Generation of GnRH-Secreting Neurons from Human Pluripotent Stem Cells

Presentation Number: LBF-084
Date of Presentation: March 6th, 2015

Hataiwan Chokechuwattanalert1, Carina Lund1, Kristiina Pulli1, Venkatram Yellapragada1, Paolo Giacobini2, Karolina Lundin1, Sanna Vuoristo1, Timo Tuuri3, Parinya Noisa1 and Taneli Raivio*1
1University of Helsinki, Helsinki, Finland, 2INSERM U1172, University of Lille 2, France, Lille, France, 3Helsinki University Central Hospital, Helsinki, Finland

Abstract

Hypothalamic gonadotropin-releasing hormone (GnRH) secretion is essential for puberty and reproductive function. We have developed a 27-day, two-step protocol to differentiate human pluripotent stem cells (hPSCs; human embryonic stem cells and induced pluripotent stem cells) into GnRH-secreting neurons. First, hPSCs were differentiated toward neuroectodermal lineage by using 10 days of dual SMAD inhibition, which led to forebrain-specific neural progenitor cell (F-NPC) identity, as shown by robust upregulation of PAX6, EMX2, FOXG1, and GSX2. Second, GnRH neuron fate specification was achieved by exposing F-NPCs to FGF8, a key ligand implicated in GnRH ontogeny. The ensuing GnRH-expressing cells were TUJ1-positive, bipolar-shaped neurons that also expressed DCC and NRP-1, similar to human fetal GnRH neurons. Importantly, hPSC-derived GnRH neurons secreted GnRH into the culture medium, a feature suggesting their neuroendocrine maturity. Taken together, our results provide a new translational tool for investigating the mechanisms of human puberty and its disorders.

 

Nothing to Disclose: HC, CL, KP, VY, PG, KL, SV, TT, PN, TR