A Platform for Repeated Electric Field Stimulation of Organotypic Spinal Cord Slice Cultures Enables Comparison of Myelin Basic Protein Signal Across Stimulation Paradigms.

BACKGROUND: Electric field (EF) stimulation is a promising strategy to promote repair in the central nervous system (CNS). The cellular mechanisms underlying CNS repair induced by multi-day subthreshold EF stimulation remain poorly understood, in part because systematic exploration of stimulation parameter space is challenging in animal models. Rat organotypic spinal cord slice cultures (OSCs) provide a physiologically relevant ex vivo model that preserves cell-to-cell interactions and tissue architecture, making them well suited for studying parameter-dependent EF effects. However, a method enabling controlled, repeatable, and biocompatible EF stimulation of OSCs has not yet been established. NEW METHOD: We developed and validated a stimulation platform and protocol for repeated EF stimulation of OSCs. The platform generated a spatially uniform transverse EF across submerged OSCs while maintaining electrochemical biocompatibility. To demonstrate biological applicability, we applied the method to lysolecithin (LPC)-induced demyelinated OSCs. RESULTS: OSCs remained viable during repeated daily 40-minute submersion and stimulation over five days. EF stimulation influenced myelin basic protein, with higher-intensity or longer-duration stimulation producing a pattern consistent with partial recovery. COMPARISON WITH EXISTING METHODS: Previous platforms have demonstrated the feasibility of applying EFs to organotypic slice cultures for short durations, but whether OSCs can tolerate repeated submersion and EF stimulation without compromised viability remains unresolved. This method addresses that gap by enabling repeated, biocompatible EF stimulation of OSCs. CONCLUSIONS: These findings establish a method for controlled EF stimulation of OSCs and highlight its utility for systematically studying EF-mediated CNS repair mechanisms.