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.%容受期宫腔液对于胚胎建立早期妊娠起着关键作用, 宫腔液内含的胞外囊泡 (extracellular vesicles, EVs) 可能是子宫内膜与胚胎之间交流的新模式.目前为止, 仍然没有一种提取方法能同时保证EVs的含量、纯度和生物活性.传统的超速梯度离心法对设备和样本量要求高, 耗时长, 因此寻求在少量样本中高效提取高纯度EVs的新方法十分重要.本研究对同一份大鼠容受期宫腔液样本分别采取PEG沉淀和亲和膜离心柱两款试剂盒进行EVs提取, 使用透射电镜、Western-blot和动态光散射进行形态、表面蛋白质和粒径分布的鉴定, 并对其内容物RNA进行了浓度和片段分布比较.结果显示, 两种试剂盒都可以成功从少量的宫腔液样本中分离得到EVs, 且效率较传统离心法大大提高, 但均无法完全区分其亚群;亲和膜离心柱提取法得到的EVs浓度更高, RNA片段集中在20～40 nt, 对于开展后续mi RNA建库测序分析具有更好效果.
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