Generation of kisspeptin-responsive GnRH neurons from human pluripotent stem cells

Ariel Poliandri, Duncan Miller, Sasha Howard, Muriel Nobles, Gerard Ruiz-Babot, Stephen Harmer, Andrew Tinker, Tristan McKay, Leonardo Guasti, Leo Dunkel

Research output: Contribution to journalArticle (Academic Journal)peer-review

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Abstract

GnRH neurons are fundamental for reproduction in all vertebrates, integrating all reproductive inputs. The inaccessibility of human GnRH-neurons has been a major impediment to studying the central control of reproduction and its disorders. Here, we report the efficient generation of kisspeptin responsive GnRH-secreting neurons by directed differentiation of human Embryonic Stem Cells and induced-Pluripotent Stem Cells derived from a Kallman Syndrome patient and a healthy family member. The protocol involves the generation of intermediate Neural Progenitor Cells (NPCs) through long-term Bone morphogenetic protein 4 inhibition, followed by terminal specification of these NPCs in media containing Fibroblast Growth Factor 8 and a NOTCH inhibitor. The resulting GnRH-expressing and -secreting neurons display a neuroendocrine gene expression pattern and present spontaneous calcium transients that can be stimulated by kisspeptin. These in vitro generated GnRH expressing cells provide a new resource for studying the molecular mechanisms underlying the development and function of GnRH neurons.

Original languageEnglish
Pages (from-to)12-22
Number of pages11
JournalMolecular and Cellular Endocrinology
Volume447
Early online date21 Feb 2017
DOIs
Publication statusPublished - 15 May 2017

Bibliographical note

Copyright © 2017 Elsevier B.V. All rights reserved.

Keywords

  • Calcium/metabolism
  • Cell Differentiation
  • Cell Line
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Gonadotropin-Releasing Hormone/metabolism
  • Human Embryonic Stem Cells/metabolism
  • Humans
  • Kisspeptins/metabolism
  • Neural Stem Cells/metabolism
  • Neurons/metabolism
  • Pluripotent Stem Cells/metabolism

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