Development of Gonadotropin-Releasing Hormone-Secreting Neurons from Human Pluripotent Stem Cells

Stem Cell Reports. 2016 Aug 9;7(2):149-57. doi: 10.1016/j.stemcr.2016.06.007. Epub 2016 Jul 14.

Abstract

Gonadotropin-releasing hormone (GnRH) neurons regulate human puberty and reproduction. Modeling their development and function in vitro would be of interest for both basic research and clinical translation. Here, we report a three-step protocol to differentiate human pluripotent stem cells (hPSCs) into GnRH-secreting neurons. Firstly, hPSCs were differentiated to FOXG1, EMX2, and PAX6 expressing anterior neural progenitor cells (NPCs) by dual SMAD inhibition. Secondly, NPCs were treated for 10 days with FGF8, which is a key ligand implicated in GnRH neuron ontogeny, and finally, the cells were matured with Notch inhibitor to bipolar TUJ1-positive neurons that robustly expressed GNRH1 and secreted GnRH decapeptide into the culture medium. The protocol was reproducible both in human embryonic stem cells and induced pluripotent stem cells, and thus provides a translational tool for investigating the mechanisms of human puberty and its disorders.

Keywords: GnRH; Kallmann syndrome; gonadotropin-releasing hormone; human embryonic stem cells; human pluripotent stem cells; hypogonadotropic hypogonadism; induced pluripotent stem cells; puberty.

MeSH terms

  • Cell Line
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Fibroblast Growth Factor 8 / pharmacology
  • Forkhead Transcription Factors / metabolism
  • Gonadotropin-Releasing Hormone / metabolism*
  • Humans
  • Nerve Tissue Proteins / metabolism
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / metabolism
  • Neurons / cytology*
  • Neurons / drug effects
  • Neurons / metabolism*
  • Nose / cytology
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • Receptors, Notch / metabolism
  • Smad Proteins / antagonists & inhibitors
  • Smad Proteins / metabolism

Substances

  • FOXG1 protein, human
  • Forkhead Transcription Factors
  • Nerve Tissue Proteins
  • Receptors, Notch
  • Smad Proteins
  • Fibroblast Growth Factor 8
  • Gonadotropin-Releasing Hormone