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Large-scale generation of human iPSC-derived neural stem cells/early neural progenitor cells and their neuronal differentiation
Open accessAbstract
Induced pluripotent stem cell (iPSC)-based technologies offer an unprecedented opportunity to perform high-throughput screening of novel drugs for neurological and neurodegenerative diseases. Such screenings require a robust and scalable method for generating large numbers of mature, differentiated neuronal cells. Currently available methods based on differentiation of embryoid bodies (EBs) or directed differentiation of adherent culture systems are either expensive or are not scalable. We developed a protocol for large-scale generation of neuronal stem cells (NSCs)/early neural progenitor cells (eNPCs) and their differentiation into neurons. Our scalable protocol allows robust and cost-effective generation of NSCs/eNPCs from iPSCs. Following culture in neurobasal medium supplemented with B27 and BDNF, NSCs/eNPCs differentiate predominantly into vesicular glutamate transporter 1 (VGLUT1) positive neurons. Targeted mass spectrometry analysis demonstrates that iPSC-derived neurons express ligand-gated channels and other synaptic proteins and whole-cell patch-clamp experiments indicate that these channels are functional. The robust and cost-effective differentiation protocol described here for large-scale generation of NSCs/eNPCs and their differentiation into neurons paves the way for automated high-throughput screening of drugs for neurological and neurodegenerative diseases.
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“Alternatively, NSCs could be differentiated from induced pluripotent stem cells (iPSCs). However, the expensive, multistep, laborious differentiation protocol is not suitable for large-scale, uniform generation of NSCs [110].”
From Extracellular vesicles as nanotheranostic platforms for targeted neurological disorder interventions (2024) · cites this paper as [110]
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Metadata
- Authors
- D’Aiuto, L., Zhi, Y., Kumar Das, D., Wilcox, M. R., Johnson, J. W., McClain, L., MacDonald, M. L., Di Maio, R., Schurdak, M. E., Piazza, P., Viggiano, L., Sweet, R., Kinchington, P. R., Bhattacharjee, A. G., Yolken, R., & Nimgaonka, V. L.
- Affiliations
- University of Pittsburgh (US)
- University of Bari Aldo Moro (IT)
- Johns Hopkins University (US)
- Corresponding author
- Nimgaonka, Vishwajit L
- ORCID
- Venue
- Organogenesis
- Publisher
- Informa UK Limited
- Published
- 2014-10-02
- Volume
- 10
- Issue
- 4
- Pages
- 365-377
- ISSN
- 1547-6278 (ISSN-L)
- 1555-8592
- PMC
- PMC4594592
- OpenAlex
- W1817943252
- Cited by
- 131
- References
- 33
- Citations by year
- 2026: 3 · 2025: 9 · 2024: 13 · 2023: 6 · 2022: 15 · 2021: 22 · 2020: 17 · 2019: 17 · 2018: 11 · 2017: 11 · 2016: 5 · 2015: 2
- FWCI
- 2.46
- Type
- journal article
- Language
- en
- Update notices
- correction (2015-08-07)
- Open access
- bronze
- doi.org
- Locations
- Organogenesis (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- CINECA IRIS Institutional Research Information System (University of Bari Aldo Moro) (submittedVersion)
- Funders
- U.S. Department of Veterans Affairs
- Indo-US Science and Technology Forum
- National Science Council
- China Scholarship Council
- National Institutes of Health (MH071533, MH045817, MH63480)
- Cancer Institute, University of Pittsburgh (P30CA047904)
- National Institute of Mental Health (MH071533, R01MH042191, 07R-1712, (MH 63480, R01MH071533, R01MH063480, MH045817, R01MH045817, T32MH016804)
- National Cancer Institute (P30CA047904)
- 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
- induced pluripotent stem cells, neural stem cells, neuronal differentiation, VGLUT1, ligand-gated ion channels, synaptic proteins, high-throughput drug screening, neurodegenerative diseases, iPSC-derived neurons
- MeSH terms
- Cell Differentiation, Cells, Cultured, Humans, Neurons, Cell Proliferation, Induced Pluripotent Stem Cells, Neural Stem Cells, Batch Cell Culture Techniques
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