Kihika, Joseph KanyiWood, SusannaRhodes, LesleySmith, Kirsty FThompson, LucyChallenger, SarahRyan, Ken G2025-01-152022-01-132025-01-1520222021-12-172045-232235027556 (pubmed)PMC8758677 (pmc)https://hdl.handle.net/10182/17963Dinoflagellates are among the most diverse group of microalgae. Many dinoflagellate species have been isolated and cultured, and these are used for scientific, industrial, pharmaceutical, and agricultural applications. Maintaining cultures is time-consuming, expensive, and there is a risk of contamination or genetic drift. Cryopreservation offers an efficient means for their long-term preservation. Cryopreservation of larger dinoflagellate species is challenging and to date there has been only limited success. In this study, we explored the effect of cryoprotectant agents (CPAs) and freezing methods on three species: Vulcanodinium rugosum, Alexandrium pacificum and Breviolum sp. A total of 12 CPAs were assessed at concentrations between 5 and 15%, as well as in combination with dimethyl sulfoxide (DMSO) and other non-penetrating CPAs. Two freezing techniques were employed: rapid freezing and controlled-rate freezing. Breviolum sp. was successfully cryopreserved using 15% DMSO. Despite exploring different CPAs and optimizing the freezing techniques, we were unable to successfully cryopreserve V. rugosum and A. pacificum. For Breviolum sp. there was higher cell viability (45.4 ± 2.2%) when using the controlled-rate freezing compared to the rapid freezing technique (10.0 ± 2.8%). This optimized cryopreservation protocol will be of benefit for the cryopreservation of other species from the family Symbiodiniaceae.12 pagesElectronicen© 2022 The Author(s).DinoflagellidaDimethyl SulfoxideDrug CombinationsCryoprotective AgentsCryopreservationCell SurvivalDose-Response Relationship, DrugDinoflagellidaDimethyl SulfoxideDrug CombinationsCryoprotective AgentsCryopreservationCell SurvivalDose-Response Relationship, DrugJournal Article10.1038/s41598-021-04227-22045-2322ANZSRC::310411 Plant and fungus systematics and taxonomyANZSRC::310802 Plant biochemistryANZSRC::310803 Plant cell and molecular biologyhttps://creativecommons.org/licenses/by/4.0/Attribution