Extracellular vesicles from bovine follicular fluid isolated by different methods have distinct characteristics and microRNA profile

• Grant number: 21/12560-7
• Support Opportunities: Scholarships in Brazil - Master
• Start date: December 01, 2022
• End date: June 16, 2024
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Juliano Coelho da Silveira
• Grantee: Gislaine dos Santos
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Associated research grant: 21/06645-0 - Extracellular vesicles as a platform for diagnostic and manipulation of the in vitro embryo production system: the next generation in animal reproduction, AP.JP2
• Associated scholarship(s): 23/00884-8 - Proteomic and lipidomic analysis of extracellular vesicles from bovine follicular fluid isolated by different methods, BE.EP.MS

Abstract

In vitro production of bovine embryos (IVP) is a technique capable of increasing the number of offspring for each donor leading to increased genetic progress of the offspring. However, the technique still have some limitations including the cell culture medium. Therefore, new technologies are being developed to mimic the in vivo produced embryos. Small extracellular vesicles (EVs) secreted by cells and found in body fluids (such as a follicular fluid and oviductal fluid) are being studied as a supplement in PIVE, as it contains bioactive materials responsible for regulating cell function. For this, several EVs isolation protocols are available in the literature. However, it is not clear whether the characteristics of EVs present in the follicular environment are different according to the isolation method. The hypothesis of this study is that small EVs from follicular fluid isolated by different methods (ultracentrifugation, size exclusion chromatography, or precipitation) present distinct morphologic characteristics and miRNA profiles. Thus, the objectives of this project are to evaluate the morphologic characteristics and miRNA contents of small VEs separated by three different isolation methodologies as well as to evaluate the supplementation effects during IVM of bovine cumulus oocyte complexes. The results obtained with this project will help to understand the variations between EVs isolated by different methodologies and their different effects when used as a supplement in the IVM of bovine cumulus oocyte complexes.