Proteomic and lipidomic analysis of extracellular vesicles from bovine follicular fluid isolated by different methods

• Grant number: 23/00884-8
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: June 26, 2023
• End date: December 11, 2023
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Juliano Coelho da Silveira
• Grantee: Gislaine dos Santos
• Supervisor: W. Andy Tao
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Institution abroad: Purdue University, United States
• Associated to the scholarship: 21/12560-7 - Extracellular vesicles from bovine follicular fluid isolated by different methods have distinct characteristics and microRNA profile, BP.MS

Abstract

Extracellular vesicles (EVs) are nanoparticles secreted by different cell types in body fluids and carry bioactive materials (lipids, proteins, miRNAs) as a means of intercellular communication. Recent studies have demonstrated the role of EVs as biomarkers or as supplements to culture medium. However, the challenge is to define an accurate and reliable protocol for the EVs isolation due to their high heterogeneity and overlapping biophysical properties. Thus, we hypothesized that EVs from bovine follicular fluid (FF) isolated by three different methodologies present distinct subtypes of EVs as well as lipids and protein contents. For this, EVs will be isolated by ultracentrifugation (UC), size exclusion chromatography (SEC; Izon Science qEV 35 nm; NZL) or precipitation (Exoquick-TC, System Biosciences; USA). All isolations will be performed with 100 µL of FF and finally resuspended in 300 µL of PBS. The lipidomic and proteomic will be realized by mass spectrometry in Purdue University (IN, EUA) in order to determine the contents of these EVs. At the end of the experiments, we hope to be able to predict the impact of each EV isolation method on experiments involving culture media supplementation or diagnosis. Moreover, we expect to have results that will shift paradigms related to the impact of EVs isolation methods in the recovery of subtypes of EVs from follicular fluid. (AU)