BioProject

GnRH neurons are fundamental for reproduction in all vertebrates ultimately integrating all reproductive inputs. More...

GnRH neurons are fundamental for reproduction in all vertebrates ultimately 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 Pluripotent Stem Cells. 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 a media containing FGF8 and a NOTCH inhibition. 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. Overall design: Hues7 embryonic stem cells (ESCs) where differentiated into Neural Progenitor Cells (NPCs). NPCs were expanded for 4 passages and finally differentiated into neurons for 21 (GnRH-EC 21) or 35 (GnRH-EC 35) days. Total RNA was extracted from ESCs, NPCs, GnRH-EC 21, and GnRH-EC 35, hybridized to an Illumina genome wide gene expression array (HT12v4) and data processed using Illumina's Genome Studio software. Less...