Isolation and characterization ​of extracellular vesicles ​from Coffea arabica L. cell cultures

Exosomes are extracellular vesicles (EVs), typically nano-sized, that play a pivotal role in intercellular signalling and communication. Plant EVs can facilitate the transfer of various molecules between cells, which are involved in physiological processes such as plant immunity, stress responses, growth and development. These properties have raised interest in biotechnological, medical and agricultural applications in recent years, as it could contribute to disclose new scenarios for managing human and plant diseases. The full potential of EVs can be harnessed by studying their biogenesis, cargo composition and functional roles, however, isolating them from whole plants or tissues presents technical challenges. Conversely, plant cell cultures are excellent models for their investigation. In this work, we isolated EVs from cell suspension cultures of Coffea arabica, a species known as a major source of high-value compounds. We obtained EVs through differential centrifugation and characterized them regarding morphometric traits, surface charge and protein composition. In particular, the latter experiments aimed to study their biogenesis through immunoblotting and proteomic analysis.