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人羊水来源外泌体的分离与鉴定。

Isolation and characterization of human amniotic fluid-derived exosomes.

机构信息

Division of Maternal-Fetal Medicine & Perinatal Research, Department of Obstetrics & Gynecology, The University of Texas Medical Branch, Galveston, TX, United States.

Division of Maternal-Fetal Medicine & Perinatal Research, Department of Obstetrics & Gynecology, The University of Texas Medical Branch, Galveston, TX, United States.

出版信息

Methods Enzymol. 2020;645:181-194. doi: 10.1016/bs.mie.2020.07.006. Epub 2020 Jul 30.

Abstract

Extracellular vesicles (EVs), specifically exosomes of 50-150nm, have emerged as important communication channels between cells and tissues and can be isolated from multiple biofluids including blood, urine and amniotic fluid. No standardized approach for exosome isolation from these biofluids has been established. This chapter outlines an optimized approach for isolating exosomes from human amniotic fluid samples. Like plasma, amniotic fluid contains many protein and cellular contaminants that requires multiple steps for cleanup. Therefore, to ensure samples contain minimal contaminants, including larger EVs, we also outline multiple methods for characterization of isolated exosomes for size, morphology and protein markers.

摘要

细胞外囊泡(EVs),特别是 50-150nm 的外泌体,已成为细胞和组织之间重要的通讯渠道,可以从多种生物体液中分离出来,包括血液、尿液和羊水。目前尚未建立从这些生物体液中分离外泌体的标准化方法。本章概述了一种从人羊水样本中分离外泌体的优化方法。与血浆一样,羊水中含有许多蛋白质和细胞污染物,需要经过多个步骤进行清理。因此,为了确保样品中包含最小的污染物,包括较大的 EVs,我们还概述了用于分离的外泌体的大小、形态和蛋白质标记物的多种表征方法。

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