摘要
Abstract
The uterine luminal fluid (ULF) during implantation period plays a vital role in the establishment of early pregnancy. Extracellular vesicles (EVs) in ULF may be a new mode of communication between en-dometrium and embryo. To this day, there is no extraction method that can ensure the content, purity and biological activity of EVs at the same time. The traditional method, ultracentrifugation, is time-consuming and has high requirement for the equipment and volume of samples. Therefore, it is important to develop a new method which can efficiently extract EVs from a small amount of sample with high purity. Herein, two commercial kits, with the methods of PEG precipitation and affinity-membrane spin column, respectively, were used to isolate EVs from the same sample of rat ULF. The morphology, specific surface proteins and particle sizes of EVs were characterized separately by transmission electron microscopy (TEM), Western-blot and dynamic light scattering (DLS). The concentration and fragment distribution of EV RNA were further analyzed. The results showed that both commercial kits can successfully isolate EVs from a small amount of ULF but not separate the subgroups of EVs. Through affinity-membrane spin column, higher concentration of EVs can be extracted and the RNA fragments in them were around 20～40 nt. Therefore, this kind of kit is a better choice for the subsequent EV mi RNA sequencing experiments.关键词
大鼠/容受期/胞外囊泡(EVs)/宫腔液(ULF)/PEG沉淀法/亲和膜离心柱Key words
rat/receptivity/extracellular vesicles (EVs)/uterine luminal fluid (ULF)/PEG precipitation/affinity-membrane spin column分类
医药卫生
