Highly Expandable Human iPS Cell-Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High-Throughput Screening

Curr Protoc Hum Genet. 2017 Jan 11;92:21.8.1-21.8.21. doi: 10.1002/cphg.33.

Abstract

Reprogramming of human somatic cells into induced pluripotent stem (iPS) cells has greatly expanded the set of research tools available to investigate the molecular and cellular mechanisms underlying central nervous system (CNS) disorders. Realizing the promise of iPS cell technology for the identification of novel therapeutic targets and for high-throughput drug screening requires implementation of methods for the large-scale production of defined CNS cell types. Here we describe a protocol for generating stable, highly expandable, iPS cell-derived CNS neural progenitor cells (NPC) using multi-dimensional fluorescence activated cell sorting (FACS) to purify NPC defined by cell surface markers. In addition, we describe a rapid, efficient, and reproducible method for generating excitatory cortical-like neurons from these NPC through inducible expression of the pro-neural transcription factor Neurogenin 2 (iNgn2-NPC). Finally, we describe methodology for the use of iNgn2-NPC for probing human neuroplasticity and mechanisms underlying CNS disorders using high-content, single-cell-level automated microscopy assays. © 2017 by John Wiley & Sons, Inc.

Keywords: drug discovery; high-content imaging; high-throughput screening; induced pluripotent stem cell; neurodegeneration; neuroplasticity; neuropsychiatric.

MeSH terms

  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Differentiation*
  • Cell Line
  • Cell Proliferation
  • Central Nervous System Diseases / drug therapy*
  • Central Nervous System Diseases / pathology*
  • High-Throughput Screening Assays*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Models, Biological*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Neural Stem Cells / cytology*
  • Neural Stem Cells / metabolism
  • Neuronal Plasticity
  • Neurons / cytology*
  • Neurons / metabolism
  • Single-Cell Analysis

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • NEUROG2 protein, human
  • Nerve Tissue Proteins