Direct Differentiation of Functional Neurons from Human Pluripotent Stem Cells (hPSCs)

Methods Mol Biol. 2021:2352:117-126. doi: 10.1007/978-1-0716-1601-7_8.

Abstract

Somatic cell nuclear transfer and in vitro induction of pluripotency in somatic cells by defined factors provided unambiguous evidence that the epigenetic state of terminally differentiated somatic cells is not static and can be reversed to a more primitive one. Inspired by these results, stem cell biologists have identified approaches to directly convert fibroblasts into induced neuronal (iN) cells, indicating that direct lineage conversions are possible between distantly related cell types. More recently, we took advantages of pro-neurogenic capacity of iN factors and developed methods to rapidly derive functionally mature neurons directly from human pluripotent stem cells (hPSCs) through a brief induction of defined transcription factors. In this chapter, we describe the detailed methods used to attain the direct conversion from hPSCs to glutamatergic and GABAergic iN cells.

Keywords: Direct differentiation; GABAergic; Glutamatergic; Induced neuronal cells; Pluripotent stem cells; Transcription factors.

MeSH terms

  • Cell Differentiation*
  • Cell Line
  • Cell Separation
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Fluorescent Antibody Technique
  • GABAergic Neurons / cytology
  • GABAergic Neurons / metabolism
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / biosynthesis
  • Genetic Vectors / genetics
  • Humans
  • Lentivirus / genetics
  • Neurogenesis
  • Neuroglia / cytology
  • Neuroglia / metabolism
  • Neurons / cytology*
  • Neurons / metabolism*
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • Transcription Factors
  • Transduction, Genetic

Substances

  • Transcription Factors