Low pH increases the yield of exosome isolation
Low pH increases the yield of exosome isolation
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DOI：
10.1016/j.bbrc.2015.03.172
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发表时间：
2015-05-22
影响因子：
3.1
通讯作者：
Kim, Manho
中科院分区：
文献类型：
--
作者：
Ban, Jae-Jun;Lee, Mijung;Kim, Manho
外泌体是由各种细胞分泌的细胞外囊泡。外泌体通过在细胞之间递送多种分子来介导细胞间通讯。癌细胞来源的外泌体似乎与肿瘤的进展和转移有关。肿瘤微环境被认为是酸性的，这种低pH控制外泌体生理学，导致肿瘤进展。尽管微环境pH对外泌体的重要性，但大多数外泌体研究都是在不考虑pH的情况下进行的。因此，需要研究不同pH下外泌体稳定性和分离产率的差异。在这项研究中，我们研究了总外泌体蛋白和RNA的产量后，孵育在酸性，中性和碱性条件培养基。在不同pH下孵育外泌体后，通过蛋白质印迹法研究代表性的外泌体标记物。结果，与中性培养基相比，在酸性培养基中孵育后，外泌体蛋白和核酸的浓度显著增加。在酸性环境中孵育后观察到较高水平的外来体标记物，包括CD9、CD63和HSP 70。另一方面，在碱性条件下孵育后未检测到外泌体蛋白、外泌体RNA和外泌体标记物。总之，我们的研究结果表明，酸性条件是外来体存在和分离的有利环境。(C)2015 Elsevier Inc. All rights reserved.
Exosomes are the extracellular vesicles secreted by various cells. Exosomes mediate intercellular communication by delivering a variety of molecules between cells. Cancer cell derived exosomes seem to be related with tumor progression and metastasis. Tumor microenvironment is thought to be acidic and this low pH controls exosome physiology, leading to tumor progression. Despite the importance of microenvironmental pH on exosome, most of exosome studies have been performed without regard to pH. Therefore, the difference of exosome stability and yield of isolation by different pH need to be studied. In this research, we investigated the yield of total exosomal protein and RNA after incubation in acidic, neutral and alkaline conditioned medium. Representative exosome markers were investigated by western blot after incubation of exosomes in different pH. As a result, the concentrations of exosomal protein and nucleic acid were significantly increased after incubation in the acidic medium compared with neutral medium. The higher levels of exosome markers including CD9, CD63 and HSP70 were observed after incubation in an acidic environment. On the other hand, no exosomal protein, exosomal RNA and exosome markers have been detected after incubation in an alkaline condition. In summary, our results indicate that the acidic condition is the favorable environment for existence and isolation of exosomes. (C) 2015 Elsevier Inc. All rights reserved.