Optimizing the Utility of Sex-Specific Organotypic Hippocampal Sliced Cultures for In Vitro Ischaemia Studies

Abstract

In vitro models of ischaemia are commonly used in neuroscience research yet can be difficult to establish in a reproducible manner due to lack of experimental details commonly reported in publications. This study describes a series of optimization steps in order to produce a reliable model of organotypic hippocampal sliced cultures used for in vitro ischaemia studies. In addition, we describe the process by which sex-specific cultures were produced. Optimization data is provided regarding the age of animals used, time course of culture maintenance in vitro and optimal time of Oxygen and Glucose Deprivation (OGD) in order to produce a sufficient level of cell death. In addition, we demonstrate the effect of the age and sex of the pups on the amount of cell death produced. This study provides detailed optimization steps required to produce a reliable and reproducible sex-specific organotypic hippocampal sliced culture model which would then be suitable for exposure to OGD in order to model in vitro ischaemia and assess potential neuroprotective compounds.