Scalable Generation of Pseudo-Unipolar Nociceptors from Human Pluripotent Stem Cells

Development of new non-addictive pain drugs requires advanced strategies to differentiate human pluripotent stem cells (hPSCs) into relevant cell types amenable for translational research. Here, we developed a highly efficient and reproducible method that differentiates hPSCs into peptidergic and non-peptidergic nociceptors. By modulating specific cell signaling pathways, hPSCs were first converted into SOX10+ neural crest cells followed by differentiation into sensory neurons with in vivo-like pseudo-unipolar morphology. Detailed molecular and cellular characterizations confirmed that hPSC-derived nociceptors displayed molecular signatures that were comparable to dorsal root ganglia (DRGs) and expressed typical neuronal markers, transcription factors, neuropeptides, and over 150 ion channels and receptors, including critical pain-relevant drug targets (e.g. TRPV1, TAC1, CALCA, NAV1.7, NAV1.8). Moreover, after confirming robust functional activities and differential response to noxious stimuli and specific drugs, a robotic cell culture system was employed for large scale-up production of human sensory neurons, urgently needed to develop nociceptor-selective analgesics.