Rapid Ngn2-induction of excitatory neurons from hiPSC-derived neural progenitor cells

Methods. 2016 May 15:101:113-24. doi: 10.1016/j.ymeth.2015.11.019. Epub 2015 Nov 25.

Abstract

Since the discovery of somatic reprogramming, human induced pluripotent stem cells (hiPSCs) have been exploited to model a variety of neurological and psychiatric disorders. Because hiPSCs represent an almost limitless source of patient-derived neurons that retain the genetic variations thought to contribute to disease etiology, they have been heralded as a patient-specific platform for high throughput drug screening. However, the utility of current protocols for generating neurons from hiPSCs remains limited by protracted differentiation timelines and heterogeneity of the neuronal phenotypes produced. Neuronal induction via the forced expression of exogenous transcription factors rapidly induces defined populations of functional neurons from fibroblasts and hiPSCs. Here, we describe an adapted protocol that accelerates maturation of functional excitatory neurons from hiPSC-derived neural progenitor cells (NPCs) via lentiviral transduction of Neurogenin 2 (using both mNgn2 and hNGN2). This methodology, relying upon a robust and scalable starting population of hiPSC NPCs, should be readily amenable to scaling for hiPSC-based high-throughput drug screening.

Keywords: Directed differentiation; Modeling neuropsychiatric disease; Neuronal induction; hiPSC; iNeuron.

Publication types

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

MeSH terms

  • Basic Helix-Loop-Helix Transcription Factors / physiology*
  • Cell Culture Techniques
  • Cells, Cultured
  • Cellular Reprogramming
  • Gene Expression
  • Humans
  • Induced Pluripotent Stem Cells / physiology*
  • Lentivirus
  • Nerve Tissue Proteins / physiology*
  • Neural Stem Cells / physiology
  • Transcriptional Activation

Substances

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