Differentiation of Inflammation-Responsive Astrocytes from Glial Progenitors Generated from Human Induced Pluripotent Stem Cells

Stem Cell Reports. 2017 Jun 6;8(6):1757-1769. doi: 10.1016/j.stemcr.2017.05.011.


Astrocyte dysfunction and neuroinflammation are detrimental features in multiple pathologies of the CNS. Therefore, the development of methods that produce functional human astrocytes represents an advance in the study of neurological diseases. Here we report an efficient method for inflammation-responsive astrocyte generation from induced pluripotent stem cells (iPSCs) and embryonic stem cells. This protocol uses an intermediate glial progenitor stage and generates functional astrocytes that show levels of glutamate uptake and calcium activation comparable with those observed in human primary astrocytes. Stimulation of stem cell-derived astrocytes with interleukin-1β or tumor necrosis factor α elicits a strong and rapid pro-inflammatory response. RNA-sequencing transcriptome profiling confirmed that similar gene expression changes occurred in iPSC-derived and primary astrocytes upon stimulation with interleukin-1β. This protocol represents an important tool for modeling in-a-dish neurological diseases with an inflammatory component, allowing for the investigation of the role of diseased astrocytes in neuronal degeneration.

Keywords: astrocytes; co-culture; disease modeling; iPSCs; neurodegenerative disorders; neuroinflammation; neuropsychiatric disorders; stem cell.

Publication types

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

MeSH terms

  • Astrocytes / cytology*
  • Astrocytes / drug effects
  • Astrocytes / metabolism
  • Calcium / metabolism
  • Cell Differentiation* / drug effects
  • Cells, Cultured
  • Coculture Techniques
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Glial Fibrillary Acidic Protein / metabolism
  • Glutamic Acid / metabolism
  • Humans
  • Hyaluronan Receptors / metabolism
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Interleukin-1beta / pharmacology
  • Leukemia Inhibitory Factor / pharmacology
  • Microscopy, Fluorescence
  • Neurons / cytology
  • Neurons / metabolism
  • Principal Component Analysis
  • RNA / chemistry
  • RNA / isolation & purification
  • RNA / metabolism
  • Sequence Analysis, RNA
  • Stem Cells / cytology*
  • Stem Cells / metabolism
  • Transcriptome
  • Tumor Necrosis Factor-alpha / pharmacology


  • Glial Fibrillary Acidic Protein
  • Hyaluronan Receptors
  • Interleukin-1beta
  • Leukemia Inhibitory Factor
  • Tumor Necrosis Factor-alpha
  • Glutamic Acid
  • RNA
  • Calcium