Doctoral School of Exact and Natural Sciences, Jagiellonian University, Krakow, Poland; Department of Endocrinology, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University, Gronostajowa 9, 30-387, Krakow, Poland.
Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Kreutzwaldi 62, 51006, Tartu, Estonia; Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, Ravila St. 14b, 50411, Tartu, Estonia.
Theriogenology. 2023 Jul 15;205:79-86. doi: 10.1016/j.theriogenology.2023.04.010. Epub 2023 Apr 14.
Extracellular vesicles (EVs) are membrane-bound nanoparticles that are released by different cell types and play a crucial role in the intercellular communication. They carry various biomolecular compounds such as DNA, RNA, proteins, and lipids. Given that EVs are a new element of the communication within the ovarian follicle, extensive research is needed to optimize method of their isolation. The aim of the study was to assess size-exclusion chromatography (SEC) as a tool for effective EVs isolation from porcine ovarian follicular fluid. The characterization of EVs was performed by nanoparticle tracking analysis, transmission electron microscopy, atomic force microscopy, mass spectrometry and Western blot. We determined EVs concentration, size distribution, zeta potential, morphology, purity, and marker proteins. Our results show that SEC is an effective method for isolation of EVs from porcine follicular fluid. They displayed predominantly exosome properties with sufficient purity and possibility for further functional analyses, including proteomics.
细胞外囊泡（EVs）是由不同细胞类型释放的膜结合纳米颗粒，在细胞间通讯中发挥着关键作用。它们携带各种生物分子化合物，如 DNA、RNA、蛋白质和脂质。鉴于 EVs 是卵巢卵泡内通讯的新元素，需要进行广泛的研究来优化其分离方法。本研究的目的是评估排阻色谱（SEC）作为从猪卵巢卵泡液中有效分离 EVs 的工具。通过纳米颗粒跟踪分析、透射电子显微镜、原子力显微镜、质谱和 Western blot 对 EVs 进行了表征。我们确定了 EVs 的浓度、粒径分布、zeta 电位、形态、纯度和标记蛋白。我们的结果表明，SEC 是从猪卵泡液中分离 EVs 的有效方法。它们表现出明显的外泌体特性，具有足够的纯度和进一步进行功能分析（包括蛋白质组学分析）的可能性。
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