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Updated: Jan 24, 2026

Establishment of Orthotopic Patient-derived Xenograft Models for Brain Tumors using a Stereotaxic Device
Published on: May 2, 2025
Protocol for Seahorse 3D Mito Stress assay in patient-derived pediatric brain tumor single neurospheres
Stefania Tocci1,2, Jessica W Tsai1,2,3,4
1Children's Hospital Los Angeles, Cancer and Blood Disease Institute, Los Angeles, California, United States.
Abstract:
Metabolism is essential for cellular functions and is often altered in cancer. While Seahorse assays are well established for measuring metabolic changes in 2D cell cultures, their application to 3D models remains challenging. Here, we present a step-by-step protocol for plating individual brain tumor neurospheres in the assay microplate to measure their bioenergetics. We provide actionable recommendations and highlight pitfalls to avoid. This approach enables exploration of dynamic metabolic changes in patient-derived brain tumor neurospheres following genetic or pharmacologic interventions.
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