Isolation of Tissue Extracellular Vesicles from the Liver
Isolation of Tissue Extracellular Vesicles from the Liver
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DOI：
10.3791/58649
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发表时间：
2019-08-01
影响因子：
1.2
通讯作者：
Patel, Tushar
中科院分区：
文献类型：
--
作者：
Ishiguro, Kaori;Yen, Irene K.;Patel, Tushar
细胞外囊泡（EV）可以从许多不同的细胞类型中释放，并在大多数（如果不是全部）体液中检测到。EV可以通过将生物活性分子如RNA或蛋白质从一个细胞穿梭到另一个细胞来参与细胞间的通讯。EV的大多数研究都是在细胞培养模型或从体液中分离的EV中进行的。人们对从组织中分离EV以研究它们对生理过程的贡献以及它们在疾病中如何改变产生了兴趣。从组织中分离具有足够产量的EV在技术上具有挑战性，因为需要在不损伤细胞的情况下进行组织解离。该方法描述了从小鼠肝组织分离EV的程序。该方法包括两步过程，首先是原位胶原酶消化，然后是差速超离心。使用胶原酶的组织灌注提供了优于肝组织的机械切割或均质化的优点，因为其增加了获得EV的产率。使用这种两步法从肝脏分离EV将有助于组织EV的研究。
Extracellular vesicles (EVs) can be released from many different cell types and detected in most, if not all, body fluids. EVs can participate in cell-to-cell communication by shuttling bioactive molecules such as RNA or protein from one cell to another. Most studies of EVs have been performed in cell culture models or in EVs isolated from body fluids. There is emerging interest in the isolation of EVs from tissues to study their contribution to physiological processes and how they are altered in disease. The isolation of EVs with sufficient yield from tissues is technically challenging because of the need for tissue dissociation without cellular damage. This method describes a procedure for the isolation of EVs from mouse liver tissue. The method involves a two-step process starting with in situ collagenase digestion followed by differential ultra-centrifugation. Tissue perfusion using collagenase provides an advantage over mechanical cutting or homogenization of liver tissue due to its increased yield of obtained EVs. The use of this two-step process to isolate EVs from the liver will be useful for the study of tissue EVs.