An Alternative Procedure for the Storage and Preservation of Vermamoeba vermiformis and Other Amoebae in the Laboratory

Abstract Free‐living amoebae (FLA), particularly Vermamoeba vermiformis and Acanthamoeba spp., have emerged as indispensable models for the study of giant viruses. These organisms provide a unique and efficient system for the isolation and replication of giant viruses, serving as known permissive hosts and thus playing a pivotal role in advancing research in this field. FLA can alternate between vegetative (trophozoite) and resistant (cyst) forms during their life cycle. Current methods for FLA storage and preservation face limitations, including prolonged excystment times and low rates of trophozoite viability. To address these challenges, these protocols provide instructions for the storage and preservation of Vermamoeba vermiformis at refrigeration temperatures, allowing feasible and rapid reactivation of cultures as an alternative to the more fastidious cyst‐based storage methods. This method is also applicable to other FLA species, such as Acanthamoeba castellanii and Acanthamoeba polyphaga © 2025 Wiley Periodicals LLC. Basic Protocol 1 : Vermamoeba vermiformis propagation Basic Protocol 2 : Preparation of Vermamoeba vermiformis stocks for cold room storage Basic Protocol 3 : Reactivation of cold‐stored Vermamoeba vermiformis