title: Development of Gonadotropin-Releasing Hormone-Secreting Neurons from Human Pluripotent Stem Cells.
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.
authors.affiliation: Faculty of Medicine, Department of Physiology, University of Helsinki, Helsinki 00014, Finland.
authors.forename: Carina
authors.initials: C
authors.lastname: Lund
authors.affiliation: Faculty of Medicine, Department of Physiology, University of Helsinki, Helsinki 00014, Finland.
authors.forename: Kristiina
authors.initials: K
authors.lastname: Pulli
authors.affiliation: Faculty of Medicine, Department of Physiology, University of Helsinki, Helsinki 00014, Finland.
authors.forename: Venkatram
authors.initials: V
authors.lastname: Yellapragada
authors.affiliation: Inserm, Jean-Pierre Aubert Research Center, Development and Plasticity of the Neuroendocrine Brain, Unité 1172, 59045 Lille Cedex, France; School of Medicine, University of Lille, Lille 59000, France.
authors.forename: Paolo
authors.initials: P
authors.lastname: Giacobini
authors.affiliation: Department of Obstetrics and Gynecology, HUCH, Helsinki 00029, Finland.
authors.forename: Karolina
authors.initials: K
authors.lastname: Lundin
authors.affiliation: Faculty of Medicine, Department of Physiology, University of Helsinki, Helsinki 00014, Finland.
authors.forename: Sanna
authors.initials: S
authors.lastname: Vuoristo
authors.affiliation: Department of Obstetrics and Gynecology, HUCH, Helsinki 00029, Finland.
authors.forename: Timo
authors.initials: T
authors.lastname: Tuuri
authors.affiliation: School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; Faculty of Medicine/Physiology, University of Helsinki, Biomedicum Helsinki, PO Box 63 (Haartmaninkatu 8), Helsinki 00014, Finland.
authors.forename: Parinya
authors.initials: P
authors.lastname: Noisa
authors.affiliation: Faculty of Medicine, Department of Physiology, University of Helsinki, Helsinki 00014, Finland; Children's Hospital, Pediatric Research Center, Helsinki University Central Hospital (HUCH), Helsinki 00029, Finland; Faculty of Medicine/Physiology, University of Helsinki, Biomedicum Helsinki, PO Box 63 (Haartmaninkatu 8), Helsinki 00014, Finland. Electronic address: taneli.raivio@helsinki.fi.
authors.forename: Taneli
authors.initials: T
authors.lastname: Raivio
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D051524:Fibroblast Growth Factor 8 / Q000494:pharmacology; D051858:Forkhead Transcription Factors / Q000378:metabolism; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D006801:Humans; D009419:Nerve Tissue Proteins / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D009666:Nose / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051880:Receptors, Notch / Q000378:metabolism; D051785:Smad Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: C090173:FOXG1 protein, human; D051858:Forkhead Transcription Factors; D009419:Nerve Tissue Proteins; D051880:Receptors, Notch; D051785:Smad Proteins; D051524:Fibroblast Growth Factor 8; D007987:Gonadotropin-Releasing Hormone
keywords: GnRH; Kallmann syndrome; gonadotropin-releasing hormone; human embryonic stem cells; human pluripotent stem cells; hypogonadotropic hypogonadism; induced pluripotent stem cells; puberty