4.1 Article

Protocol Generation of highly pure motor neurons from human induced pluripotent stem cells

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STAR PROTOCOLS
Volume 3, Issue 1, Pages -

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ELSEVIER
DOI: 10.1016/j.xpro.2022.101223

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Funding

  1. NIH [W81XWH2010186]
  2. Department of Defense (DoD) Peer Reviewed Medical Research Program (PRMRP) Discovery Award
  3. [NIH/NINDS NS112910]

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This study presents a protocol for generating neural progenitor cells and functional motor neurons from human pluripotent stem cells. The optimized induction condition allows for high yield and purity, enabling the identification of dysregulated factors in diseased neurons.
Generation of human motor neurons (MNs) overcomes the inaccessibility to patient brain tissues and greatly facilitates the research in MN-related diseases. Here, we describe a protocol for generation of neural progenitor cells (NPCs) from human induced pluripotent stem cells (hiPSCs), followed by preparation of functional MNs. The optimized induction condition with the expression of three transcription factors in a single lentiviral vector significantly improved the yield and purity, mak-ing it possible to biochemically identify dysregulated factors in diseased neurons. For complete details on the use and execution of this protocol, please refer to Ding (2021), Ding et al. (2021), and Sepehrimanesh and Ding (2020).

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