The development and optimization of an in vitro E18 rat cortex epilepsy model

Abstract

Abstract This study describes efforts to create a cell culture-based in vitro model of epilepsy, based on E18 rat cortex material. To study this model, two assays are used. The calcium-6 assay is used to measure network activity in vitro. Immunofluorescent staining is performed against a variety of cell types commonly found in the central nervous system and the cortex in efforts to characterize and quantify cells in culture. This report describes comparisons and choices made for cell seeding density, medium change protocols, culture plate coatings, medium choices, the effects of medium changes on network development, and long-term culture as part of optimization of the culturing conditions. Additionally, it compares epilepsy models commonly used in vitro including addition of convulsant drugs like 4-Aminopyridine as well as incubation of cells in low magnesium conditions. The final part of the report describes preliminary efforts to determine the effects of nutritional supplementation and ketosis inducing conditions in culture.