Isolation and characterization of exosomes from cell culture supernatants and biological fluids.
Isolation and characterization of exosomes from cell culture supernatants and biological fluids.
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DOI：
10.1002/0471143030.cb0322s30
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发表时间：
2006-04-01
影响因子：
--
通讯作者：
Clayton, Aled
中科院分区：
文献类型：
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作者：
Thery, Clotilde;Amigorena, Sebastian;Clayton, Aled
外来体是在细胞培养上清液和不同生物流体中发现的小膜囊泡。外泌体通过向内出芽进入隔室的内腔而在特定的内体群体中形成，称为多泡体（MVB）。在MVB与质膜融合后，这些内部囊泡被分泌。外来体具有一组确定的膜和胞质蛋白。外泌体的生理功能仍然是一个有争议的问题，但在各种实验系统中越来越多的结果表明它们参与多种生物过程。由于细胞培养上清液和生物液体都含有不同类型的脂质膜，因此进行高质量的外泌体纯化至关重要。本单元描述了从各种来源纯化外泌体的不同方法，并讨论了评估纯化外泌体制剂纯度和均匀性的方法。
Exosomes are small membrane vesicles found in cell culture supernatants and in different biological fluids. Exosomes form in a particular population of endosomes, called multivesicular bodies (MVBs), by inward budding into the lumen of the compartment. Upon fusion of MVBs with the plasma membrane, these internal vesicles are secreted. Exosomes possess a defined set of membrane and cytosolic proteins. The physiological function of exosomes is still a matter of debate, but increasing results in various experimental systems suggest their involvement in multiple biological processes. Because both cell-culture supernatants and biological fluids contain different types of lipid membranes, it is critical to perform high-quality exosome purification. This unit describes different approaches for exosome purification from various sources, and discusses methods to evaluate the purity and homogeneity of the purified exosome preparations.