Krishna Chaitanya Pavani, Ann Van Soom, Katrien Smits
Extracellular vesicles (EVs) serve as key communication signals among embryos. They are secreted into the embryo culture medium and hold potential as biomarkers for assessing embryo quality. However, the limited volume of embryo-conditioned medium presents challenges for EV enrichment. This chapter details an effective method for purifying and characterizing EVs from in vitro embryo cultures by using the OptiPrep™ density gradient ultracentrifugation (ODG) method. Additionally, we also provide methods for preparing EV-free embryo culture media.