Protocol for stable isotopic tracing to assess cellular lipogenic activity in induced neural stem cells
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 [<sup>13</sup>C<sub>6</sub>]-glucose and [<sup>13</sup>C<sub>5</sub>, <sup>15</sup>N<sub>2</sub>]-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.<sup>1</sup>.
Authors
- D’Alessandro Angelo
- Frank Edenhofer (ORCID: https://orcid.org/0000-0002-6489-714X)
- Christian Frezza (ORCID: https://orcid.org/0000-0002-3293-7397)
- Rosana-Bristena Ionescu (ORCID: https://orcid.org/0000-0002-4488-5467)
- Daniel Stephenson (ORCID: https://orcid.org/0000-0002-5698-3400)
- Alexandra M. Nicaise (ORCID: https://orcid.org/0000-0001-7160-5988)
- Cory M. Willis (ORCID: https://orcid.org/0000-0001-7938-7276)
- Stefano Pluchino
- Luca Peruzzotti-Jametti
- Julie A. Reisz
- Marta Suarez Cubero
- Pranathi Prasad
- Monika Dzieciatkowska
Institutions
- Universität Innsbruck (AT)
- University of Cambridge (GB)
- NIHR Imperial Biomedical Research Centre (GB)
- Cologne Excellence Cluster on Cellular Stress Responses in Aging Associated Diseases (DE)
- University of Colorado Anschutz Medical Campus (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2026-01-03
- DOI
- https://doi.org/10.1016/j.xpro.2025.104309
- Primary Topic
- RNA Research and Splicing
- Type
- article
- Field-Weighted Citation Impact
- 0.00