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Isolation and Characterization of Equine Uterine Extracellular Vesicles: A Comparative Methodological Studyстатья
Статья опубликована в высокорейтинговом журнале
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 9 апреля 2021 г.
- Авторы: Almiñana Carmen, Vegas Alba Rudolf, Tekin Muhittin, Hassan Mubbashar, Uzbekov Rustem, Fröhlich Thomas, Bollwein Heinrich, Bauersachs Stefan
- Журнал: International Journal of Molecular Sciences
- Том: 22
- Год издания: 2021
- Издательство: MDPI
- Местоположение издательства: Basel, Switzerland
- Номер статьи: 979
- DOI: 10.3390/ijms22020979
- Аннотация: Extracellular vesicles (EVs) have been identified in the uterine fluid in different species and have been pointed as key players in the embryo-maternal dialogue, maternal recognition of pregnancy and establishment of pregnancy. However, little is known about the uterine EVs in the mare. Therefore, the present study aimed at characterizing EVs from uterine lavage of cyclic mares by comparing five EVs isolation methods and the combination of them: (1) ultracentrifugation (UC); (2) concentration of lavage volume by Centricon ultrafiltration (CE); (3) the use of CE with different washing steps (phosphate-buffered salinewith or without trehalose); (4) size-exclusion chromatog-raphy with iZON-qEV columns, and (5) a combination of the methods with best results based on EVs yield, purity, and protein cargo profiles. Transmission electron microscopy and Western blot-ting confirmed the isolation of EVs by all methods but with quantitative and qualitative differences. Mass spectrometry provided differences in protein profiles between methods, number of identified proteins, and protein classes. Our results indicate that the combination of CE/trehalose/iZON/UC is an optimal method to isolate equine uterine EVs with good yield and purity that can be applied in future studies to determine the role of equine uterine EVs in embryo-maternal interactions.
- Добавил в систему: Узбеков Рустем Эдуардович
Прикрепленные файлы
| № | Имя | Описание | Имя файла | Размер | Добавлен |
|---|---|---|---|---|---|
| 1. | Alminana_et_al_2021_IJMS_20_01_2021.pdf | Alminana_et_al_2021_IJMS_20_01_2021.pdf | 4,7 МБ | 20 января 2021 [rustuzbekov] |