STAR Protocols (Sep 2024)

Protocol for evaluating mitochondrial respiration in iPSC-derived neurons by the Seahorse XF analyzer

  • Chiara Fasano,
  • Andrea Cavaliere,
  • Valeria Tiranti,
  • Camille Peron

Journal volume & issue
Vol. 5, no. 3
p. 103127

Abstract

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Summary: Here, we present a protocol describing the quantification of oxygen consumption rate (OCR) and maximal respiration rate (MRR) in living induced pluripotent stem cell (iPSC)-derived neurons using the Seahorse analyzer. We guide you through the whole process: culture amplification and seeding of neural progenitor cells (NPCs), their differentiation into neurons, and normalization of the results to cell number in the analytical phase. The assessment of cellular mitochondrial function, by analyzing mitochondrial respiration, could be useful in various diseases as well as in drug screening.For complete details on the use and execution of this protocol, please refer to Aleo et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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