Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Sep;1(9):e244.
doi: 10.1002/cpz1.244.

In Vitro Generation of Posterior Motor Neurons from Human Pluripotent Stem Cells

Matt Wind  1 , Anestis Tsakiridis  1
Affiliations
Free article

In Vitro Generation of Posterior Motor Neurons from Human Pluripotent Stem Cells

Matt Wind et al. Curr Protoc. 2021 Sep.
Free article

Abstract

The ability to generate spinal cord motor neurons from human pluripotent stem cells (hPSCs) is of great use for modelling motor neuron-based diseases and cell-replacement therapies. A key step in the design of hPSC differentiation strategies aiming to produce motor neurons involves induction of the appropriate anteroposterior (A-P) axial identity, an important factor influencing motor neuron subtype specification, functionality, and disease vulnerability. Most current protocols for induction of motor neurons from hPSCs produce predominantly cells of a mixed hindbrain/cervical axial identity marked by expression of Hox paralogous group (PG) members 1-5, but are inefficient in generating high numbers of more posterior thoracic/lumbosacral Hox PG(8-13)+ spinal cord motor neurons. Here, we describe a protocol for efficient generation of thoracic spinal cord cells and motor neurons from hPSCs. This step-wise protocol relies on the initial generation of a neuromesodermal-potent axial progenitor population, which is differentiated first to produce posterior ventral spinal cord progenitors and subsequently to produce posterior motor neurons exhibiting a predominantly thoracic axial identity. © 2021 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Differentiation of neuromesodermal progenitors Basic Protocol 2: Posterior ventral spinal cord progenitor differentiation Basic Protocol 3: Posterior motor neuron differentiation.

Keywords: human pluripotent stem cells (hPSCs); motor neurons; neuromesodermal progenitors.

PubMed Disclaimer

References

Literature Cited

    1. Amoroso, M. W., Croft, G. F., Williams, D. J., O'Keeffe, S., Carrasco, M. A., Davis, A. R., … Wichterle, H. (2013). Accelerated high-yield generation of limb-innervating motor neurons from human stem cells. Journal of Neuroscience, 33(2), 574-586. doi: 10.1523/jneurosci.0906-12.2013.
    1. Cambray, N., & Wilson, V. (2007). Two distinct sources for a population of maturing axial progenitors. Development, 134(15), 2829-2840. doi: 10.1242/dev.02877.
    1. Chambers, S. M., Fasano, C. A., Papapetrou, E. P., Tomishima, M., Sadelain, M., & Studer, L. (2009). Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nature Biotechnology, 27(3), 275-280. doi: 10.1038/nbt.1529.
    1. Cooper, F., Gentsch, G. E., Mitter, R., Bouissou, C., Healy, L., Hernandez-Rodriguez, A., … Bernardo, A. S. (2021). Rostrocaudal patterning and neural crest differentiation of human pre-neural spinal cord progenitors in vitro. bioRxiv, 2020.06.16.155564. doi: 10.1101/2020.06.16.155564.
    1. Dasen, J. S., De Camilli, A., Wang, B., Tucker, P. W., & Jessell, T. M. (2008). Hox repertoires for motor neuron diversity and connectivity gated by a single accessory factor, FoxP1. Cell, 134(2), 304-316. doi: 10.1016/j.cell.2008.06.019.

LinkOut - more resources