Senior Scientist BrainXell, Inc. Madison, Wisconsin, United States
Abstract: Chronic pain is a leading health crisis linked to reduced quality of life, mental disorders, opioid dependence, and suicide. Despite its prevalence, effective long-term therapeutic options remain elusive, as prolonged opioid treatments are highly addictive and often fatal. Preclinical drug screenings using animal models lead to high clinical trial failure rates due to genomic differences between animal and human tissues. Although human cadaver tissue offers some advantages, the limited availability and viability restrict its use. As such, human induced pluripotent stem cells (hiPSCs) and the derivatives present a scalable, physiologically relevant alternative. In this study, we developed a robust method to differentiate hiPSCs into nociceptors (NOCs) that are highly pure and functionally mature. Immunocytochemistry reveals that the cultures are >95% positive for Peripherin and β-III Tubulin, >80% positive for Brn3A, and >90% positive for NaV1.7 and NaV1.8. Electrophysiological characterization using multi-electrode arrays (MEAs) demonstrates that these NOCs are spontaneously active by day 14 and reach a stable baseline by day 25, achieving 95% active electrode yield (AEY). Functionally, these NOCs exhibit a significant increase in mean firing rate (MFR) and AEY in response to capsaicin (TRPV1 agonist, 100 nM and 67 nM). Conversely, both MFR and AEY significantly decrease in response to Huwentoxin (NaV1.7 inhibitor, 30 nM) and A803467 (NaV1.8 inhibitor, 300 nM and 150 nM). These findings demonstrate that our hiPSC-derived nociceptors are a powerful and physiologically relevant model for nociception research. Their scalability, high purity, and rapid maturation make them an ideal candidate for integration into high-throughput screening platforms, advancing the discovery and development of novel analgesics.
