2. Academic Validation
  3. Protocol for real-time assessment of mitochondrial and glycolytic ATP production in patient-derived glioma stem-like cells

Protocol for real-time assessment of mitochondrial and glycolytic ATP production in patient-derived glioma stem-like cells

  • STAR Protoc. 2024 Jun 27;5(3):103159. doi: 10.1016/j.xpro.2024.103159.
Pratibha Sharma 1,  Vinay K Puduvalli 2
Affiliations

Affiliations

  • 1 Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. Electronic address: psharma3@mdanderson.org.
  • 2 Department of Neuro-Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. Electronic address: vpuduval@mdanderson.org.
PMID: 38941182 DOI: 10.1016/j.xpro.2024.103159
Abstract

Glioma cells switch between energetic pathways to adapt and resist therapies. We present a protocol for measuring mitochondrial and glycolytic ATP rates in patient-derived glioma stem-like cells using a Seahorse XF ATP rate assay. We describe steps for growing 3D glioma stem-like cells, attaching cells to the assay plate, preparing drugs, and running the ATP rate assay. We also detail procedures for imaging viable cell numbers and normalization, with tips to overcome pitfalls in Agilent Seahorse assays.

Keywords

Cancer; Cell Biology; Cell culture; Cell-based Assays; Health Sciences; Metabolism; Neuroscience.

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