biostudies-literatureReis FCGMCTI | Conselho Nacional de Desenvolvimento Científico e TecnológicoFundação Oswaldo Cruze00080-19https://www.ebi.ac.uk/biostudies/studies/S-EPMC6429041biostudies-literatureUnknown4(2)Regular protocols for the isolation of fungal extracellular vesicles (EVs) are time-consuming, hard to reproduce, and produce low yields. In an attempt to improve the protocols used for EV isolation, we explored a model of vesicle production after growth of <i>Cryptococcus gattii</i> and <i>Cryptococcus neoformans</i> on solid media. Nanoparticle tracking analysis in combination with transmission electron microscopy revealed that <i>C. gattii</i> and <i>C. neoformans</i> produced EVs in solid media. The properties of cryptococcal vesicles varied according to the culture medium used and the EV-producing species. EV detection was reproduced with an acapsular mutant of <i>C. neoformans</i>, as well as with isolates of <i>Candida albicans</i>, <i>Histoplasma capsulatum</i>, and <i>SaccharomycemSphereA Novel Protocol for the Isolation of Fungal Extracellular Vesicles Reveals the Participation of a Putative Scramblase in Polysaccharide Export and Capsule Construction in <i>Cryptococcus gattii</i>.PMC6429041408711/2017-7301304/2017-3VPPIS-001-FIO-18-66311179/2017-7405520/2018-2440015/2018-9VPPCB-007-FIO-18-2-57Garcia AWAHonorato LAlves LRReis FCGMartins STNimrichter LRodrigues MLVainstein MHMedeiros LCStaats CCBorges BSKmetzsch LSchrank AJozefowicz LJSena BAGfalseA Novel Protocol for the Isolation of Fungal Extracellular Vesicles Reveals the Participation of a Putative Scramblase in Polysaccharide Export and Capsule Construction in <i>Cryptococcus gattii</i>.Regular protocols for the isolation of fungal extracellular vesicles (EVs) are time-consuming, hard to reproduce, and produce low yields. In an attempt to improve the protocols used for EV isolation, we explored a model of vesicle production after growth of <i>Cryptococcus gattii</i> and <i>Cryptococcus neoformans</i> on solid media. Nanoparticle tracking analysis in combination with transmission electron microscopy revealed that <i>C. gattii</i> and <i>C. neoformans</i> produced EVs in solid media. The properties of cryptococcal vesicles varied according to the culture medium used and the EV-producing species. EV detection was reproduced with an acapsular mutant of <i>C. neoformans</i>, as well as with isolates of <i>Candida albicans</i>, <i>Histoplasma capsulatum</i>, and <i>Saccharomyce2019-01-01T00:00:00Z2019 Mar2026-07-12T03:22:49.697Z2026-07-12T03:13:02.452ZS-EPMC64290413089443010.1128/mSphere.00080-19