Physiological Characterization and Transcriptomic Properties of GnRH Neurons Derived From Human Stem Cells

Area of Science:

• Neuroscience
• Developmental Biology
• Stem Cell Biology

Background:

• Gonadotropin-releasing hormone (GnRH) neurons are crucial for reproductive function.
• Generating these neurons efficiently from human stem cells is challenging.

Purpose of the Study:

• To establish an efficient method for generating GnRH neurons from human embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC).
• To characterize the physiological and transcriptomic properties of these derived GnRH neurons.

Main Methods:

• Exposure of primitive neuroepithelial cells to fibroblast growth factor 8 (FGF8) and kisspeptin.
• Generation of a mCherry-labeled GnRH human embryonic stem cell line (mCh-hESC) using CRISPR-Cas9.
• Assessment of GnRH peptide release, response to stimuli (high potassium, kisspeptin, estradiol, neurokinin B), and electrophysiological properties.
• Transcriptomic analysis of GnRH neurons derived from hESC, hiPSC, and mCh-hESC.

Main Results:

• Primitive neuroepithelial cells exposed to FGF8 and kisspeptin yielded more GnRH neurons.
• Derived GnRH (mCh-hESC) neurons exhibited pulsatile GnRH release, responded to stimuli, and generated action potentials.
• Transcriptomic analysis revealed similar developmental patterns across different stem cell-derived GnRH neurons.

Conclusions:

• Human stem cell-derived GnRH neurons provide a valuable model for studying reproductive diseases like idiopathic hypothalamic hypogonadism.
• These cells are useful for screening and testing novel contraceptive drugs.