Dissociation and reaggregation of Hydra vulgaris for studies of self-organization

Abstract

The remarkable regenerative abilities of the small cnidarian Hydra vulgaris include the capacity to reassemble itself after dissociation into individual cells. Here, we present a robust protocol for the dissociation and reaggregation of Hydra tissue that addresses many common challenges encountered during the preparation and execution of the dissociation, as well as the formation and care of regenerating cellular aggregates. Analysis of the process provides insight into the mechanisms of nervous system self-organization.For complete details on the use and execution of this protocol, please refer to Lovas and Yuste (2021).[Display omitted]•Amplify experimental Hydra and prepare for the animal’s dissociation•Dissociate Hydra into individual cells, aggregate, and monitor their reassembly•Follow Hydra’s epithelial layer segregation and neural circuit formation•A new polyp emerges from the cellular aggregate after one week of developmentPublisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.The remarkable regenerative abilities of the small cnidarian Hydra vulgaris include the capacity to reassemble itself after dissociation into individual cells. Here, we present a robust protocol for the dissociation and reaggregation of Hydra tissue that addresses many common challenges encountered during the preparation and execution of the dissociation, as well as the formation and care of regenerating cellular aggregates. Analysis of the process provides insight into the mechanisms of nervous system self-organization.