The Primary Cultures For Development Of EAN: An In Vitro Model System For Neuroprotection And Neuroregeneration Studies

The blood-brain barrier (BBB) is a highly specialized brain endothelial structure ofthe fully differentiated neurovascular system. And it is a selective barrier formed by theendothelial cells that line cerebral microvessels. In vivo, close interactions betweenendothelial cells, astrocytes and neurons contribute to the formation and function of theBBB.In the last decade more attention have been paid to in vitro models of BBBbecause of its relation to many neurodegenerative diseases and its physiologicalimportance. In this study we report an EAN (endothelial cell-astrocyte-neuron) systemusing Sprague Dawley rat cerebral capillary endothelial cells on one side of apolyethylene terephtalate filter of pore size1μm and Sprague Dawley rat astrocytes onthe other side, while Sprague Dawley rat neurons were placed at the bottom of the well.The three cell types’ morphologies were studied in both monocultures and co-culturesusing immunocytochemistry techniques. Trans-endothelial electrical resistance andpermeability measurements took place to study the characteristics and to evaluate theEAN in vitro system. Our EAN system represents a useful in vitro system that mayprove to be valuable for studying the passage of substances across the barrier as well asother aspects like physiology and pathology. Hence, this EAN in vitro system can be ofgreat benefit to increase our understanding of the role of these three cell types inneurodegenerative disorders and its potential role in neuro-protection and neuro-regeneration studies.