Pulsed electromagnetic stimulation promotes neuronal maturation by up-regulating cholesterol biosynthesis.

BACKGROUND: Stem cell therapies have emerged as transformative therapeutic strategies for neurological disorders. However, neurons derived from transplanted stem cells often exhibit low survival rates and remain in an immature state. While pulsed electromagnetic fields (PEMF) may enhance neuronal differentiation, the extent of this effect and its molecular mechanisms remain poorly characterized. METHOD: Human induced pluripotent stem cells (iPSCs) induced cortical neurons received daily PEMF stimulation (1 mT, 15 Hz, 3.75 ms pulse duration) for 7 days during differentiation. Neuronal differentiation and synaptic maturation were assessed using immunocytochemistry, qPCR, western blotting, and live-cell imaging to evaluate neurite outgrowth. Functional maturation was analyzed through calcium imaging and patch-clamp electrophysiology. Transcriptomic profiling identified key pathways involved in PEMF-modulated neuronal maturation, with the role of FDFT1-mediated cholesterol biosynthesis mechanistically validated through pharmacological inhibition and genetic knockdown. RESULT: PEMF accelerated early-stage neuronal differentiation without altering neurite outgrowth and enhanced synaptic maturation after sustained stimulation. PEMF-treated neurons displayed heightened spontaneous calcium signaling and improved functional maturation, including enhanced excitability, action potential kinetics, and voltage-gated ion channel activity. Transcriptomics revealed significant upregulation of cholesterol biosynthesis pathways, with FDFT1 (squalene synthase) as a central regulator. Pharmacological inhibition or genetic knockdown of FDFT1 abolished PEMF-induced neuronal differentiation and synaptic maturation. CONCLUSION: PEMF accelerates early-stage differentiation of human cortical neurons and enhances synaptic maturation following sustained stimulation. These effects are mechanistically linked to the activation of FDFT1-mediated cholesterol biosynthesis. This non-invasive PEMF stimulation approach represents a promising strategy to optimize stem cell-based therapies for neurological disorders.