Isolation of extracellular vesicles from insect hemolymph

Extracellular vesicles (EVs) are produced by cells of all domains of life and current research shows their involvement in intercellular transfer of information. However, a major drawback to the progress of the field relates to the fact that isolating pure EV fractions from complex biofluids is a challenging task. Isolation of EVs is often compromised by the presence of contaminating protein and lipoprotein particles, which has recently led to the establishment of guidelines for analyzing and reporting on EVs. In insects, reports on EV studies are starting to emerge, with several techniques being used without consideration of contaminant co-isolation. To address this, we optimized and validated a robust procedure for the isolation of EVs from insect hemolymph.