Organotypic brain slice culture was used to study the direct actions of relaxin on neural cells under ischemic stress. Cortical brain slices from neonatal rats were cultured for 14 days. Experimental slices were placed in a deoxygenated, glucose-free balanced salt solution (BSS) for 1 h, one group with 10−7 M H2 relaxin in the medium and the other without. Control slices were transferred to oxygenated BSS with glucose. Slices were returned to normal culture conditions and analyzed 1, 4, 8, and 24 h post-treatment using propidium iodide (PI) fluorescence to highlight cellular damage or death. The percentages of dead and dying cells in a slice were compared between groups. Relaxin treatment attenuated PI staining at 4 and 8 h postischemia, suggesting a direct action(s) on cells that improves their chance of survival during ischemia.