Derivation of dorsal spinal sensory interneurons from human pluripotent stem cells

STAR Protoc. 2021 Feb 3;2(1):100319. doi: 10.1016/j.xpro.2021.100319. eCollection 2021 Mar 19.

Abstract

We describe two differentiation protocols to derive sensory spinal interneurons (INs) from human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). In protocol 1, we use retinoic acid (RA) to induce pain, itch, and heat mediating dI4/dI6 interneurons, and in protocol 2, RA with bone morphogenetic protein 4 (RA+BMP4) is used to induce proprioceptive dI1s and mechanosensory dI3s in hPSC cultures. These protocols provide an important step toward developing therapies for regaining sensation in spinal cord injury patients. For complete details on the use and execution of this protocol, please refer to Gupta et al. (2018).

Keywords: Cell differentiation; Neuroscience; Stem cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Morphogenetic Protein 4 / pharmacology
  • Cell Differentiation / drug effects
  • Embryonic Stem Cells / cytology
  • Flow Cytometry / methods*
  • High-Throughput Nucleotide Sequencing / methods*
  • Human Embryonic Stem Cells / cytology
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Interneurons / cytology*
  • Neurons / cytology
  • Pluripotent Stem Cells / drug effects
  • Spine / cytology
  • Tretinoin / pharmacology

Substances

  • BMP4 protein, human
  • Bone Morphogenetic Protein 4
  • Tretinoin