Generation of 3D ex vivo mouse- and patient-derived glioma explant slice model for integration of confocal time-lapse imaging and spatial analysis

Development of spatial-integrative pre-clinical models is needed for glioblastoma, which are heterogenous tumors with poor prognosis. Here, we present an optimized protocol to generate three-dimensional ex vivo explant slice glioma model from orthotopic tumors, genetically engineered mouse models, and fresh patient-derived specimens. We describe a step-by-step workflow for tissue acquisition, dissection, and sectioning of 300 -mm tumor slices maintaining cell viability. The explant slice model allows the integration of confocal timelapse imaging with spatial analysis for studying migration, invasion, and tumor microenvironment, making it a valuable platform for testing effective treatment modalities.For complete details on the use and execution of this protocol, please refer to Comba et al. (2022).1

Automated summary

This article presents an optimized protocol to generate a three-dimensional ex vivo explant slice glioma model, which is suitable for studying the migration, invasion, and tumor microenvironment of glioblastoma. The model allows the integration of confocal timelapse imaging for researching effective treatment modalities.