A Guide to Generating and Using hiPSC Derived NPCs for the Study of Neurological Diseases
Open accessAbstract
Post-mortem studies of neurological diseases are not ideal for identifying the underlying causes of disease initiation, as many diseases include a long period of disease progression prior to the onset of symptoms. Because fibroblasts from patients and healthy controls can be efficiently reprogrammed into human induced pluripotent stem cells (hiPSCs), and subsequently differentiated into neural progenitor cells (NPCs) and neurons for the study of these diseases, it is now possible to recapitulate the developmental events that occurred prior to symptom onset in patients. We present a method by which to efficiently differentiate hiPSCs into NPCs, which in addition to being capable of further differentiation into functional neurons, can also be robustly passaged, freeze-thawed or transitioned to grow as neurospheres, enabling rapid genetic screening to identify the molecular factors that impact cellular phenotypes including replication, migration, oxidative stress and/or apoptosis. Patient derived hiPSC NPCs are a unique platform, ideally suited for the empirical testing of the cellular or molecular consequences of manipulating gene expression.
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Knockdown of CD44 increased lipid peroxidation in the screen, as expected because CD44v stabilizes xCT and increases cysteine uptake for GSH synthesis, inhibiting ferroptosis.
“By contrast, knockdown of CD44, whose splicing variant CD44v stabilizes the cystine/glutamate antiporter xCT at the plasma membrane and increases cysteine uptake for GSH synthesis, thereby inhibiting ferroptosis46,47, increased lipid peroxidation in our screen as expected (Fig. 2d).”
From Genome-wide CRISPRi/a screens in human neurons link lysosomal failure to ferroptosis (2021) · cites this paper as [46]
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References 27 · Cited by 58
- Authors
- Topol, A., Tran, N. N., & Brennand, K. J.
- Affiliations
- Icahn School of Medicine at Mount Sinai (US)
- Venue
- Journal of Visualized Experiments
- Publisher
- MyJove Corporation
- Published
- 2015-02-21
- Issue
- 96
- Pages
- e52495-e52495
- ISSN
- 1940-087X (ISSN-L)
- PMC
- PMC4354663
- OpenAlex
- W1980832870
- Cited by
- 58
- References
- 27
- Citations by year
- 2026: 3 · 2025: 2 · 2024: 2 · 2023: 3 · 2022: 8 · 2021: 10 · 2020: 2 · 2019: 6 · 2018: 9 · 2017: 9 · 2016: 3 · 2015: 1
- FWCI
- 3.17
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Visualized Experiments (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- New York Stem Cell Foundation
- National Institutes of Health (MH101454)
- National Institute of Mental Health (R01MH101454)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- human induced pluripotent stem cells, neural progenitor cells, iPSC-derived neurons, neurological disorders, disease modeling, fibroblast reprogramming, neurosphere differentiation, genetic screening, oxidative stress, apoptosis, cell migration
- MeSH terms
- Cell Differentiation, Cytological Techniques, Humans, Nervous System Diseases, Induced Pluripotent Stem Cells, Neural Stem Cells
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