Differential detergent sensitivity of extracellular vesicle subpopulations.

作者信息

Osteikoetxea Xabier, Sódar Barbara, Németh Andrea, Szabó-Taylor Katalin, Pálóczi Krisztina, Vukman Krisztina V, Tamási Viola, Balogh Andrea, Kittel Ágnes, Pállinger Éva, Buzás Edit Irén

机构信息

Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary.

出版信息

Org Biomol Chem. 2015 Oct 14;13(38):9775-82. doi: 10.1039/c5ob01451d.

Abstract

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.

摘要

细胞外囊泡(包括外泌体、微囊泡和凋亡小体)由于能够携带复杂信息并作为自分泌、旁分泌甚至内分泌细胞间信使,目前正迅速吸引生物学各个领域越来越多的关注。在本研究中,我们研究了基于大小的细胞外囊泡亚群对不同浓度去污剂(包括十二烷基硫酸钠、Triton X-100、吐温20和脱氧胆酸盐)的敏感性。我们确定了裂解由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对囊泡进行了表征。发现微囊泡和凋亡小体比外泌体对去污剂裂解更敏感。此外,我们发现有证据表明,在低浓度下,十二烷基硫酸钠和Triton X-100在囊泡裂解方面比脱氧胆酸盐或吐温20更有效。综上所述,我们的数据表明,将差异去污剂裂解与可调电阻脉冲传感或流式细胞术相结合,可能有助于简单快速地区分外泌体与其他细胞外囊泡亚群,以及区分囊泡结构与非囊泡结构。

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