Protocol for stable isotopic tracing to assess cellular lipogenic activity in induced neural stem cells

STAR Protocols 2026 Open access

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Authors: Rosana-Bristena Ionescu, Julie A. Reisz, Monika Ewa Dzieciatkowska, Daniel Stephenson, Alexandra M. Nicaise, Pranathi Prasad, Cory M. Willis, Marta Suarez Cubero, Luca Peruzzotti-Jametti, Frank Edenhofer, Christian Frezza, Stefano Pluchino
+1 more D’Alessandro Angelo
DOI: 10.1016/j.xpro.2025.104309
Cited by: 1
References: 1

Abstract

Here, we present a protocol to assess the lipogenic phenotype of induced neural stem cells (iNSCs) using stable isotopic tracing. We describe steps for the culture and preparation of iNSCs, labeling with [ 13 C 6 ]-glucose and [ 13 C 5 , 15 N 2 ]-glutamine, and the subsequent extraction of metabolites, lipids, and proteins from the same sample. This protocol supports single-specimen, mass spectrometry-based multi-omics workflows and is applicable to steady-state analyses, stable isotope tracing, and characterization of protein post-translational modifications. For complete details on the use and execution of this protocol, please refer to Ionescu et al. 1

Fields of Study & Subjects

RNA Research and Splicing Anesthesia and Neurotoxicity Research Neurogenesis and neuroplasticity mechanisms

Publication details

Journal / source
STAR Protocols
Publisher
Elsevier BV
Volume
7
Issue
1
Pages
104309–104309
Publication date
3 January 2026
Language
EN