Human dpsc-neuron circadian bioluminescence assay for natural compound screening

Overview

Pulp Neuro Inc offers a contract screening service built on a patient-derived dental pulp stem cell (DPSC) neuronal platform that models human circadian clock function without animal models. DPSC lines are differentiated into mature cortical-like neurons and equipped with Per2, Bmal1, or CRY1 luciferase reporters that emit bioluminescence in real time as the cellular clock oscillates. The platform has been peer-reviewed and published, demonstrating quantifiable, disease-relevant period-length defects in Prader-Willi syndrome neurons and pharmacological rescue by a small molecule, confirming that the assay responds to compound intervention. The service applies this validated circadian readout to natural ingredients and complex botanical and marine extracts, enabling partners to identify and rank circadian-active compounds for nutraceutical, cosmetic, food, beverage, or therapeutic development.

Technical specifications

Platform components:

• DPSC lines differentiated into cortical-like neurons carrying Per2, Bmal1, or CRY1 luciferase reporters
• Real-time bioluminescence recording on LumiCycle or BioTek BioSpa 8 plate readers for multi-day (5-day) kinetic measurements
• Direct compound dosing into recording media in 96-well format, minimizing assay redevelopment
• Access to a banked repository of more than 200 DPSC lines spanning 15 disorders, including Prader-Willi syndrome and other neurodevelopmental conditions
• Quantified circadian outputs including period length, amplitude, and phase shift, with built-in comparison to vehicle controls
• Mechanistic follow-up capability through CRY1 luciferase profiling and western blot assessment of CRY1 ubiquitination and protein stability, targeting the MAGEL2-USP7-CRY1 stabilization axis

Screening workflow:

• Phase 1: Curate incoming natural product or extract libraries and align with established reporter lines
• Phase 2: Primary 96-well BioSpa screen with automated period, amplitude, and phase quantification
• Phase 3: Hit validation on LumiCycle in triplicate across control and disease-relevant lines, including dose-response
• Phase 4: Mechanistic confirmation by western blot and delivery of a ranked hit list with raw traces, quantified parameters, and a written report

Technology readiness level

The assay is fully operational and validated in a peer-reviewed publication, with confirmed pharmacological responsiveness in Prader-Willi syndrome neurons. The platform is ready for immediate screening engagements, with no requirement for additional assay development. DPSC-neuron lines are already banked and characterized, and the BioSpa plate-reader format supports direct dosing of compound libraries, enabling rapid initiation of natural product screening projects.