Development of microfluidic devices for the isolation and analysis of extracellular vesicles from spent embryo culture media

• Grant number: 25/00727-5
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: June 16, 2025
• End date: June 15, 2026
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Juliano Coelho da Silveira
• Grantee: Franciele Flores Vit
• Supervisor: Ming Dao
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Institution abroad: Massachusetts Institute of Technology (MIT), United States
• Associated to the scholarship: 22/02701-5 - Generation of microfluidic devices with different geometries for in vitro fertilization, BP.PD

Abstract

The assessment of embryo quality is of paramount importance in the context of assisted reproductive technologies (ART) in order to optimize implantation and pregnancy outcomes. Extracellular vesicles (EVs) are involved in intercellular communication and have been identified as a potential biomarker for the non-invasive assessment of embryo quality. The objective of this project is to develop and compare two microfluidic devices for the isolation of extracellular vesicles (EVs) from mouse embryo culture media. One device will employ size exclusion chromatography (µSEC), while the other will utilize immunoaffinity with specific antibodies directed at EV markers. The µSEC device will be developed for the separation of EVs present in small sample volumes, while the immunoaffinity-based device will be functionalized with antibodies targeting CD63, CD81, and CD9. Extracellular vesicles (EVs) will be isolated from mouse embryo culture medium after four days of culture. The concentration, size, morphology, and RNA content of the extracellular vesicles will be analyzed using nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), flow cytometry, and RNA extraction. The intention is to evaluate the efficiency and specificity of each method for isolating embryo-derived EVs, thus contributing to advances in the non-invasive evaluation of embryos. The implementation of these microfluidic devices has the potential to improve embryo selection protocols, thus improving the results of assisted reproductive technology.