This work aimed to investigate a new approach for the realization of in vitro models of the blood-brain barrier (BBB). The attention was focused on the development of a protocol whose costs and time requirements were suitable for high-throughput analysis and studies, exploiting electrospinning technology. Firstly, porous substrates made of electrospun fibers of cellulose acetate were realized and their morphological characteristics assessed. Subsequently, a static cellular mono-culture of endothelial cells was conducted in order to obtain the in vitro model. The reliability of the model was then investigated both through the analysis of TEER and permeability values, and through confocal imaging.