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Updated: Jan 24, 2026

Establishment of Orthotopic Patient-derived Xenograft Models for Brain Tumors using a Stereotaxic Device

Establishment of Orthotopic Patient-derived Xenograft Models for Brain Tumors using a Stereotaxic Device

Published on: May 2, 2025

患者由来小児脳腫瘍単一神経球におけるSeahorse 3Dミトストレスアッセイのプロトコル

Stefania Tocci, Jessica W Tsai

bioRxiv : the preprint server for biology

|January 23, 2026

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.

Highlights:

Optimization of Seahorse assay protocol for single patient-derived brain tumor spheroidsStep-by-step instructions for successfully re-plating and transferring single brain tumor spheroidsImportance of microplate coating for the successful execution of Seahorse assaysImaging of the microplate before and after the assay allows for normalization of metabolic parameters based on neurosphere size.

キーワード:

脳腫瘍神経球代謝 Seahorseアッセイ 3D培養

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Isolation and Culture of Adult Zebrafish Brain-derived Neurospheres

Isolation and Culture of Adult Zebrafish Brain-derived Neurospheres

Published on: February 29, 2016

High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids

High-Throughput In Vitro Assay using Patient-Derived Tumor Organoids

Published on: June 14, 2021

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