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Although plant-derived extracellular vesicles have attracted increasing interest, most studies have focused on vesicles isolated from whole tissues or edible plant materials, whereas extracellular vesicles released by in vitro plant callus and suspension cell cultures remain comparatively underexplored. This review discusses plant cell cultures as controllable and potentially scalable experimental systems for investigating extracellular vesicle biogenesis, molecular cargo, isolation, characterization, and biological activity. Particular attention is given to the distinction between bona fide extracellular vesicles secreted by living plant cells and heterogeneous nanovesicle preparations generated through tissue disruption, a key issue for reproducibility and interpretation in plant EV research. We also summarize current evidence on vesicle-mediated communication, cargo modulation, engineering strategies, and potential biomedical applications. Finally, we highlight major methodological gaps, including the lack of plant-specific markers, standardized isolation workflows, quality control criteria, and harmonized reporting practices. By integrating plant cell biology, extracellular vesicle research, and plant biotechnology, in vitro plant cell cultures may provide useful model systems for studying vesicle-mediated communication and for developing biologically produced vesicles with more defined functional properties.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s00299-026-03983-7","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T09:17:08Z","timestamp":1789982228000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Extracellular vesicles released by in vitro plant cell cultures: emerging systems for vesicle-mediated communication and biomedical translation"],"prefix":"10.1007","volume":"45","author":[{"given":"Francesca Cristiana","family":"Piritore","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arianna","family":"Minoia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u00e3o","family":"Pessoa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Grazia","family":"Romanelli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca","family":"Dalle Carbonare","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1166-8033","authenticated-orcid":false,"given":"Maria Teresa","family":"Valenti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"issue":"6","key":"3983_CR1","doi-asserted-by":"publisher","first-page":"1093","DOI":"10.1039\/D2LC00006G","volume":"22","author":"CM Abreu","year":"2022","unstructured":"Abreu CM, Costa-Silva B, Reis RL, Kundu SC, Caballero D (2022) Microfluidic platforms for extracellular vesicle isolation, analysis and therapy in cancer. 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