CHARACTERIZATION OF IN VIVO AND IN VITRO EXOSOMES IN THE BOVINE SPECIE
Supporting Agencies
- Fundación Séneca 21656 / 21 y Ministerio de Ciencia e Innovación (PID 2019 106380 RB I 00 / 10 13039 501100011033)
Abstract
Recent studies have shown that extracellular vesicles may play an important role in modulating the fertilization capacity of sperm during their journey through the female reproductive tract. Extracellular vesicles (EVs), exosomes and micro vesicles, are a type of heterogeneous structures present in most body fluids, including bovine oviductal fluid. EVs contain various compounds derived from the original cell, such as proteins, lipids, mRNA, miRNA and DNA. EVs in the oviduct are produced by epithelial cells and their functions include interaction with spermatozoa, maintenance of their viability, participation in oocyte maturation and in the fertilization process.
During the in vitro fertilization process and in order to improve it by mimicking in vivo conditions, numerous researchers have used bovine oviductal epithelial cell (BOEC) cultures with remarkable improvements.
These cells produce, among others components, VEs, for this reason, in this work we have proposed a comparative study of the EVs present in the bovine oviductal fluid (OF) collected at times close to ovulation (in vivo) and those produced in BOEC cultures after 7 days of culture (in vitro) comparing the size, population distribution and protein concentration in both types. The EVs were identified by electron microscopy, their size by laser light scattering and their protein concentration by Bradford's method.
The results showed that the EVs size evaluated per intensity were similar between both experimental groups. On the other hand, differences were observed in terms of protein concentration. EVs obtained in vivo contained a greater amount of protein in their cargo than the EVs obtained in vitro.
Regarding the identification of VEs by transmission electron microscopy, only those obtained in vivo could be observed. This fact could be due to the place where they have been collected, to the method of culture of bovine oviductal epithelial cells or the shortage in their production.
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