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Articles, Cellular/Molecular

Accelerated High-Yield Generation of Limb-Innervating Motor Neurons from Human Stem Cells

Mackenzie W. Amoroso, Gist F. Croft, Damian J. Williams, Sean O'Keeffe, Monica A. Carrasco, Anne R. Davis, Laurent Roybon, Derek H. Oakley, Tom Maniatis, Christopher E. Henderson and Hynek Wichterle
Journal of Neuroscience 9 January 2013, 33 (2) 574-586; DOI: https://doi.org/10.1523/JNEUROSCI.0906-12.2013
Mackenzie W. Amoroso
1Project A.L.S./Jenifer Estess Laboratory for Stem Cell Research, New York, New York 10032;
2Departments of Pathology, Neurology, and Neuroscience, Center for Motor Neuron Biology and Disease and Columbia Stem Cell Initiative, Columbia University, New York, New York 10032;
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Gist F. Croft
1Project A.L.S./Jenifer Estess Laboratory for Stem Cell Research, New York, New York 10032;
2Departments of Pathology, Neurology, and Neuroscience, Center for Motor Neuron Biology and Disease and Columbia Stem Cell Initiative, Columbia University, New York, New York 10032;
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Damian J. Williams
3Department of Physiology and Cellular Biophysics, Columbia University, New York, New York 10032;
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Sean O'Keeffe
4Department of Biochemistry and Biophysics, Columbia University, New York, New York 10032;
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Monica A. Carrasco
4Department of Biochemistry and Biophysics, Columbia University, New York, New York 10032;
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Anne R. Davis
5Department of Obstetrics and Gynecology, Columbia University Medical Center, New York, New York 10032
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Laurent Roybon
1Project A.L.S./Jenifer Estess Laboratory for Stem Cell Research, New York, New York 10032;
2Departments of Pathology, Neurology, and Neuroscience, Center for Motor Neuron Biology and Disease and Columbia Stem Cell Initiative, Columbia University, New York, New York 10032;
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Derek H. Oakley
1Project A.L.S./Jenifer Estess Laboratory for Stem Cell Research, New York, New York 10032;
2Departments of Pathology, Neurology, and Neuroscience, Center for Motor Neuron Biology and Disease and Columbia Stem Cell Initiative, Columbia University, New York, New York 10032;
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Tom Maniatis
4Department of Biochemistry and Biophysics, Columbia University, New York, New York 10032;
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Christopher E. Henderson
1Project A.L.S./Jenifer Estess Laboratory for Stem Cell Research, New York, New York 10032;
2Departments of Pathology, Neurology, and Neuroscience, Center for Motor Neuron Biology and Disease and Columbia Stem Cell Initiative, Columbia University, New York, New York 10032;
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Hynek Wichterle
1Project A.L.S./Jenifer Estess Laboratory for Stem Cell Research, New York, New York 10032;
2Departments of Pathology, Neurology, and Neuroscience, Center for Motor Neuron Biology and Disease and Columbia Stem Cell Initiative, Columbia University, New York, New York 10032;
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Abstract

Human pluripotent stem cells are a promising source of differentiated cells for developmental studies, cell transplantation, disease modeling, and drug testing. However, their widespread use even for intensely studied cell types like spinal motor neurons is hindered by the long duration and low yields of existing protocols for in vitro differentiation and by the molecular heterogeneity of the populations generated. We report a combination of small molecules that within 3 weeks induce motor neurons at up to 50% abundance and with defined subtype identities of relevance to neurodegenerative disease. Despite their accelerated differentiation, motor neurons expressed combinations of HB9, ISL1, and column-specific markers that mirror those observed in vivo in human embryonic spinal cord. They also exhibited spontaneous and induced activity, and projected axons toward muscles when grafted into developing chick spinal cord. Strikingly, this novel protocol preferentially generates motor neurons expressing markers of limb-innervating lateral motor column motor neurons (FOXP1+/LHX3−). Access to high-yield cultures of human limb-innervating motor neuron subtypes will facilitate in-depth study of motor neuron subtype-specific properties, disease modeling, and development of large-scale cell-based screening assays.

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The Journal of Neuroscience: 33 (2)
Journal of Neuroscience
Vol. 33, Issue 2
9 Jan 2013
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Accelerated High-Yield Generation of Limb-Innervating Motor Neurons from Human Stem Cells
Mackenzie W. Amoroso, Gist F. Croft, Damian J. Williams, Sean O'Keeffe, Monica A. Carrasco, Anne R. Davis, Laurent Roybon, Derek H. Oakley, Tom Maniatis, Christopher E. Henderson, Hynek Wichterle
Journal of Neuroscience 9 January 2013, 33 (2) 574-586; DOI: 10.1523/JNEUROSCI.0906-12.2013

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Accelerated High-Yield Generation of Limb-Innervating Motor Neurons from Human Stem Cells
Mackenzie W. Amoroso, Gist F. Croft, Damian J. Williams, Sean O'Keeffe, Monica A. Carrasco, Anne R. Davis, Laurent Roybon, Derek H. Oakley, Tom Maniatis, Christopher E. Henderson, Hynek Wichterle
Journal of Neuroscience 9 January 2013, 33 (2) 574-586; DOI: 10.1523/JNEUROSCI.0906-12.2013
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