Work

Differentiation of KISS1-Expressing Cells from Human Pluripotent Stem Cells: Many Roads To Rome

Year: 2026

Type: preprint

Abstract: Introduction Kisspeptin-secreting (Kiss) neurons govern human puberty and reproduction. In the arcuate nucleus (ARC), they control pulsatile release of gonadotropin-releasing hormone (GnRH), while Kiss neurons in the preoptic area (POA) control GnRH surge. Since animal models do not fully recapitulate the human phenotype, a human model to study these neurons is crucial. Methods We differentiated human pluripotent stem cells (hPSCs) into neuron cultures using two distinct strategies: FGF8 protocol, consisting of dual SMAD inhibition (dSMADi), FGF8b, and Notch inhibition; and SHH protocol, consisting of dSMADi and SHH activation, followed by Notch inhibition. Neuron cultures obtained on day 45 were characterized at the mRNA level using RT-qPCR. Results Both strategies resulted in neuron cultures where significant KISS1 expression could be detected. SHH-derived neuron cultures expressed high NKX2-1 and the ARC markers POMC, NHLH2 , and NR5A2 , while FGF8-derived neuron cultures expressed low NKX2-1 and the anterior POA marker FOXG1 . Conclusions We provide the first ever strategies to differentiate hPSCs into neuron cultures that express KISS1 . Future studies providing in depth transcriptomic, protein, and functional characterization are needed to establish the properties of these models.

Source: bioRxiv (Cold Spring Harbor Laboratory)

DOI: 10.64898/2026.09.02.748796