Rapid and efficient generation of functional motor neurons from human pluripotent stem cells using gene delivered transcription factor codes
Open accessAbstract
Stem cell-derived motor neurons (MNs) are increasingly utilized for modeling disease in vitro and for developing cellular replacement strategies for spinal cord injury and diseases such as spinal muscular atrophy (SMA) and amyotrophic lateral sclerosis (ALS). Human embryonic stem cell (hESC) differentiation into MNs, which involves retinoic acid (RA) and activation of the sonic hedgehog (SHH) pathway is inefficient and requires up to 60 days to develop MNs with electrophysiological properties. This prolonged differentiation process has hampered the use of hESCs, in particular for high-throughput screening. We evaluated the MN gene expression profile of RA/SHH-differentiated hESCs to identify rate-limiting factors involved in MN development. Based on this analysis, we developed an adenoviral gene delivery system encoding for MN inducing transcription factors: neurogenin 2 (Ngn2), islet-1 (Isl-1), and LIM/homeobox protein 3 (Lhx3). Strikingly, delivery of these factors induced functional MNs with mature electrophysiological properties, 11-days after gene delivery, with >60-70% efficiency from hESCs and human induced pluripotent stem cells (hiPSCs). This directed programming approach significantly reduces the time required to generate electrophysiologically-active MNs by approximately 30 days in comparison to conventional differentiation techniques. Our results further exemplify the potential to use transcriptional coding for rapid and efficient production of defined cell types from hESCs and hiPSCs.
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Transcription factor driven differentiation protocols demonstrate greater reproducibility, purity, and decreased cost/generation time.
“Whilst protocols using transcription factor driven differentiation can overcome some of these challenges, demonstrating greater reproducibility, purity, and decreased cost/generation time12–14”
From Examining iPSC derived motor neuron variability and genome stability monitoring as a solution (2025) · cites this paper as [13]
Induced expression of different transcription factors yields other neuron types, such as motor neurons and inhibitory neurons.
“induced expression of different transcription factors yield other types of neurons, such as motor neurons (Hester et al., 2011; Shi et al., 2018) and inhibitory neurons (Yang et al., 2017).”
From CRISPR Interference-Based Platform for Multimodal Genetic Screens in Human iPSC-Derived Neurons (2019) · cites this paper as Hester et al., 2011
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References 50 · Cited by 183
- Authors
- Hester, M. E., Murtha, M. J., Song, S., Rao, M., Miranda, C. J., Meyer, K., Tian, J., Boulting, G., Schaffer, D. V., Zhu, M. X., Pfaff, S. L., Gage, F. H., & Kaspar, B. K.
- Affiliations
- Nationwide Children's Hospital (US)
- The Ohio State University (US)
- Howard Hughes Medical Institute (US)
- University of California, Berkeley (US)
- Salk Institute for Biological Studies (US)
- Corresponding author
- Kaspar, Brian K
- ORCID
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2011-10
- Volume
- 19
- Issue
- 10
- Pages
- 1905-1912
- ISSN
- 1525-0016 (ISSN-L)
- PMC
- PMC3188742
- OpenAlex
- W2036524825
- Cited by
- 183
- References
- 50
- Citations by year
- 2025: 6 · 2024: 5 · 2023: 12 · 2022: 11 · 2021: 12 · 2020: 13 · 2019: 13 · 2018: 11 · 2017: 11 · 2016: 18 · 2015: 23 · 2014: 16 · 2013: 17 · 2012: 11
- FWCI
- 6.24
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Howard Hughes Medical Institute
- National Institute of Neurological Disorders and Stroke (RC2NS069476, P30NS045758, RC2 NS69476-01, R01 NS6449L2-1A1, R01NS064492)
- 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
- motor neuron differentiation, human pluripotent stem cells, human induced pluripotent stem cells, adenoviral gene transfer, transcription factors, Neurogenin2, Islet1, LHX3, retinoic acid, sonic hedgehog pathway, electrophysiological maturation, spinal muscular atrophy, amyotrophic lateral sclerosis, spinal cord injury
- MeSH terms
- Cell Differentiation, Cell Line, Humans, Motor Neurons, Transcription Factors, Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, Pluripotent Stem Cells, Embryonic Stem Cells
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