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首页> 外文期刊>Journal of Extracellular Vesicles >MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA
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MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA

机译：MSC分泌至少3种EV类型，每种类型都具有独特的膜脂质，蛋白质和RNA排列

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Mesenchymal stem cell (MSC), a widely used adult stem cell candidate for regenerative medicine, has been shown to exert some of its therapeutic effects through the secretion of extracellular vesicles (EVs). These homogenously sized EVs of 100–150 ηm exhibited many exosome-like biophysical and biochemical properties and carry both proteins and RNAs. Recently, exosome-associated proteins in this MSC EV preparation were found to segregate primarily to those EVs that bind cholera toxin B chain (CTB), a GM1 ganglioside-specific ligand, and pulse-chase experiments demonstrated that these EVs have endosomal origin and carried many of the exosome-associated markers. Here, we report that only a fraction of the MSC EV proteome was found in CTB-bound EVs. Using Annexin V (AV) and Shiga toxin B subunit (ST) with affinities for phosphatidylserine and globotriaosylceramide, respectively, AV- and a ST-binding EV were identified. CTB-, AV- and ST–binding EVs all carried actin. However, the AV-binding EVs carried low or undetectable levels of the exosome-associated proteins. Only the ST-binding EVs carried RNA and EDA-containing fibronectin. Proteins in AV-binding EVs were also different from those released by apoptotic MSCs. CTB- and AV-binding activities were localized to the plasma membrane and cytoplasm of MSCs, while ST-binding activity was localized to the nucleus. Together, this study demonstrates that cells secrete many types of EVs. Specifically, MSCs secrete at least 3 types. They can be differentially isolated based on their affinities for membrane lipid-binding ligands. As the subcellular sites of the binding activities of these ligands and cargo load are different for each EV type, they are likely to have a different biogenesis pathway and possibly different functions.
机译：间充质干细胞（MSC）是一种广泛使用的再生医学成人干细胞候选物，已被证明可通过分泌细胞外囊泡（EVs）发挥其某些治疗作用。这些大小均一的100-150ηm的EV表现出许多类似外泌体的生物物理和生化特性，并携带蛋白质和RNA。最近，发现该MSC EV制剂中的外泌体相关蛋白主要与结合霍乱毒素B链（CTB），GM1神经节苷脂特异性配体的EV分离，脉冲追踪实验表明这些EV具有内体来源并携带许多与外泌体相关的标记。在这里，我们报告说，仅在CTB结合的电动汽车中发现了MSC电动汽车蛋白质组的一小部分。分别使用膜联蛋白V（AV）和志贺毒素B亚基（ST）分别与磷脂酰丝氨酸和globotriaosylceramide亲和力，鉴定出AV-和ST结合的EV。结合CTB，AV和ST的电动汽车均带有肌动蛋白。但是，与AV结合的EV携带的外泌体相关蛋白水平较低或无法检测到。仅结合ST的EV携带RNA和含有EDA的纤连蛋白。结合AV的EV中的蛋白质也不同于凋亡的MSC所释放的蛋白质。 CTB和AV绑定活动被定位到MSC的质膜和细胞质，而ST绑定活动被定位到细胞核。在一起，这项研究表明细胞分泌多种类型的电动汽车。具体而言，MSC分泌至少3种类型。可以根据它们对膜脂质结合配体的亲和力来区分它们。由于每种电动汽车类型这些配体结合活性的亚细胞位点和载货量都不同，因此它们可能具有不同的生物发生途径和可能的不同功能。

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