A High Efficient Method to Isolate Exosomes from Small Intestinal Epithelium
A High Efficient Method to Isolate Exosomes from Small Intestinal Epithelium
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从小肠上皮中分离外泌体的高效方法
DOI：
10.1007/s12033-019-00163-9
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发表时间：
2019-02
影响因子：
2.6
通讯作者：
Guoxian Ding
中科院分区：
文献类型：
--
作者：
Fan Xia;Fei Ding;Yifan Lv;Wenjuan Di;Yunlu Sheng;Guoxian Ding
肠上皮是一种重要的受体，不仅暴露于营养物质，还暴露于病原体，例如摄入的毒素、细菌菌群及其代谢物。感觉信息被传达到广泛的内分泌、神经、免疫系统，而外泌体充当细胞间通信的载体。从小肠上皮中分离外泌体作为外泌体的来源仍然更加复杂，因为它含有不同比例的源自许多不同细胞的外泌体。目前对外泌体的研究主要是使用培养细胞的上清液进行的。这是因为，在细胞培养物中，可以确定外泌体的起源，并且不含“污染”蛋白质、脂质和糖的外泌体的分离涉及大多数培养基的相对简单的组成。然而，由于几个技术问题，这在肠上皮细胞（IEC）中很难实现，包括体内生理学的重演、操作简单性、培养物随时间的稳定性和测定通量。同时，从特定细胞类型中分离外泌体仍然是一个相当大的问题，因为分离的上清液外泌体部分可能仅代表总数的一小部分，而不是反映整体情况。在此，我们提出了一种从小肠上皮细胞间隙中富集外泌体的有效方案。该方法保持了囊泡及其内容物的完整性。此外，它可能有助于更好地了解外泌体的特性并探索它们在小肠上皮细胞间通讯中的作用。
The intestinal epithelium is an important receptor that is not only exposed to nutrients but also to pathogens, such as ingested toxins, bacterial flora, and their metabolites. The sensory information is communicated to extensive endocrine, neural, immune systems, and the exosomes acts as carriers of communication from cell-to-cell. Isolation of exosomes from small intestinal epithelium remains more complex to obtain as a source of exosomes, as it contains varying proportions of exosomes derived from many different cells. Current studies on exosomes have been largely performed using supernatants of cultured cells. This is because, in a cell culture, the origin of exosomes can be determined and isolation of exosomes devoid of ‘contaminating’ proteins, lipids, and sugars involves relatively simple composition of most culture media facilitates. However, this is hard to achieve in intestinal epithelial cells (IECs) due to several technical issues, including recapitulation of in vivo physiology, operational simplicity, culture stability over time, and assay throughput. Meanwhile, separation of exosomes from a specific cell type remains to be a considerable problem, as the isolated supernatant exosomal fraction may represent only a small fraction of the total instead of reflecting the overall situation. Herein, we proposed an efficient protocol for enrichment of exosomes from the interstitial space of small intestinal epithelium. This method maintains the integrity of the vesicles as well as their contents. Also, it may help to better understand the properties of exosomes and explore their role in cell-to-cell communication of small intestinal epithelium.
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DOI：
10.1007/978-1-4939-6728-5_10
发表时间：
2017-01-01
期刊：
EXOSOMES AND MICROVESICLES: METHODS AND PROTOCOLS
影响因子：
--
作者：
Perez-Gonzalez, Rocio;Gauthier, Sebastien A.;Levy, Efrat
通讯作者：
Levy, Efrat
影响因子：
4.8
作者：
Tauro, Bow J.;Greening, David W.;Simpson, Richard J.
通讯作者：
Simpson, Richard J.
影响因子：
120.1
作者：
EL Andaloussi, Samir;Maeger, Imre;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.
影响因子：
5.6
作者：
Bu, Heng-Fu;Wang, Xiao;Tang, Yi;Koti, Viola;Tan, Xiao-Di
通讯作者：
Tan, Xiao-Di
影响因子：
24.5
作者：
Leushacke, Marc;Barker, Nick
通讯作者：
Barker, Nick