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.
Exosomes Small intestinal epithelium Extraction methods Mouse
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FX and FD designed the experiments, conducted the study, wrote and revised the manuscript. Electron microscopy analysis was performed by YFL. WD helped in NTA analysis. GD and YS gave basic advice and suggestions. All authors reviewed the manuscript.
This study was supported by the grants from the National Natural Science Foundation of China (Grant Nos. 91649122, 81870613) to Guoxian Ding.
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